Phage therapy consists of applying bacteriophages, whose natural function is to kill specific bacteria. Bacteriophages are safe, evolve together with their host, and are environmentally friendly. At ...present, the indiscriminate use of antibiotics and salt minerals (Zn
or Cu
) has caused the emergence of resistant strains that infect crops, causing difficulties and loss of food production. Phage therapy is an alternative that has shown positive results and can improve the treatments available for agriculture. However, the success of phage therapy depends on finding effective bacteriophages. This review focused on describing the potential, up to now, of applying phage therapy as an alternative treatment against bacterial diseases, with sustainable improvement in food production. We described the current isolation techniques, characterization, detection, and selection of lytic phages, highlighting the importance of complementary studies using genome analysis of the phage and its host. Finally, among these studies, we concentrated on the most relevant bacteriophages used for biocontrol of
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as agents that cause damage to crops, and affect food production around the world.
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Trichomonas vaginalis, a human urogenital tract parasite, is capable of surviving in the male microenvironment, despite of the presence of Zn2+. Concentrations > 1.6 mm of Zn2+ have a trichomonacidal ...effect; however, in the presence of ≤1.6 mm Zn2+, several trichomonad proteins are up- or down-regulated. Herein, we analyzed the proteome of a T. vaginalis male isolate (HGMN01) grown in the presence of Zn2+ and found 32 protein spots that were immunorecognized by male trichomoniasis patient serum. Using mass spectrometry (MS), the proteins were identified and compared with 23 spots that were immunorecognized in the proteome of a female isolate using the same serum. Interestingly, we found a 50-kDa metallopeptidase (TvMP50). Unexpectedly, this proteinase was immunodetected by the serum of male trichomoniasis patients but not by the female patient serum or sera from healthy men and women. We analyzed the T. vaginalis genome and localized the mp50 gene in locus TVAG_403460. Using an RT-PCR assay, we amplified a 1320-bp mp50 mRNA transcript that was expressed in the presence of Zn2+ in the HGMN01 and CNCD147 T. vaginalis isolates. According to a Western blot assay, native TvMP50 was differentially expressed in the presence of Zn2+. The TvMP50 proteolytic activity increased in the presence of Zn2+ in both isolates and was inhibited by EDTA but not by ptosyl-L-lysine chloromethyl ketone (TLCK), E64, leupeptin, or phenylmethane sulfonyl fluoride. Furthermore, the recombinant TvMP50 had proteolytic activity that was inhibited by EDTA. These data suggested that TvMP50 is immunogenic during male trichomoniasis, and Zn2+ induces its expression.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The Trichomonas vaginalis genome analysis suggested the presence of a putative deoxyhypusine synthase (TvDHS) that catalyzes the posttranslational modification of eIF-5A. Herein, we expressed and ...purified the recombinant TvDHS (rTvDHS) protein (43 kDa) and the recombinant TveIF-5A (rTveIF-5A) precursor protein (46 kDa). A 41 kDa band of the native TvDHS was recognized by western blot analysis in T. vaginalis total protein extract by a mouse polyclonal anti-rTvDHS antibody. The enzymatic activity of rTvDHS was determined by in vitro rTveIF-5A precursor modification. The modification reaction was performed by using (3H)-spermidine, and the biochemical analysis showed that rTvDHS exhibited Km value of 0.6 μM. The rTvDHS activity was inhibited by the spermidine analog, N″-guanyl-1,7-diamino-heptane (GC7). Native gel electrophoresis analysis showed two bands corresponding to an rTvDHS-rTveIF-5A complex and an intermediate form of rTveIF-5A. The two forms were subsequently separated by ion exchange chromatography to identify the hypusine residue by MS/MS analysis. Moreover, mutations in TvDHS showed that the putative HE motif present in this enzyme is involved in the hydroxylation of TveIF-5A. We observed that only hypusine-containing TveIF-5A was bound to an RNA hairpin ERE structure from the cox-2 gene, which contains the AAAUGUCACAC consensus sequence. Interestingly, 2DE-WB assays, using parasites that were grown in DAB-culture conditions and transferred to exogenous putrescine, showed the new isoform of TveIF-5A. In summary, our results indicate that T. vaginalis contains an active TvDHS capable of modifying the precursor TveIF-5A protein, which subsequently exhibits RNA binding activity.
