ABSTRACT Purpose: To evaluate the molecular evolution of endoplasmic reticulum (ER) stress during colorectal cancer carcinogenesis. Methods: Fifty-six hairless mice were divided into two groups: control (no intervention); and carcinogenesis (treated with two doses of azoxymethane at 10 mg/kg during the third and the fourth week and dextran sodium sulfate at 2.5% for seven days in the second, fifth, and eighth week). Euthanasia occurred at the fifth, 10th, 15th, and 20th week. Colons were collected, and gene expression of ER stress markers (IRE1-α, PERK, ATF6, and CHOP) was assessed via real-time quantitative polymerase chain reaction. Results: ERN1 expression was significantly higher than the control at the 10th week (1.39 ± 0.16, p 0.05) and the 20th week (1.15 ± 0.04, p 0.01). ATF6 also showed elevated expression, significantly different at the 10th week (1.71 ± 0.29, p 0.05) and the 20th week (1.14 ± 0.06, p 0.05). PERK and CHOP expressions were significantly higher than the control in the 15th (PERK = 1.30 ± 0.12, CHOP = 1.48 ± 0.23) and 20th weeks (PERK = 1.63 ± 0.20, CHOP = 1.67 ± 0.22, p 0.05). Conclusion: Upregulation of IRN1, PERK, ATF6, and CHOP demonstrates a strong ER stress response during colorectal cancer development.

Abstract Background: The study was to investigate circRBM33 in septic acute lung injury (ALI). Methods: Treatment of Murine Lung Epithelial-12 cells (MLE-12) cells was performed using 10 ng/mL Lipopolysaccharide (LPS). circRBM33, miR-15a-5p, and Enhancer of zeste homolog 1 (EZH1) were ascertained through RT-qPCR or Western blot analysis. The viability of MLE-12 cells was measured using the MTT assay, and their rate of apoptosis was ascertained through flow cytometry. B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X (Bax) were determined using Western blot analysis. Oxidative stress levels were assessed with ELISA kits, and levels of malondialdehyde(MDA) content, Superoxide Dismutase (SOD) activity, and glutathione (GSH) were detected. Dual luciferase reporter gene and RIP assays verified the targeting link between miR-15a-5p and circRBM33 or EZH1. The role of circRBM33 in ALI in vivo was determined by performing cecum ligationperforation (CLP) surgery. HE staining, W/D pulmonary edema, and histological damage scores were taken to assess the extent of lung tissue damage. ELISA was performed to determine proinflammatory factors in lung tissue and cells. Results: CircRBM33 downregulation ameliorated ALI-induced edema, apoptotic, and inflammatory reactions in mouse lung tissues. In addition, apoptosis and inflammation mediated by LPS in MLE-12 cells were ameliorated by circRBM33 downregulation, whereas miR-15a-5p knockdown or EZH1 elevation eliminated the action of silencing circRBM33. circRBM33 mediated EZH1 expression by competitive adsorption of miR-15a-5p. Conclusion: CircRBM33 improves ALI in septic mice by targeting the miR-15a-5p/EZH1 axis.

ABSTRACT Trypanosoma cruzi infection involves transmission of metacyclic trypomastigotes through injured skin or mucosa via contaminated feces from insect vectors like Triatoma dimidiata (Latreille, 1811). Currently, there is insufficient information describing the immune response to feces naturally contaminated with metacyclic trypomastigotes. Mice subcutaneously inoculated with tissue-culture derived trypomastigotes (TCT) or T. dimidiata feces containing metacyclic trypomastigotes (MT) or previously multi-exposed (ME) with feces without metacyclic trypomastigotes and then infected with feces containing metacyclic parasites or only T. dimidiata feces (F) was studied from 15 min to three months post-infection. PCR detection of parasite DNA at the inoculation site demonstrated persistence of T. cruzi DNA up to 20 days in MT and TCT but disappeared earlier in the ME test group. A rapid spread of T. cruzi DNA to regional lymph nodes was observed in all experimental groups. A lower amount of amastigote nests in the heart with concomitant intense inflammation was noticed in ME mice in comparison to the MT group. CD4 + T cell subtypes at popliteal lymph nodes shows early Th1 and Th17 responses at seven days in ME mice, whereas Th1, Th17 and Treg predominate in MT mice after three weeks, and feces induces Th1, Th17 and Treg at a later stage. Our study shows that previous exposure to feces prior to infection with T. cruzi helps control parasitism at the inoculation site and in heart tissue, and an early induction of Th1 and Th17 T cell subtypes.

