A new species of Dermadena (Digenea: Lepocreadiidae) from the stone triggerfish Pseudobalistes naufragium (Tetraodontiformes: Balistidae) in the South American Pacific Ocean.

The present paper describes a new species of Dermadena (Digenea) parasitizing Pseudobalistes naufragium in Puerto Pizarro, northern Peru, using light and scanning electronic microscopy (SEM). Additionally, molecular analysis was also performed to determine the phylogenetic affinities of Dermadena within the Lepocreadiidae. The new species is differentiated from Dermadena spatiosa, Dermadena stirlingi and Dermadena lactophrysi by presenting a curved and well-developed external seminal vesicle. Also, SEM revealed numerous dome-shaped tegument protuberances forming glandular papillae with transversal wrinkles arranged roughly in concentric rows around the acetabular region, varying in size from large at the middle of the body to small at the margin. In the molecular phylogeny, the new species formed a well-supported clade with sequences of species from the Lepocreadiidae, confirming that it belongs to this family.

A checklist of helminths associated with reptiles (Tetrapoda: Reptilia) from Peru.

An annotated checklist of the helminth parasites associated with reptiles from Peru is provided, as the result of a compilation of parasitological papers published between 1963 and January 2022 and records of species deposited in national and international collections. The list provides data on hosts, developmental stage, sites of infection, geographical distribution in Peruvian territory, code of material deposited in helminthological collections, references and taxonomic notes. The database includes records of 106 different species of helminth parasites (82 nominal species and 24 taxa identified at the generic level), the majority in the adult stage. These helminth parasites come from 18 of the 25 official Peruvian regions. Nematodes have the highest richness in number of species (79 species), followed by trematodes (17 species) and cestodes (nine species). The acanthocephalans are represented by only one species. The parasites with the highest number of records were Physaloptera retusa Rudolphi, 1819 (11 hosts), Physalopteroides venancioi (Lent, Freitas & Proença, 1946) (nine hosts), Strongyluris oscari Travassos, 1923 (seven hosts), and Parapharyngodon scleratus Travassos, 1923 (five hosts), all of which are nematodes. The 106 taxa of helminth parasites have been reported infecting 55 species of reptiles in Peru, distributed in 34 genera and 14 families. The reptile species harbouring the highest number of helminth parasites are the yellow-footed tortoise Chelonoidis denticulatus (Linnaeus) with 18 species (three trematodes and 15 nematodes), followed by the Peru desert tegu Dicrodon guttulatum Duméril & Bibron (Teiidae) with 11 species (three cestodes and eight nematodes) and the yellow-spotted Amazon River turtle Podocnemis unifilis Troschel (Podocnemididae) with 10 species (five trematodes and five nematodes). Of the 524 species of reptiles reported in Peru, only 55 (>10%) are reported as hosts of helminths representing a small proportion considering the great variety of reptile hosts that inhabit the various tropical and subtropical geographical areas of Peru.

A Mediterranean diet supplemented with extra virgin olive oil or nuts improves endothelial markers involved in blood pressure control in hypertensive women.

PURPOSE: Serum nitric oxide (NO) reduction and increased endothelin-1 (ET-1) play a pivotal role in endothelial dysfunction and hypertension. Considering that traditional Mediterranean diet (TMD) reduces blood pressure (BP), the aim of this study was to analyze whether TMD induced changes on endothelial physiology elements such as NO, ET-1 and ET-1 receptors which are involved in BP control. METHODS: Non-smoking women with moderate hypertension were submitted for 1 year to interventions promoting adherence to the TMD, one supplemented with extra virgin olive oil (EVOO) and the other with nuts versus a control low-fat diet (30 participants/group). BP, NO, ET-1 and related gene expression as well as oxidative stress biomarkers were measured. RESULTS: Serum NO and systolic BP (SBP) or diastolic BP (DBP) were negatively associated at baseline, as well as between NO and ET-1. Our findings also showed a DBP reduction with both interventions. A negative correlation was observed between changes in NO metabolites concentration and SBP or DBP after the intervention with TMD + EVOO (p = 0.033 and p = 0.044, respectively). SBP reduction was related to an impairment of serum ET-1 concentrations after the intervention with TMD + nuts (p = 0.008). We also observed changes in eNOS, caveolin 2 and ET-1 receptors gene expression which are related to NO metabolites levels and BP. CONCLUSIONS: The changes in NO and ET-1 as well as ET-1 receptors gene expression explain, at least partially, the effect of EVOO or nuts on lowering BP among hypertensive women.

