Miracle-dz/UniFAR-FTUs huggingface dataset:
EFEITOS DO AGAROL â E DO TRIGLICERIL â SOBRE A CICATRIZAÇÃO DE PELE: ESTUDO EXPERIMENTAL EM RATOS

For centuries, mankind has been incessantly researching with drugs that could accelerate the normal wound healing process. In the Hospital de Clinicas da Universidade Federal do Parana, it has been recommended that the cleansing of clean wounds with saline solution of 0.9% sodium chloride, followed by a cover of dry dressing. However, some services have empirically used Agarol and Trigliceril in the confection of dressings after cleansing. This study aimed to evaluate the evolution of cutaneous wounds treated with normal saline, Agarol and Trigliceril in rats. A group of twenty-four rats were submitted to three skin circular incisions of 1.2 cm in diameter on their backs which exposed the subcutaneous tissue. The wounds were cleaned daily with normal saline solution. This group was distributed at random within three groups: Group C, control, received a clean and dry dressing after cleansing; groups A and T received dressings with Agarol and Trigliceril respectively. The evaluation of the wounds was done on the 3rd, 7th and 14th postoperative days by histological and macroscopic studies. The results showed that on the 3rd and 14th days, no evident influence of the drugs on the healing wound was noticed. On 7th postoperative day, Agarol demonstrated to increase the amount of granulation tissue, while Trigliceril appears to decrease the neovasculature during the healing process.
Aberrant GRIA3 transcripts with multi‐exon duplications in a family with X‐linked mental retardation

Investigation of chromosomal rearrangements in patients with mental retardation (MR) is particularly informative in the search for novel genes involved in MR. We report on a family with a genomic duplication at Xq25 identified by oligo array‐CGH. Further characterization showed a partial tandem duplication of GRIA3 extending from exon 1 to exon 12. This duplication is present in two brothers with MR and on one allele in their sister with normal phenotype and completely skewed X‐chromosome inactivation. The duplication is inherited from the mother, whose cognitive level is low and X‐chromosome inactivation is random. This is the second family with partial duplication of GRIA3 associated with MR. GRIA3 expression studies in our case demonstrated a new mechanism for GRIA3 dysfunction with the presence of aberrant GRIA3 transcripts carrying multi‐exon duplications leading to a frameshift. Our study gives additional support to the implication of GRIA3 in X‐linked MR. © 2009 Wiley‐Liss, Inc.
Transforming Growth Factor-β1-induced Transcript 1 Protein, a Novel Marker for Smooth Muscle Contractile Phenotype, Is Regulated by Serum Response Factor/Myocardin Protein*

Background: TGFB1I1 is involved in vascular injury, but the regulation of TGFB1I1 expression is unknown. Results: TGFB1I1 is specifically expressed in smooth muscle cells and regulated by SRF/myocardin. Conclusion: TGFB1I1 is a novel marker for the smooth muscle contractile phenotype. Significance: This study provides the first evidence that TGFB1I1 is an SRF/myocardin-regulated smooth muscle marker and critical for maintaining the smooth muscle contractile phenotype. Serum response factor (SRF) plays a central role in regulating expression of smooth muscle-specific genes partly by associating with the potent tissue-specific cofactor myocardin. Previous studies have shown that transforming growth factor-β1-induced transcript 1 (TGFB1I1, also known as Hic-5) is a TGF-β-responsive gene and is involved in the cellular response to vascular injury, but the regulation of TGFB1I1 expression remains elusive. In this report, we demonstrated that TGFB1I1 is a novel marker for the smooth muscle contractile phenotype and is regulated by SRF/myocardin. We found that TGFB1I1 is specifically expressed in smooth muscle cells (SMCs) and in smooth muscle-rich tissues. Furthermore, TGFB1I1 expression is significantly down-regulated in a variety of models for smooth muscle phenotypic modulation. The TGFB1I1 promoter contains an evolutionarily conserved CArG element, and this element is indispensible for myocardin-induced transactivation of TGFB1I1 promoter. By oligonucleotide pulldown and chromatin immunoprecipitation assays, we found that SRF binds to this CArG element in vitro and in vivo. Ectopic expression of myocardin is sufficient to induce endogenous TGFB1I1 expression in multiple cell lines whereas knocking-down myocardin or SRF significantly attenuated TGFB1I1 expression in SMCs. Furthermore, our data demonstrated that SRF is essential for TGF-β-mediated induction of TGFB1I1. Finally, silencing of TGFB1I1 expression significantly promotes SMC proliferation. Collectively, this study provides the first evidence that TGFB1I1 is not only an SRF/myocardin-regulated smooth muscle marker but also critical for maintaining smooth muscle contractile phenotype by inhibiting smooth muscle proliferation.
Evaluation of folate metabolism gene polymorphisms as risk factors for open and closed neural tube defects