Abstract Drug addiction is one of the most important global problems. Medicinal herbs have been traditionally used in the management and treatment of opioid withdrawal syndrome and pain. The aim of this study was to determine the effect of A. dracunculus essential oil in reducing the symptoms of morphine withdrawal in mice. Male mice (25-30 g) were randomly assigned into 4 groups (n=10). Morphine-dependent groups received morphine (50 and 75 mg/kg ip; three times/day, 3 days) and a single injection of morphine (50 mg/kg) and then naloxone on the fourth day via IP injection. The control group received saline. The post-treated group received morphine for 3 days; on the fourth day, ten minutes before receiving naloxone, A. dracunculus essential oil with a dose of 75 mg/kg was injected. All groups received naloxone 2 hours after receiving the last dose of morphine; then, the morphine withdrawal symptoms were measured for 30 minutes. In the post-treated and co-treated groups, the body stretching and shaking the claw were significantly less than morphine-dependent groups (p<0/05). Also, in the post-treated and co-treated groups, the blinking, itching, and the number of standing on two legs significantly decreased compared to the morphine group. Hence, it might be concluded that A. dracunculus essential oil can significantly reduce morphine withdrawal symptoms.

Abstract To evaluate the neuropharmacological profile of new spiro thiazepinone and thiazolidinone compounds in CD1 mice after a computer-aided virtual screening based on a GABA-A/BZD site. From a library of 240 pyrazolo[1,4]thiazepin-3-ones and 39 pyrimidinyl thiazolidin-4-ones, two molecular prototypes of each series were selected by virtual screening using the rank consensus molecular docking approach, based on the GABA-A/BZD site. These compounds, coded as cpTP-0, cpTP-1, TAP-2 and cpTAP-2, were synthesised by multicomponent reactions and evaluated by neuropharmacological screening in CD1 mice (100 mg/kg, p.o.). The study revealed that cpTAP-2 exhibited a significant reduction of immobility time during the forced swimming test (FST), while it did not show any major effects in the rota-rod, open field, plus maze, tail suspension, pentylenetetrazole seizures, and barbiturate sleeping time tests. In a dose-response evaluation, cpTAP-2 reduced immobility time during forced swimming test in CD1 male mice at doses of 100 and 300 mg/kg with biologically relevant effect sizes. These results suggest that cpTAP-2 could elicit antidepressant effects possibly related to GABA-A/benzodiazepine site, although other mechanisms could be implicated.

Abstract Specific Pathogen-Free (SPF) animals are bred and maintained to exclude pathogens associated with significant morbidity or mortality, which may pose a risk to research replicability. The BALB/c strain is distributed globally and is among the most commonly used inbred strains in immunology and infectious disease research. Despite being a widely distributed bacterium that causes chronic infection, Bartonella henselae infection has not been investigated in any protocol that characterizes SPF animals. The objective of this study was to investigate the potential natural infection of laboratory animals of the BALB/c lineage by B. henselae. To achieve this, ten immunocompetent BALB/c mice were obtained directly from the bioterium and euthanized for collection of samples, including blood, skin, spleen, liver, heart, eye, kidney, intestine, esophagus, and brain. DNA was extracted using a commercial kit and tested via nested PCR for the ftsZ gene, as well as conventional PCR and qualitative real-time PCR using Sybr® Green for the citrate synthase gene (gltA), all specific reactions for B. henselae. All animals showed detection of B. henselae DNA in at least two different reactions in different tissues. The sequenced amplicons showed 100 % similarity to B. henselae. The use of mice infected by B. henselae in experiments is undesirable, as the bacteria can affect several aspects of the animal's physiology and consequently influence the results of the project, especially when subjected to immunosuppression. More studies are needed to understand and confirm the natural infection in experimental animals by Bartonella spp.. To date, no additional published reports of contamination of experimental animals by these bacteria have been identified.

Abstract Sjögren’s syndrome (SS) damages exocrine glands, and Lilium brownii var. viridulum Baker (Lilii Bulbus, LB) shows potential as a therapeutic agent. This study evaluated LB’s efficacy in alleviating xerostomia using non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice and human salivary gland acinar (NS-SV-AC) cells. In vitro, NS-SV-AC cells were treated with LB (1, 5, 10, 50, and 100 µg/mL) and 5-aza-2′-deoxycytidine (2 µM/mL) for 48 h. Cell viability, fluid secretion, and aquaporin-5 (AQP-5) expression were assessed. In vivo, thirty 20-week-old NOD/SCID mice received LB orally (100, 500, and 1,000 mg/kg) for 4 weeks, with salivary secretion rates measured. AQP-5 and M3 muscarinic acetylcholine receptor (M3R) expression and inflammatory mediator levels were determined using western blotting and enzyme-linked immunosorbent assay. Histopathological examination of salivary glands was also performed. LB significantly increased NS-SV-AC cell proliferation, fluid secretion, and AQP-5 expression. In NOD/SCID mice, LB reduced anti-SSA/Ro and anti-SSB/La antibodies, tumor necrosis factor-α, interferon-γ, and interleukin-6, while increasing AQP-5 and M3R expression. This resulted in increased salivary secretion and reduced glandular inflammation. LB extract appears promising for managing oral health by enhancing salivation, upregulating AQP-5, and modulating immune-inflammatory responses.