Effects of total dietary polyphenols on plasma nitric oxide and blood pressure in a high cardiovascular risk cohort. The PREDIMED randomized trial.

BACKGROUND AND AIM: Hypertension is one of the main cardiovascular risk factors in the elderly. The aims of this work were to evaluate if a one-year intervention with two Mediterranean diets (Med-diet) could decrease blood pressure (BP) due to a high polyphenol consumption, and if the decrease in BP was mediated by plasma nitric oxide (NO) production. METHODS AND RESULTS: An intervention substudy of 200 participants at high cardiovascular risk was carried out within the PREDIMED trial. They were randomly assigned to a low-fat control diet or to two Med-diets, one supplemented with extra virgin olive oil (Med-EVOO) and the other with nuts (Med-nuts). Anthropometrics and clinical parameters were measured at baseline and after one year of intervention, as well as BP, plasma NO and total polyphenol excretion (TPE) in urine samples. Systolic and diastolic BP decreased significantly after a one-year dietary intervention with Med-EVOO and Med-nuts. These changes were associated with a significant increase in TPE and plasma NO. Additionally, a significant positive correlation was observed between changes in urinary TPE, a biomarker of TP intake, and in plasma NO (Beta = 4.84; 95% CI: 0.57-9.10). CONCLUSIONS: TPE in spot urine sample was positively correlated with plasma NO in Med-diets supplemented with either EVOO or nuts. The statistically significant increases in plasma NO were associated with a reduction in systolic and diastolic BP levels, adding to the growing evidence that polyphenols might protect the cardiovascular system by improving the endothelial function and enhancing endothelial synthesis of NO.

Inverse association between habitual polyphenol intake and incidence of cardiovascular events in the PREDIMED study.

BACKGROUND AND AIMS: Epidemiologic and biological evidence supports an inverse association between polyphenol consumption and the risk of cardiovascular disease (CVD). However, no previous studies have prospectively evaluated the relationship between polyphenol intake and the incidence of CVD in such a comprehensive way. The aim was to evaluate the association between intakes of total polyphenol and polyphenol subgroups, and the risk of major cardiovascular events (myocardial infarction, stroke or death from cardiovascular causes) in the PREDIMED study. METHODS AND RESULTS: The present work is an observational study within the PREDIMED trial. Over an average of 4.3 years of follow-up, there were 273 confirmed cases of CVD among the 7172 participants (96.3%) who completed a validated 137-item food frequency questionnaire (FFQ) at baseline. Polyphenol consumption was calculated by matching food consumption data from the FFQ with the Phenol-Explorer database on polyphenol content of each reported food. After multivariate adjustment, a 46% reduction in risk of CVD risk was observed comparing Q5 vs. Q1 of total polyphenol intake (HR = 0.54; 95% confidence interval [CI] = 0.33-0.91; P-trend = 0.04). The polyphenols with the strongest inverse associations were flavanols (HR = 0.40; CI 0.23-0.72; P-trend = 0.003), lignans (HR = 0.51; CI 0.30-0.86; P-trend = 0.007), and hydroxybenzoic acids (HR = 0.47; CI 0.26-0.86; P-trend 0.02). CONCLUSION: Greater intake of polyphenols, especially from lignans, flavanols, and hydroxybenzoic acids, was associated with decreased CVD risk. Clinical trials are needed to confirm this effect and establish accurate dietary recommendations.

