Agomelatine, a Melatonin-Derived Drug, as a New Strategy for the Treatment of Colorectal Cancer.

The potential use of agomelatine as an alternative treatment for colorectal cancer is evaluated in this work. The effect of agomelatine was studied in an in vitro model using two cell lines with different p53 statuses (HCT-116, wild-type p53, and HCT-116 p53 null) and an in vivo xenograft model. The inhibitory effects of agomelatine and melatonin were stronger in the cells harboring the wild-type p53, although in both cell lines, the effect of agomelatine was greater than that of the melatonin. In vivo, only agomelatine was able to reduce the volumes of tumors generated by the HCT-116-p53-null cells. Both treatments induced changes in the rhythmicity of the circadian-clock genes in vitro, albeit with some differences. Agomelatine and melatonin regulated the rhythmicity of Per1-3, Cry1, Sirt1, and Prx1 in the HCT-116 cells. In these cells, agomelatine also regulated Bmal1 and Nr1d2, while melatonin changed the rhythmicity of Clock. In the HCT-116-p53-null cells, agomelatine regulated Per1-3, Cry1, Clock, Nr1d2, Sirt1, and Prx1; however, melatonin only induced changes in Clock, Bmal1, and Sirt1. The differences found in the regulation of the clock genes may explain the greater oncostatic effect of agomelatine in CRC.

PARP-1 Expression Influences Cancer Stem Cell Phenotype in Colorectal Cancer Depending on p53.

Poly(ADP-ribose) polymerase-1 (PARP-1) is a protein involved in multiple physiological processes. Elevated PARP-1 expression has been found in several tumours, being associated with stemness and tumorigenesis. In colorectal cancer (CRC), some controversy among studies has been described. In this study, we analysed the expression of PARP-1 and cancer stem cell (CSC) markers in CRC patients with different p53 status. In addition, we used an in vitro model to evaluate the influence of PARP-1 in CSC phenotype regarding p53. In CRC patients, PARP-1 expression correlated with the differentiation grade, but this association was only maintained for tumours harbouring wild-type p53. Additionally, in those tumours, PARP-1 and CSC markers were positively correlated. In mutated p53 tumours, no associations were found, but PARP-1 was an independent factor for survival. According to our in vitro model, PARP-1 regulates CSC phenotype depending on p53 status. PARP-1 overexpression in a wild type p53 context increases CSC markers and sphere forming ability. By contrast, those features were reduced in mutated p53 cells. These results could implicate that patients with elevated PARP-1 expression and wild type p53 could benefit from PARP-1 inhibition therapies, meanwhile it could have adverse effects for those carrying mutated p53 tumours.

Estrés post-traumático en la atención de emergencias y rescates / Post-traumatic stress disorder in emergency response

Introducción: Los trabajadores de emergencias están expuestos a situaciones potencialmente traumáticas, por lo que sería de interés evidenciar si existe relación entre dichos profesionales y el Trastorno de Estrés Postraumático (TEPT). Nuestro objetivo es conocer si existe evidencia de relación causal entre el desarrollo de TEPT y ser profesional de rescate, además de identificar factores de vulnerabilidad asociados e intervenciones preventivas y terapéuticas eficaces. Método: Revisión sistemática mediante búsquedas bibliográficas en MEDLINE en bases de datos PUBMED, SCOPUS, EMBASE, WOS, Cochrane Library Plus, IBECS, LILACS y CISDOC, usando términos MeSH «emergency responders» y «stress disorder, post-traumatic». Se aplicaron estándares de calidad CONSORT, STROBE y comprobamos la evidencia mediante el sistema SIGN. Resultados: Se seleccionaron 6 ECA (n=783) y 1 estudio de cohortes históricas (n=4487). Se describen como factores de riesgo más frecuentes: sexo femenino 2.93(1.42-6.07), diagnóstico de depresión o ansiedad 4.72(2.33-9.57) en cronicidad y abuso de sustancias 5.12(2.62-9.97) en empeoramiento del TEPT. No hay mejoras en las intervenciones preventivas (p=0.712-0.749) y (p=0.246-0.881). En 2 ECA se evidenció una reducción en la gravedad de los síntomas del TEPT mediante Terapia Cognitivo Conductual (TCC) (p=0.001 y 0.05). Existen 2 ECA con oxitocina que revelan impacto en regiones neurales para gestión emocional (p=0.0024-0.044). Conclusiones: Existe relación causal entre el TEPT y los trabajadores de emergencias (evidencia 2+). Como tratamiento, la TCC resulta efectiva para la reducción de síntomas de TEPT (evidencia 1+). Se mostraron como principales los factores de vulnerabilidad antes mencionados (evidencia 1+). La oxitocina mostró evidencia como aliado terapéutico (evidencia 1+). (AU)

