Impact of Growth Hormone on Microglial and Astrocytic Function.

The role of growth hormone (GH) in the central nervous system (CNS) involves neuroprotection, neuroregeneration, formation of axonal projections, control of cognition, and regulation of metabolism. As GH induces insulin-like growth factor-1 (IGF-1) expression in many tissues, differentiating the specific functions of GH and IGF-1 in the organism is a significant challenge. The actions of GH and IGF-1 in neurons have been more extensively studied than their functions in nonneuronal cells (e.g., microglial cells). Glial cells are fundamentally important to CNS function. Microglia, astrocytes, oligodendrocytes, and tanycytes are essential to the survival, differentiation, and proliferation of neurons. As the interaction of the GH/IGF-1 axis with glial cells merits further exploration, our objective for this review was to summarize and discuss the available literature regarding the genuine effects of GH on glial cells, seeking to differentiate them from the role played by IGF-1 action whenever possible.

Effects of Bradykinin B2 Receptor Ablation from Tyrosine Hydroxylase Cells on Behavioral and Motor Aspects in Male and Female Mice.

The kallikrein-kinin system is a versatile regulatory network implicated in various biological processes encompassing inflammation, nociception, blood pressure control, and central nervous system functions. Its physiological impact is mediated through G-protein-coupled transmembrane receptors, specifically the B1 and B2 receptors. Dopamine, a key catecholamine neurotransmitter widely distributed in the CNS, plays a crucial role in diverse physiological functions including motricity, reward, anxiety, fear, feeding, sleep, and arousal. Notably, the potential physical interaction between bradykinin and dopaminergic receptors has been previously documented. In this study, we aimed to explore whether B2R modulation in catecholaminergic neurons influences the dopaminergic pathway, impacting behavioral, metabolic, and motor aspects in both male and female mice. B2R ablation in tyrosine hydroxylase cells reduced the body weight and lean mass without affecting body adiposity, substrate oxidation, locomotor activity, glucose tolerance, or insulin sensitivity in mice. Moreover, a B2R deficiency in TH cells did not alter anxiety levels, exercise performance, or motor coordination in female and male mice. The concentrations of monoamines and their metabolites in the substantia nigra and cortex region were not affected in knockout mice. In essence, B2R deletion in TH cells selectively influenced the body weight and composition, leaving the behavioral and motor aspects largely unaffected.

Cervical Squamous Cell Carcinoma Diagnosis by FTIR Microspectroscopy.

Cervical cancer was considered the fourth most common cancer worldwide in 2020. In order to reduce mortality, an early diagnosis of the tumor is required. Currently, this type of cancer occurs mostly in developing countries due to the lack of vaccination and screening against the Human Papillomavirus. Thus, there is an urgent clinical need for new methods aiming at a reliable screening and an early diagnosis of precancerous and cancerous cervical lesions. Vibrational spectroscopy has provided very good results regarding the diagnosis of various tumors, particularly using Fourier transform infrared microspectroscopy, which has proved to be a promising complement to the currently used histopathological methods of cancer diagnosis. This spectroscopic technique was applied to the analysis of cryopreserved human cervical tissue samples, both squamous cell carcinoma (SCC) and non-cancer samples. A dedicated Support Vector Machine classification model was constructed in order to categorize the samples into either normal or malignant and was subsequently validated by cross-validation, with an accuracy higher than 90%.

Trans-resveratrol reduced hepatic oxidative stress in an animal model without inducing an upregulation of nuclear factor erythroid 2-related factor 2.

Trans-resveratrol, a widely used supplement for humans, aims to enhance the body's antioxidant defense. Studies suggest that it exerts anti-inflammatory and antioxidant effects by activating the nuclear factor erythroid 2-related factor 2 (Nrf2). In order to evaluate this hypothesis, LDLr(-/-) mice were fed a Western diet to induce liver inflammation and oxidative stress. One group was fed a diet containing 0.60 mg/day of trans-resveratrol (RESV), while another group received no dietary supplementation (CONT). Oxidative stress biomarkers and inflammatory cytokines were assessed in liver homogenates. It was observed that trans-resveratrol decreased hepatic oxidative stress by increasing the GSH/GSSG ratio and reducing malondialdehyde (MDA) concentration. However, the RESV group exhibited a reduction in Nrf2 relative expression compared to CONT. Additionally, trans-resveratrol supplementation reduced nuclear factor-κB (NF-κB) expression but led to an increase in IL-6, with no significant changes observed in tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) concentrations. Overall, these findings indicate that the in vivo antioxidant impact induced by trans-resveratrol supplementation in hepatic tissue did not correlate with increase of inflammatory cytokines and Nrf2 relative expression. Further exploration of alternative mechanisms, such as direct radical scavenger activity, is warranted to elucidate the antioxidant effect.