Interleukin-6 concentration changes in plasma and saliva in bisphosphonate-related osteonecrosis of the jaws.

AIM: To determine the plasma and saliva levels of IL-6 in patients with bisphosphonate-related osteonecrosis of the jaws (BRONJ) and to investigate whether there is a correlation between more advanced stages of BRONJ and levels of IL-6. MATERIAL AND METHODS: We studied three groups: group 1 consisted of 30 patients with BRONJ due to intravenous bisphosphonates (ivBP), group 2 consisted of 25 patients treated with ivBP but without BRONJ, and group 3 consisted of 15 healthy controls. In each case, we assayed plasma and saliva IL-6 samples using an ELISA test. RESULTS: Significantly, higher IL-6 values were found in both saliva and plasma in group 1 vs groups 2 and 3 (P < 0.01). Group 1 showed no differences in plasma or saliva IL-6 according to patient gender (P > 0.05), type of tumor, BRONJ location, etiology of BRONJ, or disease stage (P > 0.05). We found higher plasma and saliva IL-6 values in the more advances stages of BRONJ, although the differences were not statistically significant. CONCLUSIONS: Plasma and saliva IL-6 values were higher in our patients with BRONJ than in controls and therefore might be a useful tool for monitoring the severity of BRONJ.

Gazpacho consumption is associated with lower blood pressure and reduced hypertension in a high cardiovascular risk cohort. Cross-sectional study of the PREDIMED trial.

BACKGROUND AND AIM: Hypertension is a major public health problem and a leading cause of death and disability in both developed and developing countries, affecting one-quarter of the world's adult population. Our aim was to evaluate whether the consumption of gazpacho, a Mediterranean vegetable-based cold soup rich in phytochemicals, is associated with lower blood pressure (BP) and/or reduced prevalence of hypertension in individuals at high cardiovascular risk. METHODS AND RESULTS: We selected 3995 individuals (58% women, mean age 67 y) at high cardiovascular risk (81% hypertensive) recruited into the PREDIMED study. BP, weight, and dietary and physical activity data were collected. In multivariate linear regression analyses, after adjustment, moderate and high gazpacho consumption categories were associated with reduced mean systolic BP of -1.9 mm Hg [95% confidence interval (CI): -3.4; -0.6] and -2.6 mm Hg (CI: -4.2; -1.0), respectively, and reduced diastolic BP of -1.5 mm Hg (CI: -2.3; -0.6) and -1.9 mm Hg (CI: -2.8; -1.1). By multiple-adjusted logistic regression analysis, gazpacho consumption was associated with a lower prevalence of hypertension, with OR = 0.85 (CI: 0.73; 0.99) for each 250 g/week increase and OR = 0.73 (CI: 0.55; 0.98) for high gazpacho consumption groups compared to the no-consumption group. CONCLUSIONS: Gazpacho consumption was inversely associated with systolic and diastolic BP and prevalence of hypertension in a cross-sectional Mediterranean population at high cardiovascular risk. The association between gazpacho intake and reduction of BP is probably due to synergy among several bioactive compounds present in the vegetable ingredients used to make the recipe.

Dietary intake and major food sources of polyphenols in a Spanish population at high cardiovascular risk: the PREDIMED study.

