Luis Edgar Rodríguez-Tovar

The 4T1 cell line is a widely used model for triple-negative breast cancer (TNBC) research due to its high metastatic potential and resemblance to clinical progression. However, its biological characteristics often limit the efficiency and reproducibility of conventional nucleic acid delivery protocols. Here, we present an optimized non-viral delivery framework for efficient cell-associated uptake in 4T1 cells. Five carrier systems—Xfect RNA polymer (Xfect), polyethyleneimine (PEI), chitosan (CHI), and gold nanoparticle conjugates (AuPEI and AuCHI)—were systematically evaluated using both standard (adherent) and reverse (suspension) delivery approaches. A fluorescently labeled oligonucleotide (5′-TYE563) served as a reporter cargo at final concentrations of 5 nM and 10 nM, comparing complexation times of 15 s and 15 min. The proposed protocol introduces a rapid complexation strategy combined with reverse delivery, significantly reducing preparation time while enhancing cell-associated uptake. Under optimized conditions (10 nM, 15 s, reverse), Xfect achieved near-complete cellular uptake with minimal cytotoxicity. Crucially, the colocalization of dual-fluorescence signals enabled the simultaneous validation of metabolic viability and internalized cargo within a single field of view, bypassing the need for independent bulk cytotoxicity assays. This methodology provides a rapid, reproducible, and cost-effective framework to standardize early-stage delivery conditions for difficult-to-transfect cell lines such as 4T1.

El presente artículo de investigación se centra en la evaluación del efecto citotóxico de los fitocannabinoides cannabidiol (CBD) y cannabigerol (CBG), de manera individual y en combinación (CBD:CBG), así como en una formulación a base de zeína (Ze) con CBD y CBG en conjunto (ZeCBD:CBG). Para ello, el efecto de CBD, CBG, CBD:CBG y ZeCBD:CBG en la viabilidad celular se evaluó en la línea celular intestinal Caco-2 y se evaluó la actividad metabólica dependiente de mitocondria mediante análisis de reducción de bromuro 3-(4,5-dimetiltiazol-2-il)-2-5-difeniltetrazolio o ensayo de MTT. Además, para determinar el tratamiento más efectivo, se estimó la concentración inhibitoria al 50% (IC50) de cada tratamiento evaluado mediante la prueba de Probit. Todos los tratamientos evaluados mostraron disminuir la viabilidad celular de células Caco-2 en diferente medida, siendo ZeCBD:CBG y CBD:CBG los que mostraron los valores más bajos de IC50 (12.8 µM y 9.2 µM, respectivamente). Por lo anterior, estos resultados indican que la formulación de CBD y CBG en una matriz basada en zeína muestra un menor efecto citotóxico metabólica comparado con los cannabinoides libres en las condiciones in vitro evaluadas.

Encephalitozoon cuniculi is a zoonotic mycotic pathogen that infects rabbits. When the host becomes infected, spores in the brain parenchyma cause severe granulomatous meningoencephalitis. The mechanisms by which spores reach the CNS are poorly understood. We divided 27 white New Zealand rabbits into 2 groups: 15 infected orally with 10 ⁶ spores and 12 uninfected controls. Animals were euthanized at 30 d post-infection (dpi). Histologically, spores were present within brain blood vessels at 30 dpi. To our knowledge, the localization of free E. cuniculi spores has not been demonstrated previously within blood vessels in the brain of rabbits.

Background: Phytocannabinoids such as cannabidiol (CBD) and cannabigerol (CBG) have received increasing attention in the context of inflammatory and intestinal disorders. However, direct comparisons between their individual and combined effects, as well as the influence of delivery systems, remain limited. Objectives: This study evaluated the biological effects of free and nanoencapsulated CBD and CBG, including a cannabinoid–Eudragit L100 formulation, in an in vitro TNBS-treated intestinal cell model and an in vivo murine model of TNBS-induced colitis. Methods: Cytotoxicity and treatment-associated effects of CBD, CBG, their 1:1 combination, and a nanoencapsulated formulation were assessed in TNBS-exposed Caco-2 cells. In parallel, BALB/c mice with TNBS-induced colitis were evaluated for colonic damage and inflammatory markers. Results: CBD and CBG individually showed dose-dependent effects in Caco-2 cells, while their combined administration produced a greater effect than either compound alone at higher concentrations. The nanoencapsulated formulation preserved cellular metabolic activity following TNBS exposure. In vivo, both free combined and nanoencapsulated cannabinoids were associated with reduced epithelial damage and inflammatory alterations. Conclusions: Nanoencapsulation using Eudragit L100 modulated the biological effects of CBD and CBG in experimental models of TNBS-induced intestinal injury.

