The impacts of exposure to bisphenol A in the adult female prostate Meriones unguiculatus.

The female prostate is associated with the urogenital system and presents homology in morphological terms with the male prostate. Due to its responsiveness to endogenous hormones, this gland is under a constant risk of developing prostatic pathologies and neoplasia when exposed to certain exogenous compounds. Bisphenol A (BPA) is an endocrine disruptor found in different plastic and resin products. Studies have emphasized the effects of perinatal exposure to this compound on different hormone-responsive organs. However, there have been few studies highlighting the influence on female prostate morphology of perinatal exposure to BPA. The objective of this study was to describe the histopathological alterations caused by perinatal exposure to BPA (50 µg/kg) and 17-ß estradiol (E2) (35 µg/kg) in the prostate of adult female gerbils. The results showed that E2 and BPA induced proliferative lesions in the female prostate and acted along similar pathways by modulating steroid receptors in the epithelium. BPA was also found to be a pro-inflammatory and pro-angiogenic agent. The impacts of both agents were marked in the prostatic stroma. An increase in the thickness of the smooth muscle layer and a decrease in AR expression were observed, but no alterations in the expression of ERα and ERß, leading to estrogenic sensitivity of the prostate. However, a peculiar response of the female prostate was to diminish the collagen frequency under BPA exposure correlated to smooth muscle layer. These data therefore indicate the development of features related to estrogenic and non-estrogenic tissue repercussions by BPA perinatally exposure in gerbil female prostate.

Condyloma acuminata: An evaluation of the immune response at cellular and molecular levels.

Condyloma acuminata (CA) is a benign proliferative disease mainly affecting in non-keratinized epithelia. Most cases of CA are caused by low-risk human papillomavirus (HPV), mainly HPV 6 and 11. The aim of the current study was to highlight the candidate genes and pathways associated with immune alterations in individuals who did not spontaneously eliminate the virus and, thus, develop genital warts. Paraffin-embedded condyloma samples (n = 56) were analyzed by immunohistochemistry using antibodies against CD1a, FOXP3, CD3, CD4, CD8, and IFN-γ. The immunomarkers were chosen based on the evaluation of the innate and adaptive immune pathways using qPCR analysis of 92 immune-related genes, applying a TaqMan Array Immune Response assay in HPV 6 or HPV 11 positive samples (n = 27). Gene expression analysis revealed 31 differentially expressed genes in CA lesions. Gene expression validation revealed upregulation of GZMB, IFNG, IL12B, and IL8 and downregulation of NFATC4 and IL7 in CA samples. Immunohistochemical analysis showed increased FOXP3, IFN-γ, CD1a, and CD4 expression in CA than in the control tissue samples. In contrast, CD3 and CD8 expression was decreased in CA lesion samples. Increased levels of pro-inflammatory cytokines in HPV-positive patients compared with HPV-negative patients seem to reflect the elevated immunogenicity of HPV-positive CA lesions. Host defense against HPV begins during the early stages of the innate immune response and is followed by activation of T lymphocytes, which are mainly represented by CD4+ and regulatory T cells. The low CD8+ T cell count in CA may contribute to this recurrent behavior. Additional studies are needed to elucidate the mechanism of host defense against HPV infection in CA.

The Mongolian Gerbil as a Useful Experimental Model in Reproductive Biology.

Ultimately, the Mongolian gerbils (Meriones unguiculatus) have acquired a relevant role in biological and biomedical experiments alongside other rodents. The use of gerbils in research has been mainly oriented to physiological and pharmacological studies, with special attention to nervous, digestive, and auditory systems as well as microbiology and parasitology. Ultimately, gerbils have also been applied for studying carcinogenesis in different organs and systems, since these animals show a natural propensity to develop spontaneous proliferative lesions, especially in steroid-responsive organs. This characteristic shed light on the reproductive aspects of this rodent model regarding morphological features in male and female individuals. This review of literature summarizes the significance of this model as an alternative to the use of inbred mice and rats in reproductive experimental research, highlighting recent findings. Gerbils have contributed to the expansion of knowledge in prostate biology in male and female individuals, providing studies related to prostatic morphogenesis and neoplasia. In the testes, spermiogenesis occurs in 15 steps, differently from other experimental models. Also, the complete maturation of the testis-epididymal complex occurs between the second and third months. Mammary gland alterations related to the estrous cycle and pregnancy were described, as well as its modulation under endogenous and exogenous estrogenic compounds. The ovaries frequently present ovarian cysts. Furthermore, this organ shows predominantly interstitial steroidogenic glands in the stroma, especially at aging. Adrenal gland shows a large size compared to other animals, presenting three distinct zones with a remarkable role in steroidogenesis.

