A NuRD for all seasons.

The nucleosome-remodeling and deacetylase (NuRD) complex is an essential transcriptional regulator in all complex animals. All seven core subunits of the complex exist as multiple paralogs, raising the question of whether the complex might utilize paralog switching to achieve cell type-specific functions. We examine the evidence for this idea, making use of published quantitative proteomic data to dissect NuRD composition in 20 different tissues, as well as a large-scale CRISPR knockout screen carried out in >1000 human cancer cell lines. These data, together with recent reports, provide strong support for the idea that distinct permutations of the NuRD complex with tailored functions might regulate tissue-specific gene expression programs.

Relaxation of thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) by endothelium-derived 6-nitrodopamine

6-Nitrodopamine is a novel catecholamine released by vascular tissues, heart, and vas deferens. The aim of this study was to investigate whether 6-nitrodopamine is released from the thoracic aorta and pulmonary artery rings of marmosets (Callithrix spp.) and to evaluate the relaxing and anti-contractile actions of this catecholamine. Release of 6-nitrodopamine, dopamine, noradrenaline, and adrenaline was assessed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). The relaxations induced by 6-nitrodopamine and by the selective dopamine D2 receptor antagonist L-741,626 were evaluated on U-46619 (3 nM)-pre-contracted vessels. The effects of 6-nitrodopamine and L-741,626 on the contractions induced by electric-field stimulation (EFS), dopamine, noradrenaline, and adrenaline were also investigated. Both aorta and pulmonary artery rings exhibited endothelium-dependent release of 6-nitrodopamine, which was significantly reduced by the NO synthesis inhibitor L-NAME. Addition of 6-nitrodopamine or L-741,626 caused concentration-dependent relaxations of both vascular tissues, which were almost abolished by endothelium removal, whereas L-NAME and the soluble guanylate cyclase inhibitor ODQ had no effect on 6-nitrodopamine-induced relaxations. Additionally, pre-incubation with 6-nitrodopamine antagonized the dopamine-induced contractions, without affecting the noradrenaline- and adrenaline-induced contractions. Pre-incubation with L-741,626 antagonized the contractions induced by all catecholamines. The EFS-induced contractions were significantly increased by L-NAME, but unaffected by ODQ. Immunohistochemical assays showed no immunostaining of the neural tissue markers S-100 and calretinin in either vascular tissue. The results indicated that 6-nitrodopamine is the major catecholamine released by marmoset vascular tissues, and it acts as a potent and selective antagonist of dopamine D2-like receptors. 6-nitrodopamine release may be the major mechanism by which NO causes vasodilatation.

The role of auxiliary domains in modulating CHD4 activity suggests mechanistic commonality between enzyme families.

CHD4 is an essential, widely conserved ATP-dependent translocase that is also a broad tumour dependency. In common with other SF2-family chromatin remodelling enzymes, it alters chromatin accessibility by repositioning histone octamers. Besides the helicase and adjacent tandem chromodomains and PHD domains, CHD4 features 1000 residues of N- and C-terminal sequence with unknown structure and function. We demonstrate that these regions regulate CHD4 activity through different mechanisms. An N-terminal intrinsically disordered region (IDR) promotes remodelling integrity in a manner that depends on the composition but not sequence of the IDR. The C-terminal region harbours an auto-inhibitory region that contacts the helicase domain. Auto-inhibition is relieved by a previously unrecognized C-terminal SANT-SLIDE domain split by ~150 residues of disordered sequence, most likely by binding of this domain to substrate DNA. Our data shed light on CHD4 regulation and reveal strong mechanistic commonality between CHD family members, as well as with ISWI-family remodellers.

Unexpectedly high prevalence of primary immune deficiency in fibromyalgia: serologic features and clinical correlates.

