Connective tissue metabolism in children with chronic renal failure

To assess the characteristics of connective tissue metabolism in chronic renal failure (CRF), urinary excretion of glycosaminoglycan (GAG) fractions and hydroxyproline (HYP) was determined in ten patients with CRF and in ten age-matched healthy children. CRF was found to be associated with elevated free HYP (19.9±6.1 vs 9.8±3.6 mol/day,P<0.05) and depressed peptide HYP excretion (33.1±13.5 vs 225.2±17.7 mol/day,P<0.01), a low rate of total GAG excretion (7.0±2.4 vs 16.1±1.9 mol uronic acid/day,P<0.05) with low chondroitin 4 — sulphate + chondroitin 6 — sulphate (Ch-Ss) (14.0±9.9 vs 65.0±22.1%) and a high proportion of non-sulphated or under-sulphated fractions, i.e. hyaluronic acid + chondroitin + heparan sulphate (HA+Ch+HS) (75.3±11.4 vs 31.5±5.7%). Urinary 3-methyl-histidine (3-met-HIS) excretion and plasma essential free amino acids did not differ in the two groups. In response to haemodialysis no consistent change occurred in urinary excretion of 3-met-HIS, peptide-bound HYP, total GAG or percentage distribution of individual GAG fractions. After haemodialysis all plasma amino acids decreased significantly, and there was a significant increase in urinary excretion of free HYP (P<0.05). We conclude that the alterations in urinary excretion of total and individual GAGs observed in CRF may reflect disturbed connective tissue metabolism which does not appear to be accounted for by protein malnutrition or enhanced protein breakdown and remains uninfluenced by haemodialysis therapy.     

MKK4 and metastasis suppression: a marriage of signal transduction and metastasis research

MAP kinase kinase 4 (MKK4) is a member of the stress-activated protein kinase (SAPK) signaling cascade and is involved in the regulation of many cellular processes. We have recently demonstrated a functional role for MKK4 in the suppression of metastases. In this review, we discuss the established cellular and biochemical functions of MKK4, as well as a new function for MKK4 as a metastasis suppressor gene. Because of the importance of signaling studies to this translational work, a detailed example of the strategy and tools that can be employed to define the biochemical mechanism of MKK4-mediated metastasis suppression is presented. Finally, the potential therapeutic utility of these findings is discussed.     

Diagnostic procedure in osteodystrophia deformans Paget

Simultaneous scintigraphic and X-ray examination has been evaluated in 30 patients with Paget's disease for estimation of extension and activity of the disease. Scintigraphy produces a comprehensive picture of the pathophysiologic condition. When compared with X-rays the procedure is more economic with regards to cost, time and radiation dosage. Therefore, the use of radioisotopes for evaluation of the skeleton is strongly recommended for follow-up examination in Paget's disease.     

Hyaluronan-related limited concentration by the immature kidney

The limited renal concentration performance by the immature kidney traditionally is thought to be attributed to blunted renal response to arginine vasopressin (AVP) and medullary hypotonicity. The diminished AVP-dependent osmotic water permeability of the collecting duct is the result of decreased AVP binding and adenylate cyclase activation, and low expression of aquaporin-2 (AQP2) mRNA and low levels of AQP2 protein. Moreover, the immature kidney fails to establish deep cortico-papillary osmotic gradient because of structural immaturity, limited solute transport and increased medullary blood flow. Based on indirect clinical and experimental evidences this article puts forward a hypothesis that during perinatal period the abundant hyaluronan (HA) content in the renomedullary interstitium has a primary role in antagonizing water reabsorption and limiting concentration performance. Hydration-related alterations in renal HA appears to be mediated by antidiuretic hormone.The concept of HA-mediated renal water transport may imply that interfering selectively with renal HA metabolism may provide a new therapeutic approach to promote diuresis or antidiuresis, respectively, according to the elevation or reduction in renomedullary HA.     

Urinary endothelin excretion in the neonate: influence of maturity and perinatal pathology

