Calcium-permeable acid-sensing ion channel is a molecular target of the neurotoxic metal ion lead

Acid-sensing ion channels (ASICs) are emerging as fundamental players in the regulation of neural plasticity and in pathological conditions. Here we showed that lead (Pb2+), a well known neurotoxic metal ion, reversibly and concentration-dependently inhib

Fatty acid composition of some brown seaweeds (Phaeophyta) from the coast of Karachi

Ten species of filamentous and multicellular thalloid algae (Phaeophyta) belonging to six genera of the class Dictyophyceae were collected from different seawater habitats of Karachi, Pakistan at the northern boundary of the Arabian Sea during September 1997 and July 1998. They were extracted in chloroform: methanol, saponified, subjected to column chromatography (CC, TLC), esterified and analysed for fatty acid (FA) composition initially by gas-liquid-chromatography (GLC) and finally by gas chromatography-mass spectrometry (GC-MS). Algae of the classes Laminariophyceae and Fucophyceae (Phaeophyta) displayed only a few SCFAs, PUFAs and substituted FAs, no VLCFA, C22 UFA, CFA, DCFA and monoynoic FA, large amount of C16:0, very large quantity of C18:1, very small RCCL and FA-diversity, C18 UFAs up to four DBs, C20 UFAs up to three DBs only. They were characterized by the largest amount of C18:1, lowest degree of unsaturation of C20 UFAs, lack of C22 UFAs, the shortest RCCL and the smallest FA-diversity as compared to other phyla.

Fatty acid composition of more brown seaweeds (Phaeophyta) from the coast of Karachi

Six species of multicellular, thalloid, brown algae, belonging to five genera of the classes Laminariophyceae and Fucophyceae were collected from the coastal areas of Buleji near Karachi (Pakistan) during October 1997 and February 1998. They were extracted in chloroform: methanol, saponified, subjected to column chromatography (CC, TLC), esterified and analysed for fatty acid (FA) composition initially by gas-liquid-chromatography (GLC) and finally by gas chromatography-mass spectrometry (GC-MS). They displayed only a few SCFAs, PUFAs and substituted FAs, no VLCFA, C22 UFA, CFA, DCFA and monoynoic FA, large amount of CI6:0, very large quantity of C18:1, very small RCCL and FA-diversity, C18 UFAs up to four DBs, C20 UFAs up to three DBs only. They were characterized by the largest amount of C18:1, lowest degree of unsaturation of C20 UFAs, lack of C22 UFAs, the shortest RCCL and the smallest FA-diversity as compared to other phyla.

Determination of proximate composition, shelf life and fatty acid profiles of breaded kilka (Clupeonella cultriventris) during frozen storage