OBJECTIVES: We sought laboratory evidence of primary immune deficiency (PID), a condition known to be associated with recurrent infections and autoimmunity, in fibromyalgia (FM). We correlated laboratory findings with a clinical history of recurrent infections and reduced epidermal nerve fiber density (ENFD). METHODS: We prospectively measured serum total and subclass concentrations for IgA, IgG, IgM, IgE, and mannose-binding lectin in 72 adult FM subjects (31 "FM only;" 41 "FM+RA") and compared those results against historical controls. We also administered a novel "Lifetime History of Infections" questionnaire to all FM subjects and 40 apparently healthy, community volunteers matched for age, race, and gender. ENFD values available for 49/72 FM subjects were also correlated with immunoreactant levels. RESULTS: Of FM subjects, 96% (69/72) had ≥3 and 85% (61/72) had ≥4 of 9 immunoreactants below or within the lowermost quartile of historical normal values. Recurrent sinus infections occurred more often in "FM only" (p=0.06), and "FM+RA" subjects (p=0.02) than controls. "FM+RA" subjects had a significantly greater history of recurrent, severe non-sinus infections (p=0.04). The prevalence of total IgA deficiency was significantly greater in "FM only" than in "FM+RA" subjects (p=0.04). We also found a direct correlation between total IgA (p=0.02), IgA1 (p=0.005), and IgG1 (p=0.04) concentrations and ENFD in "FM only" subjects. CONCLUSIONS: Serologic evidence of PID in FM is common and correlates with a clinical history of recurrent sinus and non-sinus infections, and reduced ENFD. This study suggests that PID may be important to diagnostic and therapeutic considerations in FM.

Synthesis and Biological Activities of Cyclodepsipeptides of Aurilide Family from Marine Origin.

Aurilides are a class of depsipeptides occurring mainly in marine cyanobacteria. Members of the aurilide family have shown to exhibit strong cytotoxicity against various cancer cell lines. These compounds bear a pentapeptide, a polyketide, and an α-hydroxy ester subunit in their structure. A large number of remarkable studies on aurilides have emerged since 1996. This comprehensive account summarizes the biological activities and total syntheses of natural compounds of the aurilide family as well as their synthetic analogues.

ATM inhibition overcomes resistance to histone deacetylase inhibitor due to p21 induction and cell cycle arrest.

The antiproliferative effect induced by histone deactylase inhibitors (HDACi) is associated with the up-regulated expression of the cyclin-dependent kinase inhibitor p21. Paradoxically, the increased expression of p21 correlates with a reduced cell killing to the drug. The direct targeting of p21 is not feasible. An alternate approach could selectively target factors upstream or downstream of p21 that affect one or more specific aspects of p21 function. HDAC inhibitors appear to activate p21 expression via ataxia telangiectasia mutated (ATM) activity. KU60019, a specific ATM inhibitor, has shown to decrease the p21 protein levels in a concentration dependent manner. We explored the potential synergistic interaction of the ATM inhibitor with romidepsin, given the potential complementary impact around p21. A synergistic cytotoxic effect was observed in all lymphoma cell lines examined when the HDACi was combined with KU60019. The increase in apoptosis correlates with decreased expression of p21 due to the ATM inhibitor. KU60019 decreased expression of the cyclin-dependent kinase inhibitor at the transcriptional level, compromising the ability of HDACi to induce p21 and cell cycle arrest and ultimately facilitating a shift toward the apoptotic phase. Central to the increased apoptosis observed when romidepsin is combined with KU60019 is the reduced expression of p21 and the absence of a G2/M cell cycle arrest that would be exploited by the tumor cells to evade the cytotoxic effect of the HDAC inhibitor. We believe this strategy may offer a promising way to identify rational combinations for HDACi directed therapy, improving their activity in malignant disease.

Lactose drives Enterococcus expansion to promote graft-versus-host disease.

Disruption of intestinal microbial communities appears to underlie many human illnesses, but the mechanisms that promote this dysbiosis and its adverse consequences are poorly understood. In patients who received allogeneic hematopoietic cell transplantation (allo-HCT), we describe a high incidence of enterococcal expansion, which was associated with graft-versus-host disease (GVHD) and mortality. We found that Enterococcus also expands in the mouse gastrointestinal tract after allo-HCT and exacerbates disease severity in gnotobiotic models. Enterococcus growth is dependent on the disaccharide lactose, and dietary lactose depletion attenuates Enterococcus outgrowth and reduces the severity of GVHD in mice. Allo-HCT patients carrying lactose-nonabsorber genotypes showed compromised clearance of postantibiotic Enterococcus domination. We report lactose as a common nutrient that drives expansion of a commensal bacterium that exacerbates an intestinal and systemic inflammatory disease.

Bridging in vitro dissolution and in vivo exposure for acalabrutinib. Part II. A mechanistic PBPK model for IR formulation comparison, proton pump inhibitor drug interactions, and administration with acidic juices.

