Regulation of Mertk Surface Expression via ADAM17 and γ-Secretase Proteolytic Processing.

Mertk, a type I receptor tyrosine kinase and member of the TAM family of receptors, has important functions in promoting efferocytosis and resolving inflammation under physiological conditions. In recent years, Mertk has also been linked to pathophysiological roles in cancer, whereby, in several cancer types, including solid cancers and leukemia/lymphomas. Mertk contributes to oncogenic features of proliferation and cell survival as an oncogenic tyrosine kinase. In addition, Mertk expressed on macrophages, including tumor-associated macrophages, promotes immune evasion in cancer and is suggested to act akin to a myeloid checkpoint inhibitor that skews macrophages towards inhibitory phenotypes that suppress host T-cell anti-tumor immunity. In the present study, to better understand the post-translational regulation mechanisms controlling Mertk expression in monocytes/macrophages, we used a PMA-differentiated THP-1 cell model to interrogate the regulation of Mertk expression and developed a novel Mertk reporter cell line to study the intracellular trafficking of Mertk. We show that PMA treatment potently up-regulates Mertk as well as components of the ectodomain proteolytic processing platform ADAM17, whereas PMA differentially regulates the canonical Mertk ligands Gas6 and Pros1 (Gas6 is down-regulated and Pros1 is up-regulated). Under non-stimulated homeostatic conditions, Mertk in PMA-differentiated THP1 cells shows active constitutive proteolytic cleavage by the sequential activities of ADAM17 and the Presenilin/γ-secretase complex, indicating that Mertk is cleaved homeostatically by the combined sequential action of ADAM17 and γ-secretase, after which the cleaved intracellular fragment of Mertk is degraded in a proteasome-dependent mechanism. Using chimeric Flag-Mertk-EGFP-Myc reporter receptors, we confirm that inhibitors of γ-secretase and MG132, which inhibits the 26S proteasome, stabilize the intracellular fragment of Mertk without evidence of nuclear translocation. Finally, the treatment of cells with active γ-carboxylated Gas6, but not inactive Warfarin-treated non-γ-carboxylated Gas6, regulates a distinct proteolytic itinerary-involved receptor clearance and lysosomal proteolysis. Together, these results indicate that pleotropic and complex proteolytic activities regulate Mertk ectodomain cleavage as a homeostatic negative regulatory event to safeguard against the overactivation of Mertk.

Effects of Dietary Intervention on Nutritional Status in Elderly Individuals with Chronic Kidney Disease.

The prevalence of chronic kidney disease (CKD) is rising, especially in elderly individuals. The overlap between CKD and aging is associated with body composition modification, metabolic abnormalities, and malnutrition. Renal care guidelines suggest treating CKD patient with a low-protein diet according to the renal disease stage. On the other hand, geriatric care guidelines underline the need for a higher protein intake to prevent malnutrition. The challenge remains of how to reconcile a low dietary protein intake with insuring a favorable nutritional status in geriatric CKD populations. Therefore, this study aims to evaluate the effect of a low-protein adequate energy intake (LPAE) diet on nutritional risk and nutritional status among elderly CKD (stage 3-5) patients and then to assess its impact on CKD metabolic abnormalities. To this purpose, 42 subjects [age ≥ 65, CKD stage 3-5 in conservative therapy, and Geriatric Nutritional Risk Index (GNRI) ≥ 98] were recruited and the LPAE diet was prescribed. At baseline and after 6 months of the LPAE diet, the following data were collected: age, sex, biochemical parameters, anthropometric measurements, body composition, and the GNRI. According to their dietary compliance, the subjects were divided into groups: compliant and non-compliant. For the compliant group, the results obtained show no increased malnutrition risk incidence but, rather, an improvement in body composition and metabolic parameters, suggesting that the LPAE diet can provide a safe tool in geriatric CKD patients.

Hypothalamic-Ovarian axis and Adiposity Relationship in Polycystic Ovary Syndrome: Physiopathology and Therapeutic Options for the Management of Metabolic and Inflammatory Aspects.

