Miracle-dz/UniFAR-FTUs huggingface dataset:
Relação entre a lesão condral e o pico de torque após reconstrução do ligamento cruzado anterior do joelho: estudo de casos

CONTEXT: The association between anterior cruciate ligament (ACL) injury and the development of secondary osteoarthritis has been the subject of several studies. Ligament injury predisposes joint cartilage lesions, thus influencing motor control and peak torque. OBJECTIVE: To evaluate the influence of the degree of chondral lesion on peak torque of the anterior and posterior musculature of the thigh following ACL injury and reconstruction. METHOD: Six male subjects with total unilateral ACL injury were evaluated 24 hours before surgery and at two times after surgery (4.66 ± 1.03 and 15.83 ± 2.63 months, respectively). Isokinetic evaluations of knee flexion and extension were made in concentric mode, at velocities of 60o/s and 180o/s, and the variables of hamstring peak torque (HPT) and quadriceps peak torque (QPT) and hamstring/quadriceps ratio (H/Q) were analyzed. The modified Mankin histological scale was used to evaluate the macroscopic degree of chondral injury. The ANOVA statistical test (p< 0.05) was used to compare the results between the three times studied, with the Newman-Keuls post-hoc test. Spearman's correlation test was then applied to evaluate the influence of the degree of chondral injury on the peak torque of the thigh musculature. RESULTS: There were statistically significant differences in the H/Q ratio at the velocity of 60o/s (p= 0.01). The degree of chondral injury degree ranged from 1 to 5, although there was no correlation with the isokinetic findings. CONCLUSION: There was an imbalance in agonist/antagonist relationship, with regard to torque, between muscle groups of the knee following ACL reconstruction. However, the degree of secondary chondral lesion did not influence the progressive gains in this muscle groups over the study period.
Assessment of sexual dysfunction in women with Behcet’s disease

Behcet’s disease (BD) is a progressive inflammatory disease which urogenital involvement mainly consists of genital aphthous ulcers, urethritis and recurrent cystitis. We evaluated female sexual dysfunction (FSD) in women with BD and investigate whether coexistent depression has an effect on sexual functions. A total of 25 women with BD and 25 age voluntary healthy women who served as the control group were evaluated with a detailed medical and sexual history, including female sexual function index (FSFI) ques¬tionnaire and the Beck Depression Inventory (BDI). Serum steroid hormone levels were measured in all subjects. The group with BD had higher scores for BDI than the control patients (p<0.001) and the mean FSFI score was significantly lower than the score for the controls. FSD was diagnosed in 14 of 25 BD patients (56%), while 4 of 25 healthy women (16%). Women with BD scored worse on all domains of FSD than did the controls (p<0.001). In the correlation analysis, the FSFI score indicated a significant negative correlation with the BDI score (r=-0.901, p<0.001). The results of our study have shown that female patients with BD have distinct sexual dysfunction compared with healthy controls and coexistent depression has an additional negative effect on sexual function. J. Exp. Clin. Med., 2012; 29:28-32
Voltage-dependent Anion Channel 1-based Peptides Interact with Hexokinase to Prevent Its Anti-apoptotic Activity*

In brain and tumor cells, the hexokinase isoforms, HK-I and HK-II, bind to the voltage-dependent anion channel (VDAC) in the outer mitochondrial membrane. The VDAC domains interacting with these anti-apoptotic proteins were recently defined using site-directed mutagenesis. Now, we demonstrate that synthetic peptides corresponding to the VDAC1 N-terminal region and selected sequences bound specifically, in a concentration- and time-dependent manner, to immobilized HK-I, as revealed by real time surface plasmon resonance technology. The same VDAC1-based peptides also detached HK bound to brain or tumor-derived mitochondria. Moreover, expression of the VDAC1-based peptides in cells overexpressing HK-I or HK-II prevented HK-mediated protection against staurosporine-induced release of cytochrome c and subsequent cell death. One loop-shaped VDAC1-based peptide corresponding to a selected sequence and fused to a cell-penetrating peptide entered the cell and prevented the anti-apoptotic effects of HK-I and HK-II. This peptide detached mitochondrial-bound HK better than did the same peptide in its linear form. Both cell-expressed and exogenously added cell-penetrating peptide detached mitochondrial-bound HK-I-GFP. These results point to HK-I and HK-II as promoting tumor cell survival through binding to VDAC1, thereby inhibiting cytochrome c release and apoptotic cell death. Moreover, VDAC1-based peptides interfering with HK-mediated anti-apoptotic activity may potentiate the efficacy of conventional chemotherapeutic agents.
Novel insights in the role of peripheral corticotropin‐releasing factor and mast cells in stress‐induced visceral hypersensitivity

