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Breast cancer is the most diagnosed type of cancer worldwide and the second cause of death in women. Triple-negative breast cancer (TNBC) is the most aggressive, and due to the lack of specific targets, it is considered the most challenging subtype to treat and the subtype with the worst prognosis. The present study aims to determine the antitumor effect of beta-D-glucose-reduced silver nanoparticles (AgNPs-G) in a murine model of TNBC, as well as to study its effect on the tumor microenvironment. In an airbag model with 4T1 tumor cell implantation, the administration of AgNPs-G or doxorubicin showed antitumoral activity. Using immunohistochemistry it was demonstrated that treatment with AgNPs-G decreased the expression of PCNA, IDO, and GAL-3 and increased the expression of Caspase-3. In the tumor microenvironment, the treatment increased the percentage of memory T cells and innate effector cells and decreased CD4+ cells and regulatory T cells. There was also an increase in the levels of TNF-α, IFN-γ, and IL-6, while TNF-α was increased in serum. In conclusion, we suggest that AgNPs-G treatment has an antitumor effect that is demonstrated by its ability to remodel the tumor microenvironment in mice with TNBC.

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AIM: To evaluate the influence of an experimental solution of cobalt-doped F18 bioactive glass (F18Co) on tissue repair following regenerative endodontic procedure (REP) in rat molars. METHODOLOGY: The F18Co solution was prepared at a ratio of 1:5 F18Co powder to distilled water. The right or left upper first molars of 12 Wistar rats were used, where the pulps were exposed, removed, and irrigated with 2.5% sodium hypochlorite (NaOCl), followed by 17% ethylenediaminetetraacetic acid (EDTA) (5 min each). Subsequently, the molars were divided into two groups (n = 6): REP-SS and REP-F18Co, where they received a final irrigation (5 min) with saline solution (SS) or F18Co solution, respectively. Then, intracanal bleeding was induced, and the tooth was sealed. Untreated molars were used as controls (n = 3). At 21 days, the rats were euthanized, and the specimens were processed for analysis of mineralized tissue and soft tissue formation inside the root canal using haematoxylin-eosin. The presence and maturation of collagen were evaluated by Masson's trichrome and picrosirius red staining. Immunolabelling analyses of proliferating cell nuclear antigen (PCNA) and osteocalcin (OCN) were performed. The data were submitted to the Mann-Whitney U-test (p < .05). RESULTS: There was a similar formation of mineralized tissue in thickness and length in REP-SS and REP-F18Co groups (p > .05). Regarding the presence of newly formed soft tissue, most specimens of the REP-F18Co had tissue formation up to the cervical third of the canal, whilst the REP-SS specimens showed formation up to the middle third (p < .05), and there was higher maturation of collagen in REP-F18Co (p < .05). The number of PCNA-positive cells found in the apical third of the root canal was significantly higher in the F18Co group, as well as the OCN immunolabelling, which was severe in most specimens of REP-F18Co, and low in most specimens of REP-SS. CONCLUSION: The final irrigation with F18Co bioactive glass solution in REP did not influence mineralized tissue formation but induced soft tissue formation inside the root canals, with higher collagen maturation, and an increase in PCNA-positive cells and OCN immunolabelling.

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ABSTRACT: Based on the effectiveness of resveratrol and curcumin in carcinogenesis, (E)-3-(4-hydroxy-3-methoxyphenyl)-N'-((E)-4-methoxybenzylidene) acrylohydrazide (PQM-162), curcumin-resveratrol hybrid derivative, was designed by molecular hybridization using a hydrazone functionality as a spacer moiety between pharmacophoric fragments inspired by the parent compounds. OBJECTIVES: The present study aimed to evaluate the chemopreventive effects of the hybrid against pre-neoplastic lesions induced in the colon of rodents. METHODS: The doses were determined based on the reduction in DNA damage induced by doxorubicin [15 mg/kg body weight (b.w.)] in peripheral blood of Swiss mice. Doses of 8, 16, 32, and 64 mg/kg b.w. were antimutagenic. For the evaluation of pre-neoplastic lesions in the colon, Wistar rats were treated with PQM-162 at doses of 0.5, 1, and 2 mg/kg b.w. for 6 weeks using three approaches: simultaneous treatment, pre-treatment, and post-treatment. Pre-neoplastic lesions were induced with 1,2 dimethylhydrazine (160 mg/kg b.w.). KEY FINDINGS: PQM-162 reduced the formation of aberrant crypt foci in the simultaneous treatment and post-treatment. TNF-α and COX-2 mRNA levels decreased, while Nrf2 mRNA levels increased. PQM-162 also reduced the expression of COX-2, PCNA, and ß-catenin protein markers and increased Nrf2 expression. CONCLUSION: Our findings suggest a chemopreventive potential of PQM-162 in colorectal carcinogenesis, which acts on anti-inflammatory, antioxidant, and cell proliferation pathways.