Resumen Hay una necesidad de nuevas alternativas al uso medicinal de alopurinol. En este sentido, el presente estudio obtuvo un extracto de hojas de L. leucocephala al cual se le determinó su composición química, acción inhibitoria contra xantina oxidasa (XO) in vitro, interacción inhibitoria entre el extracto y alopurinol, y acción inhibitoria contra XO in vivo empleando ratones bajo administración de hipoxantina y oxonato de potasio. Compuestos polifenoles y flavonoides fueron detectados y cuantificados en el extracto de hojas. Para el extracto solo, su valor IC50 y máximo de inhibición contra XO fueron de 334.60 µg/mL y 46.4 %. La combinación a una proporción 3:1 de alopurinol y extracto obtuvo valores IC50 y waDRI de 1.35 µg/mL, 1.13 (alopurinol) y 1015.72 (extracto) para inhibir a XO, resultando en una interacción sinérgica contra XO in vitro. Donde esta misma combinación incrementó el éxito terapéutico en el modelo murino al compararse con la administración de alopurinol solo. Nuestro estudio presenta la primera evidencia para el uso de una combinación de alopurinol y extracto de L. leucocephala a una proporción 3:1 como un sustituto para la administración de alopurinol solo.

Abstract There is a need for novel alternatives to the medical use of allopurinol. In this sense, the present study obtained a leaf extract from L. leucocephala, and its chemical composition, inhibitory action against xanthine oxidase (XO) in vitro, inhibitory interaction between the extract and allopurinol, and the inhibitory action on XO in vivo using mice treated with potassium oxonate and hypoxanthine, were determined. Polyphenol and flavonoid compounds were found in the leaf extract. For the leaf extract, the IC50 and maximal values were 334.60 µg/mL and 46.4 % for the inhibition of XO. The 3:1 ratio combination of allopurinol and extract showed IC50 and waDRI values of 1.35 µg/mL, 1.13 (allopurinol) and 1015.72 (extract) to inhibit XO, resulting in a synergistic interaction against XO in vitro. This combination also enhanced the therapeutic success in the mouse model compared with allopurinol administered alone. The present study presents the first evidence for the use of an allopurinol and L. leucocephala extract combination at a 3:1 ratio, as a substitute for the administration of allopurinol alone.

SUMMARY: There is a significant correlation between regulatory functions of orexin system and depressive symptoms. Parvalbumin (PV) interneurons, the main subtype of inhibitory g-aminobutyric acid (GABA) interneurons, are involved in the regulation of depression-like behaviors too. To provide several lines of evidence regarding neural connection between two types of neurons in the depression associated nuclei, a double-label immunohistochemical staining technique was employed to reveal overlap between PV and orexin immunoreactivity in the mouse brain. Medial prefrontal cortex (mPFC) mainly consists of the cingulate cortex, area 1 (Cg1), prelimbic cortex (PrL) and infralimbic cortex (IL), in which regions contained similar obvious overlap between orexin fibers and PV somas with axons. In the basal forebrain (BF), overlap was observed in the lateral septal nucleus, ventral part (LSV), the nucleus accumbens shell (AcbSh), bed nucleus of the stria terminalis, ventral division (BSTV), ventral pallidum (VP), central amygdaloid nucleus (Ce). In the diencephalon, besides the lateral hypothalamic area (LH) and dorsomedial hypothalamic nucleus (DM), in which two regions contained overlap between orexin positive perikarya with fibers and PV immunoreactivity, different densities of orexin and PV fibers were closely intermingled in the paraventricular hypothalamic nucleus (PVN), paraventricular thalamic nucleus (PVT), arcuate hypothalamic nucleus (Arc). In the brainstem, densities of intermixed orexin and PV fibers were from high to low in the dorsal raphe nucleus (DR), laterodorsal tegmental nucleus (LDTg), periaqueductal gray (PAG), lateral parabrachial nucleus (LPB), median raphe nucleus (MnR), and orexin buttons were embedded within PV fibers, as for the ventral tegmental area (VTA), overlap was observed between PV somas with axons and orexin fibers. Hence, PV and orexin neurons and its projections jointly target several brain regions involved in regulating depression, which provides anatomic evidence supporting coordinated PV/orexin actions within these brain regions.