Evaluation of Folate Metabolism Gene Polymorphisms as Risk Factors for Open and Closed Neural Tube Defects K. Doudney, J. Grinham, J. Whittaker, S.A. Lynch, D. Thompson, G.E. Moore, A.J. Copp, N.D. E. Greene, and P. Stanier* UCL Institute of Child Health, University College London, London, UK Department of Pathology, University of Otago, Christchurch, New Zealand Department of Epidemiology and Population Health, London School of Hygiene and Tropical Medicine, London, UK National Centre for Medical Genetics, Our Lady’s Children’s Hospital Crumlin, Dublin, Ireland Department of Neurosurgery, Great Ormond Street Hospital for Children, London, UK
Hydrogen Peroxide Stimulates the Epithelial Sodium Channel through a Phosphatidylinositide 3-Kinase-dependent Pathway*

Recent studies indicate that oxidative stress mediates salt-sensitive hypertension. To test the hypothesis that the renal epithelial sodium channel (ENaC) is a target of oxidative stress, patch clamp techniques were used to determine whether ENaC in A6 distal nephron cells is regulated by hydrogen peroxide (H2O2). In the cell-attached configuration, H2O2 significantly increased ENaC open probability (Po) and single-channel current amplitude but not the unit conductance. High concentrations of exogenous H2O2 are required to elevate intracellular H2O2, probably because catalase, the enzyme that promotes the decomposition of H2O2 to H2O and O2, is highly expressed in A6 cells. The effect of H2O2 on ENaC Po was enhanced by 3-aminotriazole, a catalase inhibitor, and abolished by overexpression of catalase, indicating that intracellular H2O2 levels are critical to produce the effect. However, H2O2 did not directly activate ENaC in inside-out patches. The effects of H2O2 on ENaC Po and amiloride-sensitive Na+ current were abolished by inhibition of phosphatidylinositide 3-kinase (PI3K). Confocal microscopy data showed that H2O2 elevated phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) in the apical membrane by stimulating PI3K. Because ENaC is stimulated by PI(3,4,5)P3, these data suggest that H2O2 stimulates ENaC via PI3K-mediated increases in apical PI(3,4,5)P3.
Novel Phosphorylation-dependent Ubiquitination of Tristetraprolin by Mitogen-activated Protein Kinase/Extracellular Signal-regulated Kinase Kinase Kinase 1 (MEKK1) and Tumor Necrosis Factor Receptor-associated Factor 2 (TRAF2)*

Background: During inflammation, tristetraprolin displays a dual function as mRNA destabilizer and regulator of NF-κB signaling. Results: Tristetraprolin is inducibly hypermodified by phosphorylation and ubiquitination, affecting cell viability. Conclusion: Tristetraprolin hypermodification provokes its functional shift toward the activation of JNK signaling. Significance: The NF-κB/JNK cross-talk is central for our understanding of chronic inflammatory diseases and cancer development. Acute versus chronic inflammation is controlled by the accurate activation and regulation of interdependent signaling cascades. TNF-receptor 1 engagement concomitantly activates NF-κB and JNK signaling. The correctly timed activation of these pathways is the key to account for the balance between NF-κB-mediated cell survival and cell death, the latter fostered by prolonged JNK activation. Tristetraprolin (TTP), initially described as an mRNA destabilizing protein, acts as negative feedback regulator of the inflammatory response: it destabilizes cytokine-mRNAs but also acts as an NF-κB inhibitor by interfering with the p65/RelA nuclear import pathway. Our biochemical studies provide evidence that TTP contributes to the NF-κB/JNK balance. We find that the MAP 3-kinase MEKK1 acts as a novel TTP kinase that, together with the TNF receptor-associated factor 2 (TRAF2), constitutes not only a main determinate of the NF-κB-JNK cross-talk but also facilitates “TTP hypermodification”: MEKK1 triggers TTP phosphorylation as prerequisite for its Lys-63-linked, TRAF2-mediated ubiquitination. Consequently, TTP no longer affects NF-κB activity but promotes the activation of JNK. Based on our data, we suggest a model where upon TNFα induction, TTP transits a hypo- to hypermodified state, thereby contributing to the molecular regulation of NF-κB versus JNK signaling cascades.
Toll-like Receptor 3 (TLR3) Signaling Requires TLR4 Interactor with Leucine-rich Repeats (TRIL)*

