Assessment of Detoxification Strategies for Used Dental Implant Healing Abutments: Macroscopic and Biological Implications.

PURPOSE: Dental implant manufacturers recommend healing abutments (HA) be used for single-patient use; however, reuse on multiple patients following decontamination and sterilization is common. This study aims to evaluate four decontamination strategies utilizing enzymatic agents, available in most clinical settings, to determine the level to which biomaterial can be removed in a group of previously used HA (uHA). Secondly, to determine the degree to which the decontaminated HA are capable of inducing an inflammatory response in-vitro compared to new, never used HA. MATERIALS AND METHODS: Fifty HA were collected following 2-4 weeks of intraoral use and distributed randomly into 5 test groups (Group A-E; n = 10/group). Group A: Enzymatic cleaner foam + Autoclave; Group B: Ultrasonic bath with enzymatic cleaner + Autoclave; Group C: Prophy jet + Enzymatic cleaner foam + Autoclave; Group D: Prophy jet + ultrasonic bath with enzymatic cleaner + Autoclave; Group E: Prophy jet + Autoclave. Ten new, sterile HA served as controls (Group "Control"). Residual protein concentration was determined by a Micro BCA protein assay while HA from each group were stained with Phloxine B and macroscopically examined for the presence of debris. To examine the inflammatory potential, human primary macrophages were exposed to HA and supernatant levels of 9 cytokines/chemokines profiles were analyzed using a multiplex bead assay. RESULTS: All test groups presented with differences in the degree of visual decontamination compared to Controls, with Groups D and E displaying the most effective surface debris removaland reduced protein concentration. Of the detoxification strategies, Groups D and E removed the greatest biomaterial while least effective was Group A. However, compared to Controls, multiplex assays revealed high levels of inflammatory cytokine secretion up to 5 days from all Test Groups (A-E) irrespective of the decontamination method used. CONCLUSION: Our study found that compared to new, never used HA, decontamination of uHA utilizing enzymatic cleaners failed to reestablish inert HA surfaces and prevent an inflammatory immune response in-vitro. Clinicians should not reuse HA even after attempts to decontaminate and sterilize HA surfaces.

Validity and Applicability of the Global Leadership Initiative on Malnutrition (GLIM) Criteria in Patients Hospitalized for Acute Medical Conditions.

(1) Background: The objectives of this study were to evaluate the concurrent and predictive validity and the applicability of the global leadership initiative on malnutrition (GLIM) criteria in patients hospitalized for acute medical conditions. (2) Methods: prospective cohort study with patients hospitalized for acute medical conditions. For validation, the methodology proposed by the GLIM group of experts was used. Sensitivity and specificity values greater than 80% with respect to those for the subjective global assessment (SGA) were necessary for concurrent validation. The time necessary to complete each nutritional assessment test was determined. (3) Results: A total of 119 patients were evaluated. The SGA was applied to the entire cohort, but the GLIM criteria could not be applied to 3.4% of the patients. The sensitivity and specificity of the GLIM criteria with respect to those for the SGA to detect malnutrition were 78.0 and 86.2%, respectively. The GLIM predictive validity criterion was fulfilled because patients with malnutrition more frequently had a hospital stay >10 days (odds ratio of 2.98 (1.21-7.60)). The GLIM criteria required significantly more time for completion than did the SGA (p = 0.006). (4) Conclusion: The results of this study do not support the use of the GLIM criteria over the SGA for the diagnosis of malnutrition in patients hospitalized for acute medical conditions.

Downregulation of miRNA-26 in chronic periodontitis interferes with innate immune responses and cell migration by targeting phospholipase C beta 1.