Visceral hypersensitivity is one of the hallmarks in irritable bowel syndrome (IBS) pathophysiology. Stress is well known to affect visceral sensitivity in humans and rodents, an effect which is associated in part with alterations of intestinal epithelial permeability in rodents. Although the pathophysiology of visceral hypersensitivity is still unclear, two key factors have been identified as playing a major role in its modulation, namely peripheral corticotropin‐releasing factor (CRF) and mast cells. In a recent study in Neurogastroenterology and Motility, van den Wijngaard et al. demonstrate that the mast‐cell dependent visceral hypersensitivity observed in maternally separated rats after an acute exposure to a psychological stress can be prevented but not reversed by the peripherally restricted CRF receptor antagonist, α‐helical CRF9–41. They further show that the preventive effect of the CRF receptor antagonist is linked to a stabilization of mast cells and maintenance of the epithelial barrier at the colonic level. These data suggest that post stress mast cell activation and subsequent visceral hypersensitivity are not targeted by peripheral CRF receptor antagonists. These novel insights in the role of peripheral CRF in the modulation of stress‐induced visceral hypersensitivity add to our growing understanding of the mechanisms that may lie at the origin of visceral pain disturbances following stress and will contribute to enhance the development of drugs that may have potential therapeutic benefits for IBS patients.
Human Dicer Binds Short Single-strand and Double-strand RNA with High Affinity and Interacts with Different Regions of the Nucleic Acids*

Human Dicer is an integral component of the RNA interference pathway. Dicer processes premicro-RNA and double-strand RNA to, respectively, mature micro-RNA and short interfering RNA (siRNA) and transfers the processed products to the RNA-induced silencing complex. To better understand the factors that are important for the binding, translocation, and selective recognition of the siRNA strands, we determined the binding affinities of human Dicer for processed products (siRNA) and short single-strand RNAs (ssRNA). siRNAs and ssRNAs competitively inhibited human Dicer activity, suggesting that they are interacting with the active site of the enzyme. The dissociation constants (Kd) for unmodified siRNAs were 5-11-fold weaker compared with a 27-nucleotide double-strand RNA substrate. Chemically modified siRNAs exhibited binding affinities for Dicer comparable with the substrate. 3′-Dinucleotide overhangs in the siRNA affected the binding affinity of human Dicer for the siRNA and biased strand loading into RNA-induced silencing complex. The Kd values for the ssRNAs ranged from 3- to 40-fold weaker than the Kd for the substrate. Sequence composition of the 3′-terminal nucleotides of the ssRNAs exhibited the greatest effect on Dicer binding. Dicer cleaved substrates containing short siRNA-like double-strand regions and extended 3′ or 5′ ssRNA overhangs in the adjacent ssRNA regions. Remarkably, cleavage sites were observed consistent with the enzyme entering the substrate from the extended 3′ ssRNA terminus. These data suggest that the siRNAs and ssRNAs interact predominantly with the PAZ domain of the enzyme. Finally, the tightest binding siRNAs were also more potent inhibitors of gene expression.
Proteolytic Processing of the Epithelial Sodium Channel γ Subunit Has a Dominant Role in Channel Activation*

Maturation of the epithelial sodium channel (ENaC) involves furin-dependent cleavage at two extracellular sites within the α subunit and at a single extracellular site within the γ subunit. Channels lacking furin processing of the α subunit have very low activity. We recently identified a prostasin-dependent cleavage site (RKRK186) in the γ subunit. We also demonstrated that the tract α D206-R231, between the two furin cleavage sites in the α subunit, as well as the tract γ E144-K186, between the furin and prostasin cleavage sites in the γ subunit, are inhibitory domains. ENaC cleavage by furin, and subsequently by prostasin, leads to a stepwise increase in the open probability of the channel as a result of release of the α and γ subunit inhibitory tracts, respectively. We examined whether release of either theα orγ inhibitory tract has a dominant role in activating the channel. Co-expression of prostasin and either wild type channels or mutant channels lacking furin cleavage of the α subunit (αR205A,R208A,R231Aβγ) in Xenopus laevis oocytes led to increases in whole cell currents to similar levels. In an analogous manner and independent of the proteolytic processing of theα subunit, amiloride-sensitive currents in oocytes expressing channels carrying γ subunits with both a mutation in the furin cleavage site and a deletion of the inhibitory tract (αβγR143A,ΔE144-K186 and αR205A,R208A,R231AβγR143A, ΔE144-K186) were significantly higher than those from oocytes expressing wild type ENaC. When channels lacked the α and γ subunit inhibitory tracts, α subunit cleavage was required for channels to be fully active. Channels lacking both furin cleavage and the inhibitory tract in theγ subunit (αβγR143A,ΔE144-K186) showed a significant reduction in the efficacy of block by the syntheticα-26 inhibitory peptide representing the tract αD206-R231. Our data indicate that removal of the inhibitory tract from the γ subunit, in the absence ofα subunit cleavage, results in nearly full activation of the channel.
Esophageal distension parameters as potential biomarkers of impaired gastrointestinal function in diabetes patients