Cannabidiol (CBD), a phytocannabinoid derived from Cannabis sativa, has demonstrated therapeutic potential across various diseases, including cancer. This study evaluates the cytotoxic effects of CBD on three human cancer cell lines (HeLa, MDA-MB-231, and CaCo-2) and two non-cancerous cell lines (HaCaT and HUVEC) used as a control. Cells were treated with CBD at concentrations of 5, 10, and 20 µM for 24, 48, 72, and 96 h. Cytotoxicity was assessed using MTT assays, nuclear morphology was evaluated via DAPI staining, and cell death mechanisms were analyzed through flow cytometry with apoptosis/necrosis markers. The LC50 values at 24 h were determined as follows: HeLa (9.4 µM), MDA-MB-231 (10.3 µM), and CaCo-2 (4.3 µM). CBD treatment induced morphological changes characteristic of cell stress and death in cancer cells, observed by optical microscopy after 24, 48, 72, and 96 h of exposure. These findings highlight the potential of CBD as an adjunctive therapeutic agent for cancer treatment versus non-malignant cells.

Cannabidiol (CBD), a phytocannabinoid derived from Cannabis sativa, has demonstrated therapeutic potential across various diseases, including cancer. This study evaluates the cytotoxic effects of CBD on three human cancer cell lines (HeLa, MDA-MB-231, and CaCo-2) and two non-cancerous cell lines (HaCaT and HUVEC) used as a control. Cells were treated with CBD at concentrations of 5, 10, and 20 µM for 24, 48, 72, and 96 h. Cytotoxicity was assessed using MTT assays, nuclear morphology was evaluated via DAPI staining, and cell death mechanisms were analyzed through flow cytometry with apoptosis/necrosis markers. The LC50 values at 24 h were determined as follows: HeLa (9.4 µM), MDA-MB-231 (10.3 µM), and CaCo-2 (4.3 µM). CBD treatment induced morphological changes characteristic of cell stress and death in cancer cells, observed by optical microscopy after 24, 48, 72, and 96 h of exposure. These findings highlight the potential of CBD as an adjunctive therapeutic agent for cancer treatment versus non-malignant cells.

A short‐tailed female bat ( Carollia perspicillata ), 1 year old, died without apparent signs of the disease while on display at an amusement park in the Municipality of Montemorelos, Nuevo León, Mexico. Amphophilic nuclear inclusion bodies were observed in the intestinal epithelia, corresponding to the virus of the adenovirus genera. Although there have been no reported adenovirus cases of this bat species in Mexico, through an anatomopathological study and support from the microscopic, ultrastructural, and molecular findings of intestinal lesions, a diagnosis of enteritis caused by Adenovirus Type 2 was made. To the authors’ knowledge, the case described herein is the first report of infection by Adenovirus Type 2 in a short‐tailed bat in Mexico.

Some of the goals of meat production systems are to improve feed efficiency, reduce costs, provide proper nutrition and avoid metabolic disorders. Hence, the aim of this work was to compare a rabbit diet based on untreated sorghum (T1) with a second one that included the cereal biotransformed by the co-cultivation of two basidiomycetes (T2). Their effects on the cereal, diets, and fecal structural fiber composition, as well as on rabbit intestinal morphology, were evaluated. A completely randomized design was applied, employing 24 New Zealand rabbits (21 days old). The fiber chemical composition results indicated that the sorghum treated with the co-culture had lower amounts of neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, hemicellulose and cellulose, as did rabbits fed the T2 diet. Decreases in the NDF, ADF and cellulose contents were detected in T2-nourished rabbit excrements at day 21 (p ≤ 0.05), while lignin percentages diminished at days 35 and 49 (p ≤ 0.05). These rabbits also showed longer jejunal and cecal villi (p ≤ 0.05). The results obtained suggest a positive effect on the bioavailability of structural fibers after sorghum fungal co-fermentation, as their content decreased in rabbit fecal matter without damaging the intestinal morphology.

The effect of short- and long-term exposure to heat stress (HS) was analyzed on blood components, performance, and intestinal epithelium integrity of pigs. Eighteen pigs (36.0 ± 3.5 kg BW) were assigned to three groups: thermo-neutral (TN); 2 d exposure to HS (2dHS); and 7 d exposure to HS (7dHS). Blood chemistry and hemogram analyses were performed; small intestine samples were analyzed for mRNA expression and histology. Compared to TN, 2dHS and 7dHS pigs reduced weight gain and feed intake; weight gain was higher in 7dHS than in 2dHS pigs (p < 0.05). White blood cells, platelet, and hematocrit were affected in 2dHS and 7dHS compared to TN pigs (p < 0.05). Short- and long-term exposure to HS affected blood concentration of triglycerides, urea, total protein, and albumin (p ≤ 0.05). Villi-height and crypt-depth decreased in HS pigs (p < 0.01). Mucin-producing and apoptotic cell number increased in 7dHS compared to TN pigs (p < 0.05). Expression of tight-junction-proteins decreased in 2dHS pigs compared to TN and 7dHS pigs (p < 0.05). Short-term exposure of pigs to HS dramatically affects performance, blood components, and integrity of the small intestine epithelia; nevertheless, pigs show signs of recovery at 7 d of HS exposure.