AIM: To evaluate the potential role of miR-26 family members in periodontal pathogenesis by assessing innate immune responses to periopathic bacteria and regulation of cytoskeletal organization. MATERIALS AND METHODS: Expression of miR-26a-5p and miR-26b-5p was quantified in gingival biopsies derived from healthy and periodontally diseased subjects before and after non-surgical (scaling and root planing) therapy by RT-qPCR. Global pathway analysis and luciferase assays were performed for target identification and validation. Cytokine expression was assessed in miR-26a-5p transfected human oral keratinocytes upon stimulation with either live Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans or Pg lipopolysaccharide (LPS). Wound closure assays were performed in cells transfected with miR-26a-5p, while the impact on cytoskeletal organization was assessed by F-actin staining. RESULTS: miR-26a-5p and miR-26b-5p were downregulated in diseased gingiva and restored 4-6 weeks post-therapy to levels comparable with healthy subjects. Target validation assays identified phospholipase C beta 1 as a bona fide novel target exhibiting antagonistic expression pattern in disease and post-therapy cohorts. miR-26a-5p transfected cells secreted higher levels of cytokine/chemokines upon stimulation with periopathogens and demonstrated impaired cell migration and cytoskeletal rearrangement. CONCLUSIONS: Downregulated miR-26a-5p levels in periodontal inflammation may interfere with key cellular functions that may have significant implications for host defence and wound healing.

Used dental implant healing abutments elicit immune responses: A comparative analysis of detoxification strategies.

PURPOSE: To determine if healing abutments (HA) can be "decontaminated" using four strategies available in clinical settings and compare the detoxification efficacy by quantifying residual biomaterial and capacity to elicit an inflammatory response in-vitro. MATERIALS AND METHODS: Forty HA collected from subjects following intraoral use were randomly distributed into four test groups (A-D): A: autoclave only, B: ultrasonic bath plus autoclave, C: prophy-jet plus autoclave, and D: Scrub sponge plus autoclave. New, sterile HA: group E (Control). Residual protein concentration was determined by Micro BCA assay and stained with Phloxine B for macroscopic examination. HA were placed in human CD14+ monocyte derived-macrophage (mo-Mφ) cultures and supernatant collected at 4, 24, 48, and 5 days to analyze cytokine profiles using multiplex bead assay. RESULTS: Test groups showed visible differences in "decontamination" levels compared to control. Groups C and D showed most effective debris removal and lowest residual protein concentration. Multiplex assay showed marked induction of pro-inflammatory cytokines by groups A and B and to a significantly lower level by groups C and D. CONCLUSION: HA were not entirely "decontaminated" using common methods available relative to new, sterile HA and were capable of stimulating an immune response.

Maintenance of S-nitrosothiol homeostasis plays an important role in growth suppression of estrogen receptor-positive breast tumors.

INTRODUCTION: Protein denitrosylation by thioredoxin reductase (TrxR) is key for maintaining S-nitrosothiol (SNO) homeostasis, although its role in tumor progression is unknown. Therefore, the present study aimed to assess the role of altered SNO homeostasis in breast cancer cells. METHODS: The impairment of SNO homeostasis in breast cancer cells was achieved with the highly specific TrxR inhibitor auranofin and/or exposure to S-nitroso-L-cysteine. S-nitrosylated proteins were detected using the biotin switch assay. Estrogen receptor (ER) alpha knockdown was achieved using RNA silencing technologies and subcellular localization of ERα was analyzed by confocal microscopy. The Oncomine database was explored for TrxR1 (TXNRD1) expression in breast tumors and TrxR1, ER and p53 expression was analyzed by immunohistochemistry in a panel of breast tumors. RESULTS: The impairment of SNO homeostasis enhanced cell proliferation and survival of ER+ MCF-7 cells, but not of MDA-MB-231 (ER-, mut p53) or BT-474 (ER+, mut p53) cells. This enhanced cell growth and survival was associated with Akt, Erk1/2 phosphorylation, and augmented cyclin D1 expression and was abolished by the ER antagonist fulvestrant or the p53 specific inhibitor pifithrin-α. The specific silencing of ERα expression in MCF-7 cells also abrogated the growth effect of TrxR inhibition. Estrogenic deprivation in MCF-7 cells potentiated the pro-proliferative effect of impaired SNO homeostasis. Moreover, the subcellular distribution of ERα was altered, with a predominant nuclear localization associated with phosphorylation at Thr311 in those cells with impaired SNO homeostasis. The impairment of SNO homeostasis also expanded a cancer stem cell-like subpopulation in MCF-7 cells, as indicated by the increase of percentage of CD44+ cells and the augmented capability to form mammospheres in vitro. Notably, ER+ status in breast tumors was significantly associated with lower TXNDR1 mRNA expression and immunohistochemical studies confirmed this association, particularly when p53 abnormalities were absent. CONCLUSION: The ER status in breast cancer may dictate tumor response to different nitrosative environments. Impairment of SNO homeostasis confers survival advantages to ER+ breast tumors, and these molecular mechanisms may also participate in the development of resistance against hormonal therapies that arise in this type of mammary tumors.