Introduction: Emergency workers are exposed to potentially traumatic situations, so it would be of interest to show whether there is a relationship between these professionals and the development of Post-Traumatic Stress Disorder (PTSD). The aim is to know if there is evidence of such relationship, in addition to identify associated vulnerability factors and effective preventive and therapeutic interventions. Method: Systematic review through bibliographic research in MEDLINE in PUBMED, SCOPUS, EMBASE, WOS, Cochrane Library Plus, IBECS, LILACS and CISDOC databases, using MeSH terms «emergency responders» and «stress disorder, post-traumatic». CONSORT, STROBE quality standards were applied and we checked the evidence using the SIGN system. Results: They were selected 6 RCTs (n = 783) and 1 historical cohort study (n = 4487). The most frequent risk factors are: female sex 2.93 (1.42-6.07), diagnosis of depression or anxiety 4.72 (2.33-9.57) in chronicity and substance abuse 5.12 (2.62-9.97) in worsening of PTSD. There are no improvements in preventive interventions (p = 0.712-0.749) and (p = 0.246-0.881). In 2 RCTs, a reduction in the severity of PTSD symptoms was evidenced by Cognitive Behavioral Therapy (CBT) (p = 0.001 and 0.05). There are 2 RCTs with oxytocin that reveal an impact on neural regions for emotional management (p = 0.0024-0.044). Conclusions: There is a causal relationship between PTSD and emergency workers (evidence 2+). As a treatment, CBT is effective in reducing PTSD symptoms (evidence 1+). The aforementioned vulnerability factors were shown as the most important (evidence 1+). Oxytocin showed evidence as a therapeutic ally (evidence 1+). (AU)

Endothelin-1 as a Mediator of Heme Oxygenase-1-Induced Stemness in Colorectal Cancer: Influence of p53.

Heme oxygenase-1 (HO-1) is an antioxidant protein implicated in tumor progression, metastasis, and resistance to therapy. Elevated HO-1 expression is associated with stemness in several types of cancer, although this aspect has not yet been studied in colorectal cancer (CRC). Using an in vitro model, we demonstrated that HO-1 overexpression regulates stemness and resistance to 5-FU treatment, regardless of p53. In samples from CRC patients, HO-1 and endothelin converting enzyme-1 (ECE-1) expression correlated significantly, and p53 had no influence on this result. Carbon monoxide (CO) activated the ECE-1/endothelin-1 (ET-1) pathway, which could account for the protumoral effects of HO-1 in p53 wild-type cells, as demonstrated after treatment with bosentan (an antagonist of both ETRA and ETRB endothelin-1 receptors). Surprisingly, in cells with a non-active p53 or a mutated p53 with gain-of-function, ECE-1-produced ET-1 acted as a protective molecule, since treatment with bosentan led to increased efficiency for spheres formation and percentage of cancer stem cells (CSCs) markers. In these cells, HO-1 could activate or inactivate certain unknown routes that could induce these contrary responses after treatment with bosentan in our cell model. However more research is warranted to confirm these results. Patients carrying tumors with a high expression of both HO-1 and ECE-1 and a non-wild-type p53 should be considered for HO-1 based-therapies instead of ET-1 antagonists-based ones.

Hyalectanase Activities by the ADAMTS Metalloproteases.

