When things get hot: Thermoregulation behavior in the lizard Sceloporus aeneus at different thermal conditions.

Rising environmental temperatures have become a global threat for ectotherms, with the increasing risk of overheating promoting population declines. Flexible thermoregulatory behavior might be a plausible mechanism to mitigate the effects of extreme temperatures. We experimentally evaluated thermoregulatory behavior in the bunchgrass lizard, Sceloporus aeneus, at three different environmental temperatures (25, 35 and 45 °C) both with and without a thermal refuge. We recorded themoregulatory behaviors (body posture and movement between hot and cold patches) and compared individual lizards across all experimental temperature and shelter combinations. Behavioral thermoregulation in S. aeneus was characterized by the expression of five body postures, whose frequencies varied based on environmental temperature and microthermal conditions. Behavioral responses allowed lizards to maintain a mean body temperature <40 °C, the critical thermal maximum for temperate species, even at extreme environmental temperatures (45 °C). Although S. aeneus express an array of behavioral postures that provide an effective mechanism to cope with elevating temperatures, the presence of a thermal refuge was important to better achieve this. Together, our study offers a novel method to evaluate microhabitat preference that encompasses both behavioral observations and time-space analysis based on the ambient thermal distribution, a consideration that can aid in the formulation of more accurate predictions on ectotherm vulnerability related to increasing global environmental temperatures.

Functional evolution of the colony-stimulating factor 1 receptor (CSF1R) and its ligands in birds.

Macrophage colony-stimulating factor (CSF1 or M-CSF) and interleukin 34 (IL34) are secreted cytokines that control macrophage survival and differentiation. Both act through the CSF1 receptor (CSF1R), a type III transmembrane receptor tyrosine kinase. The functions of CSF1R and both ligands are conserved in birds. We have analyzed protein-coding sequence divergence among avian species. The intracellular tyrosine kinase domain of CSF1R was highly conserved in bird species as in mammals but the extracellular domain of avian CSF1R was more divergent in birds with multiple positively selected amino acids. Based upon crystal structures of the mammalian CSF1/IL34 receptor-ligand interfaces and structure-based alignments, we identified amino acids involved in avian receptor-ligand interactions. The contact amino acids in both CSF1 and CSF1R diverged among avian species. Ligand-binding domain swaps between chicken and zebra finch CSF1 confirmed the function of variants that confer species specificity on the interaction of CSF1 with CSF1R. Based upon genomic sequence analysis, we identified prevalent amino acid changes in the extracellular domain of CSF1R even within the chicken species that distinguished commercial broilers and layers and tropically adapted breeds. The rapid evolution in the extracellular domain of avian CSF1R suggests that at least in birds this ligand-receptor interaction is subjected to pathogen selection. We discuss this finding in the context of expression of CSF1R in antigen-sampling and antigen-presenting cells.

Co-targeting aurora kinase with PD-L1 and PI3K abrogates immune checkpoint mediated proliferation in peripheral T-cell lymphoma: a novel therapeutic strategy.

Peripheral T-cell non-Hodgkin lymphoma (PTCL) are heterogeneous, rare, and aggressive diseases mostly incurable with current cell cycle therapies. Aurora kinases (AKs) are key regulators of mitosis that drive PTCL proliferation. Alisertib (AK inhibitor) has a response rate ∼30% in relapsed and refractory PTCL (SWOG1108). Since PTCL are derived from CD4+/CD8+ cells, we hypothesized that Program Death Ligand-1 (PD-L1) expression is essential for uncontrolled proliferation. Combination of alisertib with PI3Kα (MLN1117) or pan-PI3K inhibition (PF-04691502) or vincristine (VCR) was highly synergistic in PTCL cells. Expression of PD-L1 relative to PD-1 is high in PTCL biopsies (∼9-fold higher) and cell lines. Combination of alisertib with pan-PI3K inhibition or VCR significantly reduced PD-L1, NF-κB expression and inhibited phosphorylation of AKT, ERK1/2 and AK with enhanced apoptosis. In a SCID PTCL xenograft mouse model, alisertib displayed high synergism with MLN1117. In a syngeneic PTCL mouse xenograft model alisertib demonstrated tumor growth inhibition (TGI) ∼30%, whilst anti-PD-L1 therapy alone was ineffective. Alisertib + anti-PD-L1 resulted in TGI >90% indicative of a synthetic lethal interaction. PF-04691502 + alisertib + anti-PD-L1 + VCR resulted in TGI 100%. Overall, mice tolerated the treatments well. Co-targeting AK, PI3K and PD-L1 is a rational and novel therapeutic strategy for PTCL.