Acalabrutinib (Calquence®) 100 mg (bid) has received accelerated approval by FDA for the treatment of adult patients with mantle cell lymphoma (MCL) who have received at least one prior therapy. Acalabrutinib is a substrate of PgP and CYP3A4, with a significant fraction of drug metabolized by first pass gut extraction and 25% absolute bioavailability. The absorption of acalabrutinib is affected by stomach pH, with lower pharmacokinetic exposure observed following co-administration with proton pump inhibitors. During dissolution at pH values below its highest basic pKa, the two basic moieties of acalabrutinib react with protons from the aqueous solution, leading to a higher pH at the drug surface than in the bulk solution. A batch-specific product particle size distribution (P-PSD), was derived from dissolution data using a mechanistic model that was based on the understanding of surface pH and the contribution of micelles to the dissolution rate. P-PSD values obtained for various batches of acalabrutinib products in simple buffers, or in complex fluids such as fruit juices, were successfully integrated into a physiologically based pharmacokinetic (PBPK) model developed using GastroPlus v9.0™. The integrated model allowed the prediction of clinical pharmacokinetics under normal physiological stomach pH conditions as well as following treatment with proton pump inhibitors. The model also accounted for lower pharmacokinetic exposure that was observed when acalabrutinib was co-administered with the acidic beverages, grapefruit juice, (which contains CYP3A inhibitors), and orange drink (which does not contain CYP3A inhibitors), relative to administration with water. The integration of dissolution data in the PBPK model enables mechanistic understanding and the establishment of more robust in vitro-in vivo correlations (IVIVC) under a variety of conditions. The model can then distinguish the interplay between dissolution and first pass extraction and how in vivo stomach pH, saturation of gut PgP, and saturation or inhibition of gut CYP3A4, will impact the pharmacokinetics of acalabrutinib.

Evidence of peripheral large nerve involvement in fibromyalgia: a retrospective review of EMG and nerve conduction findings in 55 FM subjects.

OBJECTIVE: Whereas small fiber neuropathy (SFN) is now a recognized part of fibromyalgia (FM), surprisingly little attention has been paid to any findings of large fiber neuropathy (LFN) in this disorder. Since 90% to 95% of FM subjects seen in our outpatient facility routinely undergo EMG and nerve conduction studies (NCS) we elected to retrospectively review the EMG/NCS results garnered from a large cohort of unselected subjects in order to describe the electrodiagnostic features of LFN in FM. METHODS: Records from 100 consecutive, unselected clinic patients meeting the 1990 ACR criteria for FM, who had undergone EMG/NCS, were reviewed. The same electromyographer tested all subjects. After exclusion of FM patients with any other clinically relevant condition that might influence EMG results (e.g., familial neural degenerative conditions, diabetes mellitus, Vitamin B-12 deficiency, etc.) fifty-five FM subjects remained: 29 subjects with "FM Only," and 26 subjects with FM+Rheumatoid Arthritis ("FM+RA"). All subjects had also undergone ankle area skin biopsy for determination of epidermal nerve fiber density (ENFD). Fourteen other subjects, without FM or RA, examined by the same electromyographer, were used as an EMG/NCS comparison group. RESULTS: Ninety percent of the "FM Only" subjects demonstrated a demyelinating and/or axonal, sensorimotor polyneuropathy, and 63% had findings of SFN (ENFD ≤7 fibers/mm), suggesting a mixed fiber neuropathy in most. Furthermore, 61% of the "FM Only" subjects showed EMG findings suggestive of non-myotomal lower extremity axonal motor denervation, most likely due to a polyneuropathy, and 41% satisfied published criteria for "possible" chronic inflammatory demyelinating polyneuropathy (CIDP). There was surprisingly little difference in the EMG/NCS findings between the "FM Only" and the "FM+RA" groups. With the exception of carpal tunnel syndrome, our EMG/NCS comparison group showed few to none of these findings. CONCLUSION: Our review of the EMG/NCS results, gleaned from the largest FM cohort yet studied with these modalities, shows that electrodiagnostic features of polyneuropathy, muscle denervation, and CIDP are common in FM. Furthermore these electrodiagnostic findings are often seen coincident with SFN, and are not significantly influenced by the presence of RA. These results, particularly when taken as a whole, suggest that EMG/NCS may be clinically useful in detecting LFN in FM and help in better understanding the etiopathogenesis of this painful disorder.

NR4A2 haploinsufficiency is associated with intellectual disability and autism spectrum disorder.