The hyalectan family is composed of the proteoglycans aggrecan, versican, brevican and neurocan. Hyalectans, also known as lecticans, are components of the extracellular matrix of different tissues and play essential roles in key biological processes including skeletal development, and they are related to the correct maintenance of the vascular and central nervous system. For instance, hyalectans participate in the organization of structures such as perineural nets and in the regulation of neurite outgrowth or brain recovery following a traumatic injury. The ADAMTS (A Disintegrin and Metalloprotease domains, with thrombospondin motifs) family consists of 19 secreted metalloproteases. These enzymes also perform important roles in the structural organization and function of the extracellular matrix through interactions with other matrix components or as a consequence of their catalytic activity. In this regard, some of their preferred substrates are the hyalectans. In fact, ADAMTSs cleave hyalectans not only as a mechanism for clearance or turnover of proteoglycans but also to generate bioactive fragments which display specific functions. In this article we review some of the physiological and pathological effects derived from cleavages of hyalectans mediated by ADAMTSs.

Screening of pesticides and veterinary drugs in small streams in the European Union by liquid chromatography high resolution mass spectrometry.

Water samples from 29 small waterways located in 10 different countries in the European Union were screened for the presence of a large number of pesticides (275) and veterinary drugs (101). Solid phase extraction was combined with liquid chromatography coupled to Orbitrap high resolution tandem mass spectrometry to quantify the levels of pesticides in the samples and to detect the presence of veterinary drugs. All the sampled European rivers and canals included in this investigation were contaminated with mixtures of pesticides and, in most of the cases, with several veterinary drugs at the time of sampling, without a clear national or regional pattern. In total, 103 different pesticides, 24 of them banned in the EU, and 21 veterinary drugs were found in the analysed samples. Herbicides were the main contributor to the total amount of pesticides found in the samples, with terbuthylazine present in all the samples. The maximum individual concentration recorded was of dimethenamid at 59.85 µg L-1. The maximum combined pesticide concentration was found in a sample from the Wulfdambeek canal, Belgium, with 94.02 µg L-1 comprised of a mixture of 70 different pesticides. European regulatory standards defining acceptable concentration levels were exceeded for at least one pesticide in 13 of the 29 samples analysed, with the neonicotinoid insecticides imidacloprid and clothianidin most frequently present above such limits. The majority of the veterinary drugs detected were antimicrobials, most being antibiotics. The ß-lactam antibiotic dicloxacillin was present in two thirds of the analysed samples. The application of this consistent research approach across Europe allowed the identification of a significant threat to the aquatic environment associated with pesticide contamination, and in some cases veterinary drugs, at the time of sampling in the water bodies tested.

Unctuous ZrO2 nanoparticles with improved functional attributes as lubricant additives.

One of the main drawbacks in the application of metal-oxide nanoparticles as lubricant additives is their poor stability in organic media, despite the good anti-wear, friction-reducing and high-load capacity properties described for these materials. In this work, we present a novel procedure to chemically cap the surface of ZrO2 nanoparticles (ZrO2NPs) with long hydrocarbon chains in order to obtain stable dispersions of ZrO2NPs in non-aqueous media without disrupting their attributes as lubricant additives. C-8, C-10 and C-16 saturated flexible chains were attached to the ZrO2NP surface and their physical and chemical characterization was performed by transmission electron microscopy, thermogravimetric analysis, attenuated total reflectance Fourier transform infrared spectroscopy, x-ray photoelectron spectroscopy and solid-state nuclear magnetic resonance. The dispersion stability of the modified ZrO2NPs in non-aqueous media was studied using static multiple light scattering. Tribological tests demonstrated that dispersions of the long-chain capped ZrO2NPs in base lubricating oils exhibited low friction coefficients and improved the anti-wear properties of the base oil when compared with the raw lubricating oil.

AA-NAT, MT1 and MT2 Correlates with Cancer Stem-Like Cell Markers in Colorectal Cancer: Study of the Influence of Stage and p53 Status of Tumors.

