A Comprehensive Study of Positive Body Image as a Predictor of Psychological Well-Being in Anorexia Nervosa.

Recent data suggest a close association between positive body image (PBI) and eating disorder recovery. Nevertheless, the specific mechanisms through which PBI may facilitate recovery from anorexia nervosa (AN) remain unknown. To advance understanding of these mechanisms, this study examined core indices of PBI within AN, exploring its association with emotion regulation and well-being outcomes. Data were collected from 159 female participants, 64 with AN diagnosis and 95 healthy controls (HCs), who completed measures of PBI (body appreciation, functionality appreciation, and body responsiveness), emotion regulation, and psychological well-being (depression, anxiety, stress, and psychological quality of life). The AN group reported lower levels of PBI and psychological well-being, along with greater difficulties in regulating emotions, relative to HCs. PBI variables significantly predicted emotion regulation and psychological well-being in AN, accounting for 36% to 72% of the variance, with body appreciation emerging as the strongest predictor. These findings lend credence to the view that PBI can serve as a catalyst for psychological health. We hypothesize that enhancing PBI can improve interoceptive awareness, which is crucial for emotion regulation and reducing maladaptive food-related coping. Emphasizing a mind-body connection in lifestyle could be a relevant element to consider for both treating and preventing AN.

S-Adenosyl-l-methionine restores brain mitochondrial membrane fluidity and GSH content improving Niemann-Pick type C disease.

Niemann-Pick type C (NPC) disease is a lysosomal storage disorder characterized by impaired motor coordination due to neurological defects and cerebellar dysfunction caused by the accumulation of cholesterol in endolysosomes. Besides the increase in lysosomal cholesterol, mitochondria are also enriched in cholesterol, which leads to decreased membrane fluidity, impaired mitochondrial function and loss of GSH, and has been shown to contribute to the progression of NPC disease. S-Adenosyl-l-methionine (SAM) regulates membrane physical properties through the generation of phosphatidylcholine (PC) from phosphatidylethanolamine (PE) methylation and functions as a GSH precursor by providing cysteine in the transsulfuration pathway. However, the role of SAM in NPC disease has not been investigated. Here we report that Npc1-/- mice exhibit decreased brain SAM levels but unchanged S-adenosyl-l-homocysteine content and lower expression of Mat2a. Brain mitochondria from Npc1-/- mice display decreased mitochondrial GSH levels and liquid chromatography-high resolution mass spectrometry analysis reveal a lower PC/PE ratio in mitochondria, contributing to increased mitochondrial membrane order. In vivo treatment of Npc1-/- mice with SAM restores SAM levels in mitochondria, resulting in increased PC/PE ratio, mitochondrial membrane fluidity and subsequent replenishment of mitochondrial GSH levels. In vivo SAM treatment improves the decline of locomotor activity, increases Purkinje cell survival in the cerebellum and extends the average and maximal life spam of Npc1-/- mice. These findings identify SAM as a potential therapeutic approach for the treatment of NPC disease.

Difficulties in emotion regulation and well-being in breast cancer.

Objective: Breast cancer is responsible for disruptive changes in women's lives, causing them to experience diverse and intense negative emotions that can affect their perception of well-being. The present study aimed to characterize difficulties in emotion regulation (ER), according to Gratz and Roemer's multidimensional assessment, in women with breast cancer and to relate them with General Well-Being and its different domains: Physical, Social/Familial, Emotional, and Functional. Method: Ninety-five Portuguese women with breast cancer aged between 32 and 75 years answered a sociodemographic and clinical questionnaire and the Portuguese versions of the Difficulties in Emotion Regulation Scale and the Functional Assessment of Cancer Therapy - General. Data were collected in an oncology public hospital. Results: In general, difficulties in ER presented negative correlations with General Well-Being and its domains. The multiple regression analysis findings indicated that two specific types of difficulties, Limited Access to ER Strategies and Lack of Emotional Clarity, play a significant role in predicting well-being, especially in the Emotional domain, which was most compromised in these patients. Conclusions: These difficulties should be approached within psycho-oncological interventions as they are essential contributors to improving emotional and general well-being and fostering psychological adaptation to breast cancer.