RESUMEN: Existe una correlación significativa entre las funciones reguladoras del sistema de orexina y los síntomas depresivos. Las interneuronas de parvalbúmina (PV), el principal subtipo de interneuronas inhibidoras del ácido g-aminobutírico (GABA), también están implicadas en la regulación de conductas similares a la depresión. Para proporcionar varias líneas de evidencia con respecto a la conexión neuronal entre dos tipos de neuronas en los núcleos asociados a la depresión, se empleó una técnica de tinción inmunohistoquímica de doble marcación para revelar la superposición entre la interneurona de PV y la inmunorreactividad de la orexina en el cerebro del ratón. La corteza prefrontal medial (mPFC) está formada principalmente por la corteza cingulada, área 1 (Cg1), corteza prelímbica (PrL) y corteza infralímbica (IL), en las que las regiones contenían una superposición similar entre las fibras de orexina y los somas de las interneuronas PV y sus axones. En el prosencéfalo basal (BF), se observó superposición en el núcleo septal lateral, parte ventral (LSV), la superficie del núcleo accumbens (AcbSh), el núcleo del lecho de la estría terminal, división ventral (BSTV), núcleo pálido ventral (VP) y núcleo amigdaloide central (Ce). En el diencéfalo, además del área hipotalámica lateral (LH) y el núcleo hipotalámico dorsomedial (DM), en los que dos regiones contenían superposición entre pericario positivo para orexina con fibras e inmunorreactividad de PV, se observaron que diferentes densidades de fibras de orexina e interneuronas PV estaban estrechamente entremezcladas en el núcleo hipotalámico paraventricular (PVN), núcleo talámico paraventricular (PVT) y núcleo hipotalámico arqueado (Arc). En el tronco encefálico, las densidades de fibras de orexina e interneuronas PV entremezcladas fueron altas a bajas en el núcleo del rafe dorsal (DR), núcleo tegmental laterodorsal (LDTg), sustancia gris periacueductal (PAG), núcleo parabranquial lateral (LPB) y núcleo del rafe medio (MnR). La orexina estaba incrustada dentro de las fibras de interneuronas PV. Respecto al área tegmental ventral (VTA), se observó superposición entre somas PV con axones y fibras de orexina. Por lo tanto, las neuronas PV y orexina y, sus proyecciones, actúan conjuntamente sobre varias regiones cerebrales implicadas en la regulación de la depresión, lo que proporciona evidencia anatómica que respalda las acciones coordinadas PV/orexina dentro de estas regiones cerebrales.

SUMMARY: Currently, the bacteriophages have been applied as the phage therapy for using as a new technology for treating many cancers including lung cancer. This recent study explored the of a double cyclic form of arginine-glycine-aspartic acid with four cysteine residues (RGD) modified phage particles carrying the tumor necrosis factor therapeutic gene to suppress tumor growth. Adult male BALB/c nude mice were injected with A549 lung cancer cells to induce xenograft tumor growth. The control animals were subcutaneously injected with PBS while M13 and RGD-TNF animals were injected with M13 (control phage) and RGD-TNF phage, respectively. At the end of experiment. the tumor mass size was measured. Removed tumor masses and lung tissues of all groups were performed for immunohistochemistry and western blot analysis against Ki67, TNF-α, IL-2, and IFNγ antibodies. The results obviously showed that RGD-TNF phage could suppress the xenograft tumor growth. Additionally, the tissues of tumor mass and lung were localized with all cancer markers. It was revealed that RGD-TNF phage could significantly decrease the expressions of Ki67, TNF-α, and IL-2, but increase that of IFNγ in tumor mass. Only Ki67 and TNF-α expressions were decreased in lung tissue of mice treated with A549 lung cancer cells. It was concluded that RGD-TNF phage is a potential phage therapy for lung cancer treatment to reduce the tumor mass and to regulate the cancer markers.

RESUMEN: En la actualidad, los bacteriófagos se han aplicado como terapia fágica para su uso como una nueva tecnología para tratar muchos tipos de cáncer, incluido el cáncer de pulmón. Este estudio reciente intentó utilizar el fago modificado con factor de necrosis tumoral (TNF) y ácido aspártico-arginina-glicina (RGD) para suprimir el crecimiento de tumores pulmonares. Se inyectaron células de cáncer de pulmón A549 a ratones desnudos BALB/c machos adultos para inducir el crecimiento de tumores de xenoinjerto. A los animales control se les inyectó por vía subcutánea PBS, mientras que a los animales M13 y RGD-TNF se les inyectó M13 (fago de control) y fago RGD-TNF, respectivamente. Al final del experimento, se midió el tamaño de la masa tumoral. Las masas tumorales extirpadas y los tejidos pulmonares de todos los grupos se sometieron a análisis de inmunohistoquímica y transferencia Western contra anticuerpos Ki67, TNF-α, IL-2 e IFNγ. Los resultados mostraron obviamente que el fago RGD-TNF podía suprimir el crecimiento de tumores de xenoinjerto. Además, se localizaron los tejidos de la masa tumoral y del pulmón con todos los marcadores de cáncer. Se reveló que el fago RGD-TNF podía disminuir significativamente las expresiones de Ki67, TNF-α e IL- 2, pero aumentar la de IFNγ en la masa tumoral. Solo las expresiones de Ki67 y TNF-α disminuyeron en el tejido pulmonar de ratones tratados con células de cáncer de pulmón A549. Se concluyó que el fago RGD-TNF es una terapia de fagos potencial para el tratamiento del cáncer de pulmón para reducir la masa tumoral y regular los marcadores de cáncer.

