Enhancing RECK Expression Through miR-21 Inhibition: A Promising Strategy for Bladder Carcinoma Control.

Bladder carcinoma (BC) is the tenth most frequent malignancy worldwide, with high morbidity and mortality rates. Despite recent treatment advances, high-grade BC and muscle-invasive BC present with significant progression and recurrence rates, urging the need for alternative treatments. The microRNA-21 (miR-21) has superexpression in many malignancies and is associated with cellular invasion and progression. One of its mechanisms of action is the regulation of RECK, a tumor suppressor gene responsible for inhibiting metalloproteinases, including MMP9. In a high-grade urothelial cancer cell line, we aimed to assess if miR-21 downregulation would promote RECK expression and decrease MMP9 expression. We also evaluated cellular migration and proliferation potential by inhibition of this pathway. In a T24 cell line, we inhibited miR-21 expression by transfection of a specific microRNA inhibitor (anti-miR-21). There were also control and scramble groups, the last with a negative microRNA transfected. After the procedure, we performed a genetic expression analysis of miR-21, RECK, and MMP9 through qPCR. Migration, proliferation, and protein expression were evaluated via wound healing assay, colony formation assay, flow cytometry, and immunofluorescence.After anti-miR-21 transfection, miR-21 expression decreased with RECK upregulation and MMP9 downregulation. The immunofluorescence assay showed a significant increase in RECK protein expression (p < 0.0001) and a decrease in MMP9 protein expression (p = 0.0101). The anti-miR-21 transfection significantly reduced cellular migration in the wound healing assay (p < 0.0001). Furthermore, in the colony formation assay, the anti-miR-21 group demonstrated reduced cellular proliferation (p = 0.0008), also revealed in the cell cycle analysis by flow cytometry (p = 0.0038). Our results corroborate the hypothesis that miR-21 is associated with BC cellular migration and proliferation, revealing its potential as a new effective treatment for this pathology.

Psychrometry in the thermal comfort diagnosis of production animals: a combination of the systematic review and methodological proposal.

Animal welfare and productive performance are compromised when animals are housed in environments which place them outside their thermal comfort zone. However, the identification of thermal stress, when based on air properties, suggests the use of outdated and generic indices. The objective of this work was to develop and validate a methodology for classifying and diagnosing heat stress in production animals based on psychrometric air relations. The model was created for broilers, pigs, dairy cattle, and laying birds, categorized into a total of 21 breeding phases. For each phase, a bibliographic search was carried out for the psychrometric parameters of the air-dry bulb temperature (AT) and relative humidity (RH)-that satisfied the animals' critical and ideal thermoneutral zones. Adding the local atmospheric pressure (AP), the parameters were used to calculate the enthalpy (h), resulting in five comfort ranges. Based on this, a decision tree was elaborated, consisting of three attributes (AT, RH, and h) and seven diagnostic classes, based on the psychrometric principles of air. The proposed methodology was used in a case study, with a database extracted from an individual shelter for calves. For the evaluation of the decision tree, two induction algorithms, ID3 and c4.5, were compared, both of which presented high accuracy and proposed simpler tree models than the one theoretically developed for the methodology. In conclusion, the methodology represents a great potential to characterize the thermal comfort of the animals, diagnose the causes of stress, and recommend possible corrective actions. The study revealed that decision trees can be adapted and simplified for each creation phase.

Are there differences in the adaptive profile of hair sheep and their crosses with wool breeds?