To cardiovascular disease and beyond: new therapeutic perspectives of statins in autoimmune diseases and cancer.

Statins have been successfully used in patients with hypercholesterolemia and cardiovascular diseases, but there is increasing evidence that they exert effects by much exceeding the lowering of cholesterol levels. Statins have antiatherosclerotic, antiinflammatory, antioxidant, immunomodulatory and antithrombotic effects. These "pleiotropic" effects stem from their inhibition of prenylation of the small GTP-binding proteins Ras and Rho, and to the disruption, or depletion, of cholesterol rich membrane micro-domains (membrane rafts). Through these pathways statins modulate immune responses by altering cytokine levels and by affecting the function of cells involved in both innate and adaptive responses. Anti-inflammatory and immunosuppressory properties of statins provide the rationale for their potential application in conditions in which the inflammation and immune response represent key pathogenic mechanisms, such as antiphospholipid syndrome, rheumatoid arthritis and systemic lupus erythematosus. Reduction of atherosclerosis progression in autoimmunity is also a very important effect. Statins pathways of action in systemic autoimmune diseases, and their potential therapeutic use are discussed in this review. The inhibition of mevalonate pathway by statins impairs modification of Ras and Rho GTPases, which play key roles in signaling pathways related to tumor formation, metastasis and cell death. There is experimental and clinical evidence that statins may improve the therapeutic outcome of anticancer drugs. Thus, this review will also discuss recent insights into the molecular mechanisms underlying the anticancer effects of statins and their assessment as promising candidates for inclusion into current therapeutic regimens for the treatment of malignant diseases.

AEE788 is a vascular endothelial growth factor receptor tyrosine kinase inhibitor with antiproliferative and proapoptotic effects in acute myeloid leukemia.

OBJECTIVE: Aberrant activation of tyrosine kinase receptors is frequently observed in acute myelogenous leukemia (AML). Moreover, activating mutations of the fms-like tyrosine kinase 3 (FLT3) receptor can be found in approximately 30% of patients, thereby representing one of the most frequent single genetic alterations in AML. AEE788, a novel dual receptor tyrosine kinase inhibitor of endothelial growth factor and vascular endothelial growth factor (VEGF), is being studied in several solid tumors with remarkable success. It is not known, however, about the efficacy of this inhibitor in the treatment of AML. Therefore, we investigated the effect of AEE788 in the treatment of three human AML cell lines and seven AML patient samples. MATERIALS AND METHODS: Cell survival in THP-1, MOLM-13, and MV4-11 cell lines (the two last harboring the FLT3/internal tandem duplication mutation) and AML blasts incubated with 0.5 to 15 microM AEE788 were quantified. We also studied the activation of VEGF/VEGF receptors loop, FLT3, and their downstream effectors (Akt, extracellular signal-regulated kinase, signal transducers and activators of transcription 5, and nuclear factor-kappaB). RESULTS: Our data showed that AEE788 was a tyrosine kinase inhibitor of FLT3 activity and had antiproliferative and proapoptotic activity in AML-derived cell lines and AML blasts that presented phosphorylation of the FLT3 receptor. Consistently, in these cells AEE788 abrogated VEGF/VEGF receptors activation and the survival signaling pathways studied. CONCLUSION: Taken together, the activity of AEE788 might represent a promising new option of targeting FLT3 for the treatment of AML.