SUMMARY: The current study aimed to demonstrate the physiological and histological effects of methotrexate (MTX) in laboratory mice's testes. 24 male laboratory mice, Mus musculus L., were used. The experimental mice were divided into three groups of equal number, with eight mice for each group. The first group of mice was injected with 0.1 ml of distilled water and adopted (as a control group), the second group was also injected with a low concentration of methotrexate (2 mg/kg), while the third group was injected with a high concentration of the drug (4 mg/kg). The mice were injected into the Intraperitoneal daily for 16 days. The results of the current study showed a significant decrease in sperm count at both doses (2,4 mg/kg) with a probability level of P <0.05 compared to the control group. The drug also caused a significant decrease in the number of normal sperm in mice treated with both doses, and an increase in the number of abnormalities of the sperm's head and tail. In addition, injecting males with the drug caused a significant reduction in the concentration of testosterone at both doses. The drug also had a clear effect on testicular tissue in general and the process spermatogenesis in particular. The results of the histological examination showed that the testicular tissue was affected in the treatment group compared with the control group, to varying degrees between the group treated with the low concentration and the high concentration. It has been observed that there are vacuoles in some seminiferous tubules, the appearance of degeneration and necrosis processes of germ cells, a decrease in the thickness of the germ mass, and a narrowing of the lumen of some seminiferous tubules. It was also observed that there was exfoliated, cytolysis, loss of germ cells, infiltration of inflammatory cells in some seminiferous tubules, and the appearance of amyloid between the seminiferous tubules, significantly in the group treated with the high concentration. The current study concluded that methotrexate causes severe damage to the testicles through a decrease in the number of sperm, distortions in their external appearance, a decrease in the level of testosterone, and a significant decrease in the process of spermatogenesis.

RESUMEN: El estudio tuvo como objetivo demostrar los efectos fisiológicos e histológicos del metotrexato (MTX) en los testículos de ratones de laboratorio. Se utilizaron 24 ratones macho, Mus musculus L. Los ratones se dividieron en tres grupos de ocho ratones. Al primer grupo de ratones - grupo control- se le inyectó 0,1 ml de agua destilada; al segundo grupo se le inyectó una concentración baja de metotrexato (2 mg/kg), mientras que al tercer grupo se le inyectó una concentración alta del fármaco (4 mg/kg). Los ratones fueron inyectados por vía intraperitoneal diariamente durante 16 días. Los resultados del estudio mostraron una disminución significativa en el recuento de espermatozoides en ambas dosis (2,4 mg/kg) con un nivel de probabilidad de P<0,05 en comparación con el grupo control. El fármaco provocó una disminución significativa en el número de espermatozoides normales en ratones tratados con ambas dosis, y un aumento en el número de anomalías de la cabeza y de la cola de los espermatozoides. Además, la inyección del fármaco provocó una reducción significativa en la concentración de testosterona en ambas dosis. El metotrexato también tuvo un claro efecto sobre el tejido testicular en general y en el proceso de espermatogénesis en particular. Los resultados del examen histológico mostraron que el tejido testicular se vio afectado en el grupo con tratamiento en comparación con el grupo control, en diversos grados entre el grupo tratado con la concentración baja y la concentración alta. Se observó que existían vacuolas en algunos túbulos seminíferos, la aparición de procesos de degeneración y necrosis de las células germinales, además de una disminución del grosor de la masa germinal y un estrechamiento del lumen de algunos túbulos seminíferos. También se observó que hubo exfoliación, citólisis, pérdida de células germinales, infiltración de células inflamatorias en algunos túbulos seminíferos y aparición de amiloide entre los túbulos seminíferos, significativamente mayor en el grupo tratado con concentración alta de metotrexato. Concluimos que el metotrexato causa daño severo a los testículos a través de una disminución en el número de espermatozoides, distorsiones en su apariencia externa, disminución en el nivel de testosterona y disminución significativa en el proceso de espermatogénesis.