This study aimed to evaluate and compare the physiological performance of different genetic groups of sheep, by physiological variables and serum hormone levels, in a hot weather environment. Thirty sheep from five genetic groups were used: Santa Inês (SI), ½ Dorper + ½ Santa Inês (DO), ½ Ilê de France + ½ Santa Inês (IF), ½ Suffolk + ½ Santa Inês (SK), and ½ Texel + ½ Santa Inês (TX). The readings and records of physiological parameters (respiratory rate (RR), rectal temperature (RT), auricular cavity temperature (ACT), and surface temperature (ST)) were carried out at 7:00 am, 1:00 pm, and 7:00 pm, in 12 non-consecutive days. The collections of blood samples for hormone analysis (triiodothyronine (T3), thyroxine (T4), and cortisol (CORT)) is in four consecutive days. The environmental conditions of the experimental period caused a thermal discomfort in the sheep, but not a state of thermal stress. The thermolysis mechanisms, sensitive (ST and ACT) and latent (RR) processes, were enough to maintain their homeostasis (RT). The results showed that crossbred breeds presented a higher metabolism and were more efficient at dissipating heat through thermolysis than the SI breed. The crossbred breeds were efficient at dissipating heat through the elevation of body surface temperature and respiratory rate, mainly SK and TX, i.e., crossbred breeds, despite the wool cover, used thermoregulatory mechanisms that promoted lower variation of RT. The analysis of variance showed significant effects (P < 0.05) to the time factor in the responses of T4 and T3, and to the breed factor in the responses of CORT, T4, and T3. We did not observe interaction between the factors to any of the hormonal variables. Therefore, we can state that the effect of time was independent of breed and vice versa. Thyroid hormones presented lower blood concentration in the mornings (4.03 ± 0.82, T4; 65.08 ± 10.6, T3), increasing their concentration in the afternoon (4.60 ± 1.03, T4; 70.16 ± 14.17, T3). The thyroid hormones presented a normal circadian rhythm, with the exception of SK. Air temperature (AT) showed greater correlation with physiological variables than enthalpy (H) did, in the experimental conditions. However, H showed correlation with T4 and T3. The adaptive profile of the genetic groups under study are different, but the IF genetic group showed better performance under environmental conditions.

Overexpression of miR-17-5p may negatively impact p300/CBP factor-associated inflammation in a hypercholesterolemic advanced prostate cancer model.

BACKGROUND: Previously, we demonstrated that cholesterol triggers the increase in p300/CBP-associated factor (PCAF), targeted by miR-17-5p. The p300, IL-6, PCAF, and miR-17-5p genes have important and contradictory roles in inflammation and prostate cancer (PCa). This study aimed to demonstrate the potential anti-inflammatory effect of miR-17-5 in an advanced PCa model with diet-induced hypercholesterolemia. METHODS AND RESULTS: In vitro, using the PC-3 cell line, we show that induction of miR-17-5p reduces p300 and PCAF expression, increases apoptosis, and decreases cell migration. Furthermore, we demonstrate that supplementing this same cell with cholesterol (2 µg/mL) triggers increased p300, IL-6, and PCAF. In vivo, after establishing the hypercholesterolemic (HCOL) model, xenografts were treated with miR-17-5p. Increased expression of this miR after intratumoral injections attenuated tumor growth in the control and HCOL animals and reduced cell proliferation. CONCLUSION: Our results demonstrate that inducing miR-17-5p expression suppresses tumor growth and inflammatory mediator expression. Further studies should be conducted to fully explore the role of miR-17-5p and the involvement of inflammatory mediators p300, PCAF, and IL-6.

Spatial modeling via geostatistics of the bed in a compost barn system: thermal performance assessments.

For all the benefits of a compost barn system to be achieved, the management, especially with the bedding, must be done correctly. One should consider the requirements regarding the revolving of the material, use of good quality organic matter, suitable ventilation, and an ideal animal stocking rate, so that the composting occurs efficiently. Given this, this work aimed to evaluate the spatial variability of thermal characteristics of rice straw bedding, using values of temperature (surface and internal) and humidity. The temperature and moisture values of the litter were recorded at intervals during the morning shift, from 6:00 am to 7:00 am and from 8:00 am to 9:00 am, and during the afternoon shift, from 3:00 pm to 4:00 pm and from 5:00 pm to 6:00 pm, considering the management of the turn-over, that is, before and after the movement of the litter. Litter variables were collected at points distributed on a 39-point grid. There were no differences between litter surface temperatures before and after turning over in the morning shift, but in the afternoon shift, the surface temperature was lower (P < 0.05) before, when compared to after turning. The internal temperature and litter moisture were higher (P < 0.05) before revolving in both shifts. Spatial dependence was verified (IDE) for the variables studied. Based on the geostatistical analysis, it is concluded that the IDE was rated as strong for surface and internal temperatures, which demonstrates heterogeneity among the points evaluated and a large spatial variability regarding bed temperatures. In the case of litter moisture, the greatest variability occurred in the morning period. The litter surface temperature values did not exceed the ambient air temperature values.

Thermoregulatory, behavioral, and productive responses and physical integrity of primiparous and multiparous cows on compost barn in Brazilian tropical conditions.