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•TvDHS catalyzes the posttranslational modification of rTveIF-5A precursor protein.•Only the mature rTveIF-5A was bound to a RNA hairpin ERE structure from cox-2 gene.•The hypusine residue in TveIF-5A was identify using a proteomic method.•Hypusine-containing TveIF-5A shows RNA-binding activity.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
The luteinizing hormone receptor (LHR) is a glycoprotein member of the G protein-coupled receptors superfamily. It participates in corpus luteum formation and ovulation in females and acts in ...testosterone synthesis and spermatogenesis in males. In this study, we extracted RNA from sheep testicles and synthetized the cDNA to amplify the gene lhr-bed. This gene consists of 762 bp and encodes 273 amino acids of the extracellular domain of LHR. The lhr-bed was cloned into pJET1.2/blunt, then subcloned into pCOLD II, and finally, transformed in E. coli BL21 (DE3) cells. Because the induced rLHR-Bed protein was found in the insoluble fraction, we followed a modified purification protocol involving induction at 25 °C, subjection to denaturing conditions, and on-column refolding to increase solubility. We confirmed rLHR-Bed expression by means of Western blot and mass spectrometry analysis. It is currently known that the structure stem-loop 5′UTR on pCOLD II vector is stable at 15 °C. We predicted and obtained RNAfold stability at 25 °C. We successfully obtained the recombinant LHR extracellular domain, with protein yields of 0.2 mg/L, and purity levels of approximately 90%, by means of a single chromatographic purification step. The method described here may be used to obtain large quantities of rLHR-Bed in the future.
El artículo estudia la relación entre el clima escolar y las posibilidades de participación, dentro de la escuela, con las actitudes cívicas y políticas, a través de la percepción de los estudiantes ...de nivel medio superior. Se entrevistó a docentes y se aplicó una serie de encuestas a estudiantes del Colegio de Bachilleres de la Ciudad de México (n=769) para construir un modelo de ecuaciones estructurales. Los resultados presentan una relación significativa con los valores cívicos reportados por los alumnos. Las entrevistas sugieren que las actividades dentro del aula son las únicas posibilidades de participación para los educandos y se identificó un discurso contradictorio de los docentes acerca de las actitudes políticas de los estudiantes.
The Yucatan Peninsula, located in southern Mexico, is a central honey-producing region with extraordinary biodiversity of melliferous plants. Approximately 900 plant species have been described as ...being a source of nectar and pollen for bees and other pollinators. They provide ecosystem services that help to keep plant biodiversity high and mitigate the effects of climate change. This study aimed to reveal the diversity of the pollen content in honey through a melissopalynological analysis of 22 honey samples collected in February–August 2021 from the north-central area of Campeche, Mexico. The extraction of pollen from the honey was carried out using standard methods for melissopalynological analysis. The honeys were classified by botanical origin to determine their floral sources and a diverse spectrum of 19 pollen types from 13 families was identified. Only eight were predominant: Milleria quinqueflora, Gymnopodium floribundum, Terminalia buceras, Amaranthus spinosus, Zea mays, Talisia floresii, Guazuma ulmifolia, and Croton icche. Our research shows the high quality of the honey analyzed and highlights the diversity and critical role of local melliferous flora and crops in beekeeping development in southern Mexico. The results in this study are useful for confirming the botanical origins of honey, generating information for designing nature conservation and agroecosystem management strategies, and increasing the knowledge of beekeepers in Campeche, Mexico.
