Exosomes isolated from IMMUNEPOTENT CRP, a hemoderivative, to accelerate diabetic wound healing.

The increasing risk of amputation due to diabetic foot ulcer calls for new therapeutic options; for that, we determined the role of IMMUNEPOTENT CRP (ICRP) and its parts in the wound healing process of superficial wounds in diabetic BALB/c mice. A potency test was performed to confirm the batch of ICRP, and then its parts were separated into pellets, supernatants, and exosomes, and another group of exosomes loaded with insulin was added. Viability and scratch healing were assessed in NIH-3T3, HUVEC, and HACAT cell lines. Diabetes was induced with streptozotocin, and wounds were made by dissecting the back skin. Treatments were topically applied, and closure was monitored; inflammatory cytokines in sera were also evaluated by flow cytometry, and histological analysis was performed by Masson's staining and immunohistochemistry for p-AKT, p-FOXO, p-P21, and p-TSC2. ICRP pellets and exosomes increased cellular viability, and exosomes and exosome-insulin accelerated scratch healing in vitro. Exosome-insulin releases insulin constantly over time in vitro. In vivo, treatments accelerated wound closure, and better performance was observed in pellet, exosome, and exosome-insulin treatments. Best collagen expression was induced by ICRP. P-AKT and p-FOXO were overexpressed in healing tissues. Inflammatory cytokines were downregulated by all treatments. In conclusion, IMMUNEPOTENT CRP components, especially exosomes, and the process of encapsulation of exosome-insulin accelerate diabetic wound healing and enhance cellular proliferation, collagen production, and inflammation modulation through the phosphorylation of components of the AKT pathway.

Antibacterial efficacy of novel bismuth-silver nanoparticles synthesis on Staphylococcus aureus and Escherichia coli infection models.

Introduction: The emergence of multi-drug-resistant bacteria is one of the main concerns in the health sector worldwide. The conventional strategies for treatment and prophylaxis against microbial infections include the use of antibiotics. However, these drugs are failing due to the increasing antimicrobial resistance. The unavailability of effective antibiotics highlights the need to discover effective alternatives to combat bacterial infections. One option is the use of metallic nanoparticles, which are toxic to some microorganisms due to their nanometric size. Methods: In this study we (1) synthesize and characterize bismuth and silver nanoparticles, (2) evaluate the antibacterial activity of NPs against Staphylococcus aureus and Escherichia coli in several infection models (in vivo models: infected wound and sepsis and in vitro model: mastitis), and we (3) determine the cytotoxic effect on several cell lines representative of the skin tissue. Results and discussion: We obtained bimetallic nanoparticles of bismuth and silver in a stable aqueous solution from a single reaction by chemical synthesis. These nanoparticles show antibacterial activity on S. aureus and E. coli in vitro without cytotoxic effects on fibroblast, endothelial vascular, and mammary epithelium cell lines. In an infected-wound mice model, antibacterial effect was observed, without effect on in vitro mastitis and sepsis models.

Unveiling the role of hydroxyapatite and hydroxyapatite/silver composite in osteoblast-like cell mineralization: An exploration through their viscoelastic properties.

Mechanical properties are becoming fundamental for advancing the comprehension of cellular processes. This study addresses the relationship between viscoelastic properties and the cellular mineralization process. Osteoblast-like cells treated with an osteogenic medium were employed for this purpose. Additionally, the study explores the impact of hydroxyapatite (HA) and hydroxyapatite/silver (HA/Ag) composite on this process. AFM relaxation experiments were conducted to extract viscoelastic parameters using the Fractional Zener (FZ) and Fractional Kelvin (FK) models. Our findings revealed that the main phases of mineralization are associated with alterations in the viscoelastic properties of osteoblast-like cells. Furthermore, HA and HA/Ag treatments significantly influenced changes in the viscoelastic properties of these cells. In particular, the HA/Ag treatment demonstrated a marked enhancement in cell fluidity, suggesting a possible role of silver in accelerating the mineralization process. Moreover, the study underscores the independence observed between fluidity and stiffness, indicating that modifications in one parameter may not necessarily correspond to changes in the other. These findings shed light on the factors involved in the cellular mineralization process and emphasize the importance of using viscoelastic properties to discern the impact of treatments on cells.