The compost barn is presented as a system which is capable of providing a suitable environment for dairy cows, but this must be assessed for different climatic regions. Few studies have been carried out evaluating the physics of the thermal environment of this system under tropical conditions. In this study, we evaluated thermoregulatory, behavioral, and productive responses and physical integrity in primiparous and multiparous cows housed in a compost barn system under tropical conditions. From a total of 121 clinically healthy dairy cows aged 3 to 6 years, 30 Girolando cows (7/8) were randomly selected, divided into two groups, according to calving order (primiparous and multiparous), body weight, lactation curve and milk production for the evaluations. Thus, group 1 (primiparous) with an average weight of 524 kg and production of 30 kg was characterized, and group 2 (multiparous) with an average weight and production of 635 kg and 36 kg, respectively. The enthalpy was higher (P < 0.05) in the internal environment of the shed at the three evaluated times (3:30 a.m., 11:30 a.m., and 6:30 p.m.), but the humidity did not vary (P > 0.05) between the internal and external environments at the evaluated times. Respiratory rate was higher (P < 0.0001) in multiparous cows at 11:30 a.m., but was similar at 3:30 a.m. and 6:30 p.m. when compared with primiparous cows. The coat surface temperature was higher (P < 0.001) at 3:30 a.m., but similar at the other two times. For the variables lameness and dirtiness, the vast majority of animals presented scores considered adequate (1 and 2), indicating that it was able to provide an ideal physical environment. Regarding animal behavior, panting (O) and lying idle (OD) were higher (P < 0.05) in multiparous cows. Multiparous cows have higher (P < 0.0001) milk production. Milk production has a negative correlation with enthalpy. The CB system was not able to provide a suitable thermal environment for the animals. Multiparous cows present higher heat stress with change in behavioral responses, especially at midday, but with higher milk production when compared to primiparous cows in compost barn under tropical conditions.

Thermal performance of a low-profile cross-ventilated freestall dairy barn with evaporative cooling pads in a hot and humid climate.

A commercial low-profile cross-ventilated system (LPCV) free-stall barn with evaporative cooling pads on the air intake side was used to assess the potential of cross-ventilation with evaporative cooling to alleviate heat stress in lactating dairy cows. A total of 5712 records (238 days × 24 h) of air temperature (AT) and relative humidity (RH) values were collected. In order to state the effectiveness of the ventilation system and potential heat stress, the temperature humidity index (THI) and enthalpy (h) were calculated outside and inside the LPCV barn. The cooling capacity of the LPCV system is greater in conditions of low humidity, and the difference between the values of the internal and external temperature reached -12 °C. The LPCV reduced exposure to heat stress, increasing the time cows spent in thermoneutral conditions. This system could reduce the occurrence of temperature above 25 °C from 33 to 5% and 11% in sections A (represents the inlet side of the barn) and D (represents the outlet side of the barn), respectively. On the other hand, the LPCV kept a high and constant rate of RH throughout the day. The average inside and outside AT were significantly different, but differences in temperature were observed across the width of the barn. Analyzing the barn by the indexes, the THI showed that the barn promoted cows more time confined under non-stressful thermal conditions (between 83.8 and 90% vs 68.7%) and stress period much lower compared to the external environment (between 10 and 16.2% vs. 31.3%). The enthalpy index showed that there was not much difference between the internal and external environments. The LPCV system is another installation alternative for raising dairy cow in a hot and humid climate.

Vaginal temperature modeling in Holstein cows and cluster analysis by the cophenetic correlation coefficient.

The objective was to establish a model for the prediction and characterization of vaginal temperature in Holstein cows, based on environmental predictors and thermal comfort indices, through cluster analysis, validation by the cophenetic correlation coefficient, and multiple regression analysis. The micrometeorological characterization of the site was carried out by recording the air temperature (Tair), the relative humidity (RH), the black globe temperature (BGT), the black globe temperature and humidity (BGHI), and dew point temperature (TDP). The recording of vaginal temperature (Tv) was performed in eight dairy cows using temperature sensors, equipped with data loggers, coupled with intravaginal devices. The data were analyzed using descriptive statistics and cluster analysis (CA) by using the hierarchical agglomerative method based on the value of the cophenetic correlation coefficient (CCC >0.70), in which representative physiological models were established, characterizing the Tv through multiple regression. In the afternoon the coefficient of variation (CV) was low for all variables, indicating homogeneity of the meteorological variables and efficiency of the ventilation system. The temperature and humidity index (THI) was mild only on the morning. There was a variation of 0.28 °C of Tv between shifts, sufficient to characterize the condition of comfort and stress of the animal, with values above 39 °C indicating animal stress. Tv showed strong correlation with BGT, Tair, TDP and RH, assuming that physiological variables, such as Tv, tend to have greater relationship with abiotic variables. Empirical models were established for estimating Tv based on the analyses performed in this study. Model 1 is recommended for TDP ranges of 14.00-21.00 °C and RH of 30-100%, while model 2 can be used for Tair situations up to 35 °C. The regression models for estimating Tv are promising for characterizing the thermal comfort of dairy cows housed in compost barn systems.