Resumo Introdução: O objetivo deste estudo foi investigar o papel do fenofibrato, um agonista do receptor ativado por proliferadores de peroxissoma-α, na lesão renal induzida por obesidade (lipotoxicidade) em camundongos submetidos à uninefrectomia. Métodos: Camundongos C57BL/6 foram submetidos a uninefrectomia e cirurgias simuladas (sham) e alimentados com dietas normocalóricas ou rica em gordura. Após 10 semanas, os camundongos obesos receberam fenofibrato a 0,02% por 10 semanas. Avaliamos função e morfologia renal, bem como níveis de mediadores inflamatórios e fibróticos e marcadores do metabolismo lipídico. Resultados: Camundongos alimentados com dieta rica em gordura desenvolveram obesidade e hiperlipidemia características, com subsequente acúmulo de lipídios e danos renais, incluindo expansão mesangial, fibrose intersticial, inflamação e proteinúria. Essas alterações foram maiores em camundongos obesos uninefrectomizados do que em camundongos obesos sham. O tratamento com fenofibrato preveniu hiperlipidemia e lesões glomerulares, reduziu o acúmulo de lipídios, melhorou a disfunção renal e atenuou a inflamação e fibrose renal. Além disso, o tratamento com fenofibrato reduziu a expressão no tecido renal do inibidor do ativador do plasminogênio-1, da proteína quimiotática de monócitos-1 e a expressão local do fator de crescimento de fibroblastos-21. Conclusão: A ativação do receptor ativado por proliferadores de peroxissoma-α pelo fenofibrato, com subsequente lipólise, atenuou lesões glomerulares e tubulointersticiais induzidas por lipotoxicidade renal, protegendo, assim, os rins de camundongos uninefrectomizados contra lesões induzidas por obesidade. Os achados do estudo sugerem uma via na ação farmacológica do fenofibrato, fornecendo insights sobre os mecanismos envolvidos no dano renal causado pela obesidade em doadores renais. Introdução peroxissomaα, peroxissomaα peroxissoma α, α (lipotoxicidade Métodos C57BL6 CBL C57BL 6 C BL C57BL/ sham (sham 1 semanas 002 0 02 0,02 lipídico Resultados características renais mesangial intersticial proteinúria disso plasminogênio1, plasminogênio1 plasminogênio 1, plasminogênio-1 monócitos1 monócitos monócitos- fibroblastos21. fibroblastos21 fibroblastos 21. 21 fibroblastos-21 Conclusão lipólise protegendo assim 00 0,0 plasminogênio- fibroblastos2 2 fibroblastos-2 0, fibroblastos-

Abstract Introduction: The objective of this study was to investigate the role of fenofibrate, a peroxisome proliferator-activated receptor-α agonist, in obesity-induced kidney damage (lipotoxicity) in mice with uninephrectomy. Methods: C57BL/6 mice underwent uninephrectomy and sham surgeries and were fed normocaloric or high-fat diets. After 10 weeks, obese mice were administered 0.02% fenofibrate for 10 weeks. Kidney function and morphology were evaluated, as well as levels of inflammatory and fibrotic mediators and lipid metabolism markers. Results: High-fat diet-fed mice developed characteristic obesity and hyperlipidemia, with subsequent renal lipid accumulation and damage, including mesangial expansion, interstitial fibrosis, inflammation, and proteinuria. These changes were greater in obese uninephrectomy mice than in obese sham mice. Fenofibrate treatment prevented hyperlipidemia and glomerular lesions, lowered lipid accumulation, ameliorated renal dysfunction, and attenuated inflammation and renal fibrosis. Furthermore, fenofibrate treatment downregulated renal tissue expression of plasminogen activator inhibitor-1, monocyte chemoattractant protein-1, and local expression of fibroblast growth factor-21. Conclusion: Peroxisome proliferator-activated receptor-α activation by fenofibrate, with subsequent lipolysis, attenuated glomerular and tubulointerstitial lesions induced by renal lipotoxicity, thus protecting the kidneys of uninephrectomy mice from obesity-induced lesions. The study findings suggest a pathway in the pharmacological action of fenofibrate, providing insight into the mechanisms involved in kidney damage caused by obesity in kidney donors. Introduction proliferatoractivated proliferator activated receptorα receptor α agonist obesityinduced lipotoxicity (lipotoxicity Methods C57BL6 CBL C57BL 6 C BL C57BL/ highfat high fat diets 1 weeks 002 0 02 0.02 evaluated markers Results Highfat High dietfed diet expansion fibrosis proteinuria dysfunction Furthermore inhibitor1, inhibitor1 inhibitor 1, inhibitor-1 protein1, protein1 protein protein-1 factor21. factor21 factor 21. 21 factor-21 Conclusion lipolysis donors 00 0.0 inhibitor- protein- factor2 2 factor-2 0. factor-