PURPOSE OF REVIEW: The goal of the present review is to address the main adiposity-related alterations in Polycystic Ovary Syndrome (PCOS) focusing on hypothalamic-pituitary-ovarian (H-P-O) axis and to provide an overview of nutraceutical and pharmacological therapeutic strategies. RECENT FINDINGS: Female reproduction is a complex and delicate interplay between neuroendocrine signals involving the H-P-O axis. Elements that disrupt the balance of these interactions can lead to metabolic and reproductive disorders, such as PCOS. This disorder includes menstrual, metabolic, and biochemical abnormalities as well as hyperandrogenism, oligo-anovulatory menstrual cycles, insulin resistance, and hyperleptinemia which share an inflammatory state with other chronic diseases. Moreover, as in a self-feeding cycle, high androgen levels in PCOS lead to visceral fat deposition, resulting in insulin resistance and hyperinsulinemia, further stimulating ovarian and adrenal androgen production. In fact, regardless of age and BMI, women with PCOS have more adipose tissue and less lean mass than healthy women. Excessive adiposity, especially visceral adiposity, is capable of affecting female reproduction through direct mechanisms compromising the luteal phase, and indirect mechanisms as metabolic alterations able to affect the function of the H-P-O axis. The intricate crosstalk between adiposity, inflammatory status and H-P-O axis function contributes to the main adiposity-related alterations in PCOS, and alongside currently available hormonal treatments, nutraceutical and pharmacological therapeutic strategies can be exploited to treat these alterations, in order to enable a more comprehensive synergistic and tailored treatment.

Adherence to antiretroviral therapy thought package-refill among HIV+ persons at "D. Cotugno" hospital, Naples, Italy.

Background: A gold-standard for the measurement of adherence to antiretroviral therapy (ART) is lacking. Aim of this study is to verify the feasibility of a package-refill-based measurement of ART at "D. Cotugno" hospital, Naples, Italy, and the factors associated to adherence. Methods: In the period January 2018-August 2020, we calculated the package-refill as the ratio between ART-packages actually withdrawn, and the ART packages needed to regularly take ART. Adherence was associated, trough a univariate e multivariate logistic regression, to demographical, behavioural and clinical factors. Results: 1140 HIV+ subjects were included. At univariate logistic regression inadequate package-refill-based adherence is associated with HIV-RNA higher than 50 copies/mmL (OR 3.77-IC95% 2.76-5.13) and with HIVRNA higher than 200 copies/mmL (OR 3.98-IC95% 2.69-5.90). Being not-Italian and Injective-drug-user are associated with low adherence, having HIV/AIDS for more than 8 years is associated with better adherence. Conclusions: Package-refill is a suitable method for measuring adherence and is associated with the condition of viral failure.

Aquatic Pollution and Risks to Biodiversity: The Example of Cocaine Effects on the Ovaries of Anguilla anguilla.

Pollution is one of the main causes of the loss of biodiversity, currently one of the most important environmental problems. Important sources of aquatic pollution are illicit drugs, whose presence in waters is closely related to human consumption; their psychoactive properties and biological activity suggest potential adverse effects on non-target organisms, such as aquatic biota. In this study, we evaluated the effect of an environmentally relevant concentration of cocaine (20 ng L−1), an illicit drug widely found in surface waters, on the ovaries of Anguilla anguilla, a species critically endangered and able to accumulate cocaine in its tissues following chronic exposure. The following parameters were evaluated: (1) the morphology of the ovaries; (2) the presence and distribution of enzymes involved in oogenesis; (3) serum cortisol, FSH, and LH levels. The eels exposed to cocaine showed a smaller follicular area and a higher percentage of connective tissue than controls (p < 0.05), as well as many previtellogenic oocytes compared with controls having numerous fully vitellogenic and early vitellogenic oocytes. In addition, the presence and location of 3ß-hydroxysteroid dehydrogenase, 17ß-hydroxysteroid dehydrogenase, and P450 aromatase differed in the two groups. Finally, cocaine exposure decreased FSH and LH levels, while it increased cortisol levels. These findings show that even a low environmental concentration of cocaine affects the ovarian morphology and activity of A. anguilla, suggesting a potential impact on reproduction in this species.

Cerebral pCREB-dependent social behavioral adversities following a short-term exposure to obesogenic diets in young hamsters.

