Cancer Progression Is not Different in Mice of Different Gender Inoculated With Cells of the Triple-Negative 4T1 Breast Cancer Model.

Background: Breast cancer in men is a rare and poorly studied disease, and its treatment is based on women breast cancer studies. However, clinical outcome is not the same in men and women. Basic studies and clinical trials in animal models provide detailed information on cancer, origin, development, cell signaling pathways, sites of metastasis, and target molecules. It is necessary to explore the biology of breast cancer in male animal models that allow observing their similarity. Methods: The triple-negative 4T1 breast cancer model was developed in both male and female mice and studied weekly during 4 weeks. For that, twenty 8-week-old female and male BALB/c mice were used. Sixteen mice (eight males and eight females) were inoculated into the second left thoracic mammary pad with 20,000 4T1 cells, resuspended in 20 µL phosphate-buffered saline (PBS). All samples were processed for immunodetection, characterized histopathologically and immunohistochemically. Results: In this work, we describe the development of a triple-negative 4T1 breast cancer model in male BALB/c mice. Breast tumors were characterized histopathologically at different time points and corresponded to a moderately differentiated invasive ductal carcinoma, estrogen receptor ER-/progesterone receptor PR-/human epidermal growth factor receptor 2 HER2-/Ki67+, with histological grade II (moderately differentiated; a solid mass with occasional duct formation and moderate to severe nuclear pleomorphism), infiltrating the adipose and muscular tissue, and metastasis to lungs. From the results, we did not observe differences in the time of tumor development, necrosis, color change of tumor tissue, and lung metastasis between male and female mice. Even though we did not find histological differences, response to treatment and molecular signaling may be different. Conclusions: The histogenesis of male breast tumors was similar to that of female BALB/c mice. The histological and immunohistochemical characteristics of male tumors also match the features reported for stage IV human breast cancer of men and women. The murine male breast cancer model described here can be a significant tool to explore the molecular mechanisms involved in male breast cancer tumorigenesis and metastasis and may bring new approaches for clinical treatment of triple-negative breast cancer in men.

Determination of histone epigenetic marks in Arabidopsis and tomato genes in the early response to Botrytis cinerea.

KEY MESSAGE: Determination of histone epigenetic marks in Arabidopsis and tomato genes in the early response to Botrytis cinerea may contribute to find biomarkers of the early detection of this devastating pathogen. Recent studies have linked epigenetic modifications with plant responses to biotic stresses. Information about specific histone marks upon necrotrophic pathogens is scarce. Here we wondered whether the altered responsiveness of specific genes in plants infected with Botrytis cinerea was associated with changes in chromatin structure. We performed a chromatin immunoprecipitation analysis that obtained differential epigenetic signature of activating marks H3K4me3, H3K9ac, and the repressor one H3K27me3 on both the promoter and the body of the highly induced PR1 in Arabidopsis plants infected with B. cinerea at 24 and 33 h after inoculation. We also determined the histone marks' profile in two differentially expressed genes in response to B. cinerea, as well as to oxidative stress, given its relevance in this infection. These are both the induced CYP71A13, which encodes a cytochrome P450 involved in camalexin synthesis, and is essential against this necrotroph and the repressed EXL7 (Exordium-like 1). We also adapted our protocol in tomato plants infected with B. cinerea. At 24 hpi, H3K4me3 level increased on the promoter and at different locations of the body of the genes induced upon B. cinerea, including DES (divinyl ethyl synthase), LoxD (lipoxygenase D), DOX1 (α-dioxygenase 1), PR2 (pathogenesis-related protein2), WRKY53 and WRKY33. The histone modifications determined herein will allow future studies on epigenetic marks and their transgenerational inheritance in plants infected with B. cinerea. In addition, the analyzed genes are potential biomarkers of B. cinerea infection that could contribute to its early detection in tomato and related crops.

Absence of Cu-Zn superoxide dismutase BCSOD1 reduces Botrytis cinerea virulence in Arabidopsis and tomato plants, revealing interplay among reactive oxygen species, callose and signalling pathways.

Plants activate responses against pathogens, including the oxidative burst. Necrotrophic pathogens can produce reactive oxygen species (ROS) that benefit the colonization process. Previously, we have demonstrated that tomato plants challenged with Botrytis cinerea accumulate ROS and callose, together with the induction of genes involved in defence, signalling and oxidative metabolism. Here, we studied the infection phenotype of the Δbcsod1 strain in both tomato and Arabidopsis plants. This mutant lacks bcsod1, which encodes Cu-Zn superoxide dismutase (SOD). This enzyme catalyses the conversion of superoxide ion ( O2-) into hydrogen peroxide (H2 O2 ). ROS play a protective role and act as signals in plants. Δbcsod1 displayed reduced virulence compared with wild-type B05.10 in both species. Plants infected with Δbcsod1 accumulated less H2 O2 and more O2- than those infected with B05.10, which is associated with an increase in the defensive polymer callose. This supports a major role of fungal SOD in H2 O2 production during the plant-pathogen interaction. The early induction of the callose synthase gene PMR4 suggested that changes in ROS altered plant defensive responses at the transcriptional level. The metabolites and genes involved in signalling and in response to oxidative stress were differentially expressed on Δbcsod1 infection, supporting the notion that plants perceive changes in ROS balance and activate defence responses. A higher O2- /H2 O2 ratio seems to be beneficial for plant protection against this necrotroph. Our results highlight the relevance of callose and the oxylipin 12-oxo-phytodienoic acid (OPDA) in the response to changes in the oxidative environment, and clarify the mechanisms that underlie the responses to Botrytis in Arabidopsis and tomato plants.

