Protective Effects of Fucoxanthin on Hydrogen Peroxide-Induced Calcification of Heart Valve Interstitial Cells

Cardiovascular diseases such as atherosclerosis and aortic valve sclerosis involve inflammatory reactions triggered by various stimuli, causing increased oxidative stress. This increased oxidative stress causes damage to the heart cells, with subsequent cell apoptosis or calcification. Currently, heart valve damage or heart valve diseases are treated by drugs or surgery. Natural antioxidant products are being investigated in related research, such as fucoxanthin (Fx), which is a marine carotenoid extracted from seaweed, with strong antioxidant, anti-inflammatory, and anti-tumor properties. This study aimed to explore the protective effect of Fx on heart valves under high oxidative stress, as well as the underlying mechanism of action. Rat heart valve interstitial cells under H₂O₂-induced oxidative stress were treated with Fx. Fx improved cell survival and reduced oxidative stress-induced DNA damage, which was assessed by cell viability analysis and staining with propidium iodide. Alizarin Red-S analysis indicated that Fx has a protective effect against calcification. Furthermore, Western blotting revealed that Fx abrogates oxidative stress-induced apoptosis via reducing the expression of apoptosis-related proteins as well as modulate Akt/ERK-related protein expression. Notably, in vivo experiments using 26 dogs treated with 60 mg/kg of Fx in combination with medical treatment for 0.5 to 2 years showed significant recovery in their echocardiographic parameters. Collectively, these in vitro and in vivo results highlight the potential of Fx to protect heart valve cells from high oxidative stress-induced damage.     

Changes of HCO3-/Cl- exchanger activity during meiotic maturation in Balb/c strain mouse oocytes and zygotes

Intracellular pH-regulatory mechanisms are acquired by growing mouse oocytes with meiotic competence, and these mechanisms become fully active when the oocytes develop to the germinal vesicle (GV) stage as shown in CF1 and Balb/c strains mice. On the other hand, there is some evidence showing that intracellular pH-regulatory mechanisms are inhibited at the stages of Metaphase I (MI) and II (MII) oocytes in the CF1 strain mouse and hamster. Since it has been shown that the intracellular pH regulatory mechanism can be functionally different among mouse strains (e.g., CF1, Balb/c), the aim of this study was to investigate the activity of HCO3-/Cl- exchanger (anion exchanger, AE), which protects cells against alkalosis during the meiotic maturation process, in the GV oocyte up to the pronuclear (PN) zygote derived from the Balb/c strain mouse. Intracellular pH (pHi) was recorded using a microspectrofluorometric technique during meiotic maturation stages. KSOM-based solutions were used as culture and recording solutions. AE activity was determined using a Cl- removal assay and was reported as the change in pHi per minute. AE activity was high in GV stage oocytes but was significantly inhibited at the MI and MII stages. AE activity was higher in the PN zygote stage. This activity was significantly inhibited in all oocyte and zygote stages by 4,4'-Diisocyanatostilbene-2,2'-disulfonic acid disodium salt. After alkalosis induction, the pHi of MI and MII stage oocytes did not completely recover; however, almost complete recovery occurred in the GV stage oocytes and PN zygotes. These results suggest that AE is inhibited during the meiotic maturation process in the Balb/c strain mouse.     

RNAi-mediated knockdown of INHBB increases apoptosis and inhibits steroidogenesis in mouse granulosa cells