The characterization of colon cancer stem cells (CSCs) may help to develop novel diagnostic and therapeutic procedures. p53 loss increases the pool of CSCs in colorectal cancer (CRC). Recent reports suggest that the oncostatic effects of melatonin could be related to its ability to kill CSCs. Although there are no data linking the loss of p53 function and melatonin synthesis or signaling in cancer, melatonin does activate the p53 tumor-suppressor pathway in this disease. In this work, we analyze whether the expression of melatonin synthesis and signaling genes are related to the expression of CSC markers and the implication of p53 status in samples from patients with CRC. Arylalkylamine N-acetyltransferase (AA-NAT), MT1, and MT2 expression decreased in tumor samples versus normal mucosa samples in mutated p53 (mtp53) tumors versus those with wild-type p53 (wtp53). Further, AA-NAT and MT2 expression were lower in advanced stages of the disease in wtp53 tumors. On the contrary, CD44 and CD66c expression was higher in tumor versus normal mucosa in wtp53 tumors. Additionally, CD44 expression was higher in advanced stages of the disease regardless of the p53 status. Patients with CD44highCD66chigh and wtp53 tumors in advanced stages showed low expression of AA-NAT and MT2 in wtp53 tumors. These results could indicate a possible interaction of these pathways in CRC.

Time-of-flight mass spectrometry assessment of fluconazole and climbazole UV and UV/H2O2 degradability: Kinetics study and transformation products elucidation.

The efficiency of UV irradiation for the removal of the antimycotic drugs fluconazole (FCZ) and climbazole (CBZ) from water samples is evaluated. Degradation experiments, at laboratory scale, were carried out with spiked aliquots of ultrapure water solutions and treated wastewater samples using low-pressure mercury lamps emitting at 254 nm. Time course of precursor pollutants and identification of arising transformation products (TPs) was performed by injection of different reaction time aliquots in a liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) system. Chemical structures of identified TPs were proposed from their full-product ion spectra, acquired using different collision energies. During UV irradiation experiments, the half-lives (t1/2) of FCZ and CBZ were similar in ultrapure water solutions and wastewater samples; however, the first species was more recalcitrant than the second one. Four TPs were identified in case of FCZ resulting from substitution of fluorine atoms by hydroxyl moieties and intramolecular cyclization with fluorine removal. CBZ interacted with UV radiation through reductive dechlorination, hydroxylation and cleavage of the ether bond; moreover, five additional primary TPs, with the same empirical formula as CBZ, were also noticed. Given the relatively long t1/2 of FCZ under direct photolysis (ca. 42 min), UV irradiation was combined with H2O2 addition to promote formation of reactive hydroxyl radicals. Under such conditions, the degradation rate of FCZ was enhanced significantly and no TPs were detected. These latter conditions allowed also the effective removal of CBZ TPs.

Identification of antimycotic drugs transformation products upon UV exposure.

The reactivity of three imidazolic, environmental persistent antimycotic drugs (clotrimazole, CTZ; ketoconazole, KTZ; and miconazole, MCZ) upon exposure to ultraviolet (UV) radiation is discussed. First, precursor compounds were immobilized in a silicone support which was further exposed to UV light at two different wavelengths: 254 and 365 nm. After solvent desorption, degradation kinetics of the precursor pharmaceuticals, identification of the arising transformation products (TPs) and evaluation of their time-course were investigated by liquid chromatography (LC) with quadrupole time-of-flight (QTOF) mass spectrometry (MS) detection. The three antimycotics displayed similar stabilities when exposed to 254 nm light; however, CTZ was significantly more stable than MCZ and KTZ when irradiated with the 365 nm lamp. TPs identified in silicone supports resulted from de-chlorination, cleavage, intra-molecular cyclization and hydroxylation reactions. Many of these species were also detected when exposing other solid matrices, such as sand and agricultural soil, previously spiked with target compounds, to UV light. The 50% estimated lethal concentration, calculated using the 48-h Daphnia magna test, for the two main TPs of CTZ and MCZ, at both wavelengths, were lower than those corresponding to the precursor drugs.

Selective extraction of antimycotic drugs from sludge samples using matrix solid-phase dispersion followed by on-line clean-up.