Toll-like receptors (TLRs) are a family of proteins that act as the primary sensors of microbial products. Many TLRs require accessory molecules in order to recognize these microbial products and initiate signal transduction cascades. We have identified TRIL (TLR4 interactor with leucine-rich repeats) as a novel modulator of TLR4 signaling showing high expression in the brain. We now show that TRIL also plays a role in TLR3 signaling. TRIL is expressed intracellularly in the astrocytoma cell line U373 and in the monocytic cell line THP1. TRIL co-localizes with the endosomal compartment. These data are consistent with a role for TRIL in TLR3 signaling and endosomal TLR4 signaling. TRIL was induced by the TLR3 ligand poly(I:C). Overexpression of TRIL enhanced cytokine production and interferon-stimulated response element (ISRE) luciferase activity following poly(I:C) stimulation in U373. TRIL interacted with TLR3, and this interaction was enhanced following poly(I:C) stimulation. Transient knockdown of TRIL with siRNA or stable knockdown using shRNA in U373 cells inhibited TLR3 signaling, reducing ISRE luciferase, RANTES, and type I interferon production. Knockdown of TRIL did not affect TLR2 signaling. Most accessory molecules identified to date, such as CD14, gp96, PRAT4a, and Unc93B, all play roles in multiple TLR signaling pathways, and we now show that this is also the case for TRIL.
Chronic treatment with sildenafil stimulates Leydig cell and testosterone secretion

The phosphodiesterase type 5 (PDE5) inhibitor, Sildenafil, is a novel, oral treatment approach for pulmonary hypertension. As Leydig cells present PDE5, this study was conducted to investigate the effects of the chronic treatment with Sildenafil (25 mg/kg) on male Swiss Webster mice steroidogenesis. After a 4‐week long experimental design, Leydig cells were analysed by morphological and immunocytochemical procedures. Serum testosterone was assayed by radioimmunoassay. Leydig cells presented noteworthy ultrastructural alterations, such as a vesicular smooth endoplasmic reticulum, large vacuoles scattered through the cytoplasm, enlarged mitochondria with discontinue cristaes and whorle membranes with vesicles at the periphery, which are typical characteristics of an activated steroid‐secreting cell. Important immunocytochemical labelling for steroidogenic acute regulatory protein, cytochrome P450 side‐chain cleavage enzyme and testosterone were detected in isolated Leydig cells. In addition, Sildenafil‐treated mice showed significant increased levels of total testosterone. The results obtained in the present study are consistent with the hypothesis that the accumulation of cyclic guanosine monophosphate by PDE5 inhibition could be involved in the androgen biosynthesis stimulation. Important clinical implications of hormonal disorders should be taken into account for patients with pulmonary hypertension.
Femur bone repair in ovariectomized rats under the local action of alendronate, hydroxyapatite and the association of alendronate and hydroxyapatite

