Maternal supply of cysteamine alleviates oxidative stress and enhances angiogenesis in porcine placenta

Background:Oxidative stress in placenta is associated with the occurrence of adverse pregnancy outcomes in sow,but there are few satisfactory treatment strategies for these conditions.This study investigated the potential of cysteamine(CS)as an antioxidant protectant for regulating the reproductive performance,redox status,and placental angiogenesis of sows.Methods:The placental oxidative stress status and vascular density of piglets with different birth weights:<1.0 kg(low birth weight,LBW)and 1.4–1.6 kg(normal birth weight,NBW)were evaluated,followed by allotting 84 sows to four treatments(n=21)and feeding them with a basal diet supplemented with 0,100,300,or 500 mg/kg of CS from d 85 of gestation to d 21 of lactation,respectively.Placenta,serum,and colostrum samples of sows or piglets were collected,and the characteristics of sows and piglets were recorded.Furthermore,the in vivo results were validated using porcine vascular endothelial cells(PVECs).Results:Compared with the NBW placentae,the LBW placentae showed increased oxidative damage and were vulnerable to angiogenesis impairment.Particularly,H2O2-induced oxidative stress prompted intracellular reactive oxygen species generation and inhibited the tube formation and migration of PVECs as well as the expression of vascular endothelial growth factor-A(VEGF-A)in vitro.However,dietary CS supplementation can alleviate oxidative stress and improve the reproductive performance of sows.Specifically,compared with the control group,dietary 100 mg/kg CS could(1)decrease the stillbirth and invalid rates,and increase both the piglet birth weight in the low yield sows and the placental efficiency;(2)increase glutathione and reduce malondialdehyde in both the serum and the colostrum of sows;(3)increase the levels of total antioxidant capacity and glutathione in LBW placentae;(4)increase the vascular density,the mRNA level of VEGF-A,and the immune-staining intensity of platelet endothelial cell adhesion molecule-1 in the LBW placentae.Furthermore,the in vitro experiment indicated that CS pre-treatment could significantly reverse the NADPH oxidase 2-ROS-mediated inactivation of signal transducer and activator of transcription-3(Stat3)signaling pathway induced by H2O2 inhibition of the proliferation,tube formation,and migration of PVECs.Meanwhile,inhibition of Stat3 significantly decreased the cell viability,tube formation and the VEGF-A protein level in CS pretreated with H_(2)O_(2)-cultured PVECs.Conclusions:The results indicated that oxidative stress and impaired angiogenesis might contribute to the occurrence of LBW piglets during pregnancy,but CS supplementation at 100 mg/kg during late gestation and lactation of sows could alleviate oxidative stress and enhance angiogenesis in placenta,thereby increasing birth weight in low yield sows and reducing stillbirth rate.The in vitro data showed that the underlying mechanism for the positive effects of CS might be related to the activation of Stat3 in PVECs.

Cysteamine increases expression and activity of H^1-K^+-ATPase of gastric mucosal cells in weaning piglets