BACKGROUND AND AIMS: Epidemiological data have shown an inverse association between the consumption of polyphenol-rich foods and the risk of cardiovascular disease or overall mortality. A comprehensive estimation of individual polyphenol intake in nutritional cohorts is needed to gain a better understanding of this association. The aim of this study was to estimate the quantitative intake of polyphenols and the major dietary sources in the PREDIMED (PREvención con DIeta MEDiterránea) cohort using individual food consumption records. METHODS AND RESULTS: The PREDIMED study is a large, parallel-group, multicentre, randomised, controlled 5-year feeding trial aimed at assessing the effects of the Mediterranean diet on the primary prevention of cardiovascular disease. A total of 7200 participants, aged 55-80 years, completed a validated 1-year food frequency questionnaire (FFQ) at baseline. Polyphenol consumption was calculated by matching food consumption data from the FFQ with the recently developed Phenol-Explorer database on polyphenol content in foods. The mean total polyphenol intake was 820 ± 323 mg day⁻¹ (443 ± 218 mg day⁻¹ of flavonoids and 304 ± 156 mg day⁻¹ of phenolic acids). Hydroxycinnamic acids were the phenolic group with the highest consumption and 5-caffeoylquinic acid was the most abundantly ingested individual polyphenol. The consumption of olives and olive oil was a differentiating factor in the phenolic profile of this Spanish population compared with other countries. CONCLUSION: In Mediterranean countries, such as Spain, the main dietary source of polyphenols is coffee and fruits, but the most important differentiating factor with respect to other countries is the consumption of polyphenols from olives and olive oil.

Total polyphenol excretion and blood pressure in subjects at high cardiovascular risk.

BACKGROUND AND AIMS: Dietary factors are critical for the prevention and treatment of hypertension, but data on the effects of specific nutrients on blood pressure (BP) are scarce. The aim of this study was to assess the relationship between total polyphenol excretion (TPE) in urine, as an objective measurement of total polyphenol intake and BP in an elderly population at high cardiovascular risk. METHODS AND RESULTS: Cross-sectional substudy of 589 high-risk participants entering in the PREDIMED trial. BP was measured and TPE was determined in urine by Folin-Ciocalteu assay. A significant positive association was observed between TPE in urine and daily intake of fruit and vegetables (F&V), coffee or wine after adjusting for potential confounders. The intake of 100 g of F&V (Beta=0.150;P<0.001) had a greater contribution to TPE than 100 mL of coffee (Beta=0.141;P=0.001), and the latter two foods contributed more than the consumption of 100 mL of wine (Beta=0.120;P=0.019). An inverse association was observed between urinary TPE and the prevalence of hypertension. Participants in the highest quartile of urinary TPE had a reduced prevalence of hypertension compared to those in the lowest quartile (Odds Ratio=0.64; 95% confidence interval 0.45 to 0.92; P=0.015). Systolic and diastolic BP were inversely associated with urinary TPE after adjustment for potential confounders (P=0.024 and P=0.003, respectively). CONCLUSIONS: Polyphenol intake, assessed via TPE in urine, was negatively associated with BP levels and prevalence of hypertension in an elderly Mediterranean population at high cardiovascular risk. Participants with the highest intake of polyphenol-rich foods showed the lowest BP measurements.

Liver gene expression in relation to hepatic steatosis and lipid secretion in two duck species.

The susceptibility to development of hepatic steatosis is known to differ between Muscovy and Pekin ducks. Although some experiments were conducted to decipher these differences, few data have been produced to analyse the role of specific genes in this process. For this purpose, expression levels of genes involved in lipid (ATP citrate lyase, malic enzyme 1, fatty acid synthase, stearoyl-CoA desaturase 1, diacylglycerol O-acyl transferase 2, microsomal triglyceride transfer protein, apolipoprotein A1, apolipoprotein B, sterol regulatory element binding factor 1, hepatocyte nuclear factor 4, choline/ethanolamine phosphotransferase 1, carnitine palmitoyl transferase 1A, peroxisome proliferator-activated receptor alpha and sterol O-acyltransferase) and carbohydrate (activating transcription factor 4 or cAMP-response element binding protein, mitochondrial malate dehydrogenase 2 and carbohydrate responsive element binding protein) metabolism and in other functions were analysed in the liver of Pekin and Muscovy ducks fed ad libitum or overfed. A specific positive effect of feeding was observed on the expression of genes involved mainly in fatty acids and TG synthesis and glycolysis, and negative effect on genes involved in beta-oxidation. Interestingly, a strong species effect was also observed on stearoyl-CoA desaturase 1 and to a lesser extent on diacylglycerol O-acyl transferase 2 expression, leading to large differences in expression levels between Pekin and Muscovy overfed ducks, which could explain the difference in lipid metabolism and steatosis ability observed between the two duck species. These results should shed light on gene expression that might underlie susceptibility to hepatic steatosis in humans.