Effect of Prenatal Iron Supplementation Adapted to Hemoglobin Levels in Early Pregnancy on Fetal and Neonatal Growth-ECLIPSES Study.

In this randomized clinical trial, we evaluated the effects of prenatal iron supplementation adapted to pregnant women's initial hemoglobin (Hb) levels on fetal growth parameters until birth in women from the Mediterranean coast of northern Spain. All (n = 791) women were iron-supplemented during pregnancy according to Hb levels at the 12th gestational week: stratum 1 (Hb: 110-130 g/L) received 40 or 80 mg iron daily; stratum 2 (Hb > 130 g/L) received 40 or 20 mg iron daily. Fetal biometric and anthropometric measurements were evaluated in the three trimesters and at birth, respectively. In stratum 1, using 80 mg/d instead of 40 mg/d increased the risk of fetal head circumference > 90th percentile (OR = 2.49, p = 0.015) at the second trimester and fetal weight (OR = 2.36, p = 0.011) and femur length (OR = 2.50, p = 0.018) < 10th percentile at the third trimester. For stratum 2, using 40 mg/d instead of 20 mg/d increased the risk of fetal head circumference > 90th percentile (OR = 3.19, p = 0.039) at the third trimester. A higher risk of delivering an LGA baby (OR = 2.35, p = 0.015) for birthweight was also observed in stratum 1 women receiving 80 mg/d. It is crucial to adjust the prenatal iron supplementation to each pregnant woman's needs, i.e., adapted to their initial Hb levels, to achieve optimal fetal development, since excessive iron doses appear to adversely influence fetal growth.

Bioconjugate based on cisplatin and bacterial exopolysaccharide with reduced side effects: A novel proposal for cancer treatment.

BACKGROUND: In the search for alternatives that attenuate the toxicity associated to oncologic treatment with cisplatin (CDDP) and considering the potential health-beneficial properties of exopolysaccharides (EPS) produced by lactic acid bacteria, it was aimed on this study to evaluate the cytotoxic, toxicologic and antitumoral efficacy of a bioconjugate based on CDDP and EPS, on the experimental tumor of sarcoma 180. METHODS: After the synthesis of the cis-[Pt(NH3)2(Cl)2] complex and of the conjugate containing Lactobacillus fermentum exopolysaccharide was tested both in vitro and in vivo for evaluating the acute toxicity. RESULTS: The antitumoral study was performed using mice transplanted with sarcoma 180. The bioconjugate showed low to medium cytotoxicity for the cell lines tested, as well moderated acute toxicity. After determining the LD50, the following experimental groups were established for the antitumor assay: Control (NaCl 0,9%), CDDP (1 mg/kg), EPS and bioconjugate composition (200 mg/kg). The bioconjugate promoted a 38% regression in tumor mass when compared to the control, and a regression of 41% when compared to CDDP. Liver histopathological analysis revealed discrete alterations in animals treated with (CDDP + EPS) when compared to control. The bioconjugate also minimized changes in the renal parenchyma resulting from the tumor. CONCLUSION: Our results indicate that when CDDP is associated with EPS, this composition was more biocompatible, showing itself as a potent chemotherapeutic agent and lower tissue toxicity.

Psychometric analysis of the body responsiveness questionnaire in the Portuguese population.

Body responsiveness refers to the tendency to be attuned to the body's needs and use interoceptive information to guide behavior. Despite its potential beneficial effect on the development of positive body image, this construct is currently understudied. To boost research in this area, we examined the factor structure, gender invariance, and psychometric properties of a Portuguese translation of the Body Responsiveness Questionnaire (BRQ). A total of 650 men and women (aged 18-80 years) completed the Portuguese BRQ. To assess its convergent validity, participants also completed measures of body appreciation, emotion regulation, depression symptoms, and psychological quality of life. Exploratory factor analysis indicated a two-factor structure of the BRQ, which was upheld using confirmatory factor analysis: "Importance of Interoceptive Awareness" (ω = .85-.87) and "Perceived Connection" between body and mind (ω = .71-.74). BRQ scores had partial scalar invariance across gender, and no significant gender differences. Convergent and known-groups validity was supported. Participants with overweight/obesity (vs. normal weight) and middle-aged adults (vs. young adults) assigned higher importance to body signals to guide behavior. The Portuguese version of the BRQ is a psychometrically sound measure of body responsiveness and it may contribute to a comprehensive assessment of positive body image to guide intervention.