Intratumoral Restoration of miR-137 Plus Cholesterol Favors Homeostasis of the miR-137/Coactivator p160/AR Axis and Negatively Modulates Tumor Progression in Advanced Prostate Cancer.

MicroRNAs (miRNAs) have gained a prominent role as biomarkers in prostate cancer (PCa). Our study aimed to evaluate the potential suppressive effect of miR-137 in a model of advanced PCa with and without diet-induced hypercholesterolemia. In vitro, PC-3 cells were treated with 50 pmol of mimic miR-137 for 24 h, and gene and protein expression levels of SRC-1, SRC-2, SRC-3, and AR were evaluated by qPCR and immunofluorescence. We also assessed migration rate, invasion, colony-forming ability, and flow cytometry assays (apoptosis and cell cycle) after 24 h of miRNA treatment. For in vivo experiments, 16 male NOD/SCID mice were used to evaluate the effect of restoring miR-137 expression together with cholesterol. The animals were fed a standard (SD) or hypercholesterolemic (HCOL) diet for 21 days. After this, we xenografted PC-3 LUC-MC6 cells into their subcutaneous tissue. Tumor volume and bioluminescence intensity were measured weekly. After the tumors reached 50 mm3, we started intratumor treatments with a miR-137 mimic, at a dose of 6 µg weekly for four weeks. Ultimately, the animals were killed, and the xenografts were resected and analyzed for gene and protein expression. The animals' serum was collected to evaluate the lipid profile. The in vitro results showed that miR-137 could inhibit the transcription and translation of the p160 family, SRC-1, SRC-2, and SRC-3, and indirectly reduce the expression of AR. After these analyses, it was determined that increased miR-137 inhibits cell migration and invasion and impacts reduced proliferation and increased apoptosis rates. The in vivo results demonstrated that tumor growth was arrested after the intratumoral restoration of miR-137, and proliferation levels were reduced in the SD and HCOL groups. Interestingly, the tumor growth retention response was more significant in the HCOL group. We conclude that miR-137 is a potential therapeutic miRNA that, in association with androgen precursors, can restore and reinstate the AR-mediated axis of transcription and transactivation of androgenic pathway homeostasis. Further studies involving the miR-137/coregulator/AR/cholesterol axis should be conducted to evaluate this miR in a clinical context.

The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer.

Prostate cancer (PCa) has a high prevalence and represents an important health problem, with an increased risk of metastasis. With the advance of CRISPR-Cas9 genome editing, new possibilities have been created for investigating PCa. The technique is effective in knockout oncogenes, reducing tumor resistance. MMP9 and miR-21 target genes are associated with PCa progression; therefore, we evaluated the MMP-9 and miR-21 targets in PCa using the CRISPR-Cas9 system. Single guide RNAs (sgRNAs) of MMP9 and miR-21 sequences were inserted into a PX-330 plasmid, and transfected in DU145 and PC-3 PCa cell lines. MMP9 and RECK expression was assessed by qPCR, WB, and IF. The miR-21 targets, integrins, BAX and mTOR, were evaluated by qPCR. Flow cytometry was performed with Annexin5, 7-AAD and Ki67 markers. Invasion assays were performed with Matrigel. The miR-21 CRISPR-Cas9-edited cells upregulated RECK, MARCKS, BTG2, and PDCD4. CDH1, ITGB3 and ITGB1 were increased in MMP9 and miR-21 CRISPR-Cas9-edited cells. Increased BAX and decreased mTOR were observed in MMP9 and miR-21 CRISPR-Cas9-edited cells. Reduced cell proliferation, increased apoptosis and low invasion in MMP9 and miR-21 edited cells was observed, compared to Scramble. CRISPR-Cas9-edited cells of miR-21 and MMP9 attenuate cell proliferation, invasion and stimulate apoptosis, impeding PCa evolution.

Treatment of a Snakebite Injury With Secondary Bacterial Infection: A Case Study.