Is there a relationship between the kinetics of lipoprotein lipase activity after a meal and the susceptibility to hepatic steatosis development in ducks?

The difference in the ability of Pekin and Muscovy ducks to develop hepatic steatosis could result from a different peripheral lipoprotein lipase (LPL) activity, which hydrolyses triacylglycerol secreted by the liver. We studied the kinetics of plasma LPL activity in response to a meal at different ages in Pekin and Muscovy ducks. For that purpose, blood samples were taken at 5, 9, 12, 13, and 14 wk of age just before and 1, 2, 4, and 8 h after a meal. To release LPL into general circulation, an i.v. injection of heparin (400 IU/kg of BW) was administered 10 min before blood collection. For that reason, different ducks per genotype were used for each point of measurement (n = 6). Plasma LPL activity measured before the meal was negatively correlated with the weight of the fatty liver measured in the same ducks at 14 wk of age (r = -0.58, P < 0.001). Plasma triacylglycerol level measured before the meal was negatively correlated with plasma LPL activity measured in the same ducks (r = -0.31, P = 0.025) and was negatively correlated with plasma LPL activity measured in the same ducks for each age and each timing (r = -0.39, P < 0.001). At 14 wk of age for Muscovy and Pekin ducks, we observed that a high plasma LPL activity (>200 IU/L of plasma) corresponded to a relatively low development of fatty liver (190 g) induced by overfeeding, whereas a low plasma LPL activity (<150 IU/L of plasma) corresponded to a high propensity to develop fatty liver (470 g). In conclusion, plasma LPL activity measured just before the meal during the rearing period could be used as a marker of hepatic steatosis development during the overfeeding period.

Gene expression and protein content in relation to intramuscular fat content in Muscovy and Pekin ducks.

Independent of their nutritional condition, Pekin ducks always exhibit higher i.m. fat content than Muscovy ducks. To understand this difference between species, the expression level of genes involved in lipid metabolism was analyzed in the pectoralis major muscle of Pekin and Muscovy ducks ad libitum-fed or overfed. The lipoprotein lipase (LPL) gene expression was not different between species and not influenced by overfeeding. The protein content for LPL was higher in Pekin ducks than in Muscovy ducks when birds were ad libitum-fed, whereas in overfed ducks, we found no difference between species. Adipocyte fatty acid-binding protein (A-FABP) gene expression and protein content were higher in Pekin ducks than in Muscovy ducks for each nutritional condition (suggesting a higher intracellular transport within i.m. adipocytes of fatty acids mainly provided by liver for this species). Overfeeding did not affect the expression of genes involved in oxidation [carnitine palmitoyl transferase 1A (CPT1A), cytochrome-c oxidase 4 (COX4), succinyl-coenzyme A:3-ketoacid coenzyme A transferase (SCOT)] but increased the expression of fatty acid synthase (FAS) involved in lipogenesis. For all nutritional conditions, Pekin duck exhibited higher expression levels of CPT1A, COX4, SCOT, and FAS than Muscovy ducks. Results for mRNA SCOT suggested that the muscles of Pekin ducks use ketone bodies as an energy source. In conclusion, i.m. lipogenesis could contribute to the i.m. fat, particularly in Pekin ducks.

Evaluation of pollutant exposure by chemical and biological markers in a Mediterranean French urban stream: a step for in situ calibration of multixenobiotic resistance transporter expression as biomarker in Chironomidae larvae.