Human Papillomavirus-Associated Oral Epithelial Dysplasia: Report of 5 Illustrative Cases from Latin America.

BACKGROUND: Human papillomavirus-associated oral epithelial dysplasia (HPV-OED) is a distinct oral epithelial disorder characterized by viral cytopathic changes caused by transcriptionally active high-risk HPV. The aim of the present study was to report 5 additional cases from Latin America. METHODS: Clinical data from five patients with HPV-OED were obtained from the archives of three oral pathology services from Brazil and Chile. All cases were submitted to morphological, p16 expression and in situ hybridization (ISH) for HPV analyses. RESULTS: Four patients were male and one patient was female, with a mean age of 55.4 years. Four patients were HIV seropositive and two were smokers. Three cases affected the buccal mucosa and commissure, one of which had an additional plaque in the soft palate, and one case each occurred on the floor of mouth and lower labial mucosa. Most cases presented as well-demarcated white plaques with a verrucous surface. One case presented multiple lesions ranging from normal to white-colored slightly elevated plaques with a cobblestone surface. Peripheral mucosal pigmentation was observed in two cases. All five cases presented with the characteristic microscopic features of HPV-OED, including severe dysplasia with numerous karyorrhectic and apoptotic cells, full-thickness "block positivity" for p16 and high Ki-67 index (> 90%) sharply demarcated from the adjacent non-dysplastic epithelium. Wide-spectrum DNA ISH-HPV was positive in 4 cases. All patients were treated with conservative surgical excision with no signs of recurrence after a mean of 39-month follow-up. CONCLUSION: This represents the first series of HPV-OED from Latin America; most cases presented as well-demarcated papillary white plaques affecting the buccal mucosa and commissure of HIV-positive middle-aged men, two of them exhibiting peripheral pigmentation caused by reactive melanocytes. The typical microscopic findings of HPV-OED were observed in all cases, which also showed strong p16 positivity in a continuous band through the full thickness of the epithelium and high Ki67.

Zonal expression of StARD1 and oxidative stress in alcoholic-related liver disease.

Alcoholic-related liver disease (ALD) is one of the leading causes of chronic liver disease and morbidity. Unfortunately, the pathogenesis of ALD is still incompletely understood. StARD1 has emerged as a key player in other etiologies of chronic liver disease, and alcohol-induced liver injury exhibits zonal distribution. Here, we report that StARD1 is predominantly expressed in perivenous (PV) zone of liver sections from mice-fed chronic and acute-on-chronic ALD models compared to periportal (PP) area and is observed as early as 10 days of alcohol feeding. Ethanol and chemical hypoxia induced the expression of StARD1 in isolated primary mouse hepatocytes. The zonal-dependent expression of StARD1 resulted in the accumulation of cholesterol in mitochondria and increased lipid peroxidation in PV hepatocytes compared to PP hepatocytes, effects that were abrogated in PV hepatocytes upon hepatocyte-specific Stard1 KO mice. Transmission electron microscopy indicated differential glycogen and lipid droplets content between PP and PV areas, and alcohol feeding decreased glycogen content in both areas while increased lipid droplets content preferentially in PV zone. Moreover, transmission electron microscopy revealed that mitochondria from PV zone exhibited reduced length with respect to PP area, and alcohol feeding increased mitochondrial number, particularly, in PV zone. Extracellular flux analysis indicated lower maximal respiration and spared respiratory capacity in control PV hepatocytes that were reversed upon alcohol feeding. These findings reveal a differential morphology and functional activity of mitochondria between PP and PV hepatocytes following alcohol feeding and that StARD1 may play a key role in the zonal-dependent liver injury characteristic of ALD.

Untreated osteoporosis and higher FRAX as risk factors for tooth loss: a 5-year prospective study.

