RESUMO
GSTs (Glutathione S-transferases) are known to catalyze the nucleophilic attack of the sulfhydryl group of reduced glutathione (GSH) on electrophilic centers of xenobiotic compounds, including insecticides and acaricides. Genome analyses of the polyphagous spider mite herbivore Tetranychus urticae (two-spotted spider mite) revealed the presence of a set of 32 genes that code for secreted proteins belonging to the GST family of enzymes. To better understand the role of these proteins in T. urticae, we have functionally characterized TuGSTd01. Moreover, we have modeled the structure of the enzyme in apo form, as well as in the form with bound inhibitor. We demonstrated that this protein is a glutathione S-transferase that can conjugate glutathione to 1-chloro-2,4-dinitrobenzene (CDNB). We have tested TuGSTd01 activity with a range of potential substrates such as cinnamic acid, cumene hydroperoxide, and allyl isothiocyanate; however, the enzyme was unable to process these compounds. Using mutagenesis, we showed that putative active site variants S11A, E66A, S67A, and R68A mutants, which were residues predicted to interact directly with GSH, have no measurable activity, and these residues are required for the enzymatic activity of TuGSTd01. There are several reports that associate some T. urticae acaricide resistance with increased activity of GSTs . However, we found that TuGSTd01 is not able to detoxify abamectin; in fact, the acaricide inhibits the enzyme with Ki = 101 µM. Therefore, we suggest that the increased GST activity observed in abamectin resistant T. urticae field populations is a part of the compensatory feedback loop. In this case, the increased production of GSTs and relatively high concentration of GSH in cells allow GSTs to maintain physiological functions despite the presence of the acaricide.
Assuntos
Acaricidas , Tetranychidae , Acaricidas/farmacologia , Animais , Glutationa Transferase/genética , Ivermectina/análogos & derivados , Tetranychidae/genéticaRESUMO
Background: Interval throwing programs are used in rehabilitation of throwing injuries, especially ulnar collateral ligament injuries. Athletes who are rehabilitating begin by throwing on flat ground progressing through increasing distances, number of throws, and intensity of throwing. If the athlete is a baseball pitcher, the flat-ground throwing phase is followed by pitching on a mound at progressively increased effort. The goal is to build back arm strength and capacity with an emphasis on proper mechanics. Purpose: To determine whether interval throwing progressively builds joint kinetics (specifically, elbow varus torque) to the level required during full-effort baseball pitching. A secondary purpose was to examine the kinematics produced during interval throwing compared to those seen during baseball pitching. Study Design: Systematic Review. Methods: Following PRISMA guidelines, PubMed, Embase, Web of Science, SPORTDiscus, and Google Scholar were systematically searched for biomechanical studies of flat-ground throwing and partial-effort pitching in baseball between 1987 and 2023. Studies that reported the biomechanics of either flat-ground throwing, or partial-effort pitching were included in this review. The AXIS tool was used to assess study quality. Results: Thirteen articles met the inclusion criteria. Ten studies were determined to be of moderate quality, while three studies were deemed high quality. Elbow varus torque during partial-effort pitching was less than during full-effort pitching. Elbow varus torque for most flat-ground throws did not exceed full-effort pitching torque. While most studies showed increased elbow varus torque with increased flat-ground throwing distance, the distance at which elbow varus torque matched or exceeded full-effort pitching elbow varus torque was not consistent.As flat-ground throwing distance increased, shoulder external rotation angle and shoulder internal rotation velocity increased. Arm slot (forearm angle above horizontal) decreased as flat-ground throwing distance increased. For varied effort pitching, shoulder external rotation angle, shoulder internal rotation velocity, elbow extension velocity, and ball velocity increased as effort increased. While the front knee extended slightly from foot contact to ball release in full-effort pitching, the front knee flexed slightly during partial-effort pitching. Conclusions: An interval throwing program progressively builds elbow varus torque up to levels produced in full-effort baseball pitching. While differences exist between interval throwing kinematics and pitching kinematics, the patterns are similar in general. Level of Evidence: 2.
RESUMO
Aflatoxins, a family of fungal secondary metabolites, are toxic and carcinogenic compounds that pose an enormous threat to global food safety and agricultural sustainability. Specifically agricultural products in African, Southeast Asian and hot and humid regions of American countries suffer most damage from aflatoxin producing molds due to the ideal climate conditions promoting their growth. Our recent studies suggest that Vibrio gazogenes (Vg), an estuarine bacterium non-pathogenic to plants and humans, can significantly inhibit aflatoxin biosynthesis in the producers. In this study, we investigated the mechanism underlying Vg-dependent aflatoxin inhibition using the prominent aflatoxin producer, Aspergillus flavus. We show that aflatoxin inhibition upon Vg treatment was associated with fungal uptake of Vg-prodigiosin, a red pigment, which was consistently visible inside fungal hyphae during treatment. The association of prodigiosin with aflatoxin inhibition was further evident as Serratia marcescens, another prodigiosin producer, significantly inhibited aflatoxin, while non-producers like Escherichia coli, Staphylococcus aureus, Vibrio harveyi, and Vibrio fischeri did not. Also, pure prodigiosin significantly inhibited aflatoxin biosynthesis. Endocytosis inhibitors, filipin and natamycin, reduced the Vg-prodigiosin uptake by the fungus leading to a significant increase in aflatoxin production, suggesting that uptake is endocytosis-dependent. The Vg treatment also reduced hyphal fusion (>98% inhibition) and branching, which are both endosome-dependent processes. Our results, therefore, collectively support our theory that Vg-associated aflatoxin inhibition is mediated by an endocytosis-dependent uptake of Vg-prodigiosin, which possibly leads to a disruption of normal endosomal functions.
RESUMO
Delta-9-tetrahydrocannabinol (THC) is the primary psychoactive compound in Cannabis, which is studied extensively for its medicinal value. A central gap in the science is the underlying mechanisms surrounding THC's therapeutic effects and the role of gut metabolite profiles. Using a mass-spectrometry based metabolomics, we show here that intraperitoneal injection of THC in C57BL/6 mice modulates metabolic profiles that have previously been identified as integral to health. Specifically, we investigated the effects of acute (single THC injection denoted here as '1X') and short -term (five THC injections on alternate days denoted as '5X') THC administration on fecal and intestinal tissue metabolite profiles. Results are consistent with the hypothesis that THC administration alters host metabolism by targeting two prominent lipid metabolism pathways: glycerophospholipid metabolism and fatty acid biosynthesis.