Immunotherapies inducing immunogenic cell death in cancer: insight of the innate immune system.

Cancer immunotherapies include monoclonal antibodies, cytokines, oncolytic viruses, cellular therapies, and other biological and synthetic immunomodulators. These are traditionally studied for their effect on the immune system's role in eliminating cancer cells. However, some of these therapies have the unique ability to directly induce cytotoxicity in cancer cells by inducing immunogenic cell death (ICD). Unlike general immune stimulation, ICD triggers specific therapy-induced cell death pathways, based on the release of damage-associated molecular patterns (DAMPs) from dying tumour cells. These activate innate pattern recognition receptors (PRRs) and subsequent adaptive immune responses, offering the promise of sustained anticancer drug efficacy and durable antitumour immune memory. Exploring how onco-immunotherapies can trigger ICD, enhances our understanding of their mechanisms and potential for combination strategies. This review explores the complexities of these immunotherapeutic approaches that induce ICD, highlighting their implications for the innate immune system, addressing challenges in cancer treatment, and emphasising the pivotal role of ICD in contemporary cancer research.

Efecto aditivo in vitro de Chlorella sorokiniana en combinación con Vincristina sobre la inhibición del crecimiento de células de cáncer de colon HT-29 / In vitro additive effect of Chlorella sorokiniana in combination withVincristine on HT-29 colon cancer cells growth inhibition

Objetivo: Evaluar el efecto in vitro de la combinación deChlorella sorokiniana con Vincristina contra el crecimiento decélulas de cáncer de colon HT-29.Material y método:Chlorella sorokiniana se cultivó enmedio López-Chuken. El efecto inhibitorio de la microalga solay en combinación con Vincristina en el crecimiento tumoral seevaluó mediante la técnica de MTT, contra células de cáncerde colon humano HT-29, y se analizó mediante el softwareSynergyFinder 2.0.Resultados: El crecimiento Chlorella sorokiniana fue cons-tante al día 28 a una temperatura de 34 oC ± 3 oC. El efectoinhibitorio de Vincristina sobre células HT-29 fue del 60% apartir de 0.0037μg/mL. La inhibición por Chlorella sorokinianafue del 60% al 80% a las concentraciones de 106-108.Además, la combinación de Vincristina/Chlorella inhibió el cre-cimiento tumoral entre 70% y 90%, siendo la concentraciónmenor de Chlorella la que mostró un mejor efecto en combi-nación con Vincristina. El análisis de los resultados enSynergyFinder mostró un score de -0.708, determinando unefecto aditivo. Conclusión:Chlorella sorokiniana presenta un efecto adi-tivo en combinación con Vincristina contra la línea de cáncerde colon humano HT-29. La suplementación de C. sorokinianaen la dieta de pacientes con cáncer de colon podría mejorarsu tratamiento y por consecuencia su recuperación (AU)

Objective:To evaluate in vitro the effect of the combina-tion of Chlorella sorokiniana with vincristine on HT-29 coloncancer cells.Material and method:Chlorella sorokininana growth wasconstant on day 28 at a temperature of 34 oC ± 3 oC. Chlorellasorokiniana was cultured in López-Chuken medium. HT-29 cellsgrowth inhibition by the microalga alone or in combination withvincristine was evaluated by the colorimetric reduction MTT as-say, and analyzed using the SynergyFinder 2.0 software. Results:The inhibitory effect of Vincristine on HT-29 cellswas 60% from 0.0037μg/mL. Tumor cells growth inhibition by106 to 108 Chlorella sorokiniana cells ranged from 60% to80%. The combination of vincristine and Chlorella inhibitedtumor cells growth from 70% to 90%, being the lower con-centration of Chlorella the one that showed a better effect incombination with vincristine. The analysis of the results inSynergyFinder showed a score of -0.708, determining an ad-ditive effect.Conclusion: Chlorella sorokiniana has an additive effect incombination with vincristine against the human colon cancerline HT-29. Supplementation of C. sorokiniana in the diet ofpatients with colon cancer may improve their treatment andrecovery (AU)

CAR-T Cell Therapy: From the Shop to Cancer Therapy.