BACKGROUND: Pit viper snakebites are challenging as they often cause tissue injury and secondary bacterial infection that may impair full recovery of the affected limb. We describe the evolution of a snakebite injury with secondary infection and the use of specialized dressings to achieve tissue repair and complete closure of the wound. CASE: Ms E., a 45-year-old woman, was bitten by a pit viper that began as a small lesion and progressed to necrosis, cellulitis, edema, and hyperemia of the perilesional skin, local inflammation, and infection. We implemented a combination of topical hydrogel therapy with calcium alginate and hydrofiber with 1.2% silver to promote autolytic debridement, combat local infection, and provide a moist wound environment. The wound required daily local treatment for 2 months due to extensive tissue damage, combined with the proteolytic action of the bothropic venom. CONCLUSION: The care of wounds caused by snakebites is a challenge for the health care team due to tissue loss resulting from the action of the venom and secondary bacterial infection. Close follow-up with the use of systemic antibiotics and topical therapies proved effective in minimizing tissue loss in this case.

Evaluation of environmental enrichment on productive and behavioral responses of finishing pigs.

Productive and behavioral responses of finishing pigs with and without access to environmental enrichment (EE) were evaluated. A total of 432 pigs, male and female, Hampshire line, initial and final weight of 22-27 kg and 110-125 kg, respectively, were studied for 112 days. The study design was in random blocks, with 6 treatments, distributed in a factorial scheme 2 × 3 (sex × treatment) and 12 repetitions/treatment, totalling 72 pens. The animals were weighed and distributed in 3 blocks-light (20.3 to 24.8 kg), medium weight (24.8 to 27.7 kg), and heavy (27.7 to 33.8 kg). The treatments were as follows: branched chains for males (T1); branched sisal ropes for males (T2), males without EE (T3); branched chains for females (T4); branched sisal ropes for females (T5) and females without EA (T6). There was no effect (P > 0.05) of interaction (enrichment × sex) and sex on performance. Daily consumption and daily weight gain were lower (P < 0.05) in the chain group compared to the sisal rope and control groups. Feed conversion was similar (P > 0.05) between treatments under study. There was no interaction between gender and enrichment for behavioral responses (P = 0.8986). However, there was an environmental enrichment (P = 0.0017; P < 0.001) and gender (P = 0.0251; P < 0.001) effect in relation to the animals' behavior in the morning and in the afternoon, respectively. Calmer animals, regardless of sex, were observed for groups without enrichment (control) and morning and afternoon periods. The animals interacted with the types of enrichment under study, given the higher probabilities of interaction with the object for males and females in the morning and afternoon shifts. It was observed, regardless of the shifts, that the control treatment for males and females was different when compared to the other enrichments for males and females. In general, the pigs' behaviors were expressed more in the afternoon when compared to the morning. It was observed that the animal interaction and aggression (afternoon shift) behaviors are more related to the control group, while object interaction is more related to the enriched treatments. The use of sisal ropes and branched chains guarantees the performance and behavior of pigs in the finishing phase, but we recommend the use of branched chains, as this enrichment increased the profitability of the farmer by reducing feed consumption, maintaining feed conversion and animal welfare, besides being a more durable material than sisal rope.

Evaluation of environmental enrichment on thermophysical responses, carcass traits, and meat quality of finishing pigs.

Environmental enrichment consists of a way to provide better living conditions for animals, but its influence on physical integrity, thermoregulatory responses, and quality of pork meat is not known. The objective of this study was to evaluate the thermoregulatory responses, lesion score, lameness, carcass traits, and meat quality of pigs with and without access to environmental enrichment (EE) in the finishing phase. A total of 432 pigs of the Hampshire lineage, males and females, with average initial and final weights between 22-27 kg and 110-125 kg respectively were evaluated. The experimental design used was in randomized blocks, with 6 treatments, distributed in a 2×3 factorial scheme (sex × environmental enrichment), with 12 repetitions per treatment, totaling 72 stalls. The treatments were branched chain for males (T1), branched sisal rope for males (T2), males without EE (T3), branched chain for females (T4), branched sisal rope for females (T5), and females without EE (T6). Weekly, two assessments of physiological data were performed, in loco, in the morning and afternoon. Lesions on the tail, ear, body, and lameness were evaluated on the 1st, 16th, 37th, 51st, 79th, 93rd, and 112th days. On the 112th day, 72 animals were slaughtered for carcass traits and meat quality analyses. Statistical analysis was performed using generalized and mixed linear models. There was no effect (p>0.05) of the interaction of the studied factors (environmental enrichment × sex × period) on the head, back, leg, and average temperature. However, there was an effect of the period factor (p<0.01), with higher temperatures in the afternoon compared to the morning. For tail injury, there was no association between injury degree and gender (p=0.56) and between injury degree and enrichment (p=0.99), and there was no interaction effect (p=0.15). For ear injury, there was no association between the degree of ear injury and gender (p=0.72), as well as for the degree of ear injury and enrichment (p=0.99) and interaction (p=0.47). There was a higher occurrence of categories 2, 3, and 4 for lesions on the body in males (p=0.0257), regardless of the EE. Water retention, carcass yield, cut yield, marbling pattern, pH, and color showed no effect for sex, enrichment, and interaction between the two factors (p>0.05). The use of environmental enrichment with sisal ropes and branched chains does not change the thermophysical responses, carcass traits, and meat quality of finishing pig.