Excessive fat and sugar intake represents a risk towards the development of different pathologies, such as obesity, diabetes, sociability and memory deficits. Although the adolescence stage is a susceptible period for these and other risks, effects of energy-dense nutrients in such an age period have not been fully investigated. In the present study, neurobehavioral alterations following a 4-week exposure to either normal diet (ND) or high-fat diet (HFD) plus normal water (NW) or liquid sugar (LS) were evaluated in young hamsters. HFD + LS and ND + LS significantly reduced food intake and water consumption, which was, in the latter group, almost completely substituted by LS. All obesogenic diets accounted for increased abdominal fat and liver weight with respect to body weight (p < 0.05-0.001). Additionally, glucose levels notably increased (p < 0.0001) together with insulin and triglycerides in HFD + LS (p < 0.001) and ND + LS (p < 0.01) while cholesterol displayed only a moderate increase (p < 0.05) in HFD + NW and HFD + LS. Animals fed with HFD and/or LS exhibited impaired social memory plus increased winning percentages (0.05 < p < 0.01) during the tube test. Interestingly, these same treatments led to a down-regulation of phosphorylated cAMP Response-Element Binding Protein (pCREB) in HFD + NW (p < 0.0001) for all areas, but rather was upregulated (p < 0.05) in ND + LS of the amygdala. Overall, in view of a brief exposure to palatable foods interfering with normal metabolic and social memory activities, the downregulation of pCREB constitutes a key indicator of neurobehavioral deficits during obesogenic diets. Compensatory mechanisms may be also occurring in the amygdala that strongly regulates emotional states via connections with other limbic areas.

Enfermedad Tromboembólica Venosa en el período grávido puerperal. Diagnóstico, tratamiento y profilaxis / Venous Thromboembolic Disease in the puerperal gravid period. Diagnosis, treatment and prophylaxis / Doença tromboembólica venosa no período gravídico puerperal. Diagnóstico, tratamento e profilaxia

Resumen: El embarazo y puerperio son situaciones asociadas fisiológicamente con un marcado aumento del riesgo de enfermedad tromboembólica venosa. El riesgo se estima entre 7 a 10 veces superior que en mujeres de igual edad y 15-35 veces superior que la población control, durante el puerperio y hasta la sexta semana post-parto. Los factores de riesgo son distintos durante el período prenatal y post-parto. Los métodos diagnósticos incluyen radiografía, angio-tomografía de tórax y centellograma pulmonar ventilación-perfusión, ecodoppler venoso y venografía por resonancia nuclear magnética. El tratamiento supone un desafío que va desde la elección de la droga anticoagulante, los cambios en la farmacocinética durante el embarazo, entre otros. Se presentan algoritmos diagnósticos y terapéuticos.

Abstract: Pregnancy and the puerperium are physiologically associated with a markedly increased risk of venous thromboembolic disease. The risk is estimated between 7 to 10 times higher than in women of the same age and 15-35 times higher than the control population, during the puerperium and up to the sixth week postpartum. Risk factors are different during the prenatal and postpartum period. Diagnostic methods include radiography, chest angio-tomography and ventilation-perfusion lung scintigraphy, venous Doppler ultrasound, and magnetic resonance venography. Treatment involves a challenge that ranges from the choice of anticoagulant drug, changes in pharmacokinetics during pregnancy, among others. Diagnostic and therapeutic algorithms are presented.

Resumo: A gravidez e o puerpério estão fisiologicamente associados a um risco acentuadamente aumentado de doença tromboembólica venosa. O risco é estimado entre 7 a 10 vezes maior do que em mulheres da mesma idade e 15 a 35 vezes maior do que na população controle, durante o puerpério e até a sexta semana pós-parto. Os fatores de risco são diferentes durante o período pré-natal e pós-parto. Os métodos de diagnóstico incluem radiografia, angiotomografia de tórax e cintilografia pulmonar de ventilação-perfusão, ultrassom Doppler venoso e venografia por ressonância magnética nuclear. O tratamento envolve um desafio que vai desde a escolha do anticoagulante, mudanças na farmacocinética durante a gravidez, entre outros. Algoritmos diagnósticos e terapêuticos são apresentados.

Alkyphenol Exposure Alters Steroidogenesis in Male Lizard Podarcis siculus.