NR4A2, a member of the nuclear receptor superfamily, is involved in modulation of target gene transcription, regulating several developmental processes such as regulation of cellular homeostasis, neuronal development, inflammation and carcinogenesis. 2q24.1 deletions are extremely rare, and only 1 patient with a de novo deletion encompassing only NR4A2 gene was reported so far. We report 3 additional patients with a de novo deletion encompassing NR4A2: 2 patients have deletions encompassing only NR4A2 gene and 1 patient has a deletion including NR4A2 and the first exon of GPD2. Our patients presented a neurodevelopmental disorder including language impairment, developmental delay, intellectual disability and/or autism spectrum disorder. We suggest that NR4A2 haploinsufficiency is implicated in neurodevelopmental disorder with high penetrance.

Apoptotic intrinsic pathway proteins predict survival in canine cutaneous mast cell tumours.

Mast cell tumours (MCTs) are the most frequent canine round cell neoplasms and show variable biological behaviours with high metastatic and recurrence rates. The disease is treated surgically and wide margins are recommended. Adjuvant chemotherapy and radiotherapy used in this disease cause DNA damage in neoplastic cells, which is aimed to induce apoptotic cell death. Resisting cell death is a hallmark of cancer, which contributes to the development and progression of tumours. The aim of this study was to investigate the expression of the proteins involved in the apoptotic intrinsic pathway and to evaluate their potential use as prognostic markers for canine cutaneous MCTs. Immunohistochemistry for BAX, BCL2, APAF1, Caspase-9, and Caspase-3 was performed in 50 canine cases of MCTs. High BAX expression was associated with higher mortality rate and shorter survival. BCL2 and APAF1 expressions offered additional prognostic information to the histopathological grading systems. The present results indicate that variations in the expression of apoptotic proteins are related to malignancy of cutaneous MCTs in dogs.

Efeito de diferentes intensidades de exercício em esteira sobre os parâmetros eletrocardiográficos de cães hígidos / Effect of different intensities of treadmill exercise on eletrocardiographic parameters of healthy dogs

Objetivou-se a padronização de testes de exercício de alta e baixa intensidades em esteira, bem como a avaliação do eletrocardiograma de cães submetidos a esses testes. Para tal fim, sete cães da raça Australian Cattle Dog e quatro da raça Border Collie clinicamente saudáveis foram submetidos a dois testes de exercício em esteira, com pelo menos sete dias de intervalo: T1 - teste de exercício de alta intensidade e curta duração, e T2 - teste de exercício de baixa intensidade e longa duração. A amplitude e a duração de ondas e intervalos foram avaliadas no momento antes do exercício (M0) e nos momentos imediatamente após o término dos testes (MPE) e 30 minutos após (M30). A frequência e o ritmo cardíacos foram avaliados antes dos testes e continuamente por 30 minutos após o término do exercício. Verificou-se diferença significativa somente para a duração do intervalo QT em M30 em T1, além de algumas arritmias, como complexos atriais e ventriculares prematuros isolados em três animais após o teste T1, e em quatro após T2. Os testes de exercício foram adequados para promover estimulação simpática nos cães, contudo não causaram alterações significativas no eletrocardiograma, provavelmente em razão do excelente condicionamento físico dos animais.(AU)

This study aimed to stardardize high and low intensity exercise tests, and evaluate the electrocardiogram of dogs submitted to these tests. Seven clinically healthy Australian Cattle dogs and four Border Collies underwent two exercise treadmill tests, with at least a seven day interval: T1 - high intensity and short duration exercise test, and T2 - low intensity and long duration exercise test. Amplitude and duration of waves and intervals were assessed at resting time before exercise (M0), at immediately after (MPE) and at 30 minutes (M30) after the end of the tests. Heart rate and cardiac rhythm were evaluated before the tests and continuously for 30 minutes after the end of exercise. There was a significant difference only for duration of the QT interval at M30 in T1, and some arrhythmias such as isolated atrial and ventricular premature complexes in three animals after T1 test, and in four dogs after T2. The exercise tests of the present study was suitable to promote sympathetic stimulation in dogs, however did not cause significant changes on the electrocardiogram probably because of the excellent physical fitness of the dogs.(AU)

Zeólitas naturais na dieta de frangos de corte / Natural zeolites in broiler diet