Fish scale artefact on an intraoral imaging receptor.

OBJECTIVES: To describe an artefact, termed the fish scale artefact, present on an intraoral imaging receptor. METHODS: Thirty brand new DIGORA Optime photostimulable phosphor (PSP) plates (Soredex/Orion Corp., Helsinki, Finland) were imaged using the dental digital quality assurance radiographic phantom (Dental Imaging Consultants LLC, San Antonio, TX). All PSP plates were scanned at the same spatial resolution (dpi) using the high resolution mode. Two evaluators assessed all 30 plates. Each evaluator assessed the 30 PSP plates separately for purposes of establishing interrater reliability, and then together in order to obtain the gold standard result. RESULTS: The fish scale artefact was detected on 46.7% of the PSP plates. The kappa coefficient for interrater reliability was 0.86 [95% CI (0.69-1.00)], indicating excellent interrater reliability. For Evaluator 1, sensitivity was 0.85 [95% CI (0.55-0.98)]; specificity was 0.94 [CI (0.71-1.00)] and overall accuracy was 0.90 [95% CI (0.73-0.98)]. For Evaluator 2, sensitivity was 1.00 [95% CI (0.75-1.00)]; specificity was 0.94 [CI (0.71-1.00)] and overall accuracy was 0.97 [95% CI (0.83-1.00)]. These results indicate excellent agreement with the gold standard for both evaluators. CONCLUSIONS: Utilizing a comprehensive quality assurance protocol, we identified a fish scale artefact inherent to the image receptor. Additional research is needed to determine if the artefact remains static over time or if it increases over time. Likewise, research to determine the potential sources contributing to an increase in the artefact is needed.

Disruption of Aneuploidy and Senescence Induced by Aurora Inhibition Promotes Intrinsic Apoptosis in Double Hit or Double Expressor Diffuse Large B-cell Lymphomas.

Double hit (DH) or double expressor (DE) diffuse large B-cell lymphomas (DLBCL) are aggressive non-Hodgkin's lymphomas (NHL) with translocations and/or overexpressions of MYC and BCL-2, which are difficult to treat. Aurora kinase (AK) inhibition with alisertib in DH/DE-DLBCL induces cell death in ∼30%, while ∼70% are aneuploid and senescent cells (AASC), a mitotic escape mechanism contributing to drug resistance. These AASCs elaborated a high metabolic rate by increased AKT/mTOR and ERK/MAPK activity via BTK signaling through the chronic active B-cell receptor (BCR) pathway. Combinations of alisertib + ibrutinib or alisertib + ibrutinib + rituximab significantly reduced AASCs with enhanced intrinsic cell death. Inhibition of AK + BTK reduced phosphorylation of AKT/mTOR and ERK-1/2, upregulated phospho-H2A-X and Chk-2 (DNA damage), reduced Bcl-6, and decreased Bcl-2 and Bcl-xL and induced apoptosis by PARP cleavage. In a DE-DLBCL SCID mouse xenograft model, ibrutinib alone was inactive, while alisertib + ibrutinib was additive with a tumor growth inhibition (TGI) rate of ∼25%. However, TGI for ibrutinib + rituximab was ∼50% to 60%. In contrast, triple therapy showed a TGI rate of >90%. Kaplan-Meier survival analysis showed that 67% of mice were alive at day 89 with triple therapy versus 20% with ibrutinib + rituximab. All treatments were well tolerated with no changes in body weights. A novel triple therapy consisting of alisertib + ibrutinib + rituximab inhibits AASCs induced by AK inhibition in DH/DE-DLBCL leading to a significant antiproliferative signal, enhanced intrinsic apoptosis and may be of therapeutic potential in these lymphomas. Mol Cancer Ther; 16(10); 2083-93. ©2017 AACR.