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This work aims to study the testicular morphology and spermatogenesis of Gymnotus carapo to provide information on their reproductive biology which is useful in managing this species as a fishing resource. The testicles were isolated and fixed in 10% formalin; subsequently, they were processed for scanning electron microscopy with conventional histological technique. To analyze the cell proliferation of germline cells and Sertoli cells, immunodetection of the proliferating cell nuclear antigen (PCNA) protein was performed. In G. carapo spermatogenesis, the spermatogenic line is organized into cysts. Spermatogonia A is characterized by more bigger and solitary cells. Spermatogonia B are smaller cells; their nucleus has a larger area concerning the cytoplasm and is grouped in tubules. Spermatocytes (I-II) are smaller than spermatogonia in the prophase of meiotic division. Spermatids are cells with dense, rounded nucleus. The sperm were found in the lumen of the tubule. By immunostaining PCNA, it was possible to observe the proliferative activity of germ line cells and Sertoli cells during the cyst reorganization phase. These results are the basis for future studies focusing on the analysis compared to females of the reproductive cycle of G. carapo.

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Liver impact of prolonged GH-treatment given to non-GH-deficient growing mice between the third and eighth week of life was evaluated in both sexes. Tissues were collected 6 h after last dose or four weeks later. Somatometric, biochemical, histological, immunohistochemical, RT-qPCR and immunoblotting determinations were performed. Five-week GH intermittent administration induced body weight gain and body and bone length increase, augmented organ weight, higher hepatocellular size and proliferation, and increased liver IGF1 gene expression. Phosphorylation of signaling mediators and expression of GH-induced proliferation-related genes was decreased in GH-treated mice liver 6h after last injection, reflecting active sensitization/desensitization cycles. In females, GH elicited EGFR expression, associated to higher EGF-induced STAT3/5 phosphorylation. Four weeks after treatment, increased organ weight concomitant to body weight gain was still observed, whereas hepatocyte enlargement reverted. However, basal signaling for critical mediators was lower in GH-treated animals and in male controls compared to female ones, suggesting signaling declination.

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The extracts of Platycladus orientalis (L.) Franco leaves have shown promising anti-cancer, anti-oxidant and anti-inflammatory potency with the traditional knowledge of healing HPV associated warts. The purpose of this research is to assess the synergistic activity of sorafenib and Platycladus orientalis (L) leaf extraction on cervical cancer cells. The cytotoxicity efficiency of different concentrations of Sorafenib and ethanol extract of Platycladus orientalis (L.) leaves were tested on HeLa cells by MTT and Trypan blue assays. The synergistic effect of the IC50 concentrations of Sorafenib and Platycladus orientalis (L.) on HeLa cell by MTT assay, and mRNA expression levels of tumor suppressor tazarotene-induced gene 3 (TIG3), proliferating cell nuclear antigen (PCNA) gene and apoptosis modulator (Bcl-2) gene by RT-PCR were evaluated with individual treatments. Combination treatment showed a relatively more expression of TIG3 and less expression of Bcl-2 and PCNA was observed. Growth factor-induced MAPKP activation was arrested by compound combination treatment, which and suppression of proliferation-induced apoptosis of cervical cancer cells. Based on the our results, the combination of sorafenib and crude leaf extract from Platycladus orientalis (L.) can effectively suppress cervical cancer cell growth, thereby providing an interesting rationale for further clinical trials and in-vivo studies.