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Currently agriculture has difficulty keeping up with the demand for food around the world, which has generated a boom in the development of sustainable alternatives for producing food and caring for ...the environment. Therefore, the present study aims to show a backyard system comprising 50 cm × 50 cm pinewood boxes where maize and peanut were tested under control and compost conditions. The experiments were carried out for nine months starting from compost production and the sowing of the crops, which were irrigated with temporary rain. The compost was produced by converting ~213 kg of organic residues into ~300 kg of mature compost. The fertilizer treatment consisted of two doses of compost (1 kg doses). The developing plants were compared between conditions in both crops. In addition, the nutritional values of the compost and compost and soil were evaluated. Interestingly, the correlation analyses of the morphological properties of the soils showed that the effects of the nutrients were positively associated with the morphology of the crops studied. Finally, the yield produced for maize was 9 kg/m2 and 6.6 kg/m2 and that for peanuts was 184 g/m2 and 73 g/m2 under compost and control conditions, respectively. We consider that the development of new alternatives for producing food in times of crisis or situations of limited resources is necessary for the development of humanity and the care of the environment.
Previously, metalloproteinase was isolated and identified from
Trichomonas vaginalis
, belonging to the aminopeptidase P-like metalloproteinase subfamily A/B, family M24 of clan MG, named TvMP50. The ...native and recombinant TvMP50 showed proteolytic activity, determined by gelatin zymogram, and a 50 kDa band, suggesting that TvMP50 is a monomeric active enzyme. This was an unexpected finding since other Xaa-Pro aminopeptidases/prolidases are active as a biological unit formed by dimers/tetramers. In this study, the evolutionary history of TvMP50 and the preliminary crystal structure of the recombinant enzyme determined at 3.4 Å resolution is reported. TvMP50 was shown to be a type of putative, eukaryotic, monomeric aminopeptidase P, and the crystallographic coordinates showed a monomer on a “pseudo-homodimer” array on the asymmetric unit that resembles the quaternary structure of the M24B dimeric family and suggests a homodimeric aminopeptidase P-like enzyme as a likely ancestor. Interestingly, TvMP50 had a modified N-terminal region compared with other Xaa-Pro aminopeptidases/prolidases with three-dimensional structures; however, the formation of the standard dimer is structurally unstable in aqueous solution, and a comparably reduced number of hydrogen bridges and lack of saline bridges were found between subunits A/B, which could explain why TvMP50 portrays monomeric functionality. Additionally, we found that the
Parabasalia
group contains two protein lineages with a “pita bread” fold; the ancestral monomeric group 1 was probably derived from an ancestral dimeric aminopeptidase P-type enzyme, and group 2 has a probable dimeric kind of ancestral eukaryotic prolidase lineage. The implications of such hypotheses are also presented.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
The zinc fingers proteins (ZNF) are the largest family of DNA binding proteins and can act as transcriptional factors in eukaryotes. ZNF are implicated in activation in response to environmental ...stimulus by biometals such as Zn
2+
. Many of these proteins have the classical C
2
H
2
zinc finger motifs (C
2
H
2
-ZNFm) of approximately 30 amino acids, where a Zn
2+
ion is coordinated by two cysteine and two histidine residues.
Trichomonas vaginalis
is a protozoan parasite than responds to environmental changes including Zn
2+
. Until now has not been described any ZNF that could be involved in the regulation of genic expression of
T. vaginalis
. Here, we characterized in silico and experimentally an annoted ZNF (TvZNF1) from
T. vaginalis
and isolated the gene,
tvznf1
encoding it. TvZNF1 have eight C
2
H
2
-ZNFm with residues that maybe involved in the structural stability of DNA binding motifs. In this work we confirmed the Zn
2+
upregulation expression of
tvznf1
gene. Recombinant TvZNF1 was able to bind to specific DNA sequences according to EMSA assay. Additionally, we demonstrated that recombinant TvZNF1 bind to MRE signature in vitro, which strongly suggests its role in transcriptional regulation, similar to the one observed for mammalian MTF-1. This result suggested a conserved mechanism of genic regulation mediated by ZNFs in
T. vaginalis
.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
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