The present study aimed production of a new product with various texture and sensory properties in chase of the impetus for increasing human consumption considering suitable resources of Kilka fish in Caspian Sea. Following deheading, gutting, and brining, common Kilka were battered in two different formulations, i.e. simple batter and tempura batter, via automated predusting machinery and then, they were fried through flash frying for 30 seconds at 170°C in sunflower oil after they were breaded with bread crumbs flour. The products were subjected to continuous freezing at -40°C and were kept at -18°C in cold storage for four months once they were packed. Chemical composition (protein, fat, moisture, and ash), fatty acid profiles (29 fatty acids), chemical indices of spoilage (peroxide value, thiobarbituric acid, free fatty acids, and volatile nitrogen), and microbial properties (total bacteria count and coliform count) were compared in fresh and breaded Kilka at various times before frying (raw breaded Kilka), after frying (zero-phase), and in various months of frozen storage (phases 1, 2, 3, and 4). Organoleptic properties of breaded Kilka (i.e. odor, taste, texture, crispiness, cohesiveness of batter) and general acceptability in the phases 0, 1, 2, 3, and 4 were evaluated. The results obtained from chemical composition and fatty acid profiles in common Kilka denoted that MUFA, PUFA, and SFA were estimated to be 36.96, 32.85, and 29.12 g / 100g lipid, respectively. Levels of ù-3 and ù-6 were 7.6 and 1.12 g / 100 gr lipid, respectively. Docosahexaonoic acid (20.79%) was the highest fatty acid in PUFA group. ù-3/ù-6 and PUFA/SFA ratios were 7.6 and 1.12, respectively. The high rates of the indices and high percentage of ù-3 fatty acid in common Kilka showed that the fish can be considered as invaluable nutritional and fishery resources and commonsensical consumption of the species may reduce the risk of cardiovascular diseases. Frying breaded Kilka affected overall fat and moisture contents so that moisture content in fried breaded Kilka decreased significantly compared to raw breaded Kilka, while it was absolutely reverse for fat content. Overall fat content in tempura batter treatment was significantly lower than that of simple batter treatment (P≤0.05). Presence of hydrocolloids, namely proteins, starch, gum, and other polysaccharides, in tempura batter may prohibit moisture evaporation and placement with oil during frying process in addition to boosting water holding capacity through confining water molecules. During frying process, fatty acids composition of breaded Kilka with various batters changed so that rates of some fatty acids such as Palmitic acid (C16:0), Stearic acid (C18:0), Oleic acid (C18:1 ù-9cis), and linoleic acid (C18:3 ù-3) increased considerably following frying; however, ù-3/ù-6, PUFA/SFA, and EPA+DHA/C16:0 ratios (Polyan index) decreased significantly after frying. ù-3/ù-6, PUFA/SFA, and EPA+DHA/C16:0 ratios in tempura batter treatment were higher than those of simple batter treatment which is an indicator of higher nutritional value of breaded Kilka with tempura batter. Significant elevations were found in peroxide, thiobarbituric acid, and free fatty acids in fried breaded Kilka samples compared to raw samples which points to fat oxidation during cooking process. Overall microorganism count and coliform count decreased following heating process. Both breaded Kilka samples were of high sanitation quality at zero-phase according to ICMSF Standard. The results acquired from organoleptic evaluation declared that odor, cohesiveness, and general acceptability indices, among others, had significant differences between the treatments (P≤0.05). In all evaluated properties, breaded Kilka with tempura batter in different phases gained higher scores than breaded Kilka with simple batter. During cold storage of various treatments of breaded Kilka, total lipid content, PUFA, MUFA, ù-3, ù- 3/ù-6, PUFA/SFA, Polyen index decreased significantly. The mentioned reductions in addition to significant elevation of spoilage indices, namely peroxide, thiobarbituric acid, and free fatty acids, during frozen storage, indicate to oxidation and enzymatic mechanism activity during frozen storage of breaded Kilka. Considering sensory evaluation at the end of the fourth month and TVB-N contents exceeded eligible rate in the fourth month, shelf life of the products during frozen storage was set to be three months at -18°C. The results obtained from statistical tests indicate to better quality of breaded Kilka processed with tempura batter compared to simple batter in terms of organoleptic evaluation, spoilage indices, and high quality of fat in various sampling phases.

Supply of Astaxanthin and its combinations through live feed (Moina micrura) enrichment affects the growth, survival and fatty acid profile of Macrobrachium rosenbergii larvae

A study was carried out with three replicates to determine the effects of feeding Moina micrura enriched with astaxanthin alone (M1) or astaxanthin in combination with either vitamin E (M2), vitamin D (M3) or Cod Liver oil (M4) on the growth, survival and fatty acid composition of giant fresh water prawn Macrobrachium rosenbergii (de Man) larvae. Growth rate was expressed as the time taken to the settlement of 95% post larvae. Maximum growth, the lowest time taken to the 95% PL settlement (38.5±0.50 days), was observed in larvae fed with M3 Moina. The highest survival rate (66.0±1.00%) was observed in those fed with M4 Moina and the second highest survival (61.0±1.00%) and growth rates (40.0±0.00 days) were shown with M2 Moina. The minimum values for both growth (42.5±0.50 days) and survival (33.0±1.50%) were observed in the group fed un-enriched Moina. Results also showed that the survival of prawn larvae increased as the quantities of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) increased in the dietary Moina. The highest levels of EPA (5.57±0.21%), DHA (3.50±0.21%) and highest total Highly Unsaturated Fatty Acids (HUFA) (13.87±0.68%) were seen in the Moina fed on astaxanthin and Cod Liver Oil (CLO). The results of the study showed that the nutritive quality of Moina, with respect to important fatty acids, can be increased by enrichment and will influence the growth, survival and the fatty acid composition of fresh water prawn larvae fed on them.