Abstract Introduction: Fructose supplementation is recognized as a risk factor for metabolic dysfunction-associated steatotic liver disease (MASLD), but its role in the progression to metabolic dysfunction-associated steatohepatitis (MASH) remains less understood. This study aimed to evaluate the effect of dietary fructose on hepatic lipid metabolism (SREBP-1c, FAS, CD36, PPAR-α, ACOX-1, and CAT-1) in a murine model of hepatic steatosis progression from MASLD to MASH, as characterized by a NAS score. Materials and Methods: Male C57BL/6 mice were fed for 20 weeks with one of the following diets: 1) control diet (CD; 10% fat, 20% protein, 70% carbohydrates); 2) CD+F (30% fructose w/v); 3) high-fat diet (HFD; 60% fat, 20% protein, 20% carbohydrates); and 4) HFD+F (30% fructose w/v). General parameters, liver histology, and expression (qPCR, immunohistochemistry) of lipid metabolism markers (SREBP-1c, FAS, CD36, PPAR-α, ACOX1, CAT-1) were evaluated. Results: Mice fed an HFD+F showed increased SREBP-1c nuclear translocation and mRNA expression of FAS and CD36, while mitochondrial β-oxidation of free fatty acids was impaired due to the inhibition of PPAR-α activity and ACOX1 mRNA levels. Conclusion: Fructose supplementation exerted a strong pro-lipogenic and antilipolytic effect on the liver of mice fed an HFD, leading to increased lipid accumulation, elevated activity, and expression of FAS and CD36, as well as impairment in mitochondrial β-oxidation of free fatty acids compared to CD-fed mice. These findings suggest that fructose-induced alterations in hepatic lipid metabolism may play a critical role in the progression from MASLD to MASH, and its impact may be influenced by the metabolic state.

Resumen Introducción: La suplementación con fructosa se reconoce como un factor de riesgo para la enfermedad hepática esteatótica asociada a la disfunción metabólica (MASLD), pero su papel en la progresión hacia la esteatohepatitis asociada a la disfunción metabólica (MASH) sigue siendo menos comprendido. Este estudio tuvo como objetivo evaluar el efecto de la fructosa dietaria en el metabolismo lipídico hepático (SREBP-1c, FAS, CD36, PPAR-α, ACOX-1 y CAT-1) en un modelo murino de progresión de la esteatosis hepática de MASLD a MASH, caracterizado por un puntaje NAS. Materiales y métodos: Se alimentaron ratones machos C57BL/6 durante 20 semanas con una de las siguientes dietas: 1) dieta control (DC; 10% grasa, 20% proteína, 70% carbohidratos); 2) DC+F (30% fructosa p/v); 3) dieta alta en grasa (DAG; 60% grasa, 20% proteína, 20% carbohidratos); y 4) DAG+F (30% fructosa p/v). Se evaluaron parámetros generales, histología hepática y expresión (qPCR, inmunohistoquímica) de los marcadores del metabolismo lipídico (SREBP-1c, FAS, CD36, PPAR-α, ACOX1, CAT-1). Resultados: Los ratones alimentados con DAG+F mostraron un aumento en la translocación nuclear de SREBP-1c y la expresión de ARNm de FAS y CD36, mientras que la β-oxidación mitocondrial de los ácidos grasos libres se vio afectada debido a la inhibición de la actividad de PPAR-α y los niveles de ARNm de ACOX1. Conclusión: La suplementación con fructosa ejerció un fuerte efecto pro-lipogénico y antilipolítico en el hígado de los ratones alimentados con DAG, lo que llevó a un aumento en la acumulación de lípidos, mayor actividad y expresión de FAS y CD36, así como una disminución en la β-oxidación mitocondrial de los ácidos grasos libres en comparación con los ratones alimentados con DC. Estos hallazgos sugieren que las alteraciones en el metabolismo lipídico hepático inducidas por la fructosa pueden desempeñar un papel crítico en la progresión de MASLD a MASH y que su impacto podría depender del estado metabólico.

Resumen Los hongos con amatoxinas son los que generan la mayor cantidad de intoxicaciones mortales por mice tismo en el planeta. Estas toxinas son producidas no solo por las especies de Amanita, como la muy conocida Amanita phalloides, sino también por otros géneros, entre los que se encuentra Lepiota. En este trabajo se expone el tratamiento de un paciente de 51 años con un peso de 79 kg, derivado al hospital luego de 36 h de haber ingerido hongos, los cuales fueron identificados como Lepiota brunneoincarnata, en nuestro conocimiento una especie hallada por primera vez en la Argentina. El paciente presentaba malestar general, con náuseas y repetidos vómitos acompañados de dolor abdominal y diarrea. Gracias a una rápida anamnesis e identificación aproximada y temprana del hongo se le colocó sonda nasogástrica y se suministró carbón activado seriado. Además de N-acetilcisteína, fitomenadiona y penicilina G EV. El paciente fue dado de alta a los 11 días de su ingreso. Este caso realza la importancia de obtener una anamnesis correcta y suficiente sobre hongos, poder rea lizar un rápido análisis de ellos e iniciar un tratamiento oportuno de la intoxicación. Queda demostrado, una vez más, la importancia de tener Centros de Información y Asesoramiento Toxicológico (CIAT) con experiencia y conocimiento de micetismos.