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.

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.

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.

SUCCOR quality: validation of ESGO quality indicators for surgical treatment of cervical cancer.

OBJECTIVE: To evaluate whether compliance with European Society of Gynaecological Oncology (ESGO) surgery quality indicators impacts disease-free survival in patients undergoing radical hysterectomy for cervical cancer. METHODS: In this retrospective cohort study, 15 ESGO quality indicators were assessed in the SUCCOR database (patients who underwent radical hysterectomy for International Federation of Gynecology and Obstetrics (FIGO) stage 2009 IB1, FIGO 2018 IB1, and IB2 cervical cancer between January 2013 and December 2014), and the final score ranged between 0 and 16 points. Centers with more than 13 points were classified as high-quality indicator compliance centers. We constructed a weighted cohort using inverse probability weighting to adjust for the variables. We compared disease-free survival and overall survival using Cox proportional hazards regression analysis in the weighted cohort. RESULTS: A total of 838 patients were included in the study. The mean number of quality indicators compliance in this cohort was 13.6 (SD 1.45). A total of 479 (57.2%) patients were operated on at high compliance centers and 359 (42.8%) patients at low compliance centers. High compliance centers performed more open surgeries (58.4% vs 36.7%, p<0.01). Women who were operated on at centers with high compliance with quality indicators had a significantly lower risk of relapse (HR=0.39; 95% CI 0.25 to 0.61; p<0.001). The association was reduced, but remained significant, after further adjustment for conization, surgical approach, and use of manipulator surgery (HR=0.48; 95% CI 0.30 to 0.75; p=0.001) and adjustment for adjuvant therapy (HR=0.47; 95% CI 0.30 to 0.74; p=0.001). Risk of death from disease was significantly lower in women operated on at centers with high adherence to quality indicators (HR=0.43; 95% CI 0.19 to 0.97; p=0.041). However, the association was not significant after adjustment for conization, surgical approach, use of manipulator surgery, and adjuvant therapy. CONCLUSIONS: Patients with early cervical cancer who underwent radical hysterectomy in centers with high compliance with ESGO quality indicators had a lower risk of recurrence and death.

SUCCOR cone study: conization before radical hysterectomy.

OBJECTIVE: To evaluate disease-free survival of cervical conization prior to radical hysterectomy in patients with stage IB1 cervical cancer (International Federation of Gynecology and Obstetrics (FIGO) 2009). METHODS: A multicenter retrospective observational cohort study was conducted including patients from the Surgery in Cervical Cancer Comparing Different Surgical Aproaches in Stage IB1 Cervical Cancer (SUCCOR) database with FIGO 2009 IB1 cervical carcinoma treated with radical hysterectomy between January 1, 2013, and December 31, 2014. We used propensity score matching to minimize the potential allocation biases arising from the retrospective design. Patients who underwent conization but were similar for other measured characteristics were matched 1:1 to patients from the non-cone group using a caliper width ≤0.2 standard deviations of the logit odds of the estimated propensity score. RESULTS: We obtained a weighted cohort of 374 patients (187 patients with prior conization and 187 non-conization patients). We found a 65% reduction in the risk of relapse for patients who had cervical conization prior to radical hysterectomy (hazard ratio (HR) 0.35, 95% confidence interval (CI) 0.16 to 0.75, p=0.007) and a 75% reduction in the risk of death for the same sample (HR 0.25, 95% CI 0.07 to 0.90, p=0.033). In addition, patients who underwent minimally invasive surgery without prior conization had a 5.63 times higher chance of relapse compared with those who had an open approach and previous conization (HR 5.63, 95% CI 1.64 to 19.3, p=0.006). Patients who underwent minimally invasive surgery with prior conization and those who underwent open surgery without prior conization showed no differences in relapse rates compared with those who underwent open surgery with prior cone biopsy (reference) (HR 1.94, 95% CI 0.49 to 7.76, p=0.349 and HR 2.94, 95% CI 0.80 to 10.86, p=0.106 respectively). CONCLUSIONS: In this retrospective study, patients undergoing cervical conization before radical hysterectomy had a significantly lower risk of relapse and death.