Additive effect of PTK787/ZK 222584, a potent inhibitor of VEGFR phosphorylation, with Idarubicin in the treatment of acute myeloid leukemia.

Acute myeloid leukemia (AML) is a disease with a poor prognosis. It has been demonstrated that AML cells express vascular endothelial growth factor (VEGF) as well as Flt-1 and KDR, resulting in an autocrine pathway for cell survival. PTK787/ZK 222584 is a new oral antiangiogenic molecule that inhibits tyrosine kinase activity of all known VEGF receptors. The present study aimed to investigate the therapeutic efficacy of combining PTK787/ZK 222584 with a chemotherapeutic agent, such as Idarubicin, for treatment of AML. We have analyzed in four AML cell lines and seven AML patient samples, cell proliferation, apoptosis, angiogenesis. and activation of several related intracellular pathways after treatment with PTK787/ZK 222584 alone or combined with Idarubicin. PTK787/ZK 222584 decreased VEGF levels and VEGF receptor phosphorylation in the AML cells showing Fms-like tyrosine kinase 3/internal tandem duplication mutation (Flt3/ITD). Both drugs, given separately, inhibited cell proliferation and promoted apoptosis. Moreover, combined treatment promoted more apoptosis and inhibition of cell proliferation than each compound administered separately in all AML cells. In conclusion, PTK787/ZK 222584 combined with Idarubicin achieved a better therapeutic efficacy than chemotherapy alone in AML cells, especially in those with Flt3/ITD, in which the combination further prevented activation of the angiogenic process.

Utilidad de la determinación de la subunidad alfa tras estímulo con TRH como indicador de persistencia y/o recurrencia tumoral en gonadotropinomas / Utility of alpha subunit determination alter thyrotropin-releasing hormone stimulation as an indicator of gonadotropinoma persistente and/or recurrence

Objetivo: Los gonadotropinomas son tumores originados en las células gonadotropas de la hipófisis anterior causales de la síntesis y la secreción de gonadotrofinas (folitropina [FSH] y lutropina [LH]). La mayoría de estos tumores tienen una producción alterada de gonadotropinas y de sus subunidades (folitropina beta, subunidad alfa y, con menos frecuencia, lutropina beta). Los gonadotropinomas pueden presentar una respuesta de la subunidad alfa de las gonadotropinas al estímulo con protirrelina (TRH) que podría diferenciar estos tumores de los no funcionantes. De igual forma, esta prueba podría ser de utilidad tras la cirugía para poderdiscernir los posibles restos tumorales respecto a los cambios posquirúrgicos. Sujetos y método: Se estudió a 24 pacientes intervenidos de macroadenoma hipofisario, de los que 14 fueron diagnosticados de gonadotropinoma en el estudio histológico. Se les practicó la prueba de lasubunidad alfa tras la administración de TRH antes y después de la cirugía.Resultados: En el estudio prequirúrgico el 50% de los gonadotropinomas tuvieron una respuesta positiva a dicha prueba y en el posquirúrgico otro 50%. El 83% de los pacientes con gonadotropinoma presentaban signos de recidiva/persistencia tumoral y/o cambios en laresonancia magnética (RM) de control posquirúrgico; el 83% de estos pacientes (41,6% del total) tuvo una respuesta positiva de la subunidad alfa tras el estímulo con TRH. En el grupo de macroadenomas no gonadotropinomas sólo un 33% tuvo respuesta positiva antes de la cirugía y otro 33%, después. En la RM practicada después de la cirugía, todos mostraban signos radiológicos compatibles con cambios inflamatorios posquirúrgicos o signos de persistencia y/o recidivatumoral. Conclusiones: Dicha prueba podría ser de ayuda en el diagnóstico diferencial de los gonadotropinomas, así como en el seguimiento y la valoración posquirúrgica de estos tumores (AU)