Composition and amino acid content of meal from some commercially less important fishes of Maharashtra

Details are given of the yield and composition of dried waste from the filleting wastes of 3 commercially less utilized fish of the Maharashtra coast (Saurida tumbil, Caranx sexfasciatus and Sphyraena jello). The amino acid composition after acid hydrolysis is detailed for the three species.

Amino acid requirements for maturation of rhesus monkey (Macacca mulatta) oocytes in culture

This study evaluated the effects of different amino acid formulations on supporting meiotic and cytoplasmic maturation of rhesus monkey (Macacca mulatta) oocytes in vitro. Five hundred and forty-six cumulus-oocyte complexes (COCs) aspirated from unstimulated adult monkey follicles (greater than or equal to 1000 mum in diameter) were cultured in either modified Connaught Medical Research Laboratories 1066 medium (mCMRL-1066) or in one of eight chemically defined media (modified basic medium 5 supplemented with 5.5 mmol glucose l(-1), 0.003 mmol pantothenic acid l(-1) and different amino acid formulations) as below: (1) modified basic medium 5 (mBM5) containing no amino acid; (2) mBM5 + 0.2 mmol glutamine l(-1); (3) mBM5 + 11 amino acids from hamster embryo culture medium 6 (HECM-6) (11 AA); (4) mBM5 + Eagle's non-essential amino acids (NEA); (5) mBM5 + NEA + 0.2 mmol glutamine l(-1); (6) mBM5 + Eagle's essential amino acids (EA) without glutamine; (7) mBM5 + EA + 0.2 mmol glutamine l(-1); (8) mBM5 + Eagle's 20 amino acids (20 AA) + 0.2 mmol glutamine l(-1); and (9) mCMRL-1066 (control). All media contained FSH, LH, oestradiol and progesterone. After maturation, mature oocytes were subjected to the same fertilization and embryo culture procedures. COCs matured in treatment 5 had greater potential to progress to metaphase II (66%; P < 0.05) than did those in treatments 1 (37.3%), 2 (48.3%)f 3 (41%), 6 (41%) and 9 (43%). Oocytes matured in treatment 8 had the best morula (53%) and blastocyst (18%) developmental responses (P<0.05). The lowest (P<0.05) morula and blastocyst developmental responses were obtained from COCs matured in treatments 1 (0%) and 6 (8%). The other media supported intermediate embryonic development (range 11-38% of morula and blastocyst). These results indicate that the choice of amino acids affects the competence of oocyte maturation and that Eagle's 20 AA with 0.2 mmol glutamine l(-1) is more efficient than the other amino acid formulations for maturation of rhesus monkey oocytes.

The study of Indian shrimp larvae feeding by enriched rotifer with vitamin C and unsaturated fatty acid (epa, dha) and its effects on growth factors, survival rate and environmental stresses

In this experiment, the feeding of Indian white shrimp larvae by unenriched rotifers (treatment 1) and enriched with highly unsaturated fatty acid (treatment 2) and highly unsaturated fatty acid along with vitamin C (treatment 3) on the growth factors, survival and resistance against salinity and formalin stress tests were studied and their differences with control treatment including newly hatched Artemia nauplii is compared. In this the study four treatments in a completely randomized design with 3 replicates per treatment were used. Farming of shrimp larvae of Zoea II to postlarvae 5 was done in 20 liter plastic bucket. Present results indicated that growth factors and survival rate of stage Zoea II to postlarvae 1 in treatments 1, 2 and 3 improve rather than control in which this case was due to optimal size rotifer rather than Artemia nauplii. Also, treatments 2 and 3 feeding with oil liver cod emulsion enriched rotifer have the highest concentration of DHA (mg/g DW) and the ratio DHA/EPA in which due to have shown the highest growth factors and a significant difference (P<0.05) with treatments 1 and control. The highest survival at stage PL1 were observed in treatment 3 that was enriched with ascorbyl palmitate in which have to the synergistic properties of vitamin C rather than treatments 2, 1 and control and showed a significant difference (P<0.05). But in stage PL5 the highest amount of growth and survival rates were related to control treatment which showed a significant difference (P<0.05) with other treatments that control has higher size rather than treatments 1, 2 and 3. Also, among experiment treatments that the two treatments 2 and 3 due to enrichment