BACKGROUND: Nonylphenol (NP) and Octylphenol (OP) are persistent and non-biodegradable environmental contaminants classified as endocrine disruptor chemicals (EDCs). These compounds are widely used in several industrial applications and present estrogen-like properties, which have extensively been studied in aquatic organisms. The present study aimed to verify the interference of these compounds alone, and in mixture, on the reproductive cycle of the male terrestrial vertebrate Podarcis siculus, focusing mainly on the steroidogenesis process. METHODS: Male lizards have been treated with different injections of both NP and OP alone and in mixture, and evaluation has been carried out using a histological approach. RESULTS: Results obtained showed that both substances are able to alter both testis histology and localization of key steroidogenic enzymes, such as 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß- hydroxysteroid dehydrogenase (17ß-HSD) and P450 aromatase. Moreover, OP exerts a preponderant effect, and the P450 aromatase represents the major target of both chemicals. CONCLUSIONS: In conclusion, NP and OP inhibit steroidogenesis, which in turn may reduce the reproductive capacity of the specimens.

OctylPhenol (OP) Alone and in Combination with NonylPhenol (NP) Alters the Structure and the Function of Thyroid Gland of the Lizard Podarcis siculus.

Different environmental contaminants disturb the thyroid system at many levels. AlkylPhenols (APs), by-products of microbial degradation of AlkylPhenol Polyethoxylates (APEOs), constitute an important class of Endocrine Disrupting Chemicals (EDCs), the two most often used environmental APs being 4-nonylphenol (4-NP) and 4-tert-octylphenol (4-t-OP). The purpose of the present study was to investigate the effects on the thyroid gland of the bioindicator Podarcis siculus of OP alone and in combination with NP. We used radioimmunoassay to determine their effects on plasma 3,3',5-triiodo-L-thyronine (T3), 3,3',5,5'-L-thyroxine (T4), thyroid-stimulating hormone (TSH), and thyrotropin-releasing hormone (TRH) levels in adult male lizards. We also investigated the impacts of AP treatments on hepatic 5'ORD (type II) deiodinase and hepatic content of T3 and T4. After OP and OP + NP administration, TRH levels increased, whereas TSH, T3, and T4 levels decreased. Lizards treated with OP and OP + NP had a higher concentration of T3 in the liver and 5'ORD (type II) activity, whereas T4 concentrations were lower than that observed in the control group. Moreover, histological examination showed that the volume of the thyroid follicles became smaller in treated lizards suggesting that that thyroid follicular epithelial cells were not functionally active following treatment. This data collectively suggest a severe interference with hypothalamus-pituitary-thyroid axis and a systemic imbalance of thyroid hormones.

Effects of environmental cocaine concentrations on COX and caspase-3 activity, GRP-78, ALT, CRP and blood glucose levels in the liver and kidney of the European eel (Anguilla anguilla).

Cocaine is one of the most widely used illicit drugs in the world, and as a result of incomplete removal by sewage treatment plants it is found in surface waters, where it represents a new potential risk for aquatic organisms. In this study we evaluated the influence of environmental concentrations of cocaine on the liver and the kidney of the European eel (Anguilla anguilla). The eels were exposed to 20 ng L-1 of cocaine for fifty days, after which, three and ten days after the interruption of cocaine exposure their livers and kidneys were compared to controls. The general morphology of the two organs was evaluated, as well as the following parameters: cytochrome oxidase (COX) and caspase-3 activities, as markers of oxidative metabolism and apoptosis activation, respectively; glucose-regulated protein (GRP)78 levels, as a marker of endoplasmic reticulum (ER)-stress; blood glucose level, as stress marker; serum levels of alanine aminotransferase (ALT), as a marker of liver injury and serum levels of C-reactive protein (CRP), as a marker of the inflammatory process. The liver showed morphologic alterations such as necrotic areas, karyolysis and pyknotic nuclei, while the kidneys had dilated glomeruli and the renal tubules showed pyknotic nuclei and karyolysis. In the kidney, the alterations persisted after the interruption of cocaine exposure. In the liver, COX and caspase-3 activities increased (COX: P = 0.01; caspase-3: P = 0.032); ten days after the interruption of cocaine exposure, COX activity returned to control levels (P = 0.06) whereas caspase-3 activity decreased further (P = 0.012); GRP78 expression increased only in post-exposure recovery specimens (three days: P = 0.007 and ten days: P = 0.008 after the interruption of cocaine exposure, respectively). In the kidney, COX and caspase-3 activities increased (COX: P = 0.02; caspase-3: P = 0.019); after the interruption of cocaine exposure, COX activity remained high (three days: P = 0.02 and ten days: P = 0.029 after the interruption of cocaine exposure, respectively) whereas caspase-3 activity returned to control values (three days: P = 0.69 and ten days: P = 0.67 after the interruption of cocaine exposure, respectively). Blood glucose and serum ALT and CRP levels increased (blood glucose: P = 0.01; ALT: P = 0.001; CRP: 0.015) and remained high also ten days after the interruption of cocaine exposure (blood glucose: P = 0.009; ALT: P = 0.0031; CRP: 0.036). These results suggest that environmental cocaine concentrations adversely affected liver and kidney of this species.