An effective and selective, modular sample preparation method for the extraction of eight antimycotic drugs, belonging to three different chemical classes, from digested sludge samples is proposed. To this end, matrix solid-phase dispersion (MSPD) was on-line connected with a cationic exchanger solid-phase extraction (SPE) cartridge. Analytes were extracted from the MSPD syringe, which contained the freeze-dried sludge sample dispersed with C18 plus a clean-up layer of primary and secondary amine (PSA) sorbent, with 10 mL of methanol. This extract flowed also through the SPE cartridge, where target compounds remained trapped while neutral interferences are released. After discarding the MSPD syringe, analytes were recovered with 10 mL of methanol (0.5% in NH3) before LC-MS/MS determination using a hybrid quadrupole time-of-flight (QTOF) mass spectrometer furnished with an electrospray ionization (ESI) source. In comparison with previously published sample preparation methodologies, the developed approach greatly simplifies sample handling and reduces attenuation of ESI ionization for sample extracts when compared to standard solutions. The obtained absolute recoveries ranged between 70 and 118%, and the limits of quantification (LOQs) of the method varied between 5 and 8 ng g(-1). Four antimycotic drugs were ubiquitous in urban sludge samples, with maximum average concentrations (above 400 ng g(-1)) corresponding to clotrimazole (CTZ). The screening capabilities of the LC-QTOF-MS system demonstrated that the developed modular extraction and purification methodology might be useful for the selective extraction of other basic drugs (e.g., sertraline, amitryptiline, and amiodarone) from sludge.

Melatonin reduces endothelin-1 expression and secretion in colon cancer cells through the inactivation of FoxO-1 and NF-κß.

Melatonin is an indoleamine that is synthesised from tryptophan under the control of the enzymes arylalkylamine N-acetyltransferase (AA-NAT) and acetylserotonin methyltransferase (ASMT). Melatonin inhibits colon cancer growth in both in vivo and in vitro models; however, a precise mechanism responsible for inhibiting tumour growth has not been clearly described. Endothelin-1 (ET-1) is a peptide that acts as a survival factor in colon cancer, inducing cell proliferation, protecting carcinoma cells from apoptosis and promoting angiogenesis. The data presented show that melatonin inhibits edn-1 mRNA expression (the first step in ET-1 synthesis), ECE-1 protein expression and the release of ET-1 from colorectal cancer cells in vitro. ET-1 levels in cultured media present a similar inhibition pattern to that of edn-1 mRNA expression despite the inhibition of ECE-1 protein after melatonin treatment, which suggests that an endopeptidase other than ECE-1 could be mainly responsible for ET-1 synthesis. The inhibition of edn-1 expression is due to an inactivation of FoxO1 and NF-κß transcription factors. FoxO1 inactivation is associated with an increased Src phosphorylation, due to elevated cAMP content and PKA activity, whereas NF-κß inactivation is associated with the blockade of Akt and ERK phosphorylation due to the inhibition of PKC activity after melatonin treatment. Melatonin also inhibits edn-1 promoter activity regulated by FoxO1 and NF-κß. Finally, a significant correlation was observed between AA-NAT and edn-1 expression downregulation in human colorectal cancer tissues. In conclusion, melatonin may be useful in treating colon carcinoma in which the activation of ET-1 plays a role in tumour growth and progression.

Variation of transaminases, HCV-RNA levels and Th1/Th2 cytokine production during the post-partum period in pregnant women with chronic hepatitis C.