Cancer is a worldwide health problem. Nevertheless, new technologies in the immunotherapy field have emerged. Chimeric antigen receptor (CAR) technology is a novel biological form to treat cancer; CAR-T cell genetic engineering has positively revolutionized cancer immunotherapy. In this paper, we review the latest developments in CAR-T in cancer treatment. We present the structure of the different generations and variants of CAR-T cells including TRUCK (T cells redirected for universal cytokine killing. We explain the approaches of the CAR-T cells manufactured ex vivo and in vivo. Moreover, we describe the limitations and areas of opportunity for this immunotherapy and the current challenges of treating hematological and solid cancer using CAR-T technology as well as its constraints and engineering approaches. We summarize other immune cells that have been using CAR technology, such as natural killer (NK), macrophages (M), and dendritic cells (DC). We conclude that CAR-T cells have the potential to treat not only cancer but other chronic diseases.

Systemic Delivery of Magnetogene Nanoparticle Vector for Gene Expression in Hypoxic Tumors.

Cancer is a disease that causes millions of deaths per year worldwide because conventional treatments have disadvantages such as unspecific tumor selectivity and unwanted toxicity. Most human solid tumors present hypoxic microenvironments and this promotes multidrug resistance. In this study, we present "Magnetogene nanoparticle vector" which takes advantage of the hypoxic microenvironment of solid tumors to increase selective gene expression in tumor cells and reduce unwanted toxicity in healthy cells; this vector was guided by a magnet to the tumor tissue. Magnetic nanoparticles (MNPs), chitosan (CS), and the pHRE-Luc plasmid with a hypoxia-inducible promoter were used to synthesize the vector called "Magnetogene nanoparticles" by ionic gelation. The hypoxic functionality of Magnetogene vector nanoparticles was confirmed in the B16F10 cell line by measuring the expression of the luciferase reporter gene under hypoxic and normoxic conditions. Also, the efficiency of the Magnetogene vector was confirmed in vivo. Magnetogene was administered by intravenous injection (IV) in the tail vein and directed through an external magnetic field at the site of tumor growth in C57Bl/6 mice. A Magnetogene vector with a size of 50 to 70 nm was directed and retained at the tumor area and gene expression was higher at the tumor site than in the others tissues, confirming the selectivity of this vector towards hypoxic tumor areas. This nanosystem, that we called the "Magnetogene vector" for systemic delivery and specific gene expression in hypoxic tumors controlled by an external magnetic designed to target hypoxic regions of tumors, can be used for cancer-specific gene therapies.

p16INK4a and pRb expression in laryngeal squamous cell carcinoma with and without infection by EBV or different genotypes of HPV: a retrospective study.

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) represents one of the principal tumors of the head and neck. Human papillomavirus (HPV) and Epstein-Barr virus (EBV) are considered risk factors for the development and the clinical prognosis of LSCC. High levels of p16INK4a are suggested as a surrogate marker of HPV or EBV infection in some head and neck tumors but in LSCC is still controversial. Furthermore, pRb expression may be considered an additional biomarker but it has not been clearly defined. This work aimed to compare the expression of pRb and p16INK4a as possible biomarkers in tumor tissues with and without infection by EBV or different genotypes of HPV from patients with LSCC. METHODS: Tumor samples from 103 patients with LSCC were previously investigated for the presence and genotypes of HPV using the INNO-LiPA line probe assay and for the infection of EBV by qPCR. p16 INK4a and pRb expression was assessed by immunohistochemistry. RESULTS: Of the 103 tumor samples, expression of p16INK4a was positive in 55 (53.4%) and of this, 32 (56.1%) were positive for HPV whereas 11 (39.3%) were EBV positive but both without a significantly difference (p > 0.05). pRb expression was positive in 78 (75.7%) and a higher frequency of this expression was observed in HPV negative samples (87.0%) (p = 0.021) and in high-risk HPV negative samples (85.2%) (p = 0.010). No difference was observed when comparing pRb expression and EBV infection status (p > 0.05). CONCLUSION: Our results support the suggestion that p16INK4a is not a reliable surrogate marker for identifying HPV or EBV infection in LSCC. On the other hand, most of our samples had pRb expression, which was more frequent in tumors without HPV, suggesting that pRb could indicate HPV negativity. However, more studies with a larger number of cases are required, including controls without LSCC and evaluating other molecular markers to determine the real role of p16INK4a and pRb in LSCC.