Reciprocal feedback inhibition of the androgen receptor and PI3K as a novel therapy for castrate-sensitive and -resistant prostate cancer.

Gain-of-function of the androgen receptor (AR) and activation of PI3K/AKT/mTOR pathway have been demonstrated to correlate with progression to castration-resistant prostate cancer (CRPC). However, inhibition of AR or PI3K/mTOR alone results in a reciprocal feedback activation. Therefore, we hypothesized that dual inhibition of the AR and PI3K/mTOR pathway might lead to a synergistic inhibition of cell growth and overcome drug resistance in CRPC. Here, we reported that androgen-depletion increased AR protein level and Akt phosphorylation at Ser473 and Thr308 in LNCaP cells. Moreover, we developed resistance cell lines of LNCaP to Enzalutamide (or MDV3100), an AR inhibitor (named as LNCaP 'MDV-R') and PF-04691502, a PI3K/mTOR inhibitor (named as LNCaP 'PF-R'). MTS analysis showed that LNCaP 'PF-R' was strongly resistant to Enzalutamide treatment, and on the other hand, LNCaP 'MDV-R' was 6-fold resistant to PF-04691502 treatment. Mechanistically, LNCaP 'MDV-R' cells had significantly reduced AR, loss of PSA and increase Akt activity in contrast with LNCaP 'PF-R' cells. Combined inhibition of PI3K/mTOR and AR pathways with a variety of small molecular inhibitors led to a synergistic suppression of cell proliferation and a profound increase of apoptosis and cell cycle arrest in both androgen-dependent LNCaP and independent CRPC 22Rv1 cell lines. In conclusion, this study provides preclinical proof-of-concept that the combination of a PI3K/mTOR inhibitor with an AR inhibitor results in a synergistic anti-tumor response in non-CRPC and CRPC models.

The development and maintenance of the mononuclear phagocyte system of the chick is controlled by signals from the macrophage colony-stimulating factor receptor.

BACKGROUND: Macrophages have many functions in development and homeostasis as well as innate immunity. Recent studies in mammals suggest that cells arising in the yolk sac give rise to self-renewing macrophage populations that persist in adult tissues. Macrophage proliferation and differentiation is controlled by macrophage colony-stimulating factor (CSF1) and interleukin 34 (IL34), both agonists of the CSF1 receptor (CSF1R). In the current manuscript we describe the origin, function and regulation of macrophages, and the role of CSF1R signaling during embryonic development, using the chick as a model. RESULTS: Based upon RNA-sequencing comparison to bone marrow-derived macrophages grown in CSF1, we show that embryonic macrophages contribute around 2% of the total embryo RNA in day 7 chick embryos, and have similar gene expression profiles to bone marrow-derived macrophages. To explore the origins of embryonic and adult macrophages, we injected Hamburger-Hamilton stage 16 to 17 chick embryos with either yolk sac-derived blood cells, or bone marrow cells from EGFP+ donors. In both cases, the transferred cells gave rise to large numbers of EGFP+ tissue macrophages in the embryo. In the case of the yolk sac, these cells were not retained in hatched birds. Conversely, bone marrow EGFP+ cells gave rise to tissue macrophages in all organs of adult birds, and regenerated CSF1-responsive marrow macrophage progenitors. Surprisingly, they did not contribute to any other hematopoietic lineage. To explore the role of CSF1 further, we injected embryonic or hatchling CSF1R-reporter transgenic birds with a novel chicken CSF1-Fc conjugate. In both cases, the treatment produced a large increase in macrophage numbers in all tissues examined. There were no apparent adverse effects of chicken CSF1-Fc on embryonic or post-hatch development, but there was an unexpected increase in bone density in the treated hatchlings. CONCLUSIONS: The data indicate that the yolk sac is not the major source of macrophages in adult birds, and that there is a macrophage-restricted, self-renewing progenitor cell in bone marrow. CSF1R is demonstrated to be limiting for macrophage development during development in ovo and post-hatch. The chicken provides a novel and tractable model to study the development of the mononuclear phagocyte system and CSF1R signaling.

Klhl31 attenuates ß-catenin dependent Wnt signaling and regulates embryo myogenesis.