This study analyses the evolution of liver disease in women with chronic hepatitis C during the third trimester of pregnancy and the post-partum period, as a natural model of immune modulation and reconstitution. Of the 122 mothers recruited to this study, 89 were HCV-RNA+ve/HIV-ve and 33 were HCV-RNA-ve/HIV-ve/HCVantibody+ve and all were tested during the third trimester of pregnancy, at delivery and post-delivery. The HCV-RNA+ve mothers were categorized as either Type-A (66%), with an increase in ALT levels in the post-partum period (>40 U/L; P<0.001) or as Type-B (34%), with no variation in ALT values. The Type-A mothers also presented a significant decrease in serum HCV-RNA levels in the post-delivery period (P<0.001) and this event was concomitant with an increase in Th1 cytokine levels (INFγ, P = 0.04; IL12, P = 0.01 and IL2, P = 0.01). On the other hand, the Type-B mothers and the HCV-RNA-ve women presented no variations in either of these parameters. However, they did present higher Th1 cytokine levels in the partum period (INFγ and IL2, P<0.05) than both the Type-A and the HCV-RNA-ve women. Cytokine levels at the moment of delivery do not constitute a risk factor associated with HCV vertical transmission. It is concluded that differences in the ALT and HCV-RNA values observed in HCV-RNA+ve women in the postpartum period might be due to different ratios of Th1 cytokine production. In the Type-B women, the high partum levels of Th1 cytokines and the absence of post-partum variation in ALT and HCV-RNA levels may be related to permanent Th1 cytokine stimulation.

Optimization of matrix solid-phase dispersion conditions for organic fungicides determination in soil samples.

A simplified sample preparation method, based on the matrix solid-phase dispersion technique, is proposed for the sensitive determination of 15 organic fungicides in vineyard soils by gas chromatography-mass spectrometry (GC-MS). Under final working conditions, sieved samples (0.5 g) were blended and dispersed with 2 g of C18 and transferred to a polypropylene syringe containing 1 g of diatomaceous earth. Analytes were recovered using 10 mL of ethyl acetate, this extract was concentrated to 1 mL and fungicides determined by GC-MS, without additional cleanup. The method provided recoveries in the range from 74 to 122% for soils with total carbon contents up to 5.5% and it allowed the use of external standard as quantification technique. Inter-day precision, given as relative standard deviations, stayed between 3 and 13%, and the limits of quantification were comprised between 0.6 and 15 ng g(-1). Several fungicides were found in the top layer of vineyard soils with the highest detection frequency and maximum concentration corresponding to iprovalicarb. Some real samples were also submitted to pressurized liquid extraction. Measured concentrations were in excellent agreement with those obtained by matrix solid-phase dispersion, which reinforces the accuracy of the latter methodology.

Gender-related invasion differences associated with mRNA expression levels of melatonin membrane receptors in colorectal cancer.

Melatonin inhibits growth and invasive capacity of colon cancer cells in vitro through its membrane (MT1 and MT2) and/or nuclear receptors (RORα). Previous studies showed that this indoleamine is present in both the normal and colon cancer at similar levels. Therefore, we analyzed MT1, MT2, and RORα expression in tumor samples versus normal mucosa (NM) from patients suffering from colorectal cancer (CRC). Given the existence of sex differences in the incidence and pathology of CRC and the involvement of steroid receptors in the oncostatic actions of melatonin in some types of cancer, we also analyzed the expression of androgen (AR) and estrogen receptor (ER) α and ERß. Finally, we conducted some experiments in colon cancer cell lines to corroborate the experiments carried out in human tumors. We found a decreased expression of MT1, MT2, AR, ERα, and ERß in tumor samples versus NM, but no changes in RORα expression in the whole cohort of patients. Classifying tumors by stage and gender, MT1, MT2, AR, ERα, and ERß expression decreased in both early stage and advanced tumors, but only in male patients. On the other hand, MT1 and MT2 expression correlated positively with AR, ERα, and ERß expression in male patients and with ERα or ERß in female patients. In vitro, the invasive capacity was higher in cells with the least expression of MT1, MT2, and AR, and nonselective MT1/MT2 agonists inhibited cell growth and invasion. These results could indicate a possible interaction of these pathways.

Factors associated with hepatic steatosis in obese children and adolescents.