AIM: To determine the in vivo andin vivo effects of cysteamine (CS) on expression and activity of H+-K+-ATPase of gastric mucosal cells in weaning piglets.METHODS: Eighteen litters of newborn Xinhuai piglets were employed in the in vivo experiment and allocated to control and treatment groups. From 12 d of age (D12), piglets in control group were fed basal diet, while the treatment group received basal diet supplemented with 120 mg/kg CS. Piglets were weaned on D35 in both groups. Six piglets from each group (n = 6) were slaughtered on D28 (one week before weaning), D35(weaning), D36.5, D38, D42, and D45 (36 h, 72 h,one week and 10 d after weaning), respectively. Semiquantitative RT-PCR was performed todetermine the levels of H+-K+-ATPase mRNA in gastric mucosa. H+-K+-ATPase activity in gastric mucosa homogenate was also determined. Gastric mucosal epithelial cells from piglets through primary cultures were used to further elucidate the effect of CS on expression and activity of H+-K+-ATPase in vivo. Cells were treated for 20 h with 0.001,0.01, and 0.1 mg/mL of CS (n = 4), respectively. The mRNA expression of H+-K+-ATPase and somatostatin (SS)as well as the H+-K+-ATPase activity were determined.RESULTS: in vivo, both mRNA expression and activity of H+-K+-ATPase in gastric mucosa of control group exhibited a trend to increase from D28 to D45, reaching a peak on D45, but did not show significant age differences. Furthermore, neither the mRNA expression nor the activity of H+-K+-ATPase was affected significantly by weaning. CS increased the mRNA expression of H+-K+-ATPase by 73%, 53%, 30% and 39% on D28(P = 0.014), D35 (P = 0.017), D42 (P = 0.013) and D45(P = 0.046), respectively. In accordance with the mRNA expression, H+-K+-ATPase activities were significantly higher in treatment group than in control group on D35(P = 0.043) and D45 (P = 0.040). In vivo, CS exhibited a dose-dependent effect on mRNA expression and activity of H+-K+-ATPase. Both H+-K+-ATPase mRNA expression and activity in gastric mucosal epithelial cells were significantly elevated after 20 h of exposure to the moderate (H+-K+-ATPase expression: P=0.03; H+-K+-ATPase activity: P = 0.014) and high concentrations (H+-K+-ATPase expression: P=0.017; H+-K+-ATPase activity:P = 0.022) of CS. Significant increases in SS mRNA expression were observed to accompany the elevation of H+-K+-ATPase expression and activity induced by the moderate (P = 0.024) and high concentrations (P = 0.022) of CS. Low concentration of CS exerted no effects either on expression and activity of H+-K+-ATPase or on SS mRNA expression in cultured gastric mucosal epithelial cells.CONCLUSION: No significant changes are observed in mRNA expression and activity of H+-K+-ATPase in gastric mucosa of piglets around weaning from D28 to D45. CS increases expression and activity of gastric H+-K+-ATPase in vivo and in vivo. SS is involved in mediating the effect of CS on gastric H+-K+-ATPase expression and activity in weaning piglets.

Effect of using ascorbic acid and cysteamine supplementation onin-vitrodevelopment of buffalo embryos

Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.

Effect of melatonin and/or cysteamine on development and vitrification of buffalo embryos

Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin (10 ng/mL) and cysteamine (50 μM) + melatonin (10 ng/mL) treatment groups. In experiment 1, previous treatments were added duringin vitromaturation and culturing of buffalo oocytes.Results:Cleavage and blastocyst rates were significantly (P<0.05) increased in melatonin treated group (70.5±0.9 and 12.8±1.0, respectively). However this effect was potentiated when combined with cysteamine (74.0±1.7 and 14.8±1.7, respectively). In experiment 2, the treatements were added in maturtaion, culturing as well as post-warming culture media. Embryos at 7 d were vitrified.Viability assessement directly after warming showed significant increase (P<0.05) in cysteamine, melatonin and their combination groups (76.8±2.8, 80.0±2.1 and 83.3±1.7, respectively) than control (65.8±2.4);but the viability after 24 h post-warming was the best in cysteamine + melatonin combination group (61.4±2.1).Conculsions: Enriching maturation, culturing and post-warming media of buffalo oocytes and embryos with melatonin and/or cysteamine have significantly beneficial effects on oocyte developmental competence as well as embryos vitrification procedure outcomes which in turn resulting in enhancement of commercial buffalo embryo production.

Effects of Dietary Supplementation with Cysteamine on Performance, Carcass Characteristics, Meat Quality and Antioxidant Status in Finishing Pigs

The current study was to investigate the effects of cysteamine （CS） on carcass characteristics, meat quality, and antioxidant status in finishing pigs. A total of 24 crossbred （PIC variety） finishing pigs （60.05±1.24 kg, 12 gilts and 12 barrows） were assigned randomly to one of the three dietary groups, with four pens/group （one gilt and one barrow per pen）. Pigs were fed with a basal diet containing 0 （control）, 70, or 140 mg/kg CS for 47 days. The results indicated that dietary CS supplementation increased （P〈0.05） the average daily gain and feed intake with optimal responses occurring at 70 mg/kg. Dietary supplementation of CS increased （P〈0.05） the dressing percentage and lean percentage of finishing pigs with optimal responses occurring at 140 mg/kg. The CS supplementation, however, had no effect （P〉0.05） on meat quality. Dietary CS supplementation increased （P〈0.05） total antioxidant capacity （T-AOC） and decreased malondialdehyde （MDA） with optimal responses occurring at 70 mg/kg. CS supplementation at both 70 mg/kg and 140 mg/kg doses improved （P〈0.05） the muscle RNA/DNA ratio. Collectively, the results suggest dietary supplementation of 70 mg/kg CS could significantly improve growth performance and antioxidant status without adverse effects on the gastrointestinal tracts in finishing pigs while the 140 mg/kg CS supplementation improved the carcass quality. However, meat quality was not affected by CS supplementation.