Background Gastrointestinal (GI) symptoms, such as nausea, vomiting, bloating, postprandial fullness, and abdominal pain, are frequent in patients with diabetes mellitus (DM). The pathogenesis is complex and multi‐factorial. To determine easy accessible and valid biomarkers for disordered GI function in DM patients, we aimed to study esophageal mechanical parameters and their relation to symptoms typically arising from the digestive tract.
Lizard tail spinal cord: a new experimental model of spinal cord injury without limb paralysis

Spinal cord injury (SCI) is a well‐known devastating lesion that sadly is very resistant to all treatment attempts. This fact has stimulated the exploration of multiple regenerative strategies that are examined at both the basic and clinical level. For laboratory research, different in vivo models are used, but each has many important limitations. The main limitation of these models is the high level of animal suffering related to the inflicted neurologic injury. It has caused a growing tendency to limit the injury, but this, in turn, produces incomplete SCI models and uncertainties in the neuroregeneration interpretation. To overcome such limitations, a new experimental SCI model is proposed. Geckos have been extensively examined as a potential animal model of SCI. Their spinal cord extends into the tail and can be transected without causing the typical neurologic consequences observed in rat models. In this study, we compared the gecko tail SCI model with the rat model of thoracic SCI. Anatomic and histologic analyses showed comparability between the gecko and rat in diameter of spinal canal and spinal cord, as well as applicability of multiple staining techniques (hematoxylin and eosin, immunostaining, and scanning and transmission electron microscopy). We tested the suitability of in vivo study with 3 prototype implants for the reconstruction of SCI: a multichannel sponge, a multilaminar tube, and a gel cylinder. These were compared with a spinal cord excision (control). A 20‐wk observation revealed no adverse effects of SCI on the animals' well‐being. The animals were easily housed and observed. Histologic analysis showed growth of nervous tissue elements on implant surface and implant cellular colonization. The study showed that the gecko SCI model can be used as a primary model for the assessment of SCI treatment methods. It provides a platform for testing multiple solutions with limited animal suffering before performing tests on mammals. Detailed results of the experimental conditions and testing techniques are provided. —Szarek, D. , Marycz, K. , Lis, A. , Zawada, Z. , Tabakow, P., Laska, J., Jarmundowicz, W. Lizard tail spinal cord: a new experimental model of spinal cord injury without limb paralysis. FASEB J. 30, 1391–1403 (2016). www.fasebj. org
Multiple rapid swallow responses segregate achalasia subtypes on high‐resolution manometry

Background Multiple rapid swallows (MRS) inhibit esophageal peristalsis and lower esophageal sphincter (LES) tone; a rebound excitatory response then results in an exaggerated peristaltic sequence. Multiple rapid swallows responses are dependent on intact inhibitory and excitatory neural function and could vary by subtype in achalasia spectrum disorders.
Bone morphogenetic protein signaling promotes morphogenesis of blood vessels, wound epidermis, and actinotrichia during fin regeneration in zebrafish