This study was aimed at semi-quantifying the membrane density of multixenobiotic resistance (MXR) transporters in Chironomidae Orthocladiinae larvae from an urban stream by ELISA assay. The relationships between the MXR transporter membrane density and limnological parameters and pollutant concentrations, 16 priority polycyclic aromatic hydrocarbons (PAHs), as per the US Environmental Protection Agency (EPA) and seven polychlorobiphenyl congeners (PCBs), were assessed. Midge larvae were collected, and limnological parameters and pollutant concentrations were measured in three sites of a French Mediterranean urban stream, two located after sewage treatment plants, and one closed to the river mouth, and in two additional sites, one on the stream tributary, and one in a non-urbanized stream located in the same region. Results show that the PAH and PCB contamination levels are different between sites and that some congener concentrations are above their threshold toxic effect level (TEL). The MXR transporter membrane density was significantly higher in larvae from the tributary, the most polluted site, as compared with larvae from the non-urbanized stream. The MXR transporter density was positively correlated with 10 of the 16 US-EPA PAH concentrations and the increase in the MXR transporter density seems to be due to the US-EPA PAH concentrations that were above their TEL. No relations with PCB concentrations or limnological parameters were found. The results suggest that the MXR transporter membrane density in Chironomidae larvae could be an interesting biological marker of PAH exposure in freshwater ecosystems.

ELISA detection of multixenobiotic resistance transporter induction in indigenous freshwater Chironomidae larvae (Diptera): A biomarker calibration step for in situ monitoring of xenobiotic exposure.

A new simple and sensitive method to distinguish chemically polluted from unpolluted situations in freshwater ecosystems is reported. For this purpose, Chironomus gr thumni larvae were collected from a polluted urban river downstream a sewage treatment plant. For the first time, ELISA assay was used to semi-quantify the multixenobiotic resistance transporters (MXR) in these small pertinent bioindicators. The use of samples immediately fixed in the field gives a delay to isolate larvae and allows multi-sampling along a longitudinal transect in a river at a given time. Results exhibit an induction of MXR proteins in larvae from the polluted river and a deinduction in larvae maintained 11days in unpolluted water. They show new evidences to use midge larvae in biomonitoring environmental programs. They answer to first biomarker calibration steps for the ongoing development of MXR transporters as a detection tool of xenobiotic impacts on bioindicator invertebrates in their freshwater habitats.

Impact of the components of metabolic syndrome on oxidative stress and enzymatic antioxidant activity in essential hypertension.

The objective of the present study was to analyze the impact of metabolic syndrome (MS) and its individual components on oxidative stress (OX) and on the activity of antioxidant enzymes of patients with essential hypertension. One hundred and eighty-seven hypertensives, 127 (61.9%) of them having criteria for MS according to the International Diabetes Federation criteria and 30 healthy normotensive subjects were included. OX status was assessed by measuring glutathione oxidized/glutathione reduced and reactive oxygen species-induced byproducts of lipid peroxidation, malondialdehyde, and DNA damage, 8-oxo-dG genomic and mitochondrial. Antioxidant enzymatic activity of Cu/Zn extracellular-superoxide dismutase (SOD) and catalase (CAT) was measured in plasma and glutathione peroxidase 1 in hemolysed erythrocytes. In mononuclear cells, total-SOD activity, CAT and glutathione peroxidase 1, were assessed as well. The OX state in both blood and peripheral mononuclear cells observed in hypertensives were not enhanced by the addition of components of the so-called MS. Likewise, the reduction in the activity of antioxidant enzymes, both extracellular and cytoplasmic, was not affected by the presence of additional components of the MS. Neither the number of components nor the individual addition of each of them, low high-density lipoprotein, triglycerides, abdominal obesity or fasting glucose, further impact in the OX abnormalities observed in those with only hypertension in absence of other components. In conclusion, the present data indicates that contribution of MS components to the OX burden generated by high blood pressure is minimal.

