The ameliorative effects of Nigella sativa, thymoquinone, and bentonite against aflatoxicosis in broilers via AFAR and Nrf2 signalling pathways, and down-regulation of caspase-3.

1. Aflatoxins (AFs) are metabolites which especially have toxic effects on proteins, and are detoxified by the aflatoxin-B1 aldehyde reductase (AFAR) pathway. In this pathway, the aldo-keto reductase family 7, member A2 (AKR7A2) enzyme, which is controlled by nucleic-related erythroid factor 2 (Nrf2), plays an active role. However, data on the efficacy of this critical pathway in broilers is limited.2. The aim of the following study was to investigate the changes in the expression levels of AKR7A2, Nrf2, and caspase-3, and the effects of Nigella sativa seeds (NS), thymoquinone (TMQ), and bentonite (BNT) in broilers exposed to AFs.3. One-hundred broilers were divided into ten groups (control (CNT); AF; NS; TMQ; BNT; AF+TMQ; AF+NS; AF+BNT; AF+BNT+NS; AF+BNT+TMQ) and fed for 28 d. AF, TMQ, NS and BNT were added to diets at levels of 2 mg/kg, 300 mg/kg, 50 g/kg and 10 g/kg respectively.4. The addition of AF to the diet decreased AKR7A2 and Nrf2 levels dramatically, but increased caspase-3 (P < 0.01). TMQ, NS and BNT additions to the diet eliminated all negative effects caused by AF (P < 0.01); and AKR7A2 and Nrf2 were further raised in TMQ and NS groups when compared to the control group. TMQ and NS showed a positive effect on detoxification parameters when given together with BNT.5. Supplementation with NS and TMQ enhanced AF detoxification via the AFAR pathway, by increasing AKR7A2 and Nrf2 levels, in addition to reducing hepatocyte apoptosis.

Urinary mRNA for the Diagnosis of Renal Allograft Rejection: The Issue of Normalization.

Urinary messenger RNA (mRNA) quantification is a promising method for noninvasive diagnosis of renal allograft rejection (AR), but the quantification of mRNAs in urine remains challenging due to degradation. RNA normalization may be warranted to overcome these issues, but the strategies of gene normalization have been poorly evaluated. Herein, we address this issue in a case-control study of 108 urine samples collected at time of allograft biopsy in kidney recipients with (n = 52) or without (n = 56) AR by comparing the diagnostic value of IP-10 and CD3ε mRNAs-two biomarkers of AR-after normalization by the total amount of RNA, normalization by one of the three widely used reference RNAs-18S, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Hypoxanthine-guanine phosphoribosyltransferase (HPRT)-or normalization using uroplakin 1A (UPK) mRNA as a possible urine-specific reference mRNA. Our results show that normalization based on the total quantity of RNA is not substantially improved by additional normalization and may even be worsened with some classical reference genes that are overexpressed during rejection. However, considering that normalization by a reference gene is necessary to ensure polymerase chain reaction (PCR) quality and reproducibility and to suppress the effect of RNA degradation, we suggest that GAPDH and UPK1A are preferable to 18S or HPRT RNA.

Early Low Urinary CXCL9 and CXCL10 Might Predict Immunological Quiescence in Clinically and Histologically Stable Kidney Recipients.

We monitored the urinary C-X-C motif chemokine (CXCL)9 and CXCL10 levels in 1722 urine samples from 300 consecutive kidney recipients collected during the first posttransplantation year and assessed their predictive value for subsequent acute rejection (AR). The trajectories of urinary CXCL10 showed an early increase at 1 month (p = 0.0005) and 3 months (p = 0.0009) in patients who subsequently developed AR. At 1 year, the AR-free allograft survival rates were 90% and 54% in patients with CXCL10:creatinine (CXCL10:Cr) levels <2.79 ng/mmoL and >2.79 ng/mmoL at 1 month, respectively (p < 0.0001), and 88% and 56% in patients with CXCL10:Cr levels <5.32 ng/mmoL and >5.32 ng/mmoL at 3 months (p < 0.0001), respectively. CXCL9:Cr levels also associate, albeit less robustly, with AR-free allograft survival. Early CXCL10:Cr levels predicted clinical and subclinical rejection and both T cell- and antibody-mediated rejection. In 222 stable patients, CXCL10:Cr at 3 months predicted AR independent of concomitant protocol biopsy results (p = 0.009). Although its positive predictive value was low, a high negative predictive value suggests that early CXCL10:Cr might predict immunological quiescence on a triple-drug calcineurin inhibitor-based immunosuppressive regimen in the first posttransplantation year, even in clinically and histologically stable patients. The clinical utility of this test will need to be addressed by dedicated prospective clinical trials.