An evaluation was made of the local action of alendronate sodium (A), hydroxyapatite (HA) and the association of both substances (A + HA), in different molar concentrations, on the femur bone repair of ovariectomized rats. Ninety‐eight animals were divided into seven groups: control (C), starch (S), alendronate 1 mol (A1), alendronate 2 mols (A2), hydroxyapatite 1 mol (HA1), hydroxyapatite 2 mols (HA2) and the association of alendronate + hydroxyapatite (A + HA). Rats weighing about 250 g were ovariectomized and 2.5‐mm diameter bone defects were made on the left femur 30 days later. Each experimental group had defects filled with appropriate material, except for group C (control). The animals were killed 7 and 21 days after surgery. Histological, histomorphometric and statistical analyses of bone neoformation in the bone defect site were performed. From the histological standpoint, the major differences occurred after 21 days. All specimens in groups C, S, HA1 and HA2 presented linear closure of the bone defect, and most animals in groups A1, A2 and A + HA showed no bone neoformation in the central area of the defect. No statistically significant difference was found among the experimental groups after 7 days; after 21 days, group HA2 presented the highest amount of neoformed bone. There was no significant difference among groups A1, A2 and A + HA in the two study periods. It was concluded that alendronate, either isolated or in association with hydroxyapatite, had an adverse effect on bone repair in this experimental model. Moreover, the hydroxyapatite used here proved to be biocompatible and osteoconductive, with group HA2 showing the best results.
The Electric Capacity of Tumors of the Breast

INTRODUCTION A suspension of biological cells or a biological tissue when placed in a conductivity cell, behaves as though it were a pure resistance in parallel with a pure capacity. This parallel capacity is the capacity of the specimen of tissue which is measured while the specific capacity is the capacity of a one centimeter cube of the tissue. While the resistance of biological tissues has been studied by many investigators, little attention has been directed to their capacity. During the course of an investigation of the capacities of various biological systems, which is at present in progress in this laboratory, certain facts regarding the capacity of tumors were noted which justified a somewhat extensive study of the practical aspects of this subject. In short, it was found that certain types of malignant tumors have a rather high capacity in comparison with benign tumors or with inactive tissues of the same or similar character. Early in the investigation it was determined that tumors of the breast constituted the most convenient and uncomplicated material for study and while numerous tumors of other tissues have been measured, only those belonging to this group will be reported here. Fifty-eight cases have been admitted to this series, which constitutes an unselected group comprising all the cases of tumors of the breast which have been operated on at the Cleveland Clinic Hospital for a period of about one year, together with sporadic cases taken at random prior to this time and a few cases, particularly of benign tumors, which were operated on at other hospitals in the city. 2
Crystal Structure of Menin Reveals Binding Site for Mixed Lineage Leukemia (MLL) Protein*

Menin is a tumor suppressor protein that is encoded by the MEN1 (multiple endocrine neoplasia 1) gene and controls cell growth in endocrine tissues. Importantly, menin also serves as a critical oncogenic cofactor of MLL (mixed lineage leukemia) fusion proteins in acute leukemias. Direct association of menin with MLL fusion proteins is required for MLL fusion protein-mediated leukemogenesis in vivo, and this interaction has been validated as a new potential therapeutic target for development of novel anti-leukemia agents. Here, we report the first crystal structure of menin homolog from Nematostella vectensis. Due to a very high sequence similarity, the Nematostella menin is a close homolog of human menin, and these two proteins likely have very similar structures. Menin is predominantly an α-helical protein with the protein core comprising three tetratricopeptide motifs that are flanked by two α-helical bundles and covered by a β-sheet motif. A very interesting feature of menin structure is the presence of a large central cavity that is highly conserved between Nematostella and human menin. By employing site-directed mutagenesis, we have demonstrated that this cavity constitutes the binding site for MLL. Our data provide a structural basis for understanding the role of menin as a tumor suppressor protein and as an oncogenic co-factor of MLL fusion proteins. It also provides essential structural information for development of inhibitors targeting the menin-MLL interaction as a novel therapeutic strategy in MLL-related leukemias.
Comparative study of the expression of metalloproteases and their inhibitors in different localizations within primary tumours and in metastatic lymph nodes of breast cancer

