Introduction: Due to the coronavirus pandemic, identifying the infected individuals has become key to limiting its spread. Virus nucleic acid real-time RT-PCR testing has become the current standard diagnostic method but high demand could lead to shortages. Therefore, we propose a detection strategy using a one-step nested RT-PCR. Methodology: The nucleotide region in the ORF1ab gene that has the greatest differences between the human coronavirus and the bat coronavirus was selected. Primers were designed after that sequence. All diagnostic primers are species-specific since the 3´ end of the sequence differs from that of other species. A primer set also creates a synthetic positive control. Amplified products were seen in a 2.5% agarose gel, as well as in an SYBR Green-Based Real-Time RT-PCR. Results: Amplification was achieved for the positive control and specific regions in both techniques. Conclusions: This new technique is flexible and easy to implement. It does not require a real-time thermocycler and can be interpreted in agarose gels, as well as adapted to quantify the viral genome. It has the advantage that if the coronavirus mutates in one of the key amplification nucleotides, at least one pair can still amplify, thanks to the four diagnostic primers.
Centruroides possanii is a recently discovered species of “striped scorpion” found in Mexico. Certain species of Centruroides are known to be toxic to mammals, leading to numerous cases of human intoxications in the country. Venom components are thought to possess therapeutic potential and/or biotechnological applications. Hence, obtaining and analyzing the secretory gland transcriptome and venom proteome of C. possanii is relevant, and that is what is described in this communication. Since this is a newly described species, first, its LD50 to mice was determined and estimated to be 659 ng/g mouse weight. Using RNA extracted from this species and preparing their corresponding cDNA fragments, a transcriptome analysis was obtained on a Genome Analyzer (Illumina) using the 76-base pair-end sequencing protocol. Via high-throughput sequencing, 19,158,736 reads were obtained and ensembled in 835,204 sequences. Of them, 28,399 transcripts were annotated with Pfam. A total of 244 complete transcripts were identified in the transcriptome of C. possanii. Of these, 109 sequences showed identity to toxins that act on ion channels, 47 enzymes, 17 protease inhibitors (PINs), 11 defense peptides (HDPs), and 60 in other components. In addition, a sample of the soluble venom obtained from this scorpion was analyzed using an Orbitrap Velos apparatus, which allowed for identification by liquid chromatography followed by mass spectrometry (LC-MS/MS) of 70 peptides and proteins: 23 toxins, 27 enzymes, 6 PINs, 3 HDPs, and 11 other components. Until now, this work has the highest number of scorpion venom components identified through omics technologies. The main novel findings described here were analyzed in comparison with the known data from the literature, and this process permitted some new insights in this field.
12 February 2024 in Journal of Nutrition and Metabolism
Moringa oleifera (MO) is a native tree of Asia and is cultivated in some areas of Mexico as part of traditional horticulture. The aim of the present study was to compare the efficacy of MO infusion vs. MO ethanolic extract for the simultaneous treatment of nonalcoholic fatty liver (NAFLD), hyperlipidemia, and hyperglycemia in a murine model fed with a high-fat diet (HFD). BALB/c mice were fed a balanced diet (healthy control) or an HFD for 6months. With this, the NAFLD model was established before starting a therapeutic intervention with MO for two months. The phytochemical analysis by nuclear magnetic resonance in 1H and 13C experiments showed signals for pyrrole alkaloids and triterpenes as the main constituents of the extract and infusion preparation. A significant reduction of SGPT, SGOT, lipids, urea, and glucose in blood among NAFLD groups treated with MO (infusion or extract) was found, when compared to the NAFLD-placebo group. Steatosis and liver inflammation were found to be decreased in the MO groups, as infusion or ethanolic extract. Infusion produced a better therapeutic effect than the extract in all parameters, except glycemic control, where the extract was better. As an additional finding, it is noteworthy that treatment with MO, particularly through infusion, resulted in improved motor activity. Moreover, a reduction in anxiety-like behavior was observed exclusively with the administration of infusion. These observations provide valuable insights into the potential broader effects of Moringa oleifera beyond the primary aim of the study.