Objective: Gonadotropinomas are adenomas of the gonadotropic cells of the anterior pituitary. These cells produce and secrete gonadotropins (follicle-stimulating hormone and luteinizing hormone). Most of these tumors show altered production of gonadotropins and their subunits (the - FSH, and, less frequently, -LH subunits). The thyrotropin-releasing hormone (TRH) stimulation test could differentiate these tumors from nonfunctioning tumors. Equally, this test could be able to distinguish between postsurgical changes and tumoral remnants after surgery. Subjects and method: We studied 24 patients with pituitary macroadenoma, 14 of who had a histological diagnosis of gonadotroph adenoma. The TRH stimulation test was performed before and after surgery. Results: Both before and after surgery, a positive result to the TRH test was obtained in 50% of gonadotropinomas. Magnetic resonance imaging (MRI)performed after surgery revealed that 83% of the patients with gonadotropinoma had signs of tumoral persistence or recurrence and/or postsurgical changes. Of these patients, 83% (41.6% of the total) showed positive (..) (AU)

Efecto de 3 modelos de dieta en la respuesta de glucosa e insulina, perfil lipídico y función endotelial en individuos con resistencia a la insulina / Effect of 3 model diets on the glucose and insulin response, lipid profile and endothelial function in insulin-resistant subjects

Introducción. Hemos estudiado el efecto de 3 modelos de dieta en la respuesta de glucosa e insulina, el perfil lipídico y la función endotelial en individuos con resistencia a la insulina. Pacientes y métodos. Once voluntarios con sobrepeso abdominal y resistencia a la insulina realizaron 3 fases de dieta en mantenimiento de peso, en un estudio aleatorizado y cruzado: una rica en grasas saturadas (SAT), otra en grasas monoinsaturadas (MUFA) y otra en hidratos de carbono (HC). Hemos determinado los valores de glucosa e insulina, el perfil lipídico y la vasodilatación endotelial mediada por el flujo con láser Doppler. Resultados. El peso corporal y el gasto energético basal no cambiaron durante las 3 fases de la dieta. El homeostasis model assessment-insulin resistance (HOMA-ir) mejoró tras la dieta rica en MUFA, comparada con las dietas altas en SAT e HC (2,32 ± 0,35; 2,72 ± 0,37, y 2,52 ± 0,37; p < 0,05). Un desayuno rico en HC incrementa los valores posprandiales de glucosa, insulina y nitrosamina, y disminuye los valores de colesterol unido a lipoproteínas de alta densidad (cHDL) comparados con las dietas MUFA y SAT. La reactividad endotelial en ayuno y posprandial tras 150 min de la ingesta fue mayor con MUFA, comparada con las dietas ricas en SAT y en HC (271,1 ± 16,63; 220,8 ± 13,6; 244,9 ± 37,7; p < 0,05; y 251,9 ± 16,3; 180,1 ± 10,6; 181,4 ±16; ANOVA; p < 0,05, respectivamente). Conclusiones. En pacientes con resistencia a la insulina, una dieta alta en MUFA mejora el HOMA-ir, disminuye los valores posprandiales de glucosa, insulina y nitrosamina, incrementa los valores de cHDL y mejora la reactividad endotelial (AU)

Introduction. We have studied the effect of 3 model diets on glucose and insulin response, lipid profile and endothelial function in insulin resistant subjects. Patients and methods. Eleven volunteers with insulin resistance and central overweight underwent 3 phases of diet on weight-maintenance, in a randomized and crossed study: a rich-saturated fat diet (SAT), another one rich in monounsaturated fat (MUFA, rich in olive oil) and another rich in carbohydrates (CH). We determined the glucose and insulin levels, the lipid profile and the half-full flow mediated endothelial vasodilation by means of Laser-Doppler. Results. Body weight and basal energy expenditure were unchanged throughout the study. The homeostasis model assessment-insulin resistance (HOMA-ir) was improved during the MUFA-rich diet as compared with SAT and CH diets (2.32 ± 0.35; 2.72 ± 0.37 and 2.52 ± 0.37; p < 0.05). During a CH rich breakfast, postprandial glucose, insulin and nitrotyrosine levels were increased and the HDL-C levels remained lowered as compared with those during MUFA and SAT diets. Endothelial reactivity, measured fasting and after 150 min of meal test, was higher with olive oil rich diet compared with SAT and CH rich diets (baseline values: 271.1 ± 16.63, 220.8 ± 13.6, 244.9 ± 37.7, ANOVA p < 0.05; postprandial values: 251.9 ± 16.3, 180.1 ± 10.6, 181.4 ± 16, ANOVA p < 0.05). Conclusions. In insulin resistant subjects a MUFA-rich diet improves HOMA-ir, decreases postprandial glucose, insulin and nitrotyrosine levels, increases HDL-C levels and improves basal and postprandial endothelial reactivity (AU)