Klhl31 is a member of the Kelch-like family in vertebrates, which are characterized by an amino-terminal broad complex tram-track, bric-a-brac/poxvirus and zinc finger (BTB/POZ) domain, carboxy-terminal Kelch repeats and a central linker region (Back domain). In developing somites Klhl31 is highly expressed in the myotome downstream of myogenic regulators (MRF), and it remains expressed in differentiated skeletal muscle. In vivo gain- and loss-of-function approaches in chick embryos reveal a role of Klhl31 in skeletal myogenesis. Targeted mis-expression of Klhl31 led to a reduced size of dermomyotome and myotome as indicated by detection of relevant myogenic markers, Pax3, Myf5, myogenin and myosin heavy chain (MF20). The knock-down of Klhl31 in developing somites, using antisense morpholinos (MO), led to an expansion of Pax3, Myf5, MyoD and myogenin expression domains and an increase in the number of mitotic cells in the dermomyotome and myotome. The mechanism underlying this phenotype was examined using complementary approaches, which show that Klhl31 interferes with ß-catenin dependent Wnt signaling. Klhl31 reduced the Wnt-mediated activation of a luciferase reporter in cultured cells. Furthermore, Klhl31 attenuated secondary axis formation in Xenopus embryos in response to Wnt1 or ß-catenin. Klhl31 mis-expression in the developing neural tube affected its dorso-ventral patterning and led to reduced dermomyotome and myotome size. Co-transfection of a Wnt3a expression vector with Klhl31 in somites or in the neural tube rescued the phenotype and restored the size of dermomyotome and myotome. Thus, Klhl31 is a novel modulator of canonical Wnt signaling, important for vertebrate myogenesis. We propose that Klhl31 acts in the myotome to support cell cycle withdrawal and differentiation.

Cell-autonomous sex differences in gene expression in chicken bone marrow-derived macrophages.

We have identified differences in gene expression in macrophages grown from the bone marrow of male and female chickens in recombinant chicken M-CSF (CSF1). Cells were profiled with or without treatment with bacterial LPS for 24 h. Approximately 600 transcripts were induced by prolonged LPS stimulation to an equal extent in the male and female macrophages. Many transcripts encoded on the Z chromosome were expressed ∼1.6-fold higher in males, reflecting a lack of dosage compensation in the homogametic sex. A smaller set of W chromosome-specific genes was expressed only in females. LPS signaling in mammals is associated with induction of type 1 IFN-responsive genes. Unexpectedly, because IFNs are encoded on the Z chromosome of chickens, unstimulated macrophages from the female birds expressed a set of known IFN-inducible genes at much higher levels than male cells under the same conditions. To confirm that these differences were not the consequence of the actions of gonadal hormones, we induced gonadal sex reversal to alter the hormonal environment of the developing chick and analyzed macrophages cultured from male, female, and female sex-reversed embryos. Gonadal sex reversal did not alter the sexually dimorphic expression of either sex-linked or IFN-responsive genes. We suggest that female birds compensate for the reduced dose of inducible IFN with a higher basal set point of IFN-responsive genes.

Novel receptor tyrosine kinase targeted combination therapies for imatinib-resistant gastrointestinal stromal tumors (GIST).

BACKGROUND: c-Kit/α-PDGFR targeted therapies are effective for gastrointestinal stromal tumors (GIST), but, >50% develop drug resistance. METHODS: RTK expression (c-Kit, c-Met, AXL, HER-1, HER-2, IGF-1R) in pre-/post-imatinib (IM) GIST patient samples (n=16) and 4 GIST cell lines were examined for RTK inhibitor activity. GIST-882 cells were cultured in IM every other day, cells collected (1 week to 6 months) and analyzed by qRT-PCR and Western blotting. RESULTS: Immunohistochemistry pre-/post-IM demonstrated continued expression of c-Kit and HER1, while a subset expressed IGF-1R, c-Met and AXL. In GIST cells (GIST-882, GIST430/654, GIST48) c-Kit, HER1 and c-Met are co-expressed. Acute IM over-express c-Kit while chronic IM, lose c-Kit and HER-1 in GIST882 cells. GIST882 and GIST430/654 cells have an IC50 0.077 and 0.59 µM to IM respectively. GIST48 have an IC50 0.66 µM to IM, 0.91 µM to amuvatinib [AMU] and 0.67 µM to erlotinib (Erl). Synergistic combinations: GIST882, AMU + Erl (CI 0.20); IM + AMU (CI 0.50), GIST430/654, IM + afatinib (CI 0.39); IM + AMU (CI 0.42), GIST48, IM + afatinib (CI 0.03); IM + AMU (CI 0.04); AMU + afatinib (CI 0.36); IM + Erl (CI 0.63). CONCLUSION: Targeting c-Kit plus HER1 or AXL/c-Met abrogates IM resistance in GIST.