2 June 2022 in Evidence-Based Complementary and Alternative Medicine
Rheumatoid arthritis is globally present in about 1% of the population. This autoinflammatory disease modifies the connective tissue, causing pain and inflammation of the joints. Over time, it causes the loss of joint cartilage and bone mass, decreasing the patient's quality of life. Treatment options now available either give symptomatic alleviation or alter the disease process. Nonetheless, adherence to chronic treatment is typically limited due to adverse effects. As a result, new therapy approaches, such as systemic administration of neutral electrolyzed saline to improve patients’ quality of life, are being investigated. The study is a randomized prospective preclinical trial with a single-blind and a 4-arm parallel group using a collagen-induced mice model to generate rheumatoid arthritis. It was carried out on 36 male BALB/c mice, with the primary outcome measure being a scoring system for histopathologic assessment. When all groups are compared, there are significant differences. In addition, the animal model was validated by the healthy group. The animals treated with neutral electrolyzed saline had much less cartilage degradation, bone erosion, pannus development, and inflammation than the placebo-treated mice. Serum IL-6 levels were evaluated in parallel with disease severity expressed as synovitis grading of the affected joints. Spearman’s rank correlation coefficient (Rs) = 0.399 ( ) between serum IL-6 levels and the synovitis grading suggests a direct correlation between IL-6 production and disease severity. An additional trial of 20 male BALB/c mice (10 treated with placebo and 10 with neutral electrolyzed saline for 30 days) showed no clinical nor histopathological evidence of adverse effects. According to histopathological and blood test results, we conclude that neutral electrolyzed saline minimizes mechanical and inflammatory damage to the joint and may be helpful as an alternative to rheumatoid arthritis therapy.
Oxidative stress results from the excessive production of reactive oxygen species (ROS), which cause cellular and molecular damage and contribute to chronic diseases. Given the recognized antioxidant potential of benzimidazole derivatives—particularly 2-mercaptobenzimidazole—this study aimed to synthesize novel organosilicon S-silylalkylthioethers (I–IV) and N-alkylsilylthioethers (1a–3f) derived from this scaffold and to evaluate their antioxidant and antibrowning properties. The S-silylalkylthioethers were obtained by reacting 2-mercaptobenzimidazole with different chloroalkylsilanes under reflux in ethanol, followed by a reaction with alkyl halides in aprotic media at room temperature to prepare the N-alkylsilylthioethers. Structural elucidation was achieved through 1D and 2D NMR and FT-IR. Antioxidant activity was assessed using DPPH, the total antioxidant capacity, and ferric-reducing assays. The results showed several derivatives with notable antioxidant responses, revealing a clear relationship between carbon chain length, logP values, organosilicon substitution patterns, and radical-scavenging efficiency. Spearman correlation analysis further confirmed that DPPH activity is inversely related to total carbon number, molecular size, molecular weight, and LogP (ρ = −0.68 to −0.73, p < 0.001) and moderately negatively correlated with N-alkyl chain length (ρ = −0.47, p = 0.027), while S-alkyl chains showed no significant effect. These findings highlight the potential of these benzimidazole–organosilicon hybrids as antioxidant candidates and demonstrate how physicochemical properties govern their reactivity and antiradical capacity.
An experiment was carried out to evaluate the effect of spring and autumn seasons on the reproductive activity of Merino Socorro Island ewes and their crosses with Pelibuey under heat stress (HS) conditions in the tropics. All ewes (n = 80) were randomly assigned to one of two breeds during the first and second periods, respectively: (1) Twenty Socorro Island Merino ewes (SIM) and (2) 20 Pelibuey Crossbred ewes (PBC). Animals were fed the same diet and given water ad libitum. All statistical analyses were performed using SAS statistical software 9.12 procedures. In both seasons, a mean of more than 80 U of maximum THI was obtained, while in spring and autumn, the minimum THI exceeded 30 and 40 U, respectively. All animals were in oestrus and ovulated in both seasons. The frequency of animals in spring during the first 48 h of oestrus expression was greater (p < 0.05) than 48–55 h but similar (p > 0.05) than 55–65 h; in autumn during the first 48 h and 48–55 h were similar (p > 0.05), but different (p < 0.05) than 55–65 h. The duration of oestrus expression was longer in the spring than in the autumn (p < 0.05). The frequency of animals was higher (p < 0.05) in SIM than in PBC ewes during the first oestrus cycle (1–17 d) and was also higher (p < 0.05) in PBC than in SIM ewes during the second oestrus cycle (18–35 d). The SIM ewes produced more (p < 0.05) progesterone (P4) than the PBC ewes. During the sampling days of the oestrus cycle, more P4 was created in autumn than in spring (p < 0.05). Both breeds showed severe HS. In the future, ewes treated under assisted reproductive programs in the tropics may improve reproductive efficiency.