Factores asociados a sobrevida a un año en pacientes postoperados de glioblastoma.

OBJECTIVE: To identify factors associated with one-year survival in postoperative glioblastoma patients at a hospital in northeastern Mexico. MATERIAL AND METHODS: Nested case-control study. Patients operated on for glioblastoma between 2016-2019 were included. Information about clinical and surgical factors was obtained, survival was calculated by Kaplan-Meier analysis. Descriptive analysis was performed with medians and ranges, and inferential analysis with χ2, Fisher and Student t test, odds ratio and 95% confidence interval. A value of p < 0.05 was considered significant. RESULTS: Sixty-two patients with glioblastoma were included, 27 (43.5%) women and 35 (56.5%) men, median age 56 years (range: 6-83). Median survival was 3.6 months (1-52), 45 (72.6%) survived less than 12 months. The factors associated with a higher survival were administration of adjuvant treatment (p < 0.001), better functional status (p = 0.001), and absence of post-surgical complications (p = 0.034). CONCLUSIONS: Most patients with glioblastoma survive less than 12 months and the factors most strongly associated with longer survival are administration of adjuvant treatment, better functional status of the patient and absence of post-surgical complications.

OBJETIVO: Identificar los factores asociados a la sobrevida a un año en pacientes postoperados de glioblastoma en un hospital del noreste de México. MATERIAL Y MÉTODOS: Estudio de casos y controles anidado en una cohorte. Se incluyeron pacientes operados de glioblastoma entre 2016 y 2019. Se obtuvo la información sobre factores clínicos y quirúrgicos, se calculó la sobrevida mediante análisis de Kaplan-Meier. El análisis descriptivo se realizó con medianas y rangos, y el inferencial con prueba de χ2, Fisher, t de Student, razón de momios e intervalo de confianza al 95%. Se consideró significativo un valor de p < 0.05. RESULTADOS: Se incluyeron 62 pacientes con glioblastoma, 27 (43.5%) mujeres y 35 (56.5%) hombres, mediana de edad de 56 años (rango: 6-83). La mediana de sobrevida fue de 3.6 meses (1-52), 45 (72.6%) sobrevivieron menos de 12 meses. Los factores asociados a mayor sobrevida fueron: administración de tratamiento adyuvante (p < 0.001), mejor estado funcional (p = 0.001) y ausencia de complicaciones posquirúrgicas (p = 0.034). CONCLUSIONES: La mayoría de los pacientes con glioblastoma sobreviven menos de 12 meses y los factores más fuertemente asociados a mayor sobrevida son administración de tratamiento adyuvante, mejor estado funcional del paciente y ausencia de complicaciones posquirúrgicas.

Antitumoral and Immunogenic Capacity of ß-D-Glucose-Reduced Silver Nanoparticles in Breast Cancer.

Immunogenic cell death (ICD) is a type of cell death capable of stimulating immunity against cancer through danger signals that lead to an adaptive immune response. Silver nanoparticles (AgNPs) have been shown to have a cytotoxic effect on cancer cells; however, their mechanism of action is not fully understood. The present study synthesized, characterized, and evaluated the cytotoxic effect of beta-D-glucose-reduced AgNPs (AgNPs-G) against breast cancer (BC) cells in vitro; and assess the immunogenicity of cell death in vitro and in vivo. The results showed that AgNPs-G induce cell death in a dose-dependent manner on BC cell lines. In addition, AgNPs show antiproliferative effects by interfering with the cell cycle. Regarding the detection of damage-associated molecular patterns (DAMPs), it was found that treatment with AgNPs-G induces calreticulin exposure and the release of HSP70, HSP90, HMGB1, and ATP. In vivo, prophylactic vaccination did not prevent tumor establishment; however, tumor weight was significantly lower in AgNPs-G vaccinated mice, while the survival rate increased. In conclusion, we have developed a new method for the synthesis of AgNPs-G, with in vitro antitumor cytotoxic activity on BC cells, accompanied by the release of DAMPs. In vivo, immunization with AgNPs-G failed to induce a complete immune response in mice. Consequently, additional studies are needed to elucidate the mechanism of cell death that leads to the design of strategies and combinations with clinical efficacy.