Studies on metastasic lesions from human carcinomas are scarce. Therefore there is a need for such studies to identify the expression of the biological factors that will help in the assessment of the natural history of breast cancer. Here an immunohistochemical study was performed using tissue arrays and specific antibodies against matrix metalloproteinases (MMPs)‐1, 2, 7, 9, 11, 13, 14 and tissue inhibitors of metalloproteases (TIMPs)‐1, 2 and 3 in 39 patients with breast cancer. Specimens from 39 patients with node‐positive carcinomas were examined and the analysis was performed at the central core of the tumour, at the invasive front, and in the metastasic axillary lymph nodes (MALNs). Global expression of MMP‐1, 7 and 14, TIMP‐1, and 3, were significantly higher at the centre of the tumour compared with the invasive front or the MALNs. Significantly higher expression of MMP‐7 and 14, and TIMP‐3, by fibroblast‐like cells and mononuclear inflammatory cells (MICs) was seen in MALNs. In addition, in the tumour centre, the expression of MMP‐11 and TIMP‐1 and 2 by MICs, as well as TIMP‐2 expression by fibroblast‐like cells, were associated significantly with the occurrence of distant metastasis. In contrast, TIMP‐3 expression by tumour cells or by fibroblast‐like cells in this same tumour locations, as well as TIMP‐1 expression by fibroblast‐like cells at the invasive front, were associated significantly with poor prognosis. However, the expression of all of these biological factors in MALNs was not associated with the development of distant metastasis. Our data suggest that there is prognostic relevance to the expression of MMPs and TIMPs in the stromal cells of primary tumours, rather than to the expression of these enzymes in MALNs.
Structural Basis of M3 Muscarinic Receptor Dimer/Oligomer Formation*

Class A G protein-coupled receptors (GPCRs) are known to form dimers and/or oligomeric arrays in vitro and in vivo. These complexes are thought to play important roles in modulating class A GPCR function. Many studies suggest that residues located on the “outer” (lipid-facing) surface of the transmembrane (TM) receptor core are critically involved in the formation of class A receptor dimers (oligomers). However, no clear consensus has emerged regarding the identity of the TM helices or TM subsegments involved in this process. To shed light on this issue, we have used the M3 muscarinic acetylcholine receptor (M3R), a prototypic class A GPCR, as a model system. Using a comprehensive and unbiased approach, we subjected all outward-facing residues (70 amino acids total) of the TM helical bundle (TM1–7) of the M3R to systematic alanine substitution mutagenesis. We then characterized the resulting mutant receptors in radioligand binding and functional studies and determined their ability to form dimers (oligomers) in bioluminescence resonance energy transfer saturation assays. We found that M3R/M3R interactions are not dependent on the presence of one specific structural motif but involve the outer surfaces of multiple TM subsegments (TM1–5 and -7) located within the central and endofacial portions of the TM receptor core. Moreover, we demonstrated that the outward-facing surfaces of most TM helices play critical roles in proper receptor folding and/or function. Guided by the bioluminescence resonance energy transfer data, molecular modeling studies suggested the existence of multiple dimeric/oligomeric M3R arrangements, which may exist in a dynamic equilibrium. Given the high structural homology found among all class A GPCRs, our results should be of considerable general relevance.
Alterations induced by chronic lead exposure on the cells of circadian pacemaker of developing rats

Lead (Pb) exposure alters the temporal organization of several physiological and behavioural processes in which the suprachiasmatic nucleus (SCN) of the hypothalamus plays a fundamental role. In this study, we evaluated the effects of chronic early Pb exposure (CePbe) on the morphology, cellular density and relative optical density (OD) in the cells of the SCN of male rats. Female Wistar rats were exposed during gestation and lactation to a Pb solution containing 320 ppm of Pb acetate through drinking water. After weaning, the pups were maintained with the same drinking water until sacrificed at 90 days of age. Pb levels in the blood, hypothalamus, hippocampus and prefrontal cortex were significantly increased in the experimental group. Chronic early Pb exposure induced a significant increase in the minor and major axes and somatic area of vasoactive intestinal polypeptide (VIP)‐ and vasopressin (VP)‐immunoreactive neurons. The density of VIP‐, VP‐ and glial fibrillary acidic protein (GFAP)‐immunoreactive cells showed a significant decrease in the experimental group. OD analysis showed a significant increase in VIP neurons of the experimental group. The results showed that CePbe induced alterations in the cells of the SCN, as evidenced by modifications in soma morphology, cellular density and OD in circadian pacemaker cells. These findings provide a morphological and cellular basis for deficits in circadian rhythms documented in Pb‐exposed animals.