INTRODUCTION: Studies have shown that an impaired bone condition, represented by osteoporosis and increased fracture risk, may potentially aggravate periodontal disease and, consequently, the risk of tooth loss. This 5-year prospective study aimed to investigate whether systemic bone condition represents risk factor for tooth loss due to periodontal disease amongst elderly women. MATERIAL AND METHODS: Seventy-four participants, aged ≥ 65 years, who attended the 5-years recall for periodontal evaluation were involved. Baseline exposures were osteoporosis and fracture risk probabilities (FRAX). Women were grouped according to bone mineral density (BMD) and years of bone treatment for osteoporosis. The primary outcome at a 5-year follow-up was the number of tooth loss due to periodontal disease. Periodontitis staging and grading, and causes of tooth loss were recorded. RESULTS: The multivariate Poisson regression models showed that women with untreated/shortly treated osteoporosis were 4 times more likely to present higher number of tooth loss due to periodontal disease than those with normal BMD or treated for ≥ 3 years (risk ratio (RR) = 4.00, 95% CI 1.40-11.27). Higher FRAX was also linked to tooth loss (RR = 1.25, 95% CI 1.02-1.53). Receiver-operating characteristic (ROC) curve suggested that women with history of ≥ 1 tooth losses have higher chances of worse major FRAX (sensitivity = 72.2%; specificity = 72.2%). CONCLUSION: In this 5-year study, higher FRAX and untreated osteoporosis were risk factors for tooth loss. Women with normal BMD or treated for osteoporosis for ≥ 3 years did not show increased risk. Management of skeletal conditions should be emphasized with periodontal care for the prevention of tooth loss in elderly women.

Oral clinical findings and intensive care unit prognostic scores.

OBJECTIVE: Hospitalisation in intensive care unit (ICU) may cause changes in oral environment, which may influence patients' health status. The aim of this study was to evaluate the frequency of intraoral and extraoral findings observed during ICU admission, and to verify if there is an association with clinical prognosis scores. METHODS: Data regarding clinical characteristics of patients hospitalised in an ICU were collected from medical records. The prognostic scores Sepsis Related Organ Failure Assessment (SOFA) and Simplified Acute Physiology Score (SAPS 3) were estimated with data collected from admission and SOFA on the day of the oral examination as well. Data on oral mucosa lesions, saliva, dental condition and oral hygiene were evaluated during oral examinations. RESULTS: The association of oral findings with prognostic scores was statistically verified. The majority (92.2%) of the 170 evaluated patients showed extraoral or intraoral findings during ICU admission. The most frequent findings were chapped and crusted lips, coated tongue, pale mucosa, haemorrhagic lesions, candidiasis, depapillated tongue and traumatic lesions. There were significant higher prognostic scores in the presence of the following extraoral and intraoral findings: crusted and ulcerated lips, haemorrhagic lesions, jaundice, spontaneous oral bleeding, coated and depapillated tongue. Median SAPS 3 was higher in patients with poor oral hygiene. CONCLUSIONS: Oral findings were frequent in the population of patients hospitalised in the ICU and some of them were associated with worse prognostic scores. Routine oral examinations must be performed in hospitalised patients from ICUs for detection of oral markers of worse clinical prognosis.

Civic engagement among foreign-born and native-born older adults living in Europe: a SHARE-based analysis.

Civic engagement is one of the cornerstones of participatory democracy and fundamental to preventing old-age social exclusion. Even though civic engagement late-in-life has received considerable attention, there is a lacuna of research on older migrants' civic engagement. This study aims therefore to examine potential predictors of civic engagement in terms of formal volunteering and participation in political organisations among foreign-born and native-born older adults in Europe. Attention is hereby given to how socio-structural resources and social capital are associated with civic engagement, and whether these associations differ between foreign-born and native-born. Data from wave 7 of the Survey of Health, Ageing and Retirement in Europe [n = 74,150; 5710 of them are foreign-born] were used in multivariable logistic regression analyses. Results show that socio-structural and social capital variables are positively associated with volunteering and participation in political organisations, both in native-born and foreign-born older adults. The study also suggests that place of birth (in Europe vs. outside Europe) and age-upon-migration play a role in predicting civic engagement among foreign-born older adults, and are therefore features worth considering when studying older migrants' civic engagement.

Differences among a Portuguese cohort of BRCA pathogenic/likely pathogenic variants carriers choosing risk-reducing mastectomy or intensive breast surveillance.