Inflammatory repercussions in female steroid responsive glands after perinatal exposure to bisphenol A and 17-ß estradiol.

The mammary gland (MG) and female prostate are plastic reproductive organs which are highly responsive to hormones. Thus, endocrine disruptors, such as bisphenol A (BPA) and exogenous estrogens, negatively affect glandular homeostasis. In addition to previously described alterations, changes in inflammatory markers expression also trigger the development of a microenvironment that contributes to tumor progression. The current work aimed to evaluate the inflammatory responses of the MG and prostate gland to BPA (50 µg/kg) and 17-ß estradiol (35 µg/kg) exposure during the perinatal window of susceptibility. The results showed that at 6 months of age there was an increase in the number of phospho-STAT3 (P-STAT3) positive cells in the female prostate from animals perinatally exposed to 50 µg/kg BPA daily. In addition, the number of macrophages increased in these animals in comparison with nonexposed animals, as shown by the F4/80 marker. Despite an increase in the incidence of lobuloalveolar and intraductal hyperplasia, the MG did not show any difference in the expression of the four inflammatory markers evaluated: tumor necrosis factor-α, COX-2, P-STAT3, and F4/80. Analysis of both glands from the same animal led to the conclusion that exposure to endocrine disruptors during the perinatal window of susceptibility leads to different inflammatory responses in different reproductive organs. As the prostate is more susceptible to these inflammatory mechanisms, it is reasonable to affirm that possible neoplastic alterations in this organ are related to changes in the inflammatory pattern of the stroma, a characteristic that is not evident in the MG.

Combined photodynamic therapy with chloroaluminum phthalocyanine and doxorubicin nanoemulsions in breast cancer model.

Breast cancer is the most common neoplasm among women but thanks to innovative therapies, patients' prognosis has considerably improved. In this aspect, nanotechnology has been applied for cancer therapy aiming to reduce its usual side effects. In this study we aimed to evaluate the effects of nanoemulsions containing photosensitizer and chemotherapeutic agents associated with photodynamic therapy in a breast cancer in vivo model. Our results showed that synergistic treatments in which chloroaluminum phthalocyanine (NE-Pc) administered together with Doxorubicin (Dox) in the presence of laser irradiation (NE-PcDoxo + PDT) led to a reduction of 4 T1 induced breast cancer in mice, decline of tumor VEGF expression, increase in Caspase-3 expression, tissue necrosis and massive decrease in proliferative cells, as shown by Ki67 immunostaining. Furthermore, this associated treatment induced overexpression of apoptotic genes ABL1, CD70, CRADD, FASL, and NME5 and a reduction in expression of anticancer drug target genes CDK2, ERBB2, FIGF, IGF2, PARP4 and PGR. These results validate this treatment as a promising alternative to improve the currently applied anticancer strategies.

Perinatal exposure to bisphenol A impacts in the mammary gland morphology of adult Mongolian gerbils.

The endocrine disruptive effects caused by bisphenol A (BPA) are well known. Despite this, to date, evaluation of its long term effects is limited, meaning that there is still much to be unveiled in terms of alterations caused by perinatal exposure to BPA. Our aim was to determine if perinatal exposure to two different doses of BPA causes long term morphological and molecular alteration effects in the mammary gland (MG). We evaluated MG from Mongolian gerbil offspring exposed perinatally (during gestation and lactation) to 50 or 5000 µg/kg/day BPA. At 90 days of age the animals were subjected to a single dose of N-nitroso-N-methylurea in order to mimic a carcinogenic environment. At 6 months of age, animals in estrous were euthanized for morphological evaluation of the MGs. The MG architecture presented considerable changes in terms of detached epithelial cells, inflammation, glandular hyperplasia, and collagen fiber deposition. Furthermore, a higher index of epithelial cell proliferation was detected in comparison to the intact control group. In addition, we verified a higher molecular expression of EZH2 in the vehicle treated group, indicating that corn oil applied alone can alter the expression of this epigenetic biomarker. In conclusion, BPA perinatal exposure promotes significant changes in glandular cytoarchitecture and increases glandular epithelium proliferation rate, leading to the retention of stem-like properties. This event could compromise the fate and differentiation potential of mammary epithelium.

Impact of perinatal bisphenol A and 17ß estradiol exposure: Comparing hormone receptor response.