Inhibins are members of the TGFβ superfamily and act as suppressors of follicle stimulating hormone (FSH) secretion from pituitary glands via a negative feedback mechanism to regulate folliculogenesis. In this study, the INHBB gene was knocked down by three RNAi-Ready pSIREN-RetroQ-ZsGreen vector- mediated recombinant plasmids to explore the effects of INHBB silencing on granulosa cell (GC) cell cycle, apoptosis and steroid production in vitro. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and ELISA were performed to evaluate the role of INHBB in the mouse GC cell cycle, apoptosis and steroid production in vitro. The results showed that the relative mRNA and protein expression of INHBB in mouse GCs can be significantly reduced by RNAi with pshRNA-B1, pshRNA-B2 and pshRNA-B3 plasmids, with pshRNA-B3 having the best knockdown efficiency. Downregulation of the expression of INHBB significantly arrests cells in the G1 phase of the cell cycle and increases the apoptosis rate in GCs. This was further confirmed by downregulation of the protein expressions of Cyclin D1, Cyclin E and Bcl2, while the protein expression of Bax was upregulated. In addition, specific downregulation of INHBB markedly decreased the concentration of estradiol and progesterone, which was further validated by the decrease in the mRNA levels of CYP19A1and CYP11A1. These findings suggest that inhibin βB is important in the regulation of apoptosis and cell cycle progression in granulosa cells. Furthermore, the inhibin βB subunit has a role in the regulation of steroid hormone biosynthesis. Evidence is accumulating to support the concept that inhibin βB is physiologically essential for early folliculogenesis in the mouse.     

Rapid detection of <Emphasis Type="Italic">Peste des Petits Ruminants</Emphasis> (PPR) virus antigen in Sudan by agar gel precipitation (AGPT) and haemagglutination (HA) Tests

AGPT and HA tests were employed for rapid diagnosis of PPRV infection in sheep and goats in Sudan. Forty lymph nodes and spleen samples from suspected cases of PPR in both sheep and goats were examined by AGPT and HA tests for detection of PPRV antigen. Viral antigen was detected from (77.5%) of the samples tested by AGPT and (92.5%) tested by HA test. The results of both tests revealed that HA test was more sensitive than AGPT for detection of PPRV antigen (Kappa statistics 0.4366). Another advantage of the HA test over AGPT was that it can differentiate PPRV from RPV. Thus the HA test represents a quick, easy, simple, cheap and reliable confirmatory test for the diagnosis of PPR and differential diagnosis of PPRV and RPV. The HA test was carried out using chicken, goat and pig RBCs. Chicken RBCs were found to be the most sensitive for detection of PPRV antigen, followed by goat then pig RBCs. The HA time when using chicken RBCs was 20–25 minutes, using goat RBCs was 25–30 minutes and using pig RBCs was 40–45 minutes. The distribution of PPR infection in four different regions of Sudan was investigated.     

Effects of NaCl salinity on some leaf nutrient concentrations,non-photochemical quenching and the efficiency of the PSII photochemistry of two Iranian pomegranate varieties under greenhouse and field conditions: Preliminary results

The present research was conducted to study the responses of ‘Malas–e–Saveh’ (M) and ‘Shishe–Kab’ (Sh) Iranian pomegranates to sodium chloride (NaCl) stress under greenhouse and field conditions. Treatments included waters electrical conductivity (EC = 1.5, 3, 6, 9 and 12 dS m^?1 for greenhouse) and (EC = 1.05 as control, 4.61 and 7.46 dS m^?1 for field studies). Interactive effects of salinity × variety indicated the highest chlorophyll and leaf potassium concentration, and the lowest leaf chloride and sodium in control under greenhouse study. Non-photochemical quenching, effective quantum yield of photochemical energy conversion in PSII reduced under the highest salinity level in field, however, basal quantum yield of non-photochemical processes in PSII increased in the highest salinity. Sodium and chloride increased with increased in salinity. Calcium, magnesium and iron significantly decreased with increased in salinity. It seems that there are differences between pomegranate cultivars and Malas-e-Saveh is more tolerant compared with Shishe Kab.     

Effect of dietary protein level on the reproductive performance of Nile tilapia, Oreochromis niloticus (L.)