PURPOSE: Women with BRCA1 and BRCA2 (BRCA1/2) pathogenic/likely pathogenic (P/LP) variants have a higher risk to develop breast and ovarian cancer. In structured high-risk clinics, risk-reducing measures are adopted. This study aimed at characterizing these women and identify factors that may have influenced their choice between risk reduction mastectomy (RRM) and intensive breast surveillance (IBS). METHODS: This study reviewed retrospectively 187 clinical records of affected and unaffected women with P/LP variants of the BRCA1/2 genes, from 2007 to 2022, of which 50 chose RRM, while 137 chose IBS. The research focused on personal and family history and tumor characteristics and their relation with the preventive option chosen. RESULTS: Among women with personal history of breast cancer, a higher proportion opted for RRM compared to those asymptomatic (34.2% vs 21.3%, p = 0.049), with younger age determining the option for RRM (38.5 years vs 44.0 years, p < 0.001). Among women with personal history of ovarian cancer, a higher proportion opted for RRM compared to those without that history (62.5% vs 25.1%, p = 0.033), with younger age determining the option for RRM (42.6 years vs 62.7 years, p = 0.009). Women who had bilateral salpingo-oophorectomy were more likely to choose RRM than those who did not (37.3% vs 18.3%, p = 0.003). Family history was not associated with preventive option (33.3% vs 25.3, p = 0.346). CONCLUSIONS: The decision for the preventive option is multifactorial. In our study, personal history of breast or ovarian cancer, younger age at diagnosis, and previous bilateral salpingo-oophorectomy were associated with the choice of RRM. Family history was not associated with the preventive option.

Mitochondrial cholesterol: Metabolism and impact on redox biology and disease.

Cholesterol is a crucial component of membrane bilayers by regulating their structural and functional properties. Cholesterol traffics to different cellular compartments including mitochondria, whose cholesterol content is low compared to other cell membranes. Despite the limited availability of cholesterol in the inner mitochondrial membrane (IMM), the metabolism of cholesterol in the IMM plays important physiological roles, acting as the precursor for the synthesis of steroid hormones and neurosteroids in steroidogenic tissues and specific neurons, respectively, or the synthesis of bile acids through an alternative pathway in the liver. Accumulation of cholesterol in mitochondria above physiological levels has a negative impact on mitochondrial function through several mechanisms, including the limitation of crucial antioxidant defenses, such as the glutathione redox cycle, increased generation of reactive oxygen species and consequent oxidative modification of cardiolipin, and defective assembly of respiratory supercomplexes. These adverse consequences of increased mitochondrial cholesterol trafficking trigger the onset of oxidative stress and cell death, and, ultimately, contribute to the development of diverse diseases, including metabolic liver diseases (i.e. fatty liver disease and liver cancer), as well as lysosomal disorders (i.e. Niemann-Pick type C disease) and neurodegenerative diseases (i.e. Alzheimer's disease). In this review, we summarize the metabolism and regulation of mitochondrial cholesterol and its potential impact on liver and neurodegenerative diseases.

Oral manifestations of sporotrichosis: A neglected disease.

Sporotrichosis is an uncommon subacute or chronic infection caused by Sporothrix spp. In some urban areas of Latin America, sporotrichosis has been considered an emergent cosmopolitan disease of zoonotic transmission by domestic cats. There are four different clinical forms of the disease: fixed cutaneous, lymphocutaneous, multifocal or disseminated cutaneous, and extracutaneous. The oral mucosa is rarely involved, usually as unspecified chronic ulcers in the context of multifocal or disseminated cutaneous form of systemic sporotrichosis. Microscopical features include chronic granulomatous inflammation containing microabscesses and fungal hyphae positive for Periodic acid Schiff and silver-based stains. The diagnosis of sporotrichosis is usually based on culture detection and strict correlation of clinical, microscopical and laboratorial data. We herein contribute with two additional illustrative cases of oral manifestation of sporotrichosis in immunocompromised patients from an endemic urban area from Rio de Janeiro-Brazil. Key words:Sporotrichosis, ulcer, oral cavity, immunosuppression.

A Comparison of the Stability of Refined Edible Vegetable Oils under Frying Conditions: Multivariate Fingerprinting Approach.