OBJECTIVES: Obesity is associated with high prevalence of hepatic steatosis. We speculate that determinant factors of susceptibility to hepatic steatosis in obesity could differ between children and adolescents. PATIENTS AND METHODS: Blood biochemical parameters, systemic oxidative stress markers, proinflammatory cytokines, and adipokine levels were determined in 157 obese children and adolescents. The subjects were divided into 2 groups: children and adolescents, identified as such in accordance with Tanner stage and the measured level of dehydroepiandrosterone sulphate. Steatosis was evaluated by ultrasonography in 127 subjects. RESULTS: Steatosis prevalence was 44.8%. In the "children" group, those with hepatic steatosis presented higher levels of erythrocyte oxidised glutathione (GSSG) and resistin, lower levels of high-density lipoprotein (HDL) cholesterol, and lower enzymatic activities of erythrocyte glutathione reductase (GRd) and glutathione oxidase (GPx). In the "adolescents" group, those with hepatic steatosis presented higher values for body mass index z score (BMIz), insulin, peptide C, homeostatic model assessment index (HOMA-IR), alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides, GSSG, and leptin. These subjects also presented lower values for soluble leptin receptor, GRd, and GPx. In the "children" group, the only independent factor of steatosis was a decrease in GRd activity (odds ratio [OR] 0.165, 95% CI 0.03-0.84, P = 0.030). Moreover, in the "adolescent" group, the independent factors were higher for GSSG (OR 6.8, 95% CI 1.6-28.7, P = 0.010) and HOMA-IR (OR 1.9, 95% CI 1.17-3.1, P = 0.009). CONCLUSIONS: Factors associated with hepatic steatosis differ between obese children and adolescents. Oxidative stress is seen to be the main process in children, whereas in adolescents oxidative stress and insulin resistance are significant factors for steatosis.

Importance of host genetic factors HLA and IL28B as predictors of response to pegylated interferon and ribavirin.

OBJECTIVES: Viral factors are considered the best predictors of response to treatment for chronic hepatitis C (CHC), but genetic factors are known to have an important role in this respect. This paper investigates the relationships among the host genetic factors HLA and IL28B, viral factors, and the outcome of combination therapy. METHODS: A multicenter retrospective cohort of 428 previously untreated CHC patients was treated with pegylated interferon/ribavirin (pegIFN/RBV) for 48 weeks. In all, 378 (88%) of these patients were genotype 1 or 4, and 50 (12%) were genotype 2 or 3. RESULTS: Multivariate logistic regression showed the rs12979860 CC genotype (adjusted odds ratio (aOR)=4.3, 95% confidence interval (95% CI): 2.6-7), the HLA-DQB1*0301 allele (aOR=2.08, 95% CI: 1.2-3.5) and age, viral genotype, and viral load levels to be significantly associated with sustained virological response (SVR). When the variable rs12979860 was eliminated, the area under the receiver operating characteristic (ROC) curve (AUC) decreased significantly (0.76 vs. 0.69; P=0.03). AUC values derived from viral factors were lower than those corresponding to host genetic factors (0.67 vs. 0.72, respectively; P=0.04). The HLA-DQB1*0301 and A*0201 alleles were associated with rs12979860 CC genotype and SVR (P<0.0001). CONCLUSIONS: The HLA-DQB1*0301 allele and IL28B genotype are factors that are associated independently with SVR. There is a synergism between the HLA-DQB1*0301 and HLA-A*0201 alleles with polymorphism rs12979860 CC, which increases the SVR rate. IL28B genotype is the best predictor of SVR.

Genetic variation in interleukin 28B with respect to vertical transmission of hepatitis C virus and spontaneous clearance in HCV-infected children.