Hormonal regulation controls mammary gland (MG) development. Therefore some hormone-related factors can disrupt the early phases of MGs development, making the gland more susceptible to long term modifications in its response to circulating hormones. Endocrine disruptors, such as bisphenol A (BPA), are able to cause alterations in hormone receptor expression, leading to changes in the cell proliferation index, which may expose the tissue to neoplastic alterations. Thus, we evaluated the variations in hormone receptor expression in the MG of 6-month old Mongolian gerbils exposed to BPA and 17ß estradiol during the perinatal period. Receptors for estrogen alpha (ERα), beta (ERß), progesterone (PGR), prolactin (PRL-R), and co-localization of connexin 43 (Cx43) and ERα in gerbils were analyzed, and serum concentrations of estradiol and progesterone were assessed. No alterations in body, liver, and ovary-uterus complex weights were observed. However, there was an increase in epithelial ERα expression in the 17ß estradiol (E2) group and in PGR in the BPA group. Although immunohistochemistry did not show alterations in ERß expression, western blotting revealed a decrease in this protein in the BPA group. PRL-R was more present in epithelial cells in the vehicle control (VC), E2, and BPA groups in comparison to the intact control group. Cx43 was more frequent in E2 and BPA groups, suggesting a protective response from the gland against possible malignancy. Serum concentration of estradiol reduced in VC, E2, and BPA groups, confirming that alterations also impacts steroid levels. Consequently, perinatal exposure to BPA and the reference endogenous estrogen, 17ß estradiol, are able to increase the tendency of endocrine disruption in MG in a long term manner, since repercussions are observed even 6 months after exposure.

Glucose homeostasis in rats treated with 4-vinylcyclohexene diepoxide is not worsened by dexamethasone treatment.

4-vinilcyclohexene diepoxide (4-VCD) causes premature ovarian failure and may result in estrogen deficiency, characterizing the transition to estropause in rodents (equivalent to menopause in women). Estropause/menopause is associated with metabolic derangements such as glucose intolerance and insulin resistance. Glucocorticoids (GCs) are known to exert diabetogenic effects. Thus, we aimed to investigate whether rats with premature ovarian failure are more prone to the diabetogenic effects of GC. For this, immature female rats received daily injections of 4-VCD [160mg/kg body weight (b.w.), intraperitoneally (i.p.)] for 15 consecutive days, whereas control rats received vehicle. After 168days of the completion of 4-VCD administration, rats were divided into 4 groups: CTL-received daily injections of saline (1mL/kg, b.w., i.p.) for 5days; DEX-received daily injections of dexamethasone (1mg/kg, b.w., i.p.) for 5days; VCD-treated as CTL group; VCD+DEX-treated as DEX group. Experiments and euthanasia occurred one day after the last dexamethasone injection. 4-VCD-treated rats exhibited ovary hypotrophy and reduced number of preantral follicles (p<0.05). Premature ovarian failure had no impact on the body weight gain or food intake, but both were reduced by the effects of dexamethasone. The increase in blood glucose, plasma insulin and triacylglycerol levels as well as the reduction in insulin sensitivity caused by dexamethasone treatment was not exacerbated in the VCD+DEX group of rats. Premature ovarian failure did change neither the hepatic content of glycogen and triacylglycerol nor the glycerol release from perigonadal adipose tissue. Glucose intolerance was observed in the VCD group after an ipGTT (p<0.05), but not after an oral glucose challenge. Glucose intolerance and compensatory pancreatic ß-cell mass caused by GC were not modified by ovarian failure in the VCD+DEX group. We conclude that reduced ovarian function has no major implications on the diabetogenic effects promoted by GC treatment, indicating that other factors related to aging may make rats more vulnerable to GC side effects on glucose metabolism.

Biometric hoof evaluation of athletic horses of show jumping, barrel, long rope and polo modalities / Avaliação biométrica do casco de cavalos de esporte das modalidades hipismo, tambor, laço comprido e pólo

This study aimed to evaluate, through biometry, the forelimb hoof of horses participating in show jumping, barrel, long rope and polo competitions. Thirty subjects were assessed in relation to each competition (total of 120 animals). The linear measurements (cm) included the dorsal length of the toe; medial and lateral lengths of the quarter; medial and lateral heights of the quarter; lateral and medial lengths of the heel; medial and lateral heights of the heel; hoof length; hoof width; frog length; and frog width. The following angles () were measured: toe angle, pastern angle, heel angle and shoulder palette. The length of the horseshoe, coronet circumference and body weight were also assessed. With the use of hoof biometric evaluation was possible to identify the imbalances of forelimb in athletic horses and the most common were broken-backward hoof angle, with 96.7% of the animals showing this in the right forelimb (RFL) and 95.8% in the left forelimb (LFL); and contracted heels, with 95.0% in the RFL and 87.6% in the LFL. The competition type in which greatest numbers of hoof balance abnormalities were shown was the long rope, followed by the barrel. There were high frequencies of medial/lateral imbalance in all the sports. We conclude that animals used in functional tests have a high incidence of hoof balance abnormalities in the forelimbs.