Polythiophenes and polythiophene-based composites in amperometric sensing.

This overview of polythiophene-based materials provides a critical examination of meaningful examples of applications of similar electrode materials in electroanalysis. The advantages arising from the use of polythiophene derivatives in such an applicative context is discussed by considering the organic conductive material as such, and as one of the components of hybrid materials. The rationale at the basis of the combination of two or even more individual components into a hybrid material is discussed with reference to the active electrode processes and the consequent possible improvements of the electroanalytical performance. In this respect, study cases are presented considering different analytes chosen among those that are most frequently reported within the classes of organics and inorganics. The use of a polythiophene matrix to stably fix biological elements at the electrode surface for the development of catalytic biosensors and genosensors is also discussed. Finally, a few possible lines along which the next research in the field could be fruitfully pursued are outlined. Furthermore, the work still to be done to exploit the possibilities offered by novel products of organic synthesis, even along paths already traced in other fields of electrochemistry, is discussed.

Therapeutic mTOR inhibition in autosomal dominant polycystic kidney disease: What is the appropriate serum level?

Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited renal disease, and sirolimus, a mammalian target of rapamycin (mTOR) inhibitor, has been shown to significantly retard cyst expansion in animal models. The optimal therapeutic dose of sirolimus is not yet defined. Here, we report the history of a previously unknown ADPKD deceased donor whose kidneys were engrafted in two different recipients. One of the two received an immunosuppressive regimen based on sirolimus for 5 years while the other did not. After transplantation, both patients developed severe transplant cystic disease. Donor DNA sequence identified a new hypomorphic mutation in PKD1. The rate of cyst growth was identical in the two patients regardless of the treatment. While sirolimus treatment reduced the activation of mTOR in peripheral blood mononuclear cells, it failed to prevent mTOR activation in kidney tubular cells, this could account for the inefficiency of treatment on cyst growth. Together, our results suggest that the dose of sirolimus required to inhibit mTOR varies according to the tissue.

Reduction of renal mass is lethal in mice lacking vimentin. Role of endothelin-nitric oxide imbalance.

Modulation of vascular tone by chemical and mechanical stimuli is a crucial adaptive phenomenon which involves cytoskeleton elements. Disruption, by homologous recombination, of the gene encoding vimentin, a class III intermediate filament protein mainly expressed in vascular cells, was reported to result in apparently normal phenotype under physiological conditions. In this study, we evaluated whether the lack of vimentin affects vascular adaptation to pathological situations, such as reduction of renal mass, a pathological condition which usually results in immediate and sustained vasodilation of the renal vascular bed. Ablation of 3/4 of renal mass was constantly lethal within 72 h in mice lacking vimentin (Vim-/-), whereas no lethality was observed in wild-type littermates. Death in Vim-/- mice resulted from end-stage renal failure. Kidneys from Vim-/- mice synthesized more endothelin, but less nitric oxide (NO), than kidneys from normal animals. In vitro, renal resistance arteries from Vim-/- mice were selectively more sensitive to endothelin, less responsive to NO-dependent vasodilators, and exhibited an impaired flow (shear stress)- induced vasodilation, which is NO dependent, as compared with those from normal littermates. Finally, in vivo administration of bosentan, an endothelin receptor antagonist, totally prevented lethality in Vim-/- mice. These results suggest that vimentin plays a key role in the modulation of vascular tone, possibly via the tuning of endothelin-nitric oxide balance.