Burkholderia cepacia Complex Bacteria: a Feared Contamination Risk in Water-Based Pharmaceutical Products.

Burkholderia cepacia (formerly Pseudomonas cepacia) was once thought to be a single bacterial species but has expanded to the Burkholderia cepacia complex (Bcc), comprising 24 closely related opportunistic pathogenic species. These bacteria have a widespread environmental distribution, an extraordinary metabolic versatility, a complex genome with three chromosomes, and a high capacity for rapid mutation and adaptation. Additionally, they present an inherent resistance to antibiotics and antiseptics, as well as the abilities to survive under nutrient-limited conditions and to metabolize the organic matter present in oligotrophic aquatic environments, even using certain antimicrobials as carbon sources. These traits constitute the reason that Bcc bacteria are considered feared contaminants of aqueous pharmaceutical and personal care products and the frequent reason behind nonsterile product recalls. Contamination with Bcc has caused numerous nosocomial outbreaks in health care facilities, presenting a health threat, particularly for patients with cystic fibrosis and chronic granulomatous disease and for immunocompromised individuals. This review addresses the role of Bcc bacteria as a potential public health problem, the mechanisms behind their success as contaminants of pharmaceutical products, particularly in the presence of biocides, the difficulties encountered in their detection, and the preventive measures applied during manufacturing processes to control contamination with these objectionable microorganisms. A summary of Bcc-related outbreaks in different clinical settings, due to contamination of diverse types of pharmaceutical products, is provided.

Radical hysterectomy in early cervical cancer in Europe: characteristics, outcomes and evaluation of ESGO quality indicators.

INTRODUCTION: Comprehensive updated information on cervical cancer surgical treatment in Europe is scarce. OBJECTIVE: To evaluate baseline characteristics of women with early cervical cancer and to analyze the outcomes of the ESGO quality indicators after radical hysterectomy in the SUCCOR database. METHODS: The SUCCOR database consisted of 1272 patients who underwent radical hysterectomy for stage IB1 cervical cancer (FIGO 2009) between January 2013 and December 2014. After exclusion criteria, the final sample included 1156 patients. This study first described the clinical, surgical, pathological, and follow-up variables of this population and then analyzed the outcomes (disease-free survival and overall survival) after radical hysterectomy. Surgical-related ESGO quality indicators were assessed and the accomplishment of the stated recommendations was verified. RESULTS: The mean age of the patients was 47.1 years (SD 10.8), with a mean body mass index of 25.4 kg/m2 (SD 4.9). A total of 423 (36.6%) patients had a previous cone biopsy. Tumor size (clinical examination) <2 cm was observed in 667 (57.7%) patients. The most frequent histology type was squamous carcinoma (794 (68.7%) patients), and positive lymph nodes were found in 143 (12.4%) patients. A total of 633 (54.8%) patients were operated by open abdominal surgery. Intra-operative complications occurred in 108 (9.3%) patients, and post-operative complications during the first month occurred in 249 (21.5%) patients, with bladder dysfunction as the most frequent event (119 (10.3%) patients). Clavien-Dindo grade III or higher complication occurred in 56 (4.8%) patients. A total of 510 (44.1%) patients received adjuvant therapy. After a median follow-up of 58 months (range 0-84), the 5-year disease-free survival was 88.3%, and the overall survival was 94.9%. In our population, 10 of the 11 surgical-related quality indicators currently recommended by ESGO were fully fulfilled 5 years before its implementation. CONCLUSIONS: In this European cohort, the rate of adjuvant therapy after radical hysterectomy is higher than for most similar patients reported in the literature. The majority of centers were already following the European recommendations even 5 years prior to the ESGO quality indicator implementations.

Temporal characterization of the insulin resistance during puberty in mice.