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Este estudo avaliou a influência da fibrina rica em plaquetas injetável (i-PRF) no reparo após procedimento endodôntico regenerativo (REP) em molares imaturos de ratos. Molares superiores direito ou esquerdo de 18 ratos foram divididos em grupos (n = 6): SI ­ foi removido o tecido pulpar do canal mesial, feita irrigação com hipoclorito de sódio e ácido etilenodiaminotetracético, e induzido o sangramento intracanal (SI); iPRF ­ após protocolo de irrigação, foi inserido no interior do canal radicular a i-PRF, sem indução do SI; i-PRF-SI ­ foi realizado o tratamento como no grupo i-PRF, e então, induzido o SI. Ao término, os dentes foram selados. Para produzir a i-PRF, outros 3 animais foram utilizados para a coleta sanguínea intracardíaca, e o sangue foi centrifugado (1200 rpm, 8 min). Aos 21 dias, os animais foram eutanasiados e as peças preparadas para análise histológica e imunohistoquímica para antígeno nuclear de proliferação celular (PCNA) e osteocalcina (OCN). Teste estatístico foi aplicado (p < 0,05). Houve formação de tecido mineralizado em comprimento ou espessura da raiz em todos os grupos (p > 0,05). A formação de tecido conjuntivo nos canais ocorreu até terço médio na maior parte dos espécimes de SI, e até terço cervical em i-PRF e i-PRF-SI (p < 0,05). Houve células semelhantes a odontoblastos no terço apical de metade dos espécimes de i-PRF, e terços apical e médio na maior parte dos espécimes de i-PRF-SI; estas células não foram encontradas no grupo SI (p < 0,05). Os espécimes dos grupos i-PRF e i-PRF-SI tiveram significante número de células positivas para PCNA (p < 0,05) e maior imunomarcação de OCN, principalmente o grupo i-PRF-SI quando comparado ao grupo SI (p < 0,05). Conclui-se que i-PRF auxiliou o processo de reparo após REP em ratos, induzindo formação de tecido conjuntivo nos canais radiculares, presença de células semelhantes a odontoblastos e células positivas para PCNA, e imunomarcação de OCN, principalmente quando associada ao SI.

This study evaluated the influence of injectable platelet-rich fibrin (i-PRF) on repair after regenerative endodontic procedure (REP) in immature molars of rats. Right or left upper molars of 18 rats were divided into groups (n = 6): IB ­ the pulp tissue of the mesial canal was removed, irrigation with sodium hypochlorite and ethylenediaminetetraacetic acid was performed, and intracanal bleeding (IB) was induced; i-PRF ­ after the irrigation protocol, i-PRF was inserted inside the root canal, without inducing of IB; i-PRF-IB ­ the treatment was carried out as in the i-PRF group and then the IB was induced. At the end, the teeth were sealed. To produce the i-PRF, another 3 animals were used for intracardiac blood collection, and the blood was centrifuged (1200 rpm, 8 min). After 21 days, the animals were euthanized and the samples were prepared for histological and immunohistochemical analysis for proliferating cell nuclear antigen (PCNA) and osteocalcin (OCN). Statistical test was applied (P < 0.05). There was formation of mineralized tissue in root length or thickness in all groups (P > 0.05). The formation of connective tissue in the canals occurred up to the medium third in most IB specimens, and up to the cervical third in i-PRF and iPRF-IB (P < 0.05). There were odontoblast-like cells in the apical third of half of the PRF specimens, and apical and medium thirds in most of the i-PRF specimens; these cells were not found in the IB group (P < 0.05). The i-PRF and i-PRF-IB group had significant higher number of PCNA-positive cells (P < 0.05), and higher OCN immunolabeling, mainly i-PRF-IB compared to IB group (P < 0.05). It is concluded that i-PRF helped the repair process after REP in rats, inducing connective tissue formation in root canals, presence of odontoblast-like and PCNA-positive cells, and OCN immunolabeling, mainly when associated with IB.