Targeted expression of a dominant-negative EGF-R in the kidney reduces tubulo-interstitial lesions after renal injury.

The role of EGF in the evolution of renal lesions after injury is still controversial. To determine whether the EGF expression is beneficial or detrimental, we generated transgenic mice expressing a COOH-terminal-truncated EGF-R under the control of the kidney-specific type 1 gamma-glutamyl transpeptidase promoter. As expected, the transgene was expressed exclusively at the basolateral membrane of proximal tubular cells. Under basal conditions, transgenic mice showed normal renal morphology and function. Infusion of EGF to transgenic animals revealed that the mutant receptor behaved in a dominant-negative manner and prevented EGF-signaled EGF-R autophosphorylation. We next evaluated the impact of transgene expression on the development of renal lesions in two models of renal injury. After 75% reduction of renal mass, tubular dilations were less severe in transgenic mice than in wild-type animals. After prolonged renal ischemia, tubular atrophy and interstitial fibrosis were reduced in transgenic mice as compared with wild-type mice. The beneficial effect of the transgene included a reduction of tubular cell proliferation, interstitial collagen accumulation, and mononuclear cell infiltration. In conclusion, functional inactivation of the EGF-R in renal proximal tubular cells reduced tubulo-interstitial lesions after renal injury. These data suggest that blocking the EGF pathway may be a therapeutic strategy to reduce the progression of chronic renal failure.

Impaired flow-induced dilation in mesenteric resistance arteries from mice lacking vimentin.

The intermediate filament vimentin might play a key role in vascular resistance to mechanical stress. We investigated the responses to pressure (tensile stress) and flow (shear stress) of mesenteric resistance arteries perfused in vitro from vimentin knockout mice. Arteries were isolated from homozygous (Vim-/-, n = 14) or heterozygous vimentin-null mice (Vim+/-, n = 5) and from wild-type littermates (Vim+/+, n = 9). Passive arterial diameter (175+/-15 micron in Vim+/+ at 100 mmHg) and myogenic tone were not affected by the absence of vimentin. Flow-induced (0-150 microl/min) dilation (e. g., 19+/-3 micron dilation at 150 mmHg in Vim+/+) was significantly attenuated in Vim-/- mice (13+/-2 micron dilation, P < 0.01). Acute blockade of nitric oxide synthesis (NG-nitro- L-arginine, 10 microM) significantly decreased flow-induced dilation in both groups, whereas acute blockade of prostaglandin synthesis (indomethacin, 10 microM) had no significant effect. Mean blood pressure, in vivo mesenteric blood flow and diameter, and mesenteric artery media thickness or media to lumen ratio were not affected by the absence of vimentin. Thus, the absence of vimentin decreased selectively the response of resistance arteries to flow, suggesting a role for vimentin in the mechanotransduction of shear stress.

An Overview of Advanced SILAC-Labeling Strategies for Quantitative Proteomics.

Comparative, quantitative mass spectrometry of proteins provides great insight to protein abundance and function, but some molecular characteristics related to protein dynamics are not so easily obtained. Because the metabolic incorporation of stable amino acid isotopes allows the extraction of distinct temporal and spatial aspects of protein dynamics, the SILAC methodology is uniquely suited to be adapted for advanced labeling strategies. New SILAC strategies have emerged that allow deeper foraging into the complexity of cellular proteomes. Here, we review a few advanced SILAC-labeling strategies that have been published during last the years. Among them, different subsaturating-labeling as well as dual-labeling schemes are most prominent for a range of analyses including those of neuronal proteomes, secretion, or cell-cell-induced stimulations. These recent developments suggest that much more information can be gained from proteomic analyses if the labeling strategies are specifically tailored toward the experimental design.

Acute pyelonephritis as a cause of hyponatremia/hyperkalemia in young infants with urinary tract malformations.