Deficiencia congénita de Proteína C asociada a polimorfismo C677T del gen de la 5,10-metiltetrahidrofolato reductasa, a propósito de un caso / Congenital deficiency of protein C associated with C677T polymorphism of the gene from the 5,10-methyltetrahydrofolate reductase. Case report

Se presenta el caso clínico de un recién nacido con trombofilia hereditaria por deficiencia congénita de proteína C asociada a polimorfismo C677T del gen de la 5,10-metiltetrahidrofolato reductasa.

We present a clinical case of a newborn with inherited thrombophilia by congenital deficiency of protein C associated with the gene from the 5, 10-methyltetrahydrofolate reductase C677T polymorphism.

Visualisation of chicken macrophages using transgenic reporter genes: insights into the development of the avian macrophage lineage.

We have generated the first transgenic chickens in which reporter genes are expressed in a specific immune cell lineage, based upon control elements of the colony stimulating factor 1 receptor (CSF1R) locus. The Fms intronic regulatory element (FIRE) within CSF1R is shown to be highly conserved in amniotes and absolutely required for myeloid-restricted expression of fluorescent reporter genes. As in mammals, CSF1R-reporter genes were specifically expressed at high levels in cells of the macrophage lineage and at a much lower level in granulocytes. The cell lineage specificity of reporter gene expression was confirmed by demonstration of coincident expression with the endogenous CSF1R protein. In transgenic birds, expression of the reporter gene provided a defined marker for macrophage-lineage cells, identifying the earliest stages in the yolk sac, throughout embryonic development and in all adult tissues. The reporter genes permit detailed and dynamic visualisation of embryonic chicken macrophages. Chicken embryonic macrophages are not recruited to incisional wounds, but are able to recognise and phagocytose microbial antigens.

Alisertib added to rituximab and vincristine is synthetic lethal and potentially curative in mice with aggressive DLBCL co-overexpressing MYC and BCL2.

Pearson correlation coefficient for expression analysis of the Lymphoma/Leukemia Molecular Profiling Project (LLMPP) demonstrated Aurora A and B are highly correlated with MYC in DLBCL and mantle cell lymphoma (MCL), while both Auroras correlate with BCL2 only in DLBCL. Auroras are up-regulated by MYC dysregulation with associated aneuploidy and resistance to microtubule targeted agents such as vincristine. Myc and Bcl2 are differentially expressed in U-2932, TMD-8, OCI-Ly10 and Granta-519, but only U-2932 cells over-express mutated p53. Alisertib [MLN8237 or M], a highly selective small molecule inhibitor of Aurora A kinase, was synergistic with vincristine [VCR] and rituximab [R] for inhibition of cell proliferation, abrogation of cell cycle checkpoints and enhanced apoptosis versus single agent or doublet therapy. A DLBCL (U-2932) mouse model showed tumor growth inhibition (TGI) of ∼ 10-20% (p = 0.001) for M, VCR and M-VCR respectively, while R alone showed ∼ 50% TGI (p = 0.001). M-R and VCR-R led to tumor regression [TR], but relapsed 10 days after discontinuing therapy. In contrast, M-VCR-R demonstrated TR with no relapse >40 days after stopping therapy with a Kaplan-Meier survival of 100%. Genes that are modulated by M-VCR-R (CENP-C, Auroras) play a role in centromere-kinetochore function in an attempt to maintain mitosis in the presence of synthetic lethality. Together, our data suggest that the interaction between alisertib plus VCR plus rituximab is synergistic and synthetic lethal in Myc and Bcl-2 co-expressing DLBCL. Alisertib plus vincristine plus rituximab [M-VCR-R] may represent a new strategy for DLBCL therapy.