Utility of alpha subunit determination after thyrotropin-releasing hormone stimulation as an indicator of gonadotropinoma persistence and/or recurrence.

OBJECTIVE: Gonadotropinomas are adenomas of the gonadotropic cells of the anterior pituitary. These cells produce and secrete gonadotropins (follicle-stimulating hormone and luteinizing hormone). Most of these tumors show altered production of gonadotropins and their subunits (the p-FSH, a and, less frequently, p-LH subunits). The thyrotropin-releasing hormone (TRH) stimulation test could differentiate these tumors from nonfunctioning tumors. Equally, this test could be able to distinguish between postsurgical changes and tumoral remnants after surgery. SUBJECTS AND METHOD: We studied 24 patients with pituitary macroadenoma, 14 of who had a histological diagnosis of gonadotroph adenoma. The TRH stimulation test was performed before and after surgery. RESULTS: Both before and after surgery, a positive result to the TRH test was obtained in 50% of gonadotropinomas. Magnetic resonance imaging (MRI) performed after surgery revealed that 83% of the patients with gonadotropinoma had signs of tumoral persistence or recurrence and/or postsurgical changes. Of these patients, 83% (41.6% of the total) showed positive a subunit stimulation after the TRH test. In the group of non-gonadotropinoma macroadenomas, only 33% had a positive result before surgery and another 33% had a positive result after surgery. In the MRI performed after surgery, all showed tumoral persistence/recurrence or postsurgical inflammatory changes. CONCLUSIONS: This test could be useful in the differential diagnosis of gonadotropinomas as well as in the follow-upand postsurgical evaluation of these tumors.

Distribution and determinants of adiponectin, resistin and ghrelin in a randomly selected healthy population.

OBJECTIVE: Adiponectin, resistin, ghrelin and the IGF-I system seem to play an important role in the regulation of body composition throughout life, but the mechanisms are not well understood. The aim of our study was to analyse the distribution among sexes and all decades of the adult life of adiponectin, resistin and ghrelin and their relationship with anthropometric, body composition parameters and the IGF-I system. SUBJECTS: One hundred and thirty-four men and 127 healthy women were included in the study. MEASUREMENTS: Plasma concentration of adiponectin, resistin, ghrelin, total IGF-I, free IGF-I and IGFBP-3 were determined in all subjects. Body composition was evaluated by bioelectrical impedance. RESULTS: Resistin and ghrelin were not affected by age. Plasma adiponectin correlated negatively with age, body mass index (BMI), waist-to-hip ratio (WHR), waist circumference (WC), fat mass (FM) and body fat (BF) in men. Adiponectin correlated negatively with WHR and positively with free IGF-I in women. Resistin correlated positively with BMI and WC only in men, and ghrelin correlated positively with WC, BMI and FM and negatively with free IGF-I in men. In multiple regression analysis adiponectin remained associated with WHR (beta=-0.19, P=0.01) in women. Resistin was positively associated with BMI (beta=0.30, P=0.003) in women and ghrelin was negatively related to free IGF-I (beta=-0.158, P=0.019) in men. CONCLUSIONS: Plasma adiponectin declines with age and is negatively associated with FM in men. Our data suggest the existence of a positive correlation of adiponectin and the IGF-I axis in women and of an inverse relationship between ghrelin and the IGF-I system in men.