Environmental enrichment in finishing pigs: does it promote any changes in bone biomechanics?

The resistance of pigs' bone structure was evaluated for the first time, reared with and without environmental enrichment (EE) in the finishing phase using techniques in bone biomechanics; 432 swine from the Hampshire breed, being males and females, with initial body weight between 22 and 27 kg and final body weight between 110 and 125 kg were evaluated for 112 days. The experimental design was in randomized blocks, with 6 treatments, distributed in a 2 × 3 factorial scheme (sex × conditions in creation), with 12 repetitions/treatment, totaling 72 pens. The treatments were as follows: branched chain for males (T1), sisal branched string for males (T2), males without EE (T3), branched chain for females (T4), sisal branched string for females (T5), and females without EE (T6). At the end of the experimental period, all animals were slaughtered in an industrial slaughterhouse, having their femur bones collected for bone biomechanics analysis. There was no effect (P > 0.05) of the interaction (enrichment × sex) and individual factors for bone weight. There was a tendency (P = 0.08) of the interaction for flexion force, being higher in males enriched with branched ropes and chains. For breaking stress, there was an interaction effect (P = 0.04), being the females without EE the ones showing the lowest breaking stress, favoring bone fragility. The use of branched ropes and chains strengthens the bone structure in swine in the termination phase (110-125 kg BW - 183-190 days in age), being an important strategy used to meet the animal welfare requirements.

Cholesterol Triggers Nuclear Co-Association of Androgen Receptor, p160 Steroid Coactivators, and p300/CBP-Associated Factor Leading to Androgenic Axis Transactivation in Castration-Resistant Prostate Cancer.

BACKGROUND/AIMS: Cholesterol modulates intratumoral androgenic signaling in prostate cancer; however, the molecular mechanisms underlying these changes in castration-resistant prostate cancer (CRPC) are not fully elucidated. Herein, we investigated the effect of cholesterol on androgen receptor (AR) coactivators expression and tumorigenesis in vitro and in vivo. METHODS: Herein, we monitored the expression of AR coactivators (SRC-1, 2, 3 and PCAF) genes in PC-3 cells exposed to 2µg/mL of cholesterol for 8 hours by qPCR. We also performed cell migration at 0, 8, 24, 48 and 72h and flow cytometry assays (viability, apoptosis, and cell cycle) after a 24h exposure. Immunofluorescence assay was performed to evaluate the protein expression of the AR coactivators. Additionally, in vivo experiments were conducted using 22 male NOD/SCID mice. Mice were fed a standard (Control) or hypercholesterolemic (HCOL) diet for 21 days and then subcutaneously implanted with PC-3 cells. The tumor volume was calculated every two days, and after four weeks, the tumors were resected, weighed, and the serum lipid profile was measured. We also measured the intratumoral lipid profile and AR coactivators gene and protein expression by qPCR and Western Blot, respectively. Intratumor testosterone and dihydrotestosterone (DHT) concentrations were determined using ELISA. RESULTS: Cholesterol up-regulated the gene expression of coactivators SRC-1, SRC-2, SRC-3and PCAF, increasing AR expression in PC-3 cells. Next, cholesterol-supplemented PC-3 cells exhibited increased cell migration and altered cell cycle phases, leading to changes in proliferation and reduced apoptosis. We found that SRC-1, SRC-2, SRC-3 and PCAF proteins co-localized in the nucleus of cholesterol-supplemented cells and co-associate with AR. In the in vivo model, the hypercholesterolemic (HCOL) group displayed higher serum total and intratumoral cholesterol levels, increased testosterone and dihydrotestosterone concentrations, and up-regulated AR coactivator expression. The tumor volume of the HCOL group was significantly higher than the control group. CONCLUSION: Our findings revealed that increased nuclear translocation of the coactivators leads to up-regulated AR gene and protein expression, potentially influencing tumor progression. Studies targeting cholesterol-modulated changes in AR coactivator expression may provide insights into the molecular mechanisms associated with the CRPC phenotype.