[Effect of oxidative stress in patients with chronic renal failure]. / Efecto del tratamiento con hemodiálisis sobre el estrés oxidativo en pacientes con insuficiencia renal crónica.

BACKGROUND: Cardiovascular disease remains the single most common cause of excess morbidity and mortality in end-stage renal disease (ESRD) patients and the traditional risk factors can't explain the high incidence of these events. New "non-traditional" risk factors are analysed in uremic patients and the increased oxidative stress is postulated to be an important contributor to uremic cardiovascular risk. METHODS: In order to evaluate the effects of the hemodialysis treatment, a complete oxidative stress study was performed in fifteen uremic patients. Representative antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), together with oxidized/reduced glutathione ratio (GSSG/GSH) and other oxidation indicators including malondialdehyde (MDA) and 8-oxo-2'-deoxyguanosine (8-oxo-dG), were analysed to assess oxidative stress status in normal control volunteers and in uremic patients treated with hemodialysis (HD). In the latter group blood samples were taken prior and after HD to evaluate the effect of the session of HD over the oxidative markers. RESULTS: Low levels of antioxidant enzyme activities were observed in the uremic patients as compared with normal control subjects. HD treatment results in a significant recovery of these enzyme activities but remain lower as compared with control values. Levels of GSSG and GSH concentrations were increased and reduced respectively in uremic patients. These differences were even higher before the HD and were reduced upon treatment to levels closer to those observed in controls. MDA levels and 8-oxo-dG levels were also increased in uremic patients with the highest values observed in the pre-treated HD group. Even though HD treatment decreases the levels of oxidation products in mononuclear cells of uremic patients the values of the control group are not reached. CONCLUSIONS: Our results suggest that hemodialysis by itself could correct the oxidative status in these patients. The possible mechanisms involved in the oxidative stress changes with the hemodialysis treatment will be discussed below.

Hospital clinical trial: Homeopathy (Agraphis nutans 5CH, Thuya occidentalis 5CH, Kalium muriaticum 9CH and Arsenicum iodatum 9CH) as adjuvant, in children with otitis media with effusion.

BACKGROUND: Otitis media with effusion (OME) is the most common cause of paediatric hearing loss. No single treatment has proved its effectiveness. There is a lack of evidence-based medicine studies in the area of homeopathy. METHOD: A prospective randomized, double blinded interventional placebo control study was conducted. Patients, from 2 months to 12 years, with OME diagnosed by pneumatic otoscopy (PNO) and tympanometry, were randomized into two groups. Both groups received aerosol therapy (mucolytics and corticosteroids). In addition, the experimental group (EG) received homeopathy (Agraphis nutans 5CH, Thuya Occidentalis 5CH, Kalium muriaticum 9CH and Arsenicum iodatum), and the placebo group (PG) placebo, both of them for 3 months. Patients were evaluated by PNO examination and tympanometry at baseline, at 45 and 90 days. RESULTS: 97 patients were enrolled. In the EG, 61.9% of individuals were cured (PNO went from negative in the 1st visit to positive in the 3rd visit) compared with 56.8% of patients treated with placebo. 4.8% of patients in the EG suffered a recurrence (positive PNO in the 2nd visit changed to negative in the 3rd visit) while 11.4% did in the PG. No significant difference was found. Adverse events were distributed similarly, except in the case of upper respiratory tract infections, which were less frequent in EG (3 vs. 13, p: 0.009). CONCLUSION: The homeopathic scheme used as adjuvant treatment cannot be claimed to be an effective treatment in children with OME. TRIAL REGISTRATION: EUDRACT number: 2011-006086-17, PROTOCOL code: 55005646.

Different hydrolytic efficiencies of adipose tissue lipoprotein lipase on very-low-density lipoprotein subfractions separated by heparin-Sepharose chromatography.