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Este estudo avaliou os efeitos de uma solução experimental de vidro bioativo F18 dopado com cobalto (F18Co) no reparo tecidual após procedimento endodôntico regenerativo (REP, do inglês regenerative endodontic procedure) em molares de ratos. A solução de F18Co foi preparada misturando o pó de F18Co com água destilada na proporção de 1:5. Os primeiros molares superiores direito ou esquerdo de 12 ratos Wistar foram utilizados, nos quais as polpas foram expostas, removidas e os canais irrigados com hipoclorito de sódio (NaOCl) 2,5%, seguido de ácido etilenodiaminotetraacético (EDTA) 17% (5 min cada). Em seguida, os molares foram divididos em dois grupos (n = 6): REP-SS e REP-F18Co, nos quais receberam irrigação final (5 min) com solução salina (SS) ou solução de F18Co, respectivamente. Em seguida, foi induzido sangramento intracanal e o dente foi selado. Molares não tratados foram usados como controles (n = 3). Após 21 dias, os ratos foram eutanasiados e as peças processadas para análise da formação de tecido mineralizado e tecido mole dentro do canal radicular, pela técnica de hematoxilinaeosina. A presença e maturação do colágeno foram avaliadas por coloração de tricrômio de Masson e picrosírius red. Análises da imunomarcação do antígeno nuclear de proliferação celular (PCNA) e osteocalcina (OCN) foram realizadas. Os dados foram submetidos ao teste de Mann-Whitney U (p < 0,05). Houve formação semelhante de tecido mineralizado em espessura e comprimento nos grupos REP-SS e REP-F18Co (p > 0,05). Em relação à presença de tecido neoformado no interior dos canais radiculares, a maioria dos espécimes de REP-F18Co apresentou formação de tecido até o terço cervical do canal radicular, enquanto do grupo REP-SS, até o terço médio (p < 0,05), e houve maior maturação colágena em REP-F18Co (p < 0,05). O número de células positivas para PCNA encontradas no terço apical do canal radicular foi significativamente maior em REP-F18Co, assim como a imunomarcação de OCN, que foi severa na maior parte dos espécimes do grupo REP-F18Co, e leve em REPSS. Conclui-se que a irrigação final com solução de biovidro F18Co em REP não influenciou a formação de tecido mineralizado, mas induziu a formação de tecido conjuntivo no interior dos canais radiculares, com maior maturação colágena, aumento no número de células positivas para PCNA e na imunomarcação de OCN.

This study evaluated the influence of an experimental solution of cobalt-doped F18 bioactive glass (F18Co) on tissue repair following regenerative endodontic procedure (REP) in rat molars. The F18Co solution was prepared at a ratio of 1:5 F18Co powder to distilled water. The right or left upper first molars of 12 Wistar rats were used, where the pulps were exposed, removed, and irrigated with 2.5% sodium hypochlorite (NaOCl), followed by 17% ethylenediaminetetraacetic acid (EDTA) (5 min each). Subsequently, the molars were divided into two groups (n = 6): REP-SS and REP F18Co, where they received a final irrigation (5 min) with saline solution (SS) or F18Co solution, respectively. Then, intracanal bleeding was induced, and the tooth was sealed. Untreated molars were used as controls (n = 3). At 21 days, the rats were euthanized, and the specimens were processed for analysis of mineralized tissue and soft tissue formation inside the root canal using haematoxylin-eosin. The presence and maturation of collagen was evaluated by Masson's trichrome and picrosirius red staining. Immunolabeling analyses of proliferating cell nuclear antigen (PCNA) and osteocalcin (OCN) were performed. The data were submitted to the Mann-Whitney U test (P < 0.05). There was similar formation of mineralized tissue in thickness and length in REP-SS and REP-F18Co groups (P > 0.05). Regarding the presence of newly formed soft tissue, most specimens of the REP-F18Co had tissue formation up to the cervical third of the canal, while the REP-SS specimens showed formation up to the middle third (P < 0.05), and there was higher maturation collagen in REP-F18Co (P < 0.05). The number of PCNA-positive cells found in the apical third of the root canal was significantly higher in the F18Co group, as well as the OCN immunolabeling, which was severe in most specimens of REP-F18Co, and low in most specimens of REP SS. In conclusion, the final irrigation with F18Co bioactive glass solution in REP did not influence mineralized tissue formation but induced soft tissue formation inside the root canals, with higher collagen maturation, and an increase in PCNA-positive cells and OCN immunolabeling