Abstract Mushrooms containing amatoxins generate the high est number of fatal mycete poisonings on the planet. These toxins are produced not only by Amanita species, such as the well-known Amanita phalloides, but also by other genera, including Lepiota. In this work we report the treatment of a 51-year-old male patient weighing 79 kg who was referred to the hospital after 36 h of ingesting mushrooms. The mushrooms were identified as Lepiota brunneoincarnata, found for the first time in Argentina. The patient presented general malaise, nausea and repeated vomiting, abdominal pain, and diarrhea. Thanks to a quick anamnesis and early and accurate identifica tion of the fungus, he was administered a nasogastric tube and serial activated charcoal. Additionally, N-ace tylcysteine, phytomenadione and penicillin G EV were administered. The patient was discharged 11 days after admission. This case highlights the importance of obtain ing a correct and sufficient anamnesis on fungi, enabling rapid analysis of them, and initiation of timely treatment for intoxication. Once again, the importance of having Toxicological Information and Advice Centers (CIAT) with experience and knowledge of micetisms is demonstrated.

SUMMARY: Marein is a flavonoid compound that reduces blood glucose and lipids and has a protective effect in diabetes. However, the effect and mechanism(s) of marein on renal endothelial-mesenchymal transition in diabetic kidney disease (DKD) have not been elucidated. In this study, single-cell sequencing data on DKD were analyzed using a bioinformation method, and the data underwent reduced dimension clustering. It was found that endothelial cells could be divided into five subclusters. The developmental sequence of the subclusters was 0, 1, 4, 2, and 3, of which subcluster 3 had the most interstitial phenotype.The expression of mesenchymal marker protein:Vimentin(VIM), Fibronectin(FN1), and fibroblast growth factor receptor 1 (FGFR1) increased with the conversion of subclusters. In db/db mice aged 13-14 weeks, which develop DKD complications after 8-12 weeks of age, marein reduced blood levels of glucose, creatinine, and urea nitrogen, improved structural damage in kidney tissue, and reduced collagen deposition and the expression of FN1 and VIM. Marein also up-regulated autophagy marker:Light chain 3II/I(LC3II/I) and decreased FGFR1 expression in renal tissue. In an endothelial-mesenchymal transition model, a high glucose level induced a phenotypic change in human umbilical vein endothelial cells. Marein decreased endothelial cell migration, improved endothelial cell morphology, and decreased the expression of VIM and FN1. The use of the FGFR1 inhibitor, AZD4547, and autophagy inhibitor, 3-Methyladenine(3-MA), further demonstrated the inhibitory effect of marein on high glucose-induced endothelial-mesenchymal transition by reducing FGFR1 expression and up-regulating the autophagy marker protein, LC3II/I. In conclusion, this study suggests that marein has a protective effect on renal endothelial- mesenchymal transition in DKD, which may be mediated by inducing autophagy and down-regulating FGFR1 expression.

RESUMEN: La mareína es un compuesto flavonoide que reduce la glucosa y los lípidos en sangre y tiene un efecto protector en la diabetes. Sin embargo, no se han dilucidado el efecto y los mecanismos de la mareína sobre la transición endotelial- mesenquimatosa renal en la enfermedad renal diabética (ERD). En este estudio, los datos de secuenciación unicelular sobre DKD se analizaron utilizando un método de bioinformación y los datos se sometieron a una agrupación de dimensiones reducidas. Se descubrió que las células endoteliales podían dividirse en cinco subgrupos. La secuencia de desarrollo de los subgrupos fue 0, 1, 4, 2 y 3, de los cuales el subgrupo 3 tenía el fenotipo más intersticial. La expresión de la proteína marcadora mesenquimatosa: vimentina (VIM), fibronectina (FN1) y receptor del factor de crecimiento de fibroblastos. 1 (FGFR1) aumentó con la conversión de subgrupos. En ratones db/db de 13 a 14 semanas de edad, que desarrollan complicaciones de DKD después de las 8 a 12 semanas de edad, la mareína redujo los niveles sanguíneos de glucosa, creatinina y nitrógeno ureico, mejoró el daño estructural en el tejido renal y redujo la deposición y expresión de colágeno de FN1 y VIM. Marein también aumentó el marcador de autofagia: Cadena ligera 3II/I (LC3II/I) y disminuyó la expresión de FGFR1 en el tejido renal. En un modelo de transición endotelial-mesenquimal, un nivel alto de glucosa indujo un cambio fenotípico en las células endoteliales de la vena umbilical humana. Marein disminuyó la migración de células endoteliales, mejoró la morfología de las células endoteliales y disminuyó la expresión de VIM y FN1. El uso del inhibidor de FGFR1, AZD4547, y del inhibidor de la autofagia, 3-metiladenina (3-MA), demostró aún más el efecto inhibidor de la mareína en la transición endotelial-mesenquimal inducida por niveles altos de glucosa al reducir la expresión de FGFR1 y regular positivamente la proteína marcadora de autofagia. , LC3II/I. En conclusión, este estudio sugiere que la mareína tiene un efecto protector sobre la transición endotelial-mesenquimatosa renal en la ERC, que puede estar mediada por la inducción de autofagia y la regulación negativa de la expresión de FGFR1.