Human very-low-density lipoproteins (VLDL) were subfractionated by heparin-Sepharose chromatography into an unbound (A) and three bound (B, C and D) populations at increasing ionic strengths. Subfractions were characterized regarding their chemical composition and efficiency of triacylglycerol hydrolysis by rat adipose tissue LPL. The triacylglycerol content decreased, whereas the cholesterol and protein contents increased from subfractions A and B to subfraction D. VLDL-D showed the highest apo E/apo C ratio, though all the subfractions contained appreciable apo E. Appearance of VLDL-A resulted from exceeding the binding capacity of the column, since practically all its particles eluted at positions of bound VLDL under re-chromatography. Subfractions B, C and D stimulated LPL activity on emulsified tri[14C]oleoylglycerol to a similar extent, indicating that their apo C-II content was equally effective activating LPL. Incubation of tri[14C]oleoylglycerol labeled VLDL subfractions with fat pad pieces in the presence or absence of heparin resulted in greater hydrolysis and fatty acid uptake for VLDL-B and -C than for VLDL-D, a pattern observed over a wide range of LPL activities in the media. We conclude: (1) any VLDL particle can interact with heparin, which is consistent with the presence of apo E in all the subfractions, and (2) triacylglycerols in apo E-rich VLDL are less efficiently hydrolyzed by LPL than those in apo E-poor particles. We propose that richness in apo E impairs LPL action upon VLDL and decreases the rate of delivery of fatty acids to peripheral tissues.

The production of free radicals during the autoxidation of cysteine and their effect on isolated rat hepatocytes.

Autoxidizing cysteine has been shown to produce thiyl and hydroxyl radicals. Hydrogen peroxide increased the yield of both radicals which was inhibited by catalase but stimulated by copper/zinc superoxide dismutase. This effect is due to increased hydrogen peroxide production by copper/zinc superoxide dismutase as a result of superoxide dismutation. The production of superoxide radicals could not be detected probably because of its low reactivity, however, measurement of oxygen uptake and reduction of ferricytochrome c by autoxidizing cysteine clearly implicate the involvement of super oxide radicals. The production of hydroxyl radicals is postulated to proceed through a fenton reaction, however, this may not necessarily be metal ion controlled. Autoxidizing cysteine disrupts the integrity of hepatocytes causing release of glutathione, adenosine triphosphate and lactate dehydrogenase indicating that it is of little use as a therapeutic agent.

Effect of metal ion catalyzed oxidation of rifamycin SV on cell viability and metabolic performance of isolated rat hepatocytes.

The effect of rifamycin SV on metabolic performance and cell viability was studied using isolated hepatocytes from fed, starved and glutathione (GSH) depleted rats. The relationships between GSH depletion, nutritional status of the cells, glucose metabolism, lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) production in the presence of rifamycin SV and transition metal ions was investigated. Glucose metabolism was impaired in isolated hepatocytes from both fed and starved animals, the effect is dependent on the rifamycin SV concentration and is enhanced by copper (II). Oxygen consumption by isolated hepatocytes from starved rats was also increased by copper (II) and a partial inhibition due to catalase was observed. Cellular GSH levels which decrease with increasing the rifamycin SV concentration were almost depleted in the presence of copper (II). A correlation between GSH depletion and LDH leakage was observed in fed and starved cells. Catalase induced a slight inhibition of the impairment of gluconeogenesis, GSH depletion and LDH leakage in starved hepatocytes incubated with rifamycin SV, iron (II) and copper (II) salts. Lipid peroxidation measured as MDA production by isolated hepatocytes was also augmented by rifamycin SV and copper (II), especially in hepatic cells isolated from starved and GSH depleted rats. Higher cytotoxicity was observed in isolated hepatocytes from fasted animals when compared with fed or GSH depleted animals. It seems likely that in addition to GSH level, there are other factors which may have an influence on the susceptibility of hepatic cells towards xenobiotic induced cytotoxicity.