The effects of five dietary protein levels (25%, 30%, 35%, 40% and 45%) on the individual spawning frequency and the egg production of 135 tagged female Nile tilapia, Oreochromis niloticus (L.), were studied from 8 May to 19 November 1996 in outdoor concrete tanks. Virtually identical spawning patterns were found in all treatments, but there was a great deal of variation in the frequency of spawning and egg production. Overall, individual spawning frequency varied from one to 14 and individual egg production from 31 to 2828 eggs per spawning. The average number of spawnings and average number of eggs per spawning for fish receiving dietary protein levels of 25%, 30%, 35%, 40% and 45% were 8.0 ±1.6, 8.4 ± 2.2, 8.5 ± 2.7, 8.4 ± 2.5 and 9.4 ± 2.1 spawnings, and 1167 ± 305, 1082 ± 410, 1288 ± 324, 1145 ± 389 and 1328 ± 311 eggs per spawning, respectively. Fish receiving 45% dietary protein spawned more frequently than fish receiving 25% dietary protein. The total number of eggs produced per female was significantly higher for females fed 45% protein feed than females receiving 25% and 30% protein feeds. No definite trend was found in the number of eggs produced per spawning and the number of eggs produced per gram in females fed at different protein levels. Based on weekly checking, the time interval between spawnings varied from 7 to 77 days, and mean spawning intervals ranged from 15.8 to 17.1 days. Sixty per cent of females spawned after an interval 14 days, 15% after 21 days, 13.6% after 7 days, 7.2% after 28 days, 1.8% after 35 days and 1.0% after 42 days, and the time interval was 49-77 days for the rest of the females (1.5%). In all treatments, maximum spawning activity was recorded from May to August, and thereafter, it gradually decreased and no spawning females were found in November.     

Effects of supplemental dietary l‐carnitine and ractopamine on the performance of juvenile rainbow trout,Oncorhynchus mykiss

This study was carried out to investigate a possible protein‐sparing action of l ‐carnitine and ractopamine in rainbow trout, Oncorhynchus mykiss. An 8‐week feeding trial was carried out to evaluate the effects of supplementation of three levels of l ‐carnitine (0, 1 and 2 g kg^?1) and two levels of ractopamine (0 and 10 mg kg^?1) on growth performance, fillet fatty acid compositions and blood biochemical parameters in a 3 × 2 factorial experimental design. Ractopamine and 1 g kg^?1 carnitine improved the specific growth rate (1.03% and 1.05% day^?1), feed conversion ratio (FCR, 1.3 and 1.29), protein efficiency ratio (PER, 1.88 and 1.85) of fish and crude protein (73.5 and 73.8) content of fish fillet. l ‐carnitine and ractopamine increased the levels of albumin, total protein and globulin in the serum of fish. Apart from eicosapentaenoic acid and docosahexaenoic acid, other fatty acids of fish fillet were increased by ractopamine, while total saturated fatty acids were almost intact. However, the total n‐3 poly unsaturated fatty acids were reduced by l ‐carnitine supplementation (P<0.05). The present study showed that 1 g kg^?1l ‐carnitine and 10 mg kg^?1 ractopamine each can improve the performance of rainbow trout and their combination in diet could enhance the protein level and change the fatty acids profile in fillet muscle.     

Influence of Degree of Hydrolysis on Chemical Composition,Functional Properties,and Antioxidant Activities of Chinese Sturgeon (Acipenser sinensis) Hydrolysates Obtained by Using Alcalase 2.4L

Protein hydrolysates from Chinese sturgeon were prepared using Alcalase 2.4L enzyme. Under the optimum conditions (enzyme–substrate ratio of 3.5%, pH of 8.5, and temperature of 55°C), the degree of hydrolysis (DH) was 13.8%, 16.7%, and 19.1% after 1, 3, and 6 h, respectively. The contents of crude protein and amino acid increased at DH of 19.1% to 86.97% and 78.29%, respectively. There was an obvious increase in the low-molecular-weight peptides, which could enhance the hydrolysate’s functional properties such as solubility, representing more than 90% at different pH levels. The obtained protein hydrolysates revealed good emulsification properties and high oil absorption. Furthermore, good antioxidant activities such as 1, 1-diphenyl-2-picrylhydrazyl (DPPH) inhibition, ferric reducing power, and ferrous ion (Fe²⁺⁾ chelating ability were attained depending on the solution concentration. The findings indicate that the functional and antioxidant properties of protein hydrolysates could be useful in many applications of the food industry.