Adiponectin inhibits leptin-induced oncogenic signalling in oesophageal cancer cells by activation of PTP1B.

Obesity is characterised by hyperleptinaemia and hypoadiponectinaemia and these metabolic abnormalities may contribute to the progression of several obesity-associated cancers including oesophageal adenocarcinoma (OAC). We have examined the effects of leptin and adiponectin on OE33 OAC cells. Leptin stimulated proliferation, invasion and migration and inhibited apoptosis in a STAT3-dependant manner. Leptin-stimulated MMP-2 secretion in a partly STAT3-dependent manner and MMP-9 secretion via a STAT3-independent pathway. Adiponectin inhibited leptin-induced proliferation, migration, invasion, MMP secretion and reduced the anti-apoptotic effects: these effects of adiponectin were ameliorated by both a non-specific tyrosine phosphatase inhibitor and a specific PTP1B inhibitor. Adiponectin reduced leptin-stimulated JAK2 activation and STAT3 transcriptional activity in a PTP1B-sensitive manner and adiponectin increased both PTP1B protein and activity. We conclude that adiponectin restrains leptin-induced signalling and pro-carcinogenic behaviour by inhibiting the early events in leptin-induced signal transduction by activating PTP1B. Relative adiponectin deficiency in obesity may contribute to the promotion of OAC.

Production and characterisation of a monoclonal antibody that recognises the chicken CSF1 receptor and confirms that expression is restricted to macrophage-lineage cells.

Macrophages contribute to innate and acquired immunity as well as many aspects of homeostasis and development. Studies of macrophage biology and function in birds have been hampered by a lack of definitive cell surface markers. As in mammals, avian macrophages proliferate and differentiate in response to CSF1 and IL34, acting through the shared receptor, CSF1R. CSF1R mRNA expression in the chicken is restricted to macrophages and their progenitors. To expedite studies of avian macrophage biology, we produced an avian CSF1R-Fc chimeric protein and generated a monoclonal antibody (designated ROS-AV170) against the chicken CSF1R using the chimeric protein as immunogen. Specific binding of ROS-AV170 to CSF1R was confirmed by FACS, ELISA and immunohistochemistry on tissue sections. CSF1 down-regulated cell surface expression of the CSF1R detected with ROS-AV170, but the antibody did not block CSF1 signalling. Expression of CSF1R was detected on the surface of bone marrow progenitors only after culture in the absence of CSF1, and was induced during macrophage differentiation. Constitutive surface expression of CSF1R distinguished monocytes from other myeloid cells, including heterophils and thrombocytes. This antibody will therefore be of considerable utility for the study of chicken macrophage biology.

[Perception of pain during sigmoidoscopy flexible as an additional diagnostic method for irritable bowel syndrome]. / Percepción del dolor durante la sigmoidoscopia flexible como método diagnóstico adicional de síndrome de intestino irritable.

INTRODUCTION: Visceral hypersensitivity has been proposed as a biological marker of Irritable bowel syndrome (IBS). OBJECTIVE: To evaluate the pain perception during sigmoidoscopy using a visual analog scale of pain in patients with or without IBS, and to assess the pain perception as diagnostic criteria for IBS. We further assessed the sensitivity, specificity and diagnostic efficiency of pain scores to diagnose IBS. METHODS: A prospective case-control study in patients who underwent sigmoidoscopy for the evaluation of gastrointestinal symptoms. All patients completed Rome III criteria questionnaires and divided into two groups: IBS and non-IBS. All participants reported pain scores on visual analog scales after of study. Differences were evaluated. We calculated a receiver-operator characteristic curve (ROC), sensitivity, specificity and diagnostic efficiency. RESULTS: We analyzed 20 patients with IBS and 20 controls. The pain scores were higher in IBS patients compared with non-IBS patients (median, 52.5 vs. 27.5, p = 0.006). The area under the curve was 0.84, at pain score level of ≥ 40 mm with a sensitivity, specificity and diagnostic efficiency of 85%, 75% and 80%, respectively. CONCLUSIONS: The degree of pain perception was higher in IBS patients than in non-IBS patients during sigmoidoscopy. A score of pain perception in ≥ 40 mm may predict the diagnosis of IBS with good sensitivity (85%) and specificity (75%).