Challenges for cysteamine stabilization,quantification,and biological effects improvement

The aminothiol cysteamine,derived from coenzyme A degradation in mammalian cells,presents several biological applications.However,the bitter taste and sickening odor,chemical instability,hygroscopicity,and poor pharmacokinetic profile of cysteamine limit its efficacy.The use of encapsulation systems is a good methodology to overcome these undesirable properties and improve the pharmacokinetic behavior of cysteamine.Besides,the conjugation of cysteamine to the surface of nanoparticles is generally proposed to improve the intra-oral delivery of cyclodextrin-drug inclusion complexes,as well as to enhance the colorimetric detection of compounds by a gold nanoparticle aggregation method.On the other hand,the detection and quantification of cysteamine is a challenging mission due to the lack of a chromophore in its structure and its susceptibility to oxidation before or during the analysis.Derivatization agents are therefore applied for the quantification of this molecule.To our knowledge,the derivatization techniques and the encapsulation systems used for cysteamine delivery were not reviewed previously.Thus,this review aims to compile all the data on these methods as well as to provide an overview of the various biological applications of cysteamine focusing on its skin application.

Effect of Cysteamine-preparation on Dairy Performance of Dairy Holstein Friesian Cows

Thirty Holstein Friesian cows with similar weight, age, calving number, lactating length, fat percentage, and same physiology condition were selected and randomly divided into 3 groups: group I, group II and group III, 20 g and 40 g Cysteamine /(cow·day) was supplemented into the normal diet for group I and group II, respectively; group III was used as control. Results showed that, group I and group II produced 4.86 % and 6.88 % more milk than that of control (P<0.05); Meanwhile, FCM (3.5 % fat) of group I and group II was increased by 4.98 % (P<0.01) and 6.56 % (P<0.01). Change of the milk fat among the three groups were not significant(P<0.05). Average daily milk protein yield significantly was increased by 4.14 % and 2.76 % respectively (P<0.05), though the milk protein content was slightly reduced. Moreover, somatic cells of the cow in the three groups were in normal physiological range in this trial.

Remediation of the Contaminated Soils by Washing with an Aqueous Cysteamine Lixiviant

In order to decrease the content of heavy metals in the crops soils, a novel method based on using an aqueous solution bearing cysteamine as the key ingredient was studied to extract the polluted heavy metals including Cd, Cu, Zn, Ni and Pb. By using the single-factor method, remediation-related technical index were screened and they are, respectively, applied to the solid material whereby the heavy metals are released and extracted from the solid material. The biomass solution residues remaining in the solid material after the heavy metal extraction procedure is rapidly biodegradable, so that no objectionable traces remain in the solid materials or soils.

Adjunction of Avidin to a Cysteamine Self-Assembled Monolayer for Impedimetric Immunosensor

In this work an impedimetric immunosensor based on affinity immobilization method of a biotin labelled anti-human IgG antibody, used as a model system, was reported. The experimental procedure involves the growth of a self-assembled monolayer of a thiol (cysteamine) carrying terminal amine groups on gold electrodes. Glutarardehyde, a homobifunctional cross-linker, was used as a coupling reagent for the covalent linking of avidin to the amine groups of cysteamine. The attachment of the biotin labeled antibodies (anti-Human IgG) to the subsequent modified gold electrode was achieved by affinity interactions tacking advantage of the strong avidin-biotin bridge. The stepwise assembly process of the electrode was interrogated by means of cyclic voltammetry, electrochemical impedance spectroscopy and contact angle measurements. The response of the antibody modified electrode to their target antigens was investigated in the presence of BSA (bovin serum albumin) in order to alleviate non-specific adsorption problems. A proposed electrical model was used to analyse the experimental data. The resulting immunosensor has a linear dynamic range of 100 - 900 ng?ml–1 of antigen and a detection limit of 100 ng?ml–1.