Role of dioxygenase α-DOX2 and SA in basal response and in hexanoic acid-induced resistance of tomato (Solanum lycopersicum) plants against Botrytis cinerea.

Resistance of tomato (Solanum Lycopersicum) to the fungal pathogen Botrytis cinerea requires complex interplay between hormonal signalling. In this study, we explored the involvement of new oxylipins in the tomato basal and induced response to this necrotroph through the functional analysis of the tomato α-dioxygenase2 (α-DOX2)-deficient mutant divaricata. We also investigated the role of SA in the defence response against this necrotrophic fungus using SA-deficient tomato nahG plants. The plants lacking dioxigenase α-DOX2, which catalyses oxylipins production from fatty acids, were more susceptible to Botrytis, and hexanoic acid-induced resistance (Hx-IR) was impaired; hence α-DOX2 is required for both tomato defence and the enhanced protection conferred by natural inducer hexanoic acid (Hx) against B. cinerea. The divaricata plants accumulated less pathogen-induced callose and presented lower levels of jasmonic acid (JA) and 12-oxo-phytodienoic acid (OPDA) upon infection if compared to the wild type. Glutathion-S-transferase (GST) gene expression decreased and ROS production significantly increased in Botrytis-infected divaricata plants. These results indicate that absence of α-DOX2 influences the hormonal changes, oxidative burst and callose deposition that occur upon Botrytis infection in tomato. The study of SA-deficient nahG tomato plants showed that the plants with low SA levels displayed increased resistance to Botrytis, but were unable to display Hx-IR. This supports the involvement of SA in Hx-IR. NaghG plants displayed reduced callose and ROS accumulation upon infection and an increased GST expression. This reflects a positive relationship between SA and these defensive mechanisms in tomato. Finally, Hx boosted the pathogen-induced callose in nahG plants, suggesting that this priming mechanism is SA-independent. Our results support the involvement of the oxylipins pathway and SA in tomato response to Botrytis, probably through complex crosstalk of the hormonal balance with callose and ROS accumulation, and reinforce the role of the oxidative stress in the outcome of the plant-Botrytis interaction.

Absence of endo-1,4-ß-glucanase KOR1 alters the jasmonate-dependent defence response to Pseudomonas syringae in Arabidopsis.

During plant-pathogen interactions, the plant cell wall forms part of active defence against invaders. In recent years, cell wall-editing enzymes, associated with growth and development, have been related to plant susceptibility or resistance. Our previous work identified a role for several tomato and Arabidopsis endo-1,4-ß-glucanases (EGs) in plant-pathogen interactions. Here we studied the response of the Arabidopsis thaliana T-DNA insertion mutant lacking EG Korrigan1 (KOR1) infected with Pseudomonas syringae. KOR1 is predicted to be an EG which is thought to participate in cellulose biosynthesis. We found that kor1-1 plants were more susceptible to P. syringae, and displayed severe disease symptoms and enhanced bacterial growth if compared to Wassilewskija (Ws) wild-type plants. Hormonal and gene expression analyses revealed that the jasmonic acid (JA) pathway was activated more in kor1-1 plants with an increase in the JA-biosynthesis gene LOX3 and a greater accumulation of JA. Upon infection the accumulation of JA and JA-isoleucine (JA-Ile) was higher than in wild-type plants and increased the induction of LOX3 and the JA-responsive PDF1.2 gene. In addition, the increase of salicylic acid (SA) in healthy and infected kor1-1 may reflect the complex interaction between JA and SA, which results in the more susceptible phenotype displayed by the infected mutant plants. Callose deposition was enhanced in infected kor1-1 and an increase in pathogen-induced hydrogen peroxide took place. The susceptible phenotype displayed by KOR1-deficient plants was coronatine-independent. No significant changes were detected in the hormonal profile of the kor1-1 plants infected by coronatine-deficient P. syringae cmaA, which supports that absence of EG KOR1 alters per se the plant response to infection. We previously reported increased resistance of kor1-1 to B. cinerea, hence, the lack of this EG alters cell wall properties and plant responses in such a way that benefits P. syringae colonisation but restricts B. cinerea invasion.

CYP3A4 expression in breast cancer and its association with risk factors in Mexican women.