Percepción del dolor durante la sigmoidoscopia flexible como método diagnóstico adicional de síndrome de intestino irritable / Perception of pain during the flexible sigmoidoscopy as additional diagnostic method of irritable bowel syndrome

INTRODUCCIÓN: ante la ausencia de medidas objetivas para realizar el diagnóstico de Síndrome de Intestino Irritable (SII) se plantea el uso de la hipersensibilidad visceral como marcador biológico de la enfermedad. OBJETIVO: evaluar la percepción de dolor durante la sigmoidoscopia flexible mediante el uso de una escala analógica visual del dolor en pacientes con SII, además de valorar la percepción del dolor como criterio diagnóstico de ayuda al SII mediante la sensibilidad, especificidad y eficiencia diagnóstica de un valor de corte. METODOLOGÍA: se realizó un estudio prospectivo, tipo casos y controles, en pacientes con indicación para estudio sigmoidoscópico con y sin SII, para valorar la percepción del dolor después del examen mediante el empleo de una escala analógica visual. Se evaluaron las diferencias y se confeccionó una curva ROC, además de establecer la sensibilidad, especificidad y eficiencia diagnóstica. RESULTADOS: Se analizaron 20 pacientes con SII y 20 controles. El score de percepción del dolor fue mayor en los pacientes SII comparados con los pacientes no SII (mediana, 52.5 vs 27.5, p=0.006). El área bajo la curva fue de 0.84, determinándose un punto de corte en ≥ 40mm con una sensibilidad, especificidad y eficiencia diagnóstica de 85%, 75% y 80% respectivamente. CONCLUSIONES: Los pacientes con SII experimentan mayor percepción del dolor durante la sigmoidoscopia. Un valor de percepción del dolor en ≥ 40mm puede predecir el diagnóstico del SII con una buena sensibilidad (85%) y especificidad (75%).

INTRODUCTION: Visceral hypersensitivity has been proposed as a biological marker of Irritable bowel syndrome (IBS). OBJECTIVE: To evaluate the pain perception during sigmoidoscopy using a visual analog scale of pain in patients with or without IBS, and to asess the pain perception as diagnostic criteria for IBS. We further assessed the sensitivity, specificity and diagnostic efficiency of pain scores to diagnose IBS. METHODS: A prospective case-control study in patients who underwent sigmoidoscopy for the evaluation of gastrointestinal symptoms. All patients completed Rome III criteria questionnaries and divided into two groups: IBS and non-IBS. All participants reported pain scores on visual analog scales after of study. Differences were evaluated. We calculated a receiver-operator characteristic curve (ROC), sensitivity, specificity and diagnostic efficiency. RESULTS: We analyzed 20 patients with IBS and 20 controls. The pain scores were higher in IBS patients compared with non-IBS patients (median, 52.5 vs. 27.5, p = 0.006). The area under the curve was 0.84, at pain score level of ≥ 40 mm with a sensivity, specificity and diagnostic efficiency of 85%, 75% ando 80%, respectively. CONCLUSIONS: The degree of pain perception was higher in IBS patients than in non-IBS patients during sigmoidoscopy. A score of pain perception in ≥ 40 mm may predict the diagnosis of IBS with good sensivity (85%) and specificity (75%).

Aurora A inhibitor (MLN8237) plus vincristine plus rituximab is synthetic lethal and a potential curative therapy in aggressive B-cell non-Hodgkin lymphoma.