Objectives. The onset of puberty in humans is followed by an increase in insulin resistance and this transient phenomenon decreases at the end of the puberty. However, the insulin resistance during puberty has not been described in mice. Thus, in the present study we performed a temporal characterization of the development of insulin resistance during puberty in male and female C57BL/6 mice.Methods. From the fourth week of life male (n=18) and female (n=32) C57BL/6 wild-type mice were weekly subjected to insulin tolerance tests until the seventh week of life. Blood glucose levels were determined using a glucose meter through samples collected from the tail tip. Vaginal opening was assessed daily in female mice. Preputial separation was determined in a subgroup of males.Results. We observed a transient increase in the area under the curve of the insulin tolerance tests and in basal glycemia in female mice at the time of vaginal opening (between the fourth and fifth week of life) compared with previous and subsequent weeks of pubertal development. In contrast, male mice show no changes in insulin sensitivity during puberty.Conclusions. Our findings demonstrate that the insulin resistance at puberty can also be observed in female mice and this peak occurs at the time of vaginal opening. Our temporal characterization can be used as a reference for future studies that aim to study glucose homeostasis during puberty in rodents.

SUCCOR study: an international European cohort observational study comparing minimally invasive surgery versus open abdominal radical hysterectomy in patients with stage IB1 cervical cancer.

BACKGROUND: Minimally invasive surgery in cervical cancer has demonstrated in recent publications worse outcomes than open surgery. The primary objective of the SUCCOR study, a European, multicenter, retrospective, observational cohort study was to evaluate disease-free survival in patients with stage IB1 (FIGO 2009) cervical cancer undergoing open vs minimally invasive radical hysterectomy. As a secondary objective, we aimed to investigate the association between protective surgical maneuvers and the risk of relapse. METHODS: We obtained data from 1272 patients that underwent a radical hysterectomy by open or minimally invasive surgery for stage IB1 cervical cancer (FIGO 2009) from January 2013 to December 2014. After applying all the inclusion-exclusion criteria, we used an inverse probability weighting to construct a weighted cohort of 693 patients to compare outcomes (minimally invasive surgery vs open). The first endpoint compared disease-free survival at 4.5 years in both groups. Secondary endpoints compared overall survival among groups and the impact of the use of a uterine manipulator and protective closure of the colpotomy over the tumor in the minimally invasive surgery group. RESULTS: Mean age was 48.3 years (range; 23-83) while the mean BMI was 25.7 kg/m2 (range; 15-49). The risk of recurrence for patients who underwent minimally invasive surgery was twice as high as that in the open surgery group (HR, 2.07; 95% CI, 1.35 to 3.15; P=0.001). Similarly, the risk of death was 2.42-times higher than in the open surgery group (HR, 2.45; 95% CI, 1.30 to 4.60, P=0.005). Patients that underwent minimally invasive surgery using a uterine manipulator had a 2.76-times higher hazard of relapse (HR, 2.76; 95% CI, 1.75 to 4.33; P<0.001) and those without the use of a uterine manipulator had similar disease-free-survival to the open surgery group (HR, 1.58; 95% CI, 0.79 to 3.15; P=0.20). Moreover, patients that underwent minimally invasive surgery with protective vaginal closure had similar rates of relapse to those who underwent open surgery (HR, 0.63; 95% CI, 0.15 to 2.59; P<0.52). CONCLUSIONS: Minimally invasive surgery in cervical cancer increased the risk of relapse and death compared with open surgery. In this study, avoiding the uterine manipulator and using maneuvers to avoid tumor spread at the time of colpotomy in minimally invasive surgery was associated with similar outcomes to open surgery. Further prospective studies are warranted.

Protective effects of beet (Beta vulgaris) leaves extract against oxidative stress in endothelial cells in vitro.

Beetroot is an herb used worldwide as a food product, raw material for food industry, ethanol production and source of food coloring. Beet leaves are an unconventional food with antioxidant properties, which might neutralize reactive oxygen species (ROS) induced by oxidized Low-Density Lipoprotein (LDL) present in dyslipidemias. This study aimed to elucidate the effects of beet leaves on the suppression of LDL oxidative processes. Beet leaves extract was produced, characterized, and tested for its antioxidant capacity using endothelial cells in vitro. A model of human umbilical vein endothelial cells was used in various tests, including viability assay, molecular analysis of antioxidant genes, ROS labeling, and macrophage adhesion assay. The extract improved the antioxidative protection of endothelial cells against different agents including oxidized LDL-cholesterol and H2 O2 . It acted on ROS directly due to its high content of natural antioxidants, but also due to the activation and improvement of cellular defenses such as Superoxide dismutase 1, Superoxide dismutase 2, and catalase. The inhibition of LDL-mediated oxidative effects on endothelial cells may turn this unconventional food a functional food with great potential for phytotherapy of atherosclerosis as an adjuvant for medicinal treatments.