Zebrafish fin regeneration involves initial formation of the wound epidermis and the blastema, followed by tissue morphogenesis. The mechanisms coordinating differentiation of distinct tissues of the regenerate are poorly understood. Here, we applied pharmacologic and transgenic approaches to address the role of bone morphogenetic protein (BMP) signaling during fin restoration. To map the BMP transcriptional activity, we analyzed the expression of the evolutionarily conserved direct phospho‐Smad1 target gene, id1, and its homologs id2a and id3. This analysis revealed the BMP activity in the distal blastema, wound epidermis, osteoblasts, and blood vessels of the regenerate. Blocking the BMP function with a selective chemical inhibitor of BMP type I receptors, DMH1, suppressed id1 and id3 expression and arrested regeneration after blastema formation. We identified several previously uncharacterized functions of BMP during fin regeneration. Specifically, BMP signaling is required for remodeling of plexus into structured blood vessels in the rapidly growing regenerate. It organizes the wound epithelium by triggering wnt5b expression and promoting Collagen XIV‐A deposition into the basement membrane. BMP represents the first known signaling that induces actinotrichia formation in the regenerate. Our data reveal a multifaceted role of BMP for coordinated morphogenesis of distinct tissues during regeneration of a complex vertebrate appendage. —Thorimbert, V. , König, D. , Marro, J., Ruggiero, F. , Jaźwińska, A. Bone morphogenetic protein signaling promotes morphogenesis of blood vessels, wound epidermis, and actinotrichia during fin regeneration in zebrafish. FASEB J. 29, 4299‐4312 (2015). www.fasebj. org
Mst2 and Lats Kinases Regulate Apoptotic Function of Yes Kinase-associated Protein (YAP)*

The Hippo pathway in Drosophila controls the size and shape of organs. In the fly, activation of this pathway conveys growth-inhibitory signals and promotes apoptosis in epithelial cells. We “reconstituted” the Hippo pathway in a human epithelial cell line and showed that, in contrast to flies, the activation of this pathway results in anti-apoptotic signals. We have shown that in human embryonic kidney (HEK) 293 cells, the complex formation between transcriptional co-activators YAPs (Yes kinase-associated proteins) and Lats kinases requires the intact WW domains of YAPs, as well as intact Pro-Pro-AA-Tyr (where AA is any amino acid) motifs in Lats kinases. These kinases cooperate with the upstream Mst2 kinase to phosphorylate YAPs at Ser-127. Overexpression of YAP2 in HEK293 cells promoted apoptosis, whereas the Mst2/Lats1-induced phosphorylation of YAP partially rescued the cells from apoptotic death. Apoptotic signaling of YAP2 was mediated via stabilization of p73, which formed a complex with YAP2. All components of the Hippo pathway that we studied were localized in the cytoplasm, with the exception of YAP, which also localized in the nucleus. The localization of YAP2 in the nucleus was negatively controlled by the Lats1 kinase. Our apoptotic “readout” of the Hippo pathway in embryonic kidney cells represents a useful experimental system for the identification of the putative upstream receptor, membrane protein, or extracellular factor that initiates an entire signaling cascade and ultimately controls the size of organs.
Expression of endothelial phosphorylated caveolin‐1 is increased in brain injury

Aims: Increased endothelial caveolae leading to transcytosis of plasma proteins is associated with blood–brain barrier (BBB) breakdown and cerebral oedema in brain injury. Increased expression of caveolin‐1α (Cav‐1), an integral caveolar membrane protein, was reported in endothelium of arterioles and veins with BBB breakdown to fibronectin post injury. In this study the phosphorylation state of Cav‐1 and its association with BBB breakdown was determined in the rat cortical cold injury model over a period of days 0.5–6 post lesion. Methods: Expression of phosphorylated Cav‐1 was determined by immunoblotting and dual labelling immunofluorescence for phosphorylated caveolin‐1 and fibronectin, a marker of BBB breakdown. A phospho‐specific monoclonal antibody that selectively recognizes only tyrosine 14‐phosphorylated Cav‐1 (PY14Cav‐1) was used. Results: Immunoblots showed constitutive expression of PY14Cav‐1 in cortex of control rats and a significant increase in PY14Cav‐1 expression at the lesion site up to day 4 post lesion. PY14Cav‐1 immunostaining was observed in the endothelium of lesion vessels at days 0.5–4 post lesion, in neutrophils at days 0.5 and 2 and in macrophages at day 6 post lesion. Dual labelling showed that 100% of vessels with BBB breakdown to fibronectin showed endothelial PY14Cav‐1 on day 0.5, the percentage decreasing to 62% on day 4. On day 6, none of the vessels showed endothelial phosphorylated Cav‐1. Conclusions: The presence of phosphorylated Cav‐1 in endothelium of vessels showing BBB breakdown suggests that phosphorylated Cav‐1 signalling may be one of the factors associated with early BBB breakdown and brain oedema in brain injury.
COMMD1 Forms Oligomeric Complexes Targeted to the Endocytic Membranes via Specific Interactions with Phosphatidylinositol 4,5-Bisphosphate*