PURPOSE: Aurora A and B are oncogenic serine/threonine kinases that regulate mitosis. Overexpression of Auroras promotes resistance to microtubule-targeted agents. We investigated mechanistic synergy by inhibiting the mitotic spindle apparatus in the presence of MLN8237 [M], an Aurora A inhibitor with either vincristine [MV] or docetaxel [MD] in aggressive B-cell non-Hodgkin lymphoma (B-NHL). The addition of rituximab [R] to MV or MD was evaluated for synthetic lethality. EXPERIMENTAL DESIGN: Aggressive B-NHL cell subtypes were evaluated in vitro and in vivo for target modulation and anti-NHL activity with single agents, doublets, and triplets by analyzing cell proliferation, apoptosis, tumor growth, survival, and mechanisms of response/relapse by gene expression profiling with protein validation. RESULTS: MV is synergistic whereas MD is additive for cell proliferation inhibition in B-NHL cell culture models. Addition of rituximab to MV is superior to MD, but both significantly induce apoptosis compared with doublet therapy. Mouse xenograft models of mantle cell lymphoma showed modest single-agent activity for MLN8237, rituximab, docetaxel, and vincristine with tumor growth inhibition (TGI) of approximately 10% to 15%. Of the doublets, MV caused tumor regression, whereas TGI was observed with MD (approximately 55%-60%) and MR (approximately 25%-50%), respectively. Although MV caused tumor regression, mice relapsed 20 days after stopping therapy. In contrast, MVR was curative, whereas MDR led to TGI of approximately 85%. Proliferation cell nuclear antigen, Aurora B, cyclin B1, cyclin D1, and Bcl-2 proteins of harvested tumors confirmed response and resistance to therapy. CONCLUSIONS: Addition of rituximab to MV is a novel therapeutic strategy for aggressive B-NHL and warrants clinical trial evaluation.

Evidence of bioactivation of halomethanes and its relation to oxidative stress response in Chirostoma riojai, an endangered fish from a polluted lake in Mexico.

Halomethanes (HMs) are produced autochthonously in water bodies through the action of ultraviolet light in the presence of HM precursors, such as dissolved organic carbon and halogens. In mammals, toxic effects induced by HMs are diverse and include oxidative stress, which is also induced by divalent and polyvalent metals; however, in fish little information is available on HM metabolism and its possible consequences at the population level. In the present study, high CYP 2E1 and GST theta-like activities were found in viscera of the Toluca silverside Chirostoma riojai from Lake Zumpango (LZ; central Mexico). Formaldehyde, one of the HM metabolites, was correlated with CYP 2E1 activity and also induced lipid peroxidation in viscera. Hepatic CYP 2E1 activity was correlated with GST theta-like activity, suggesting the coupling of both pathways of HM bioactivation and its consequent oxidative damage. Sediment metals, among others, were also responsible for oxidative stress, particularly iron, lead, arsenic and manganese. However, under normal environmental conditions, the antioxidant enzymes of this species sustain catalysis adapted to oxidative stress. Findings suggest that this fish species apparently has mechanisms of adaptation and recovery that enable it to confront toxic agents of natural origin, such as metals and other substances formed through natural processes, e.g., HMs. This has allowed C. riojai to colonize LZ despite the high sensitivity of this species to xenobiotics of anthropogenic origin.

Frizzled-10 promotes sensory neuron development in Xenopus embryos.

Formation of the vertebrate nervous system requires coordinated cell-cell interactions, intracellular signalling events, gene transcription, and morphogenetic cell movements. Wnt signalling has been involved in regulating a wide variety of biological processes such as embryonic patterning, cell proliferation, cell polarity, motility, and the specification of cell fate. Wnt ligands associate with their receptors, members of the frizzled family (Fz). In Xenopus, five members of the frizzled family are expressed in the early nervous system. We have investigated the role of Xenopus frizzled-10 (Fz10) in neural development. We show that Fz10 is expressed in the dorsal neural ectoderm and neural folds in the region where primary sensory neurons develop. Fz10 mediates canonical Wnt signalling and interacts with Wnt1 and Wnt8 but not Wnt3a as shown in synergy assays. We find that Fz10 is required for the late stages of sensory neuron differentiation. Overexpression of Fz10 in Xenopus leads to an increase in the number of sensory neurons. Loss of Fz10 function using morpholinos inhibits the development of sensory neurons in Xenopus at later stages of neurogenesis and this can be rescued by co-injection of modified Fz10B and beta-catenin. In mouse P19 cells induced by retinoic acid to undergo neural differentiation, overexpression of Xenopus Fz10 leads to an increase in the number of neurons generated while siRNA knockdown of endogenous mouse Fz10 inhibits neurogenesis. Thus we propose Fz10 mediates Wnt1 signalling to determine sensory neural differentiation in Xenopus in vivo and in mouse cell culture.