Endocrine disrupting effects of copper and cadmium in the oocytes of the Antarctic Emerald rockcod Trematomus bernacchii.

Antarctica has long been considered a continent free from anthropic interference. Unfortunately, recent evidence indicate that metal contamination has gone so far and that its effects are still unknown. For this reason, in the present work, the potential endocrine disrupting effect of two highly polluting metals, copper and cadmium, were examined in the Antarctic teleost Trematomus bernacchii. After a 10 days waterborne exposure, ovarian metal uptake was determined by atomic absorption; in parallel, classical histological approaches were adopted to determine the effects on oocyte morphology, carbohydrate composition and presence and localization of progesterone and estrogen receptors. Results show that both metals induce oocyte degeneration in about one third of the previtellogenic oocytes, no matter the stage of development. In apparently healthy oocytes, changes in cytoplasm, cortical alveoli and/or chorion carbohydrates composition are observed. Cadmium but not copper also induces significant changes in the localization of progesterone and beta-estrogen receptors, a result that well correlates with the observed increase in ovarian metals concentrations. In conclusion, the acute modifications detected are suggestive of a significantly impaired fecundity and of a marked endocrine disrupting effects of copper and cadmium in this teleost species.

Axl and Mertk Receptors Cooperate to Promote Breast Cancer Progression by Combined Oncogenic Signaling and Evasion of Host Antitumor Immunity.

Despite the promising clinical benefit of targeted and immune checkpoint blocking therapeutics, current strategies have limited success in breast cancer, indicating that additional inhibitory pathways are required to complement existing therapeutics. TAM receptors (Tyro-3, Axl, and Mertk) are often correlated with poor prognosis because of their capacities to sustain an immunosuppressive environment. Here, we ablate Axl on tumor cells using CRISPR/Cas9 gene editing, and by targeting Mertk in the tumor microenvironment (TME), we observed distinct functions of TAM as oncogenic kinases, as well as inhibitory immune receptors. Depletion of Axl suppressed cell intrinsic oncogenic properties, decreased tumor growth, reduced the incidence of lung metastasis and increased overall survival of mice when injected into mammary fat pad of syngeneic mice, and demonstrated synergy when combined with anti-PD-1 therapy. Blockade of Mertk function on macrophages decreased efferocytosis, altered the cytokine milieu, and resulted in suppressed macrophage gene expression patterns. Mertk-knockout mice or treatment with anti-Mertk-neutralizing mAb also altered the cellular immune profile, resulting in a more inflamed tumor environment with enhanced T-cell infiltration into tumors and T-cell-mediated cytotoxicity. The antitumor activity from Mertk inhibition was abrogated by depletion of cytotoxic CD8α T cells by using anti-CD8α mAb or by transplantation of tumor cells into B6.CB17-Prkdc SCID mice. Our data indicate that targeting Axl expressed on tumor cells and Mertk in the TME is predicted to have a combinatorial benefit to enhance current immunotherapies and that Axl and Mertk have distinct functional activities that impair host antitumor response. SIGNIFICANCE: This study demonstrates how TAM receptors act both as oncogenic tyrosine kinases and as receptors that mediate immune evasion in cancer progression.

[Iron deficiency in reproductive age university women at the School of Medicine, Uruguay]. / Deficiencia de hierro en mujeres universitarias en edad reproductiva en la Facultad de Medicina, Uruguay.

Introduction: Disorders of iron metabolism are very common pathological conditions. Iron deficiency, with or without anemia, is estimated to affect more than 2 billion people.The aim of this study was to determine the prevalence of iron deficiency and anemia and their predisposing factors in a group of premenopausal women, university students of the School of Medicine of the University of the Republic in Uruguay. Methods: An observational cross-sectional study was carried out, including women of reproductive age, university students of the Faculty of Medicine. They were interviewed in order to collect clinical data and monthly menstrual volume was recorded through a pictogram. A hemogram was performed and ferritin levels were determined. Results: 196 women aged from 18 to 37 years were included. The prevalence of iron deficiency was 8.7% (n = 17) and the prevalence of anemia was 2.1% (n = 4). The presence of iron deficiency was associated with a lower consumption of red meat (p = 0.024), a higher menstrual volume (p = 0.018) and a higher frequency of abnormal uterine bleeding (p = 0.019). Conclusions: This study shows the high frequency of iron deficiency in healthy women in relation to abnormal uterine bleeding and low consumption of red meat, which raises the need to implement programs that promote educational measures in order to promote early consultation and avoid anemia and iron deficiency in these women of reproductive age.