Obstructive uropathy causes tubular resistance to aldosterone and severe metabolic imbalance may be precipitated by an episode of pyelonephritis. In the last 3 years we investigated 52 episodes of pyelonephritis (positive urine culture, elevated C reactive protein, fever, elevated neutrophil count) in 50 children between 15 days and 15 months of age. Ultrasonography voiding cystography and renal scintiscan were performed in all cases and i.v. urography in some. A salt-losing syndrome with hyponatremia and hyperkalemia (Na < 125 meq/liter; K > 6.3 meq/liter) was observed in 17 infants < 3 months, accompanied by plasma aldosterone concentration of 5000 to 23,000 pg/ml (normal value, < 1000 pg/ml). All these children had a severe urinary tract (UT) malformation (ureteropelvic junction stenosis in 7 cases, vesicoureteral reflux in 7, posterior urethral valves in 2, double system in 1). Thirteen infants < 3 months, 7 with no urinary tract malformations, did not have electrolyte imbalance. Pyelonephritis was diagnosed in 20 other patients ages 4 to 15 months, including 16 with severe UT malformations; 4 had normal UTs. We conclude that a salt-losing syndrome with tubular resistance to aldosterone can occur during pyelonephritis in young infants with congenital UT malformation, that the risk diminishes considerably or disappears after 3 months of age and that in the absence of UT malformation pyelonephritis does not cause acute sodium loss of clinical relevance.

Catch-up growth in children with chronic renal failure treated with long-term enteral nutrition.

Growth retardation commonly complicates chronic renal failure in children. Although the etiology of this growth impairment is multifactorial, inadequate nutrition is considered an important cause in infants and young children. An "aggressive" nutritional approach has been repeatedly suggested in children with early onset chronic renal failure and poor feeding habits, but the possibility of inducing catch-up growth by energy supplementation is still controversial. The nutritional effects of a long-term, home-based enteral feeding program were studied in two infants and three children with moderate to severe chronic renal failure and impaired growth associated with persistent anorexia. In all patients, renal failure had developed during the first year of life due to congenital diseases. Enteral feeding was performed at home, during the night, through a silicone rubber nasogastric tube. The treatment lasted for 1 year. The energy intake ranged between 101% and 116% of the recommended dietary allowance (RDA), and the protein intake between 96% and 113% of the RDA in all patients but one, in whom proteins were restricted to 75% of the RDA. All children showed a substantial improvement in deviation score for both weight (mean increase +1.76), height (mean increase +1.52) and in the general metabolic condition, irrespective of age, severity of osteodystrophy, or degree of renal failure. The treatment was well tolerated and, apart from a few episodes of vomiting, no complications arose during the treatment. Tube feeding may be an effective therapeutic option for overcoming malnutrition when chronic renal failure is associated with persistent anorexia. In infants and young children, growth retardation can be opposed and catch-up growth obtained.

[Growth factors. Role in the progression of renal lesions]. / Facteurs de croissance. Rôle dans la progression des lésions rénales.

FROM PATHOPHYSIOLOGY TO THERAPEUTICS: Nephrologists are faced with the continuing problem of helping patients avoid the onset or retard the development of end-stage renal failure. Despite the treatments available, the risk is still high for patients and the cost a heavy burden for the public health budget. These facts underline the importance of a detailed understanding of the mechanisms leading to the destruction of renal parenchyma in order to develop therapeutic strategies capable of slowing the inevitable progression of kidney lesions. GROWTH FACTORS: It is currently recognized that a major reduction in the number of functional nephrons, whatever the initial cause, leads in itself to a progressive deterioration of healthy nephrons and finally to complete destruction of the kidney. The underlying mechanisms remain largely unknown. One possible mechanism would involve an overexpression of several growth factors in the damaged renal parenchyma. We present in this review experimental data obtained with various approaches, including pharmacological and/or dietetic modulations and the establishment of transgenic mouse lines, to demonstrate the key role played by growth factors in the progression of renal lesions. The pathways followed by these growth factors in the process of renal destruction as well as certain elements leading to their overexpression are also discussed.