Lower levels of CXCL-8 and IL-2 on admission as predictors of early adverse reactions to Bothrops antivenom in the Brazilian Amazon.

Snakebite envenomings are considered a global health problem. The specific therapy for these envenomings consists of administering animal-derived antivenoms aiming to neutralize the venom toxins. Antivenoms have been used effectively to treat snakebites for more than a century; however, their administration may result in early and/or late adverse reactions. The present study presents the prevalence of early adverse reactions (EARs) towards Bothrops antivenom therapy in a health tertiary unit in the Brazilian Amazon and explores if specific plasma cytokines and chemokines from envenomed patients could be used as predictors of EARs. A cohort of patients bitten by Bothrops atrox was followed-up at the Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD), from 2014 to 2016. Patients were treated with the Brazilian Bothrops antivenom and CXCL-8, CCL-5, CXCL-9, CCL-2, CXCL-10, IL-6, TNF, IL-2, IL-10, IFN-y, IL-4, and IL-17A were evaluated in patients' plasma samples before and after antivenom administration. From the total of patients (n = 186), mostly were male (82.3%), inhabiting rural areas (87.1%), with an average age of 35 years. Most of the patients (83.8%) were admitted to the hospital within 6 h after the accident, 26 (14%) reported having suffered a previous snakebite, and 97 (52.1%) received between 7 and 9 antivenom vials. The frequency of antivenom-induced EARs was 11.8% (22), resulting mostly of mild reactions. Urticaria was the major EAR manifestation (46.4%). Interestingly, CXCL-8 and IL-2 showed significantly lower levels in patients who progressed to EARs, although IL-2 levels might not represent biological relevance due the small magnitude difference between groups. This study reveals that CXCL-8 and IL-2 could play a role in the onset of EARs in pit viper envenomings.

Enhancing the Luminescence of Eu(III) Complexes with the Ruthenocene Organometallic Unit as Ancillary Ligand.

Five novel Eu(III)-ß-diketonate complexes containing ruthenocene ancillary ligands (1,1'-bis(diphenylphosphoryl)ruthenocene─RcBPO) were synthesized and characterized. The coordination compounds presented the general formula [Eu(ß-dik)3(RcBPO)], where ß-dik stands for 2-thenoyltrifluoroacetonate (tta), 3-benzoyl-1,1,1-trifluoroacetone (btf), 2-dibenzoylmethanate (dbm), 2-acetyl-1,3-indandionate (aind), and 2-benzoyl-1,3-indandionate (bind), and RcBPO stands for 1,1'-bis(diphenylphosphoryl)ruthenocene. The [Eu(aind)3(RcBPO)] complex crystallizes in a monoclinic Cc non-centrosymmetric space group with the europium site environment, assuming a bicapped trigonal prism coordination polyhedron with the symmetry point group close to C2v. Photoluminescent properties for the solid-state samples were described in terms of excitation, emission, lifetime decay curves, and intrinsic and overall quantum yields. The replacement of the two coordinated H2O molecules by the RcBPO ancillary ligand leads to great enhancements of the overall quantum yields (QEuL), with the minimum increment by a factor of 5 for the case of [Eu(btf)3(RcBPO)] and the maximum enhancement of 270 times for the case of the [Eu(dbm)3(RcBPO)] complex. In addition, theoretical calculations were carried out to model the spectroscopic properties of the investigated compounds. To obtain theoretical Judd-Ofelt parameters (Ωλ, λ = 2, 4, and 6) and intramolecular energy transfer rates, the JOYSpectra web platform was employed using the structure obtained from density functional theory calculations. Hence, a rate equation model provided theoretical overall quantum yields, which are in great agreement with measured data.

CCL-2 and CXCL-8: Potential Prognostic Biomarkers of Acute Kidney Injury after a Bothrops atrox Snakebite.