Putative Wound Healing Induction Functions of Exosomes Isolated from IMMUNEPOTENT CRP.

Chronic wounds in diabetic patients can take months or years to heal, representing a great cost for the healthcare sector and impacts on patients' lifestyles. Therefore, new effective treatment alternatives are needed to accelerate the healing process. Exosomes are nanovesicles involved in the modulation of signaling pathways that can be produced by any cell and can exert functions similar to the cell of origin. For this reason, IMMUNEPOTENT CRP, which is a bovine spleen leukocyte extract, was analyzed to identify the proteins present and is proposed as a source of exosomes. The exosomes were isolated through ultracentrifugation and shape-size, characterized by atomic force microscopy. The protein content in IMMUNEPOTENT CRP was characterized by EV-trap coupled to liquid chromatography. The in silico analyses for biological pathways, tissue specificity, and transcription factor inducement were performed in GOrilla ontology, Panther ontology, Metascape, and Reactome. It was observed that IMMUNEPOTENT CRP contains diverse peptides. The peptide-containing exosomes had an average size of 60 nm, and exomeres of 30 nm. They had biological activity capable of modulating the wound healing process, through inflammation modulation and the activation of signaling pathways such as PIP3-AKT, as well as other pathways activated by FOXE genes related to specificity in the skin tissue.

IMMUNEPOTENT CRP increases intracellular calcium through ER-calcium channels, leading to ROS production and cell death in breast cancer and leukemic cell lines.

IMMUNEPOTENT CRP (ICRP) is an immunotherapy that induces cell death in cancer cell lines. However, the molecular mechanisms of death are not completely elucidated. Here, we evaluated the implication of intracellular Ca2+ augmentation in the cell death induced by ICRP on T-ALL and breast cancer cell lines. Cell death induction and the molecular characteristics of cell death were evaluated in T-ALL and breast cancer cell lines by assessing autophagosome formation, ROS production, loss of mitochondrial membrane potential, ER stress and intracellular Ca2+ levels. We assessed the involvement of extracellular Ca2+, and the implication of the ER-receptors, IP3R and RyR, in the cell death induced by ICRP, by using an extracellular calcium chelator and pharmacological inhibitors. Our results show that ICRP increases intracellular Ca2+ levels as the first step of the cell death mechanism that provokes ROS production and loss of mitochondrial membrane potential. In addition, blocking the IP3 and ryanodine receptors inhibited ER-Ca2+ release, ROS production and ICRP-induced cell death. Taken together our results demonstrate that ICRP triggers intracellular Ca2+-increase leading to different regulated cell death modalities in T-ALL and breast cancer cell lines. See also Figure 1(Fig. 1).

In Vitro Biological Activity and Lymphoma Cell Growth Inhibition by Selected Mexican Medicinal Plants.

Cancer is a major health problem with significant morbidity and mortality. In addition, plants are a source of metabolites with diverse biological properties, including antitumor potential. In this study, we investigated the in vitro murine lymphoma L5178Y-R cell growth inhibition, human peripheral blood mononuclear cells (PBMC) toxicity and proliferation, and antioxidant, hemolytic, and anti-hemolytic activities of methanol extracts from 15 plants of traditional use in Mexico. Justicia spicigera caused the highest tumor cell growth inhibition with a half maximal inhibitory concentration (IC50) of 29.10 µg/mL and a selectivity index >34.36 compared with those of PBMC, whereas Mimosa tenuiflora showed the highest lymphoproliferative activity from 200 µg/mL compared with that induced by concanavalin A. In addition, M. tenuiflora showed an antioxidant effect (IC50 = 2.86 µg/mL) higher than that of ascorbic acid. Regarding the hemolytic and anti-hemolytic activity, all extracts presented significant anti-hemolytic activity. The extract of J. spicigera is emerging as a possible source of effective antineoplastic compounds.

Design, Synthesis, and Evaluation of Peptides Derived from L1 Protein Against Bovine Papillomavirus-1/2 Identified Along Mexico's Cattle Export Route.