Vitrification of cat ovarian tissue: Does fragment size matters?

The aim of the present study was to investigate whether or not the size of the ovarian fragment influences its resistance to cryostorage. For that purpose, ovaries were collected from 34 queens (various breeds, age 1-5 year) by routine ovariectomy, transported to the laboratory and then sectioned in different sizes (3 mm × 3 mm × 3 mm, 5 mm × 3 mm × 3 mm and 7 mm × 3 mm × 3 mm) and randomly assigned to a control (GC3, GC5 and GC7, respectively) or vitrified (GV3, GV5 and GV7, respectively) groups. Vitrified-warmed fragments were evaluated by histomorphology and immunohistochemistry (for apoptotic rates by using cleaved caspase-3). Histological examination reveals that 72.97% of the follicles in GV3 and 72.58% in GV5 were normal while only 42.86% of the follicles in GV7. The main morphological alteration presented in all groups was a detachment of the epithelial cells. Similarly, immunohistochemistry evaluation using caspase 3 revealed a small proportion of apoptotic cells in GV3 (8.43%) while in GV7 30.43% of the cells expressed cleaved caspase-3. These findings indicate that fragments sectioned in 3 mm × 3 mm × 3 mm (27 mm3 ) seem more adequate for perfusion of the cryoprotectant, causing less damage to the cell after vitrification-warming.

Induction of labour vs. spontaneous vaginal delivery in twin pregnancy after 36 weeks of gestation.

Time for delivery and delivery pathway in twin pregnancies are still in great debate. Our study goal was to compare the characteristics of delivery and maternal-foetal outcome in uncomplicated near-term twin pregnancies undergoing labour induction and those with spontaneous labour. We found no statistical differences in patients with twin pregnancies who underwent labour induction and those with spontaneous labour regarding the history of previous caesarean delivery, parity, pregnancy achieved by assisted reproductive techniques (ART), chorionicity and cervical dilation at the admission as well as maternal and neonatal morbidity, and admission to the neonatal intensive care unit. There were significant differences in the caesarean section rate (60.6 vs. 33.3%, p < .05) and the time interval between delivery of the first and second foetus (9.8 vs. 11.7 min, p = .024). There was an increased incidence of caesarean section after the induction of labour. However, it appears to be a safe option.

Different interactomes for p70-S6K1 and p54-S6K2 revealed by proteomic analysis.

S6Ks are major effectors of the mTOR (mammalian target of rapamycin) pathway, signaling for increased protein synthesis and cell growth in response to insulin, AMP/ATP levels, and amino acids. Deregulation of this pathway has been related to disorders and diseases associated with metabolism, such as obesity, diabetes, and cancer. S6K family is composed of two main members, S6K1 and S6K2, which comprise different isoforms resulted from alternative splicing or alternative start codon use. Although important molecular functions have been associated with p70-S6K1, the most extensively studied isoform, the S6K2 counterpart lacks information. In the present study, we performed immunoprecipitation assays followed by mass spectrometry (MS) analysis of FLAG-tagged p70-S6K1 and p54-S6K2 interactomes, after expression in HEK293 cells. Protein lists were submitted to CRAPome (Contaminant Repository for Affinity Purification) and SAINT (Significance Analysis of INTeractome) analysis, which allowed the identification of high-scoring interactions. By a comparative approach, p70-S6K1 interacting proteins were predominantly related to "cytoskeleton" and "stress response," whereas p54-S6K2 interactome was more associated to "transcription," "splicing," and "ribosome biogenesis." Moreover, we have found evidences for new targets or regulators of the S6K protein family, such as proteins NCL, NPM1, eIF2α, XRCC6, PARP1, and ILF2/ILF3 complex. This study provides new information about the interacting networks of S6Ks, which may contribute for future approaches to a better understanding of the mTOR/S6K pathway.

S6Ks isoforms contribute to viability, migration, docetaxel resistance and tumor formation of prostate cancer cells.