In the Brazilian Amazon, the snake Bothrops atrox is the primary cause of snakebites. B. atrox (BaV) venom can cause systemic pathophysiological changes such as acute kidney injury (AKI), which leads to the production of chemokines and cytokines in response to the envenomation. These soluble immunological molecules act by modulating the inflammatory response; however, the mechanisms associated with the development of AKI are still poorly understood. Here, we characterize the profile of these soluble immunological molecules as possible predictive biomarkers of the development of AKI. The study involved 34 patients who had been victims of snakebites by Bothrops sp. These were categorized into two groups according to the development of AKI (AKI(-)/AKI(+)), using healthy donors as the control (HD). Peripheral blood samples were collected at three-time points: before antivenom administration (T0) and at 24 and 48 hours after antivenom (T1 and T2, respectively). The soluble immunological molecules (CXCL-8, CCL-5, CXCL-9, CCL-2, CXCL-10, IL-6, TNF, IL-2, IL-10, IFN-γ, IL-4, and IL-17A) were quantified using cytometric bead array. Our results demonstrated an increase in CXCL-9, CXCL-10, IL-6, IL-2, IL-10, and IL-17A molecules in the groups of patients who suffered Bothrops snakebites (AKI(-) and AKI(+)) before antivenom administration, when compared to HD. In the AKI(+) group, levels of CXCL-8 and CCL-2 molecules were elevated on admission and progressively decreased during the clinical evolution of patients after antivenom administration. In addition, in the signature analysis, these were produced exclusively by the group AKI(+) at T0. Thus, these chemokines may be related to the initiation and extension of AKI after envenomation by Bothrops and present themselves as two potential biomarkers of AKI at T0.

Effect of distance and daily periods on heat-stressed pigs and pre-slaughter losses in a semiarid region.

Effective planning animal transport is essential to safeguard animal welfare and reduce production losses. Environmental conditions, specifically extreme temperatures in combination with ranges of relative humidity are highlighted as one of the main risk factors for production losses during transport (e.g., fatalities). The majority of research evaluating both welfare and production impacts of pig transport have been primarily undertaken in Europe and North America, which cover a relatively limited range of distinct climates (e.g., temperate, sub-arctic, etc.). As a result, research on pig transport in semi-arid conditions is lacking. In this study, we evaluated the effects of both distance (short, 30 km; and long, 170 km) and transport daily periods (morning, (05:00-11:00); afternoon (12:00-17:00); and night, (23:00-04:00)) on the preslaughter losses and heat stress of pigs in commercial transport in a semiarid region. Across 19 journeys of standard slaughter-weight pig loads (124.0 ± 2.8 kg), 684 focal animals (36 per journey) were evaluated. For each journey, the load's thermal profile (THIadj and enthalpy) and physiological responses of individual pigs were recorded. On arrival at designated slaughterhouses, the percentage pig of non-ambulatory non-injured (NANI), non-ambulatory injured (NAI), death on arrival (DOA), and total losses were recorded. Short journeys in the afternoon were shown to be more detrimental to the thermal comfort of pigs, with higher rectal temperatures recorded. The highest percentage of total losses and DOA occurred in afternoon journeys, irrespective of distance, followed by the morning, with the lowest losses observed in pigs transported at night. Additionally, total losses and DOA were further exacerbated by journey distance, with higher rates observed in short journeys. Higher percentage averages of NANI and NAI were observed in shorter journeys, but daily periods effects were only observed for NANI. These results further demonstrate the welfare and production loss risks associated with journey distance and time of day (representing varying environmental conditions) during road transport of pigs, whilst providing novel data in semiarid conditions. Careful and effective planning for pig transportation is essential to minimize heat stress and production losses. Consideration of the thermal environment on the day of travel, as well as providing flexibility to adjust travel times (e.g., early morning or evening), should help to mitigate risks of heat stress and production losses during pig transport.

Can increased expression of miR-Let-7c reduce the transition potential of high-grade urothelial carcinoma?

BACKGROUND: Bladder cancer is the leading transitional cell carcinoma affecting men and women with high morbidity and mortality rates, justifying the need to develop new molecular target therapies using microRNAs. This study aimed to evaluate the behavior of the T24 cell line after transfection with miR-Let-7c precursor mimic through invasion, migration, apoptosis, and cell cycle assays. METHODS AND RESULTS: T24 cell was transfected with the Let-7c mimic and its respective control and evaluated after 24 h. The expression levels of miR-Let-7c were analyzed by qPCR. We performed wound healing, Matrigel and flow cytometry, apoptosis, and cell cycle assays to determine its effect on cellular processes. Cells transfected with miR-Let-7c showed increased apoptosis rates (p = 0.019), decreased migration 24 h (p = 0.031) and 48 h (p = 0.0006), invasion potential (p = 0.0007), and cell proliferation (p = 0.002). CONCLUSIONS: Our results demonstrate that miR-Let-7c can act in different pathways of the carcinogenic cellular processes of muscle-invasive urothelial carcinoma cells, inhibiting cell proliferation and increasing apoptosis levels, consequently limiting their invasion potential. However, further studies should be carried out better to elucidate this microRNA's role in high-grade urothelial carcinomas and unveil which targets this microRNA may present, which are intrinsically related to the cancer survival pathways.