Introduction: Bovine papillomatosis affects animal health and represents one of the greatest economic losses in the livestock sector. New control and prevention methods to protect the livestock industry from this disease are necessary. The aim of this study was to evaluate a candidate peptide for antibody production against bovine papillomavirus (BPV). Material and Methods: A total of 64 cattle underwent wart excision among 5,485 cattle distributed over 2 to 4 farms per state and 12 farms in total in the four Mexican states of Tabasco, Chiapas, Veracruz, and Nuevo León. The prevalence of bovine papillomatosis per farm was calculated by wart visualisation. The warts were genotyped by PCR and sequenced, then a phylogenetic tree was built using MEGA X software. A synthetic peptide was designed in the ABCpred, Bepipred 2.0, Bepipred IDBT, Bepitope, LBtope, and MHC II predictor online server software's based on the C-terminal region of the L1 protein. Mice antibody production was induced by subcutaneous immunisation with 50 µg of synthetic peptide and evaluated by indirect ELISA. Results: The prevalence of BPV was higher in Tabasco, Chiapas, and Veracruz. Bovine papillomaviruses 1 and 2 were found in all representative samples. A phylogenetic tree showed that Mexican sequences were located in exclusive clades yet were highly related to international ones. The peptide immunisation induced antibody titres of 1 : 10,000/1 : 1,000,000 against synthetic peptide and whole wart lysate (WWL), respectively. Conclusion: Co-infections of BPV-1 and -2 were found in all four states. Immunisation of BALB/C mice with BPV-1/2-derived synthetic peptide based on the C-terminal region of the major viral capsid protein L1 induced the production of specific antibodies able to recognise BPV-1/2 viral particles from bovine WWL.

The bovine dialyzable leukocyte extract, immunepotent CRP, synergically enhances cyclophosphamide-induced breast cancer cell death, through a caspase-independent mechanism.

Breast cancer (BC) is one of the leading causes of cancer death worldwide. Cyclophosphamide (CTX) remains a mainstay in cancer therapy despite harmful adverse effects and cell death-resistances. To face this, combinational therapy of chemotherapies and immunotherapies has been proposed. IMMUNEPOTENT CRP (ICRP) is an immunotherapy that has cytotoxic effects in several cancer cells without affecting peripheral blood mononuclear cells (PBMC) and CD3+ cells. The aim of this study was to evaluate cytotoxicity, the type of cytotoxic effect, and several features involved in cell death induced by the combination of CTX with ICRP (ICRP+CTX) in breast cancer cells as well as their effect on healthy cells. For this purpose, human and murine breast cancer cells, MCF-7, MDA-MB-231 and 4T1, or PBMC were treated for 24 hours with ICRP, CTX or ICRP+CTX in different combination ratios for the assessment of cell death. Flow cytometry and microscopy were used to determine biochemical and morphological characteristics of cell death. Assays showed that ICRP in combination with CTX induce potentiated cell death manifested with morphological changes, loss of mitochondrial membrane potential, reactive oxygen species (ROS) production, and caspase activation. In addition, it was determined that ICRP+CTX-cell death is caspase-independent in all the breast cancer cells assessed. On the other hand, ICRP did not affect CTX-cytotoxicity in PBMC. For all the above, we can propose that the combination of ICRP with CTX an effective combination therapy, promoting their use even in tumoral cells with defects on proteins implicated in the apoptotic pathway.

Development of a new generation of miniemulsion based on cottonseed oil with α-tocopherol and ZnO and evaluation of its adjuvant activity.