O objetivo deste trabalho foi avaliar o efeito da inclusão de 0,5% da zeólita natural clinoptilolita na dieta de frangos de corte sobre o consumo de água, ração e características das excretas. Foi conduzido um ensaio em gaiolas metabólicas, com frangos de corte de linhagem comercial, no período de 14 a 23 dias de idade. O experimento foi constituído de dois grupos de aves, sendo que um recebeu dieta sem inclusão de zeólitas (controle) e outro dieta com inclusão de 0,5% de zeólita (clinoptilolita) em rações isonutritivas, em um delineamento inteiramente ao acaso, com 10 repetições de 10 aves. Avaliou-se o consumo de água e de ração, nas excretas, pH, teor de nitrogênio total e umidade. A inclusão de 0,5% de zeólitas naturais na dieta não alterou (P>0,05) o consumo de água e de ração e o teor de nitrogênio das excretas, entretanto, reduziu (P<0,05) o pH e a umidade das excretas. Conclui-se que a inclusão de 0,5% de zeólitas naturais na dieta de frangos de corte reduz o pH e a umidade das excretas e pode ser utilizada como aditivo alimentar sem prejudicar o consumo de água e de ração.(AU)

The aim of this study was to evaluate the addition of 0.5% natural zeolites (clinoptilolite) to the diet of broilers and their effect on water and feed intake and excreta characteristics. The test was carried out in metabolic cages, with commercial line broilers, in the period from 14 to 23 days old. The experiment was conducted with two groups of broilers: one group received diet without zeolite inclusion (control) and the other received a diet with 0.5% inclusion of zeolite (clinoptilolite) in balanced diets, in a completely randomized design, with 10 replicates of 10 broilers. Water and feed intake, pH, total nitrogen, and moisture of excreta were evaluated. The inclusion of 0.5% of natural zeolites in the diet did not change (P>0.05) the water and feed intake and total nitrogen of excreta, nevertheless, it decreased (P<0.05) the pH and moisture of excreta. We conclude that the addition of 0.5% natural zeolites to the diet of broilers decreases pH and moisture of excreta and can be used as feed additive without compromising water and feed intake.(AU)

Increased expression of tissue inhibitor of metalloproteinase-1 correlates with improved outcome in canine cutaneous mast cell tumours.

Canine mast cell tumour (MCT) is a biologically heterogeneous disease. The extracellular matrix degradation promoted by matrix metalloproteinases (MMPs) has been studied in an attempt to elucidate the mechanisms involved in the biological behaviour of tumours. The aim of this study was to characterize the expression of MMP-2 and -9 and tissue inhibitors of metalloproteinase (TIMP)-1 and -2 in canine cutaneous MCTs and to evaluate their prognostic values. Immunohistochemical staining for MMP-2, MMP-9, TIMP-2 and TIMP-1 was performed in 46 canine cases of MCTs. TIMP-1 expression showed an independent prognostic value for post-surgical survival and disease-related mortality. Dogs with MCTs showing less than 22.9% mast cell TIMP-1 positivity were more prone to die because of the disease and had a shorter post-surgical survival. This article suggests the involvement of TIMP-1 in MCT progression, by contributing to a good outcome in patients with MCTs.

Immunohistochemical Expression of the Pluripotency Factor OCT4 in Canine Mast Cell Tumours.

Cancer stem cells (CSCs) are related to malignancy and resistance to chemotherapy in several tumours. OCT4 is a 'pluripotency factor' that is expressed by these cells. The aim of the present study was to investigate OCT4 expression in canine cutaneous mast cell tumours (MCTs) by means of immunohistochemistry. Twenty-eight cases were evaluated and showed variable immunolabelling patterns. The dogs were treated by surgery alone and followed up for a minimum of 180 days. No significant difference was found between histopathological grades and similar results were obtained for mortality due to the disease and post-surgical survival. These preliminary results suggest that OCT4 expression is not a precise prognostic indicator for canine MCT.

The Role and Importance of Small Fiber Neuropathy in Fibromyalgia Pain.

Serious investigators of fibromyalgia (FM) realize the profound implications of finding features of small fiber neuropathy (SFN) in this disorder. For the first time, an easily reproducible and generally agreed upon, peripheral tissue lesion has been reported from multiple investigative centers. Understanding how this discovery relates to other features of FM, and how one might utilize it to better comprehend, and care for, afflicted patients' painful complaints remains a challenge, however. In this article we review how the SFN seen in FM may be placed in context, and suggest how such a tissue abnormality might be used to better understand the pathophysiology of FM, and plan for its effective treatment. We also suggest how finding SFN in FM implies the need for continued focused research within the area of neuropathic disease in FM.