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SUMMARY: Acrylamide (AA) is a widely used chemical and an important monomer in various industrial and laboratory processes. In addition, AA is formed during processing of starchy food at high temperature. The aim of our study was to examine effects of subchronic AA treatment on adult rat liver using histological, stereological and biochemical methods. Adult male Wistar rats were treated with AA at doses of 25 mg/kg b.w. and 50 mg/kg b.w. for three weeks. Stereological analysis showed decrease of volume density of hepatocyte cytoplasm, and increase of volume density of hepatocyte nuclei and nucleocytoplasmic ratio in AA50mg group. Immunohistochemical analysis of the liver sections showed that treatment with AA50mg increase the percentage of PCNA positive cells, while the percentage of caspase 3 positive cells was not affected by AA. PAS-staining showed that glycogen content in hepatocytes was not affected by AA. Serological examination revealed increase of lipid peroxidation in AA50mg group, while total protein concentration, protein thiol group level, as well as, paraoxonase 1 activity were not changed in AA-exposed animals. Stereological and immunohistochemical analyses of adult liver sections suggest increase of proliferation in AA50mg group, while increase of lipid peroxidation in serum of AA50mg group indicates oxidative stress induction.

La acrilamida (AA) es un químico ampliamente utilizado y un monómero importante en varios procesos industriales y de laboratorio. Además, la AA se forma durante el procesamiento de alimentos ricos en almidón a altas temperaturas. El objetivo de nuestro estudio fue examinar los efectos del tratamiento con AA subcrónica en el hígado de rata adulta utilizando métodos histológicos, estereológicos y bioquímicos. Se trataron ratas Wistar macho adultas con AA a dosis de 25 mg/kg p.v. y 50 mg/kg de peso corporal por tres semanas. El análisis estereológico mostró una disminución de la densidad del volumen del citoplasma de los hepatocitos y un aumento de la densidad del volumen de los núcleos de los hepatocitos y la relación nucleocitoplasmática en el grupo de 50 mg de AA. El análisis inmunohistoquímico de las secciones de hígado mostró que el tratamiento con 50 mg de AA aumentó el porcentaje de células positivas para PCNA, mientras que el porcentaje de células positivas para caspasa 3 no se vio afectado por AA. La tinción con PAS mostró que el contenido de glucógeno en los hepatocitos no se vio afectado por AA. El examen serológico reveló un aumento de la peroxidación de lípidos en el grupo de 50 mg de AA, mientras que la concentración de proteína total, el nivel del grupo tiol de proteína y la actividad de paraoxonasa 1 no cambiaron en los animales expuestos a AA. Los análisis estereológicos e inmunohistoquímicos de secciones de hígado adulto sugieren un aumento de la proliferación en el grupo AA50 mg, mientras que el aumento de la peroxidación lipídica en suero del grupo AA50 mg indica inducción de estrés oxidativo.

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Leukemia is the most common childhood malignancy in Mexico, representing more than 50% of all childhood cancers. Although treatment leads to a survival of up to 90% in developing countries, in our country, it is less than 65%. Additionally, ~30% of patients relapse with poor prognosis. Alternative splicing plays an important role in transcriptome diversity and cellular biology. This mechanism promotes an increase in the assortment of proteins with potentially distinct functions from a single gene. The proliferating cell nuclear antigen (PCNA) gene encodes two transcripts for the same protein of 261 amino acids, which is associated with several important cellular processes and with several types of cancer. However, the diversity of the transcript variants expressed in this condition is not clear. Then, we used microarray gene expression to identify changes in the exon expression level of PCNA. The data were validated using RT-PCR and Sanger sequencing, and three additional transcripts (PCNA_V3, PCNA_V4, and PCNA_V5) were identified. Computational analyses were used to determine the potential proteins resulting, their structure, and interactions with PCNA native protein and themselves. Additionally, the PCNA transcript variants were inhibited using specific siRNA, determining that their inhibition contributes to the malignant characteristics in vitro. Finally, we quantified the PCNA transcript variants in acute lymphoblastic leukemia samples and identified their expression in this disease. Based on the clinical characteristics, we determined that PCNA_V2 and PCNA_V4 are expressed at significantly low levels in relapsed B-ALL patients. We conclude that the low expression of PCNA_V2 and PCNA_V4 could be a potential molecular marker of relapse in acute lymphoblastic leukemia patients.

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