BACKGROUND: In Mexico, breast cancer (BCa) is the leading type of cancer in women. Cytochrome P450 (CYP450) is a superfamily of major oxidative enzymes that metabolize carcinogens and many antineoplastic drugs. In addition, these enzymes have influence on tumor development and tumor response to therapy. In this report, we analyzed the protein expression in patients with BCa and in healthy women. Links with some clinic-pathological characteristic were also assessed. MATERIALS AND METHODS: Immunohistochemical analyses were conducted on 48 sets of human breast tumors and normal breast tissues enrolled in Hospital Militar de Especialidades de la Mujer y Neonatologia and Hospital Central Militar, respectively, during the time period from 2010 to 2011. Informed consent was obtained from all participants. Statistical analysis was performed using χ2 or Fisher exact tests to estimate associations and the Mann Whitney U test for comparison of group means. RESULTS: We found a significant CYP3A4 overexpression in BCa stroma and gland regions in comparison with healthy tissue. A significant association between protein expression with smoking, alcoholism and hormonal contraceptives use was also observed. Additionally, we observed estrogen receptor (ER) and progesterone receptor (PR) positive association in BCa. CONCLUSIONS: We suggest that CYP3A4 expression promotes BCa development and can be used in the prediction of tumor response to different treatments. One therapeutic approach may thus be to block CYP3A4 function.

Hexanoic acid protects tomato plants against Botrytis cinerea by priming defence responses and reducing oxidative stress.

Treatment with the resistance priming inducer hexanoic acid (Hx) protects tomato plants from Botrytis cinerea by activating defence responses. To investigate the molecular mechanisms underlying hexanoic acid-induced resistance (Hx-IR), we compared the expression profiles of three different conditions: Botrytis-infected plants (Inf), Hx-treated plants (Hx) and Hx-treated + infected plants (Hx+Inf). The microarray analysis at 24 h post-inoculation showed that Hx and Hx+Inf plants exhibited the differential expression and priming of many Botrytis-induced genes. Interestingly, we found that the activation by Hx of other genes was not altered by the fungus at this time point. These genes may be considered to be specific targets of the Hx priming effect and may help to elucidate its mechanisms of action. It is noteworthy that, in Hx and Hx+Inf plants, there was up-regulation of proteinase inhibitor genes, DNA-binding factors, enzymes involved in plant hormone signalling and synthesis, and, remarkably, the genes involved in oxidative stress. Given the relevance of the oxidative burst occurring in plant-pathogen interactions, the effect of Hx on this process was studied in depth. We showed by specific staining that reactive oxygen species (ROS) accumulation in Hx+Inf plants was reduced and more restricted around infection sites. In addition, these plants showed higher ratios of reduced to oxidized glutathione and ascorbate, and normal levels of antioxidant activities. The results obtained indicate that Hx protects tomato plants from B. cinerea by regulating and priming Botrytis-specific and non-specific genes, preventing the harmful effects of oxidative stress produced by infection.

[Malignant phyllodes tumor with differentiation to liposarcoma. A report of a case and bibliographic review]. / Tumor filoideo maligno con diferenciación a liposarcoma. Informe de un caso y revisión de la bibliografía.

Phyllodes tumor represents less than 1% of breast tumors and 2 to 3% of the fibroepithelial tumors. The malignancy characteristics are the excessive growth and the stromal atypia with numerous mitosis. The stromal component of the phyllodes tumor has potential to cause metaplasia to bone tissue, cartilage, and to smooth and striated muscle, including their respective malignant neoplasms. The liposarcomatous differentiation of phyllodes tumor is extremely rare. We present the case of an unusual breast tumor diagnosed as phyllodes tumor, with sarcomatous changes on its stroma, which consists of pleomorfic and myxoid liposarcoma. We also comment some generalities about clinical and diagnostic aspects, as well as anatomopathological, therapeutic, and prognostic characteristics related to the current literature.

Informe de marcadores serológicos de las hepatitis virales en el Hospital Central Militar (1992-1994) / Report on serologic markers for viral hepatitis at the central military hospital (1992-1994)

Se comunican los resultados de un estudio retrospectivo de marcadores serológicos para hepatitis viral realizado en el laboratorio de inmunología del departamento de patología clínica del Hospital Central Militar en el periodo comprendido de junio de 1992 a junio de 1994. Se realizaron 614 estudios en 537 pacientes de uno y otro sexo y de todas las edades. Los marcadores fueron diagnósticos para la infección aguda en 34.37 por ciento, de los cuales fueron 12.8 por ciento positivos para anti-HVA IgM, 10.42 por ciento positivos para anti-HBe, 11.07 por ciento positivos para anti-HC y 0.32 por ciento positivos para anti-HD. La seroprevalencia de hepatitis "A" con anti-HAV IgG fue de 90.8 por ciento, se observó disminución de anti-HC durante el último año (16.4 por ciento entre 1992-1993 a 8.26 por ciento entre 1993-1994). La prevalencia de anti-HBe se presenta sin cambio en el mismo periodo. La hepatitis "D" coexistió en coinfección con hepatitis "B" en los dos casos