Introducción: Los trastornos del metabolismo del hierro son condiciones patológicas muy frecuentes. La deficiencia de hierro, con o sin anemia, se estima que afecta a más de 2 billones de personas. El objetivo de este estudio fue determinar la prevalencia de deficiencia de hierro y anemia y los factores predisponentes en un grupo de mujeres premenopáusicas, estudiantes universitarias de la Facultad de Medicina de la Universidad de la República en Uruguay. Métodos: Se realizó un estudio observacional de corte transversal donde se incluyeron mujeres en edad reproductiva estudiantes universitarias de la Facultad de Medicina. Se les realizó una entrevista con el objetivo de recabar los datos clínicos y se registró el volumen menstrual mensual a través de un pictograma. Se realizó un hemograma y se determinaron los niveles de ferritina. Resultados: Se incluyeron 196 mujeres de 18 a 37 años. La prevalencia de ferropenia fue 8.7% (n=17) y la prevalencia de anemia fue de 2.1% (n=4). La presencia de ferropenia se asoció a un menor consumo de carne roja (p=0.024), a un mayor volumen menstrual (p=0.018) y a una mayor frecuencia de sangrado uterino anormal (p=0,019). Conclusión: Este estudio pone de manifiesto la frecuencia elevada de deficiencia de hierro en mujeres sanas en relación con sangrado uterino anormal y bajo consumo de carne roja, lo que plantea la necesidad de implementar programas que promuevan medidas educativas a fin de promover la consulta precoz y evitar la anemia y ferropenia en estas mujeres en edad reproductiva.

Spermatogenesis and regulatory factors in the wall lizard Podarcis sicula.

Spermatogenesis is an extraordinarily complex process, regulated by several factors, which leads to the differentiation of spermatogonia into spermatozoa. Among vertebrates, several reports have been focused on the lizard Podarcis sicula, a seasonal breeder and a good model for the study of reproductive processes. The goal of this review is to resume all the available data about systemic and above all local control factors involved in the control of P. sicula testicular activity. During the seasonal reproductive cycle, the variation of the expression levels of these factors determines significant variations that induce the activation or blocking of spermatogenesis. The data supplied in this review, in addition to analyze the current literature regarding the main actors of Podarcis sicula spermatogenesis, will hopefully provide a basic model that can be used for further studies on the intratesticular interaction between molecular factors that control spermatogenesis.

Adrenal gland response to endocrine disrupting chemicals in fishes, amphibians and reptiles: A comparative overview.

The adrenal gland is an essential component of the body stress response; it is formed by two portions: a steroidogenic and a chromaffin tissue. Despite the anatomy of adrenal gland is different among classes of vertebrates, the hormones produced are almost the same. During stress, these hormones contribute to body homeostasis and maintenance of ion balance. The adrenal gland is very sensitive to toxic compounds, many of which behave like endocrine-disruptor chemicals (EDCs). They contribute to alter the endocrine system in wildlife and humans and are considered as possible responsible of the decline of several vertebrate ectotherms. Considering that EDCs regularly can be found in all environmental matrices, the aim of this review is to collect information about the impact of these chemical compounds on the adrenal gland of fishes, amphibians and reptiles. In particular, this review shows the different behavior of these "sentinel species" when they are exposed to stress condition. The data supplied in this review can help to further elucidate the role of EDCs and their harmful impact on the survival of these vertebrates.

Effects of alkylphenols mixture on the adrenal gland of the lizard Podarcis sicula.