Objetivou-se avaliar, através da biometria, os cascos dos membros anteriores de equinos participantes de provas de Hipismo Clássico, Tambor, Laço Comprido e Pólo. Em cada modalidade foram avaliados 30 indivíduos totalizando 120 animais. As mensurações lineares (cm) incluíram o comprimento dorsal da pinça, comprimentos lateral e medial dos quartos, altura medial e lateral dos quartos, comprimento lateral e medial dos talões, alturas lateral e medial dos talões, comprimento do casco, largura do casco, comprimento da ranilha, largura da ranilha, e as angulares (graus) foram relativas ao ângulo do casco medido pela pinça (podogoniometria), ângulo da quartela, ângulo dos talões, ângulo das paletas. O comprimento da ferradura, circunferência da coroa do casco e peso corporal também foram avaliados. Através da biometria do casco foi possível avaliar os desequilíbrios do membro anterior de animais atletas e os mais comuns foram: quebra do eixo podal, com 96,7% dos animais apresentando essa alteração no membro torácico direito (MTD) e 95,8% no membro torácico esquerdo (MTE); talões contraídos com 95,0% no MTD e 87,6% no MTE. A modalidade com os maiores desequilíbrios foi a de Laço Comprido, seguida pelos animais de Tambor. Foi encontrada uma alta frequência de desequilíbrio médio lateral em todas as modalidades desportivas. Conclui-se que animais utilizados em provas funcionais apresentam uma alta incidência de desequilíbrios podal nos membros torácicos.

Biometric hoof evaluation of athletic horses of show jumping, barrel, long rope and polo modalities / Avaliação biométrica do casco de cavalos de esporte das modalidades hipismo, tambor, laço comprido e pólo

This study aimed to evaluate, through biometry, the forelimb hoof of horses participating in show jumping, barrel, long rope and polo competitions. Thirty subjects were assessed in relation to each competition (total of 120 animals). The linear measurements (cm) included the dorsal length of the toe; medial and lateral lengths of the quarter; medial and lateral heights of the quarter; lateral and medial lengths of the heel; medial and lateral heights of the heel; hoof length; hoof width; frog length; and frog width. The following angles () were measured: toe angle, pastern angle, heel angle and shoulder palette. The length of the horseshoe, coronet circumference and body weight were also assessed. With the use of hoof biometric evaluation was possible to identify the imbalances of forelimb in athletic horses and the most common were broken-backward hoof angle, with 96.7% of the animals showing this in the right forelimb (RFL) and 95.8% in the left forelimb (LFL); and contracted heels, with 95.0% in the RFL and 87.6% in the LFL. The competition type in which greatest numbers of hoof balance abnormalities were shown was the long rope, followed by the barrel. There were high frequencies of medial/lateral imbalance in all the sports. We conclude that animals used in functional tests have a high incidence of hoof balance abnormalities in the forelimbs.(AU)

Objetivou-se avaliar, através da biometria, os cascos dos membros anteriores de equinos participantes de provas de Hipismo Clássico, Tambor, Laço Comprido e Pólo. Em cada modalidade foram avaliados 30 indivíduos totalizando 120 animais. As mensurações lineares (cm) incluíram o comprimento dorsal da pinça, comprimentos lateral e medial dos quartos, altura medial e lateral dos quartos, comprimento lateral e medial dos talões, alturas lateral e medial dos talões, comprimento do casco, largura do casco, comprimento da ranilha, largura da ranilha, e as angulares (graus) foram relativas ao ângulo do casco medido pela pinça (podogoniometria), ângulo da quartela, ângulo dos talões, ângulo das paletas. O comprimento da ferradura, circunferência da coroa do casco e peso corporal também foram avaliados. Através da biometria do casco foi possível avaliar os desequilíbrios do membro anterior de animais atletas e os mais comuns foram: quebra do eixo podal, com 96,7% dos animais apresentando essa alteração no membro torácico direito (MTD) e 95,8% no membro torácico esquerdo (MTE); talões contraídos com 95,0% no MTD e 87,6% no MTE. A modalidade com os maiores desequilíbrios foi a de Laço Comprido, seguida pelos animais de Tambor. Foi encontrada uma alta frequência de desequilíbrio médio lateral em todas as modalidades desportivas. Conclui-se que animais utilizados em provas funcionais apresentam uma alta incidência de desequilíbrios podal nos membros torácicos.(AU)