BACKGROUND: The S6 Kinase (S6K) proteins are some of the main downstream effectors of the mammalian Target Of Rapamycin (mTOR) and act as key regulators of protein synthesis and cell growth. S6K is overexpressed in a variety of human tumors and is correlated to poor prognosis in prostate cancer. Due to the current urgency to identify factors involved in prostate cancer progression, we aimed to reveal the cellular functions of three S6K isoforms-p70-S6K1, p85-S6K1 and p54-S6K2-in prostate cancer, as well as their potential as therapeutic targets. METHODS: In this study we performed S6K knockdown and overexpression and investigated its role in prostate cancer cell proliferation, colony formation, viability, migration and resistance to docetaxel treatment. In addition, we measured tumor growth in Nude mice injected with PC3 cells overexpressing S6K isoforms and tested the efficacy of a new available S6K1 inhibitor in vitro. RESULTS: S6Ks overexpression enhanced PC3-luc cell line viability, migration, resistance to docetaxel and tumor formation in Nude mice. Only S6K2 knockdown rendered prostate cancer cells more sensitive to docetaxel. S6K1 inhibitor PF-4708671 was particularly effective for reducing migration and proliferation of PC3 cell line. CONCLUSIONS: These findings demonstrate that S6Ks play an important role in prostate cancer progression, enhancing cell viability, migration and chemotherapy resistance, and place both S6K1 and S6K2 as a potential targets in advanced prostate cancer. We also provide evidence that S6K1 inhibitor PF-4708671 may be considered as a potential drug for prostate cancer treatment.

Monochorionic versus dichorionic twins: Are obstetric outcomes always different?

This prospective cohort study compared obstetric, perinatal and postpartum outcomes of monochorionic diamniotic (n = 228) versus (vs.) dichorionic (n = 598) twin pregnancies. Statistical analysis was performed using software SPSS® v19.0.0.2. Chi square, Fischer's exact, Student's t and Mann-Withney tests were applied. Obstetrical complications rates were 85.5% vs. 75.1% (p < 0.01). Differences were found in preterm premature rupture of membranes (26.3% vs. 19.3%, p < 0.05) and intrauterine growth restriction (19.7% vs. 10.5%, p < 0.01). Twin-to-twin transfusion syndrome (TTTS) occurred in 7.9% of monochorionic pregnancies. Vaginal delivery occurred in 47.4% vs. 43.1%. Monochorionic pregnancies had earlier gestational ages at delivery and subsequently lower birthweights (p < 0.01). There was no difference in Apgar scores. Admission rate of at least one of the newborns in intensive care unit (NICU) was 50% vs. 38.9% (p < 0.05). Postpartum complications were similar. These results were the same excluding TTTS cases, except for admission in NICU (46.8% vs. 34.9%, p > 0.05). Analysing only the uncomplicated pregnancies (33 vs. 149), there were no differences in perinatal outcomes. We conclude that monochorionic pregnancies had higher rates of obstetrical complications, which were independent of TTTS occurrence in our sample. However, considering only the uncomplicated pregnancies till delivery, there were no significant differences in perinatal outcomes.

EEG epilepsy seizure prediction: the post-processing stage as a chronology.

Almost one-third of epileptic patients fail to achieve seizure control through anti-epileptic drug administration. In the scarcity of completely controlling a patient's epilepsy, seizure prediction plays a significant role in clinical management and providing new therapeutic options such as warning or intervention devices. Seizure prediction algorithms aim to identify the preictal period that Electroencephalogram (EEG) signals can capture. However, this period is associated with substantial heterogeneity, varying among patients or even between seizures from the same patient. The present work proposes a patient-specific seizure prediction algorithm using post-processing techniques to explore the existence of a set of chronological events of brain activity that precedes epileptic seizures. The study was conducted with 37 patients with Temporal Lobe Epilepsy (TLE) from the EPILEPSIAE database. The designed methodology combines univariate linear features with a classifier based on Support Vector Machines (SVM) and two post-processing techniques to handle pre-seizure temporality in an easily explainable way, employing knowledge from network theory. In the Chronological Firing Power approach, we considered the preictal as a sequence of three brain activity events separated in time. In the Cumulative Firing Power approach, we assumed the preictal period as a sequence of three overlapping events. These methodologies were compared with a control approach based on the typical machine learning pipeline. We considered a Seizure Prediction horizon (SPH) of 5 mins and analyzed several values for the Seizure Occurrence Period (SOP) duration, between 10 and 55 mins. Our results showed that the Cumulative Firing Power approach may improve the seizure prediction performance. This new strategy performed above chance for 62% of patients, whereas the control approach only validated 49% of its models.