Alkylphenols (AP) are widespread environmental compounds belonging to the large family of substances known as Endocrine Disrupting Chemicals (EDCs). The present study was carried out to assess the effects of Octylphenol (OP) alone and in combination with Nonylphenol (NP) on the hypothalamus-pituitary-adrenal gland (HPA) axis of the lizard Podarcis sicula. Lizards are good bioindicators due to their features such as wide distribution, large population and good sensitivity to contaminants. Results obtained showed a time and dose-dependent stimulation of the HPA together with a high variation of both catecholamine plasma levels and greater vascularization and hypertrophy of steroidogenic cord of adrenal gland after both OP and OP + NP treatments. Interestingly, the OP + NP mixture treatment has provoked a state of stress of the adrenal gland which in fact appeared to be characterized by the presence of a marked macrophage infiltration which can be seen especially close to the connective capsule surrounding the gland. This macrophage infiltration could be an evidence of a particularly pronounced inflammatory state to indicate, probably, an animal's response to a non-physiological situation.

Emergence of BCR-ABL1 Chronic Myeloid Leukemia in a JAK2-V617F Polycythemia Vera.

Emergence of a new chronic myeloid neoplasm in the setting of a previous one, or their concomitant appearance seems to be a rare event, but plenty of cases have been reported. We describe the case of a patient with JAK2-V617F polycythemia vera, which looses JAK2 clone and develops overt BCR-ABL1 chronic myeloid leukemia after 6 years. Once treatment with tyrosine kinase inhibitors controls BCR-ABL1 clone, JAK2 clone arises again. In this report, we review the literature and discuss the clonal relationship of this event in light of the new molecular data.

Immunolocalization of aromatase P450 in the epididymis of Podarcis sicula and Rattus rattus.

The goal of this study was to evaluate P450 aromatase localization in the epididymis of two different vertebrates: the lizard Podarcis sicula, a seasonal breeder, and Rattus rattus, a continuous breeder. P450 aromatase is a key enzyme involved in the local control of spermatogenesis and steroidogenesis and we proved for the first time that this enzyme is represented in the epididymis of both P. sicula and R. rattus. In details, P450 aromatase was well represented in epithelial and myoid cells and in the connective tissue of P. sicula epididymis during the reproductive period; instead, during autumnal resumption this enzyme was absent in the connective tissue. During the non-reproductive period, P450 aromatase was localized only in myoid cells of P. sicula epididymis, whereas in R. rattus it was localized both in myoid cells and connective tissue. Our findings, the first on the epididymis aromatase localization in the vertebrates, suggest a possible role of P450 aromatase in the control of male genital tract function, particularly in sperm maturation.

Intrauterine exposure to diethylhexyl phthalate disrupts gap junctions in the fetal rat testis.

Fetal exposure to certain phthalate esters can disrupt testis development in rodents and lead to male reproductive disorders, but with a causal link less certain in humans. Di(2-ethylhexyl) phthalate (DEHP) is one of the most common phthalates found in the environment and in rodents it is known to induce serious testis toxicity, as well as male reproductive disorders including cryptorchidism, hypospadias, impaired spermatogenesis and reduced fertility. In this study, we show that perinatal DEHP exposure disrupts gap junction localization in fetal and postnatal rat testis and correlate these findings to morphological changes. The protein Connexin 43 (CX43), normally expressed strongly in testicular gap junctions, was markedly downregulated in Leydig cells of DEHP-exposed fetal testes. In the postnatal testes, CX43 expression was recovered in the DEHP-exposed animals, even though Leydig cell clusters and malformed cords with intratubular Leydig cells were still present.

Nonylphenol acts on prostate adenocarcinoma cells via estrogen molecular pathways.

Estrogens play a role in the patho-physiology of the prostate. In the present work we studied the effects of nonylphenol (NP), a xenoestrogen, on human adenocarcinoma prostate cells (LNCaP). In order to understand molecular and cellular involvement, we observed the effects on cell cycle and we investigated the expression and the cellular localization of estrogen receptors and gene expression of cyclin D1, ki-67, c-myc, IL-8, IL-1ß. We performed the same experiments with 17ß-estradiol (E2), the most abundant estrogen circulating in nonpregnant humans in order to compare these two different substances. We demonstrated the ability of 1 × 10-10 M NP to induce proliferation of LNCaP, S-phase progression, increase of ERα expression and its translocation from the cytoplasm to the nucleus. Moreover, we observed an up-regulation of key target genes involved in cell cycle and inflammation process. Particularly, after NP treatment, IL-8 and IL-1ß mRNA levels are increased more than 50% indicating a major NP involvement in inflammation processes than E2. These data suggest the proliferative effects of NP on prostate adenocarcinoma cells and highlight some aspects of molecular pathways involved in prostate responses to NP.