The present study was undertaken to establish the developmental pattern of urinary endothelin-1 (ET-1) excretion and to define its possible role in mediating pathophysiological changes related to perinatal asphyxia/infection and dopamine treatment. Urinary ET-1 levels were measured by radioimmunoassay in 7 full-term neonates (mean gestational age 39.3 weeks) on days 1, 3 and 5, and in 9 pre-term neonates (mean gestational age 30.8 weeks) on days 1, 3, 5, 7 and weekly thereafter for 5 consecutive, weeks. The results were compared with those of three age-groups of 30 normal children (4–8 years, 9–12 years and 13–18 years); each group, consisted of 10 children. The influence of severe cardiopulmonary distress (n=16, mean gestational age 33.9 weeks, post-natal age 3.3 days) and dopamine administration in a dose of 2 g/min per kg (n=10, mean gestational and post-natal ages 32.1 weeks and 5.6 days, respectively) were also studied. In full-term infants, ET-1 concentration fell from 34.3±1.8 pmol/l on day 1 to 21.5±1.5 pmol/l on day 5 (P<0.01). In premature infants its absolute value and its post-natal fall were similar in the 1st week and no further change occurred in weeks 2–5; it stabilized at levels between 17.1±2.2 and 16.7±1.7 pmol/l. These concentrations tended to be lower than those of 25.5±1.3, 23.0±1.0 and 26.2±0.7 pmol/l measured in three groups of older children. During the 1st week, daily ET-1 excretion remained unchanged in term infants (3.1±1.0 vs. 3.7±1.5 pmol/m² per day), but there was a significant increase from 6.5±1.0 to 12.4±0.7 pmol/m² per day (P<0.01) in premature infants. During weeks 2–5, preterm infants excreted more ET-1 than older children (P<0.01). In response to perinatal ashphyxia/infection and dopamine therapy, urinary ET-1 excretion markedly rose and there was a significant positive correlation between urine flow rate and ET-1 excretion (P<0.001). We conclude that ET-1 concentration rather than excretion rate may have a role in mediating the changes in renal functions that occur soon after birth. The pathophysiological significance of the flow-dependent increase in urinary ET-1 excretion needs to be further studied.     

Ammonium chloride metabolic acidosis and the activity of renin-angiotensin-aldosterone system in children

The present study was undertaken to assess the effects of acute metabolic acidosis on the activity of the renin-angiotensin-aldosterone system in 12 children with a mean age of 8.9 years who underwent NH₄Cl loading test. Ammonium chloride was given in a dose of 0.15 g/kg per day for 3 consecutive days to evaluate renal acidification. Prior to and following NH₄Cl administration blood acid-base parameters, plasma and urine electrolytes, creatinine and aldosterone concentrations as well as plasma renin activity (PRA), urine flow rate and net H⁺ excretion were measured. Ammonium chloride administration significantly depressed blood pH (P<0.05), bicarbonate (P<0.01) and base excess (P<0.01) and resulted in a slight, but significant elevation of plasma potassium concentration (P<0.05). Furthermore, NH₄Cl ingestion induced a marked increase in urine flow rate (P<0.01) and urinary sodium, potassium and chloride excretion (P<0.01). In response to NH₄Cl metabolic acidosis, PRA doubled (4.72±1.18 vs 8.13±1.02 ng/ml per hour,P0.05) and there was a nearly fourfold increase in plasma aldosterone level (0.49±0.12 vs 1.52±0.24 ng/ml,P<0.01) and in urinary aldosterone excretion (19.2±4.3 vs 71.8±13.8 g/day,P<0.01). The elevated aldosterone production observed in this study is assumed to be mediated by the combined effect of sodium and water diuresis-related increased PRA, hyperkalaemia and the direct stimulation of adrenal steroidogenesis by metabolic acidosis.     

Low Au-content CoAu electrodes for environmental applications

Six cobalt gold (CoAu) electrodes were prepared by electroless deposition using different gold-containing solutions (acidic and weakly acidic) and different Au deposition times. Characterization of CoAu electrodes was done by scanning electron microscopy with energy-dispersive X-ray spectroscopy, N₂-sorption, and X-ray powder diffraction techniques. The possibility of using the prepared electrodes in environmental applications, i.e., for the electrochemical sensing of a trace amount of arsenic(iii) in weakly alkaline media was assessed. Employing the CoAu electrode (prepared by immersing Co/Cu into 1 mM HAuCl₄ (pH 1.8) at 30 °C for 30 s) under optimized conditions (deposition potential −0.7 V and deposition time of 60 s), a low limit of detection of 2.16 ppb was obtained. Finally, this CoAu electrode showed activity for arsenic oxidation in the presence of Cu(ii) as a model interferent as well as in real samples. Furthermore, the use of CoAu electrode as an anode in fuel cells, namely, direct borohydride – hydrogen peroxide fuel cells was also assessed. A peak power density of 191 mW cm⁻² was attained at 25 °C for DBHPFC with CoAu anode at a current density of 201 mA cm⁻² and cell voltage of 0.95 V, respectively. The peak power density further increased with the increase of the operating temperature to 55 °C.

A low Au-content CoAu electrode prepared by simple electroless deposition outperforms a pure Au electrode for versatile environmental applications: As(iii) sensing in water or as electrodes in direct borohydride-hydrogen peroxide fuel cells.     