The stability of highly consumed vegetable refined oils after discontinuous frying of potatoes was compared. Both those vegetable oils containing additives and those that did not were considered. Vegetable oil samples were evaluated using refractive index, anisidine and peroxide values, UV absorbance and dielectric constant-based determination of the content of total polar compounds. Chemical changes caused over the frying time were monitored and multivariate modelling of the data was carried out. A new gas chromatographic-mass spectroscopy method was intended to record a fingerprint of both polar and non-polar compound fractions. Multivariate models of chromatographic fingerprints were also developed, and the results obtained from both approaches were verified to be statistically similar. In addition, multivariate modelling also allows to differentiate among vegetable oils according to oxidation performance. Indeed, it was initially observed that olive oils presented the highest natural thermo-oxidative stability compared to other seed oils, although it should be noted that these differences were not significant when regarding olive pomace oils and seed oils containing synthetic additives.

Neprilysin-dependent neuropeptide Y cleavage in the liver promotes fibrosis by blocking NPY-receptor 1.

Development of liver fibrosis is paralleled by contraction of hepatic stellate cells (HSCs), the main profibrotic hepatic cells. Yet, little is known about the interplay of neprilysin (NEP) and its substrate neuropeptide Y (NPY), a potent enhancer of contraction, in liver fibrosis. We demonstrate that HSCs are the source of NEP. Importantly, NPY originates majorly from the splanchnic region and is cleaved by NEP in order to terminate contraction. Interestingly, NEP deficiency (Nep-/-) showed less fibrosis but portal hypertension upon liver injury in two different fibrosis models in mice. We demonstrate the incremental benefit of Nep-/- in addition to AT1R blocker (ARB) or ACE inhibitors for fibrosis and portal hypertension. Finally, oral administration of Entresto, a combination of ARB and NEP inhibitor, decreased hepatic fibrosis and portal pressure in mice. These results provide a mechanistic rationale for translation of NEP-AT1R-blockade in human liver fibrosis and portal hypertension.

Dietary and genetic disruption of hepatic methionine metabolism induce acid sphingomyelinase to promote steatohepatitis.

Alcoholic (ASH) and nonalcoholic. (NASH).steatohepatitis are advanced.stages.of.fatty.liver.disease.Methionine adenosyltransferase 1A (MAT1A) plays a key role in hepatic methionine metabolism and germline Mat1a deletion in mice promotes NASH. Acid sphingomyelinase (ASMase) triggers hepatocellular apoptosis and liver fibrosis and has been shown to downregulate MAT1A expression in the context of fulminant liver failure. Given the role of ASMase in steatohepatitis development, we investigated the status of ASMase in Mat1a-/- mice and the regulation of ASMase by SAM/SAH. Consistent with its role in NASH, Mat1a-/- mice fed a choline-deficient (CD) diet exhibited macrosteatosis, inflammation, fibrosis and liver injury as well as reduced total and mitochondrial GSH levels. Our data uncovered an increased basal expression and activity of ASMase but not neutral SMase in Mat1a-/- mice, which further increased upon CD feeding. Interestingly, adenovirus-mediated shRNA expression targeting ASMase reduced ASMase activity and protected Mat1a-/- mice against CD diet-induced NASH. Similar results were observed in CD fed Mat1a-/- mice by pharmacological inhibition of ASMase with amitriptyline. Moreover, Mat1a/ASMase double knockout mice were resistant to CD-induced NASH. ASMase knockdown protected wild type mice against NASH induced by feeding a diet deficient in methionine and choline. Furthermore, Mat1a-/- mice developed acute-on-chronic ASH and this outcome was ameliorated by amitriptyline treatment. In vitro data in primary mouse hepatocytes revealed that decreased SAM/SAH ratio increased ASMase mRNA level and activity. MAT1A and ASMase mRNA levels exhibited an inverse correlation in liver samples from patients with ASH and NASH. Thus, disruption of methionine metabolism sensitizes to steatohepatitis by ASMase activation via decreased SAM/SAH. These findings imply that MAT1A deletion and ASMase activation engage in a self-sustained loop of relevance for steatohepatitis.