Background: Emulsions have been widely used as immunological adjuvants. But the use of materials derived from plants such as cottonseed oil, alpha-tocopherol, or minerals such as zinc, as well as their use at the nanometric scale has been little explored. In this study, we develop a new miniemulsion and evaluated its antioxidant and phagocytic capacity, as well as parameters related to immune response stimulation by cytokine expression and antibodies production in a mice model. Methods: Formulated CN (cottonseed oil miniemulsion) and CNZ (cottonseed oil miniemulsion whit zinc oxide nanoparticles) miniemulsions were characterized by scanning electronic microscopy SEM, DLS and FT-IR. In murine macrophages, splenocytes and thymocytes primary cultures safety and cytotoxicity were determined by MTT. In macrophages the antioxidant and phagocytic capacity was evaluated. In BALB/c mice, the stimulation of the immune system was determined by the expression of cytokines and the production of antibodies. Results: The CN and CNZ presented stability for 90 days. Immediately after preparation, the CN presented a higher particle size (543.1 nm) than CNZ (320 nm). FT-IR demonstrated the correct nanoparticle synthesis by the absence of sulfate groups. CN and CNZ (1.25 to 10 µL/mL) had no toxic effect on macrophages (p = 0.108), splenocytes (p = 0.413), and thymocytes (p = 0.923). All CN and CNZ doses tested induced nitric oxide and antioxidants production in dose dependent manner when compared with control. CN-ovalbumin and CNZ-ovalbumin treatments in femoral subcutaneous tissue area showed inflammation with higher leukocyte infiltration compared with FCA. The intraperitoneal administration with CN, CNZ, and FCA showed a higher total intraperitoneal cells recruitment (CD14+) after 24 h of inoculation than control (p = 0.0001). CN and CNZ increased the phagocyte capacity with respect to untreated macrophages in the Candida albicans-phagocytosis assay. The evaluation of residual CFU indicated that only CN significantly decreased (p = 0.004) this value at 3 h. By other side, only CN increased (p = 0.002) the nitric oxide production. CNZ stimulated a major INFγ secretion compared with FCA at day 7. A major IL-2 secretion was observed at days 7 and 14, stimulated with CN and CNZ. Both miniemulsions did not affect the antibody isotypes production (IgG1, IgG2a, IgG3, IgA and IgM) at days 7, 14, 28, and 42. CN induced a significant IgG production against OVA, but lesser than FCA. Conclusions: The two new miniemulsions with adjuvant and antioxidant capacity, were capable of generating leukocyte infiltration and increased cytokines and antibodies production.

Effect of Culture Conditions of Lophocereus marginatus Endophytic Fungi on Yield and Anticancer and Antioxidant Activities.

Culture conditions affect the production of secondary metabolites in endophytic fungi. Therefore, the aim of the present study was to evaluate the yield and anticancer and antioxidant activity of endophytic fungi extracts from the cactus Lophocereus marginatus, under different culture conditions. The strains Penicillium citrinum, Aspergillus versicolor, Metarhizium anisopliae, and Cladosporium sp. were fermented in different culture media (potato dextrose agar, Czapeck broth, and malt broth), types of inoculums (spore or mycelium), and shaking conditions (150 rpm or static) for one week. Methanol extracts were obtained from mycelia, which was followed by determining their yields and evaluating their effect on L5178Y-R murine lymphoma cells growth and human peripheral blood mononuclear cells (PBMCs) viability, using the 3-[4,5dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide reduction colorimetric assay. In addition, antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl test. We determined the half-maximal inhibitory concentration (IC50) values of tumor cell growth inhibition, the selectivity index (SI), and the antioxidant activity, as compared with the healthy cells control. The best yields were obtained with the Czapeck broth medium in all the evaluated strains, reaching values of 50.3%. Of the 48 extracts evaluated, only seven significantly (p < 0.01) inhibited tumor cell growth (IC50 < 250 µg/mL). A. versicolor extract showed the highest anticancer activity, after culturing spores (IC50 = 49.62 µg/mL; SI = 15.8) or mycelium (IC50 = 69.67 µg/mL; SI = 12.2) in malt broth, under static conditions. Extracts did not present significant antioxidant activity. In conclusion, we showed that culture conditions influenced the anticancer activity of L. marginatus endophytic fungi.

Methodologies and models for measuring viscoelastic properties of cancer cells: Towards a universal classification.

Different methods and several physical models exist to study cell viscoelasticity with the atomic force microscope (AFM). In search of a robust mechanical classification of cells through AFM, in this work, viscoelastic parameters of the cancer cell lines MDA-MB-231, DU-145, and MG-63 are obtained using two methodologies; through force-distance and force-relaxation curves. Four mechanical models were applied to fit the curves. The results show that both methodologies agree qualitatively on the parameters that quantify elasticity but disagree on the parameters that account for energy dissipation. The Fractional Zener (FZ) model represents well the information given by the Solid Linear Standard and Generalized Maxwell models. The Fractional Kelvin (FK) model concentrates the viscoelastic information mainly in two parameters, which could be an advantage over the other models. Therefore, the FZ and FK models are proposed as the basis for the classification of cancer cells. However, more research using these models is needed to obtain a broader view of the meaning of each parameter and to be able to establish a relationship between the parameters and the cellular components.