The Preventive Effects of Different Doses of Glucomannan on Experimental Aflatoxicosis in Japanese Quails

ABSTRACT This experimental study was performed to investigate whether there is a protective effect of different doses of Glucomannan using against aflatoxicosis in Japanese quail, and pathological changes and relative organ weights were compared. In the experiment, 60 one-day old male Japanese quails were used as divided into six different groups. Experimental groups were designated as Control(C), aflatoxin(A), glucomannan(GM), 2-fold dose of glucomannan(2GM), aflatoxin+glucomannan(A+GM) and aflatoxin+2-fold dose of glucomannan(A+2GM). While control group quails fed the standard ration as ad libitum, other groups were fed with the administrations additionally to standard diet respectively; 2mg/kg of aflatoxin to group A, 1g/kg of glucomannan to group GM, 2g/kg of glucomannan to group 2GM, 2mg/kg of aflatoxin and 1g/kg glucomannan to group A+GM, 2mg/kg of aflatoxin and 2g/kg glucomannan to group A+2GM. All quails were euthanized at day 21 of the study and organs, (liver, spleen, kidney, thymus and bursa of Fabricius) were removed, weighed and subjected to routine histopathological procedures. Although any important macroscopic changes were not observed in the C, GM and 2GM groups, significant pathological changes were found in the groups of A, A+GM and A+2GM. In the A+GM group, the partial reduction in the severity of microscopic lesions were seen in liver, bursa of Fabricius, thymus and spleen, however a significant reduction in severity of lesions was noticed in A+2GM group. As a result of the study, 2g/kg of glucomannan has been found pathologically to be more effective than 1g/kg glucomannan in terms of the protection against aflatoxicosis by giving orally.(AU)

The Preventive Effects of Different Doses of Glucomannan on Experimental Aflatoxicosis in Japanese Quails

ABSTRACT This experimental study was performed to investigate whether there is a protective effect of different doses of Glucomannan using against aflatoxicosis in Japanese quail, and pathological changes and relative organ weights were compared. In the experiment, 60 one-day old male Japanese quails were used as divided into six different groups. Experimental groups were designated as Control(C), aflatoxin(A), glucomannan(GM), 2-fold dose of glucomannan(2GM), aflatoxin+glucomannan(A+GM) and aflatoxin+2-fold dose of glucomannan(A+2GM). While control group quails fed the standard ration as ad libitum, other groups were fed with the administrations additionally to standard diet respectively; 2mg/kg of aflatoxin to group A, 1g/kg of glucomannan to group GM, 2g/kg of glucomannan to group 2GM, 2mg/kg of aflatoxin and 1g/kg glucomannan to group A+GM, 2mg/kg of aflatoxin and 2g/kg glucomannan to group A+2GM. All quails were euthanized at day 21 of the study and organs, (liver, spleen, kidney, thymus and bursa of Fabricius) were removed, weighed and subjected to routine histopathological procedures. Although any important macroscopic changes were not observed in the C, GM and 2GM groups, significant pathological changes were found in the groups of A, A+GM and A+2GM. In the A+GM group, the partial reduction in the severity of microscopic lesions were seen in liver, bursa of Fabricius, thymus and spleen, however a significant reduction in severity of lesions was noticed in A+2GM group. As a result of the study, 2g/kg of glucomannan has been found pathologically to be more effective than 1g/kg glucomannan in terms of the protection against aflatoxicosis by giving orally.

Classification of red wines by chemometric analysis of voltammetric signals from PEDOT-modified electrodes.

Nine different types of Italian red wines of four different varieties were analysed, without any sample pre-treatments, by voltammetric techniques using a poly(3,4-ethylenedioxythiophene)-modified electrode. The data matrices consisting of the currents measured at different potentials, by repeated Cyclic Voltammetry or Differential Pulse Voltammetry, are submitted to chemometric analysis. After explorative tests based on Principal Component Analysis, Partial Least Squares-Discriminant Analysis classification models are built both for the training and for the test sets. To this aim, different classification strategies are adopted, considering the responses from the two techniques either separately or joined together to form a data matrix including the whole voltammetric information.