The non-selective Rho-kinase inhibitors Y-27632 and Y-33075 decrease contraction but increase migration in murine and human hepatic stellate cells.

BACKGROUND: The Rho-kinase ROCK II plays a major role in the activation of hepatic stellate cells (HSC), which are the key profibrotic and contractile cells contributing to the development of chronic liver disease. Inhibition of ROCK II ultimately blocks the phosphorylation of the myosin light chain (MLC) and thus inhibits stress fibre assembly and cell contraction. We investigated the effects of the ROCK inhibitors Y-33075 as well as Y-27632 in murine and human hepatic stellate cells. METHODS: Primary isolated HSC from FVB/NJ mice and the immortalized human HSC line TWNT-4 were culture-activated and incubated with Y-27632 and Y-33075 (10nM to 10µM) for 24h. Protein expression levels were analyzed by Western Blots and transcriptional levels of pro-fibrotic markers and proliferative markers were evaluated using real-time qPCR. Migration was investigated by wound-healing assay. Proliferation was assessed by BrdU assay. Contraction of HSC was measured using 3D collagen matrices after incubation with Y-27632 or Y-33075 in different doses. RESULTS: Both Rho-kinase inhibitors, Y-27632 and Y-33075, reduced contraction, fibrogenesis and proliferation in activated primary mouse HSC (FVB/NJ) and human HSC line (TWNT-4) significantly. Y-33075 demonstrated a 10-times increased potency compared to Y-27632. Surprisingly, both inhibitors mediated a substantial and unexpected increase in migration of HSC in FVB/NJ. CONCLUSION: ROCK inhibition by the tested compounds decreased contraction but increased migration. Y-33075 proved more potent than Y27632 in the inhibition of contraction of HSCs and should be further evaluated in chronic liver disease.

Role of synthetical amynoquinone ethyl 2-(1,4-dioxo-1,4-dihydronaphthalen-2-ylamino) acetate in inhibition of Ehrlich's tumor.

Quinones are naturally or synthetically occurring secondary metabolites that have various bio-dynamics, highlighting their antitumor potential. This has been explored through their selective cytotoxicity, and studies in medicinal chemistry about the relation between biological activity versus chemical structure may lead to the solution of the toxicity problems associated with quinones. In this context, the antitumor effect of a synthetic naphthoquinone, named Ethyl 2-(1,4-Dioxo-1,4-Dihydronaphthalen-2-Ylamino) Acetate, was tested using mice transplanted with Ehrlich ascitic tumor as an experimental model. The acute toxicity test was performed using 30 mice that received the aminoquinone at doses of 100, 200, 300, and 600 mg/kg. After evaluation of the clinical findings in the spontaneous activity tests, the LD50 calculation for the test substance showed low levels of toxicity at doses lower than 244.11 ± 23.29 mg/kg. Thus, three experimental groups were established, where animals transplanted with tumor cells received NaCl vehicle solution (control, n = 6), and the others were treated with 71.7 mg/kg of Methotrexate (n = 6) or 20 mg/kg of Aminoquinone (n = 6). All administrations were intraperitoneal, in a single dose. Three days after the implantation of the tumor cells the animals were weighed daily and evaluated for tumor biometry and development. The treatments occurred five days after the implantation of the tumor cells and were extended for 7 more days. At the end of the 12-day experimental period, all animals were euthanized for biochemical and histopathological analyses of the tumors and vital organs. The spontaneous activity test showed that the amount of responses associated with the nervous system tends to increase with the increase in dosage, highlighting the excitatory effect on the central nervous system in almost all dosages employed, followed by depressant activities on this system. There was a significant tumor reduction, both in animals treated with methotrexate (71.7 %) and in those treated with aminoquinone (91.6 %) in the control group. There was no significant difference in tumor volume between the animals treated with aminoquinone or methotrexate. The histopathological analysis revealed that in both treatments there were fewer mitoses in the tumor mass compared to the control group. However, there was apparent toxicity to the liver, heart, and left kidney in the treatment with methotrexate compared to aminoquinone. The significant capacity for tumor reduction presented by aminoquinone allows pointing it as a promising alternative for the development of a more efficient drug to control tumor development, being necessary for the development of new studies to deepen the knowledge about its mechanisms of action.