Molecular epidemiology and drug resistance of Mycobacterium tuberculosis in a tertiary care hospital in northeastern Mexico.

INTRODUCTION: Tuberculosis (TB) is a re-emerging disease considered a public health concern. In the present study, we analyzed the epidemiology and drug resistance of Mycobacterium tuberculosis strains isolated from patients with pulmonary TB. METHODOLOGY: Mycobacterium tuberculosis isolates (n = 190) were obtained from patients with pulmonary TB admitted to Dr. José Eleuterio González University Hospital (UH). Each M. tuberculosis isolate was analyzed by spoligotyping (spacer oligonucleotide typing) and MIRU-VNTR (Mycobacterial Interspersed Repetitive Units-Variable Number Tandem Repeat). Drug resistance was evaluated using the Anyplex™ II MTB/MDR/XDR assay. RESULTS: The predominant spoligotypes observed were X1 (SIT 119, n = 46), T1 (SIT 53, n = 40), H3 (SIT 50, n = 13), Beijing (SIT 1, n = 11), and EAI2-Manila (SIT 19, n = 8). MIRU-VNTR analysis showed that the locus QUB-26 had the highest allelic variability. The observed drug resistance included monoresistance to rifampicin (2.6%; n = 5), isoniazid (3.2%; n = 6), and fluoroquinolones (1.6%; n = 3) as well as multidrug resistance (5.3%; n = 10). All of the Beijing strains were susceptible. Regarding comorbidities, 13.7% (26/190) of the patients were co-infected with TB and HIV (TB+HIV+), and 31.6% (55/190) had TB along with diabetes (TB + diabetes). CONCLUSIONS: The most prevalent lineages were X1 (SIT 119; 24.3%) and T1 (SIT 53; 21%). An alarming proportion (12.6%) of M. tuberculosis isolates presented drug resistance. To effectively manage TB, continuous surveillance of regional strain dissemination, drug resistance profiles, and TB-associated comorbidities is crucial.

Antitumor efficacy of silver nanoparticles reduced with ß-D-glucose as neoadjuvant therapy to prevent tumor relapse in a mouse model of breast cancer.

Introduction: Neoadjuvant therapy constitutes a valuable modality for diminishing tumor volume prior to surgical resection. Nonetheless, its application encounters limitations in the context of recurrent tumors, which manifest resistance to conventional treatments. Silver nanoparticles (AgNPs) have emerged as a promising alternative for cancer treatment owing to their cytotoxic effects. Methods: Cellular viability was assessed by Alamar blue assay in 4T1 breast cancer cell line. Silver biodistribution was detected by an inductively coupled plasma optical emission spectrometer in an in vivo mice model. For neoadjuvant evaluation, mice were randomized and treated intratumoral with AgNPs-G or intraperitoneally with doxorubicin (DOX) as a control. Recurrence was determined after 170 days by counting lung metastatic nodules (dyed with Bouin solution) with histological confirmation by H&E. Masson's stain, Ki67 immunohistochemistry, and a TUNEL assay were performed in lungs from treated mice. Results: AgNPs-G reduced 4T1 cell viability and in an ex vivo assay the AgNPs-G decreased the tumor cell viability. After intravenous administration of AgNPs-G were detected in different organs. After intratumor administration, AgNPs-G are retained. The AgNPs-G treatment significantly reduced tumor volume before its surgical resection. AgNPs-G reduced the development of lung metastatic nodules and the expression of Ki67. TUNEL assay indicated that AgNPs-G didn't induce apoptosis. Conclusions: We concluded that intratumor administration of AgNPs-G reduced tumor volume before surgical resection, alongside a reduction in lung metastatic nodules, and Ki67 expression. These findings provide valuable insights into the AgNPs-G potential for intratumor and neoadjuvant cancer therapies. However, further research is needed to explore their full potential and optimize their use in clinical settings.