Cocktails of Mycotoxins,Phytoestrogens, and Other Secondary Metabolites in Diets of Dairy Cows in Austria: Inferences from Diet Composition and Geo-Climatic Factors

Dairy production is a pivotal economic sector of Austrian and European agriculture. Dietary toxins and endocrine disruptors of natural origin such as mycotoxins and phytoestrogens can affect animal health, reproduction, and productivity. This study characterized the profile of a wide spectrum of fungal, plant, and unspecific secondary metabolites, including regulated, emerging, and modified mycotoxins, phytoestrogens, and cyanogenic glucosides, in complete diets of lactating cows from 100 Austrian dairy farms. To achieve this, a validated multi-metabolite liquid chromatography/electrospray ionization–tandem mass spectrometric (LC/ESI–MS/MS) method was employed, detecting 155 of >800 tested metabolites. Additionally, the most influential dietary and geo-climatic factors related to the dietary mycotoxin contamination of Austrian dairy cattle were recognized. We evidenced that the diets of Austrian dairy cows presented ubiquitous contamination with mixtures of mycotoxins and phytoestrogens. Metabolites derived from Fusarium spp. presented the highest concentrations, were the most recurrent, and had the highest diversity among the detected fungal compounds. Zearalenone, deoxynivalenol, and fumonisin B1 were the most frequently occurring mycotoxins considered in the EU legislation, with detection frequencies >70%. Among the investigated dietary factors, inclusion of maize silage (MS) and straw in the diets was the most influential factor in contamination with Fusarium-derived and other fungal toxins and metabolites, and temperature was the most influential among the geo-climatic factors.     

Fungal Species and Multi-Mycotoxin Associated with Post-Harvest Sorghum (Sorghum bicolor (L.) Moench) Grain in Eastern Ethiopia

Sorghum is the main staple food crop in developing countries, including Ethiopia. However, sorghum grain quantity and quality are affected by contaminating fungi both under field and post-harvest stage. The aim of the current study was to assessed fungal species and multi-mycotoxins associated with sorghum grain in post-harvest samples collected from eastern Ethiopia. Fungal genera of Aspergillus, Alternaria, Bipolaris, Fusarium, Mucor, Penicillium, and Rhizoctonia were recovered in the infected grain. A liquid chromatography-tandem mass spectrometric (LC-MS/MS) was used for quantification of multiple mycotoxins/fungal metabolites. Overall, 94 metabolites were detected and grouped into eight categories. All metabolites were detected either in one or more samples. Among major mycotoxins and derivatives, deoxynivalenol (137 μg/kg), zearalenone (121 μg/kg), ochratoxin A (115 μg/kg), and fumonisin B₁ (112 μg/kg) were detected with maximum concentrations, while aflatoxin B₁ had relatively lower concentrations (23.6 μg/kg). Different emerging mycotoxins were also detected, with tenuazonic acid (1515 μg/kg) occurring at the maximum concentration among Alternaria metabolites. Fusaric acid (2786 μg/kg) from Fusarium metabolites and kojic acid (4584 μg/kg) were detected with the maximum concentration among Fusarium and Aspergillus metabolites, respectively. Unspecific metabolites were recognized with neoechinulin A (1996 μg/kg) at the maximum concentration, followed by cyclo (L-Pro-L-Tyr) (574 μg/kg) and cyclo (L-Pro-L-Val) (410 μg/kg). Moreover, metabolites form other fungal genera and bacterial metabolites were also detected at varying levels. Apparently, the study revealed that sorghum grains collected across those districts were significantly contaminated with co-occurrences of several mycotoxins. Farmers should be the main target groups to be trained on the improved management of sorghum production.     

The Role of Nitrogen Fertilization on the Occurrence of Regulated,Modified and Emerging Mycotoxins and Fungal Metabolites in Maize Kernels

The European Food Safety Authority is currently evaluating the risks related to the presence of emerging mycotoxins in food and feeds. The aim of this study was to investigate the role of soil fertility, resulting from different nitrogen fertilization rates, on the contamination of regulated mycotoxins and emerging fungal metabolites in maize grains. The trial was carried out in the 2012–2013 growing seasons as part of a long-term (20-year) experimental platform area in North-West Italy, where five different N rates, ranging from 0 to 400 kg N ha⁻¹, were applied to maize each year. Maize samples were analyzed by means of a dilute-and-shoot multi-mycotoxin LC-MS/MS method, and more than 25 of the most abundant mycotoxins and fungal metabolites were detected. Contamination by fumonisins and other fungal metabolites produced by Fusarium spp. of the section Liseola was observed to have increased in soils that showed a poor fertility status. On the other hand, an overload of nitrogen fertilization was generally associated with higher deoxynivalenol and zearalenone contamination in maize kernels, as well as a higher risk of other fungal metabolites produced by Fusarium spp. sections Discolor and Roseum. A balanced application of N fertilizer, in accordance with maize uptake, generally appears to be the best solution to guarantee an overall lower contamination by regulated mycotoxins and emerging fungal metabolites.