UNLABELLED: The vertical transmission of hepatitis C virus (HCV-VT) is a major route of HCV infection in children, but the risk factors remain incompletely understood. This study analyzed the role of interleukin 28B (IL28B) in HCV-VT and in the spontaneous clearance of HCV among infected infants. Between 1991 and 2009, 145 mothers were recruited for this study: 100 were HCV-RNA+ve / human immunodeficiency virus negative (HIV-ve), with 128 children, and 33 were HCV-RNA-ve/HCV antibody+ve, with 43 children. The infants were tested for HCV-RNA at birth and at regular intervals until the age of 6 years. IL28B (single nucleotide polymorphism rs12979860) was determined in the mothers and children. HCV-VT was assumed when children presented HCV-RNA+ve in two subsequent blood samples. HCV-VT-infected infants were categorized as: (1) transient viremia with posterior HCV-RNA-ve and without serum-conversion; (2) persistent infection with serum-conversion. Of the 31 mothers with CC polymorphism, 19 (61%) were HCV-RNA+ve, whereas among the 68 mothers with non-CC polymorphism, 56 (82%) were HCV-RNA+ve. In all, 26 of 128 (20%) infants born to the HCV-RNA+ve mothers acquired HCV infection, but only 9 (7%) were chronically infected. The rate of HCV-VT was higher among the mothers with higher HCV viremia. No HCV-VT was detected in the HCV-RNA-ve women. Neither the mothers' nor the childrens' IL-28 status was associated with an increased risk of HCV-VT. The factors influencing viral clearance among the infected children were genotype non-1 and genotype CC of IL28B. In logistic regression, child CC polymorphism was the only predictor of HCV-clearance in HCV genotype-1. CONCLUSION: High maternal viral load is the only predictive factor of HCV-VT. IL28B plays no role in HCV-VT, but IL28B CC child polymorphism is associated independently with the spontaneous clearance of HCV genotype-1 among infected children.

Hepatic expression of adiponectin receptors increases with non-alcoholic fatty liver disease progression in morbid obesity in correlation with glutathione peroxidase 1.

BACKGROUND: The prevalence of non-alcoholic fatty liver disease (NAFLD) in obesity is very high. The role of adiponectin receptors in NAFLD progression remains still unclear. We speculate that changes in the hepatic expression levels of the two adiponectin receptors may be associated with the expression of oxidative stress-related genes. METHODS: We studied 60 morbidly obese patients with NAFLD, who underwent liver biopsy at the time of bariatric surgery. We measured the hepatic messenger-RNA concentration of adiponectin receptors (ADIPOR1 and ADIPOR2), glutathione peroxidase 1 (GPx1), glutathione reductase (GRd) and inducible oxide nitric synthase. Additionally, biochemical parameters and oxidative stress markers were determined in blood samples. According to the Kleiner score, the patients were divided into two groups: group 1 (25 patients without steatohepatitis) and group 2 (25 patients with probable steatohepatitis and ten patients with steatohepatitis). RESULTS: The messenger-RNA concentration of all genes analysed in the study was higher among the patients in group 2. However, no differences in blood oxidative stress markers were observed. Strong correlations were found among the expression levels of ADIPOR1, ADIPOR2 and GPx1. The multivariate analysis showed that the only independent variable associated with NAFLD progression was the increase in GPx1 expression levels. CONCLUSIONS: NAFLD progression in morbid obesity is associated with increase in hepatic adiponectin receptor and oxidative stress-related genes. The linear correlations suggest that ADIPOR1, ADIPOR2 and GPx1 share key molecular factors in the regulation of the genetic expressions.

Quasispecies as predictive factor of rapid, early and sustained virological responses in chronic hepatitis C, genotype 1, treated with peginterferon-ribavirin.

BACKGROUND: The relationship between the complexity of the hypervariable region 1 quasispecies of HCV and responsiveness to therapy is not completely clear. OBJECTIVE: To investigate the importance of quasispecies as a predictive factor of rapid (RVR), early (EVR) and sustained (SVR) virologic response. METHODS: Prospective analysis of 82 patients with chronic hepatitis C, genotype 1, treated with peginterferon alfa-2a and ribavirin. Quasispecies (SSCP), HCV-RNA and viral load were determined at baseline and 2, 4, 8 and 12 weeks after beginning treatment. RESULTS: Less fibrosis and lower serum GGT activity were the only predictive factors for EVR. SVR predictive factors were age < or =40 years, viral load < or =600,000IU/mL, and quasispecies < or =5 bands. By logistic regression analysis, the independent factors determining SVR were age (P=0.011), viral load (P=0.027), and quasispecies (P=0.010). Quasispecies and viral load were not predictive factors of RVR. During treatment, quasispecies decreased rapidly in the SVR group. In non-EVR patients, quasispecies were slightly lower up to 8 weeks and then increased. CONCLUSIONS: Quasispecies are an important predictive factor for SVR, but are no better predictors than viral load. Quasispecies are not predictive factors for RVR or EVR.