Copper metabolism Murr1 domain 1 (COMMD1) is a 21-kDa protein involved in copper export from the liver, NF-κB signaling, HIV infection, and sodium transport. The precise function of COMMD and the mechanism through which COMMD1 performs its multiple roles are not understood. Recombinant COMMD1 is a soluble protein, yet in cells COMMD1 is largely seen as targeted to cellular membranes. Using co-localization with organelle markers and cell fractionation, we determined that COMMD1 is located in the vesicles of the endocytic pathway, whereas little COMMD1 is detected in either the trans-Golgi network or lysosomes. The mechanism of COMMD1 recruitment to cell membranes was investigated using lipidspotted arrays and liposomes. COMMD1 specifically binds phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) in the absence of other proteins and does not bind structural lipids; the phosphorylation of PtdIns at position 4 is essential for COMMD1 binding. Proteolytic sensitivity and molecular modeling experiments identified two distinct domains in the structure of COMMD1. The C-terminal domain appears sufficient for lipid binding, because both the full-length and C-terminal domain proteins bind to PtdIns(4,5)P2. In native conditions, endogenous COMMD1 forms large oligomeric complexes both in the cytosol and at the membrane; interaction with PtdIns(4,5)P2 increases the stability of oligomers. Altogether, our results suggest that COMMD1 is a scaffold protein in a distinct sub-compartment of endocytic pathway and offer first clues to its role as a regulator of structurally unrelated membrane transporters.
Leishmania‐encoded orthologs of macrophage migration inhibitory factor regulate host immunity to promote parasite persistence

Leishmania major encodes 2 orthologs of the cytokine macrophage migration inhibitory factor (MIF), whose functions in parasite growth or in the host‐parasite interaction are unknown. To determine the importance of Leishmania‐encoded MIF, both LmMIF genes were removed to produce an mif–/– strain of L. major. This mutant strain replicated normally in vitro but had a 2‐fold increased susceptibility to clearance by macrophages. Mice infected with mif–/– L. major, when compared to the wild‐type strain, also showed a 3‐fold reduction in parasite burden. Microarray and functional analyses revealed a reduced ability of mif–/– L. major to activate antigen‐presenting cells, resulting in a 2‐fold reduction in T‐cell priming. In addition, there was a reduction in inflammation and effector CD4 T‐cell formation in mif–/– L. major‐infected mice when compared to mice infected with wild‐type L. major. Notably, effector CD4 T cells that developed during infection with mif–/– L. major demonstrated statistically significant differences in markers of functional exhaustion, including increased expression of IFN‐γ and IL‐7R, reduced expression of programmed death‐1, and decreased apoptosis. These data support a role for LmMIF in promoting parasite persistence by manipulating the host response to increase the exhaustion and depletion of protective CD4 T cells. —Holowka, T. , Castilho, T. M. , Baeza Garcia, A. , Sun, T. , McMahon‐Pratt, D. , Bucala, R. Leishmania‐encoded orthologs of macrophage migration inhibitory factor regulate host immunity to promote parasite persistence. FASEB J. 30, 2249–2265 (2016). www.fasebj. org
Mechanism of MutS Searching for DNA Mismatches and Signaling Repair*

DNA mismatch repair is initiated by the recognition of mismatches by MutS proteins. The mechanism by which MutS searches for and recognizes mismatches and subsequently signals repair remains poorly understood. We used single-molecule analyses of atomic force microscopy images of MutS-DNA complexes, coupled with biochemical assays, to determine the distributions of conformational states, the DNA binding affinities, and the ATPase activities of wild type and two mutants of MutS, with alanine substitutions in the conserved Phe-Xaa-Glu mismatch recognition motif. We find that on homoduplex DNA, the conserved Glu, but not the Phe, facilitates MutS-induced DNA bending, whereas at mismatches, both Phe and Glu promote the formation of an unbent conformation. The data reveal an unusual role for the Phe residue in that it promotes the unbending, not bending, of DNA at mismatch sites. In addition, formation of the specific unbent MutS-DNA conformation at mismatches appears to be required for the inhibition of ATP hydrolysis by MutS that signals initiation of repair. These results provide a structural explanation for the mechanism by which MutS searches for and recognizes mismatches and for the observed phenotypes of mutants with substitutions in the Phe-Xaa-Glu motif.