Voltammetric determination of cysteamine at multiwalled carbon nanotubes paste electrode in the presence of isoproterenol as a mediator

A sensitive electrochemical sensor for the determination of cysteamine （CA） was developed using a modified multiwall carbon nanotube paste electrode （MWCNTPE） with isoproterenol （ISPT） as a mediator. This modified electrode showed very high electrocatalytic activity for the anodic oxidation of CA. Under the optimized conditions, the electrocatalytic peak current showed a linear relationship with CA concentration in the range of 0.3-450.0 μmol/L with a detection limit of 0.09 μmol/L CA. The modified electrode was used for the determination of CA in real samples such as urine and drug samples.

Simultaneous determination of cysteamine and folic acid in pharmaceutical and biological samples using modified multiwall carbon nanotube paste electrode

A carbon paste electrode（CPE） chemically modified with multiwall carbon nanotubes and ferrocene（FC） was used as a selective electrochemical sensor for the simultaneous determination of trace amounts of cysteamine（CA） and folic acid（FA）.This modified electrode showed very efficient electrocatalytic activity for the anodic oxidation of CA.The peak current of differential pulse voltammograms of CA and FA increased linearly with their concentration in the ranges of 0.7-200μmol/L CA and 5.0- 700μmol/L FA.The detection limits for CA and FA were 0.3μmol/L and 2.0μmoI/L,respectively.The diffusion coefficient（D） and transfer coefficient（α） of CA were also determined.These conditions are sufficient to allow determination of CA and FA both individually and simultaneously.

REACTIONS OF HYPOCRELLIN B ON MERCAPTO COMPOUNDS Ⅱ: REACTIONS OF HYPOCRELLIN B ON CYSTEINE OR CYSTEAMINE

Ⅰ. INTRODUCTIONThe target of the photodynamic action of the new Chinese photothrapeutic agent- hypocrellins is the cell membrane. This photodynamic action causes the reduction in quantity of mercapto groups in membrane proteins. But the mechanism of

Changes of somatostatin content in some brain areas of rats during oxygen－induced convulsion

The content of somatostatin(SS) in hippocampus,striatum and frontal cortex tissues of rats exposed to 600 kpa hyperbaric oxygen was determined by means of radioimmunoassay. Initial time of convulsion, severity of convulsion and survival time of rats with convulsion exposed to 700 kPa hyperbaric oxygen after intraperitoneal injection of cysteamine (CSH) or intracerebroventricular injection of anti-somatostatin serum(ASS) were also observed. The results showed that the content of SS in hippocampus and striatum tissues increased remarkably when rats were at near-convulsion ; by the time the rats developed convulsion,it had a significant increase in all brain areas observed. Intraperitoneal injection of CSH or intracerebroventricular injection of ASS could delay initial time of convulsion (ITC),prolong survival time (ST) and reduce severity of convulsion (SOC). These results suggest that SS might play a role in oxygen-induced convulsion and be one of the endogenous agents which caused oxygeninduced convulsion.

The reaction of cinnamaldehyde and cinnam(o)yl derivatives with thiols

Spurred by the alleged relevance of the thia-Michael reaction in the bioactivity of various classes of cinnam(o)yl natural products and by the development of a quick NMR assay to study this reaction, we have carried out a systematic study of the "native" reactivity of these compounds with dodecanethiol and cysteamine as models, respectively, of simple thiols and reactive protein thiols that can benefit from iminium ion catalysis in Michael reactions. Cinnamoyl esters and amides, as well as cinnamyl ketones and oximes, did not show any reactivity with the two probe thiols, while cinnamaldehyde(1a) reacted with cysteamine to afford a mixture of a thiazoline derivative and compounds of multiple addition, and with aliphatic thiols to give a single bis-dithioacetal(6). Chalchones and their vinylogous C5-curcuminoid derivatives were the only cinnamoyl derivatives that gave a thiaMichael reaction. From a mechanistic standpoint, loss of conjugation in the adduct might underlie the lack of a native Michael reactivity. This property is restored by the presence of another conjugating group on the carbonyl, as in chalcones and C5-curcuminoids. A critical mechanistic revision of the chemical and biomedical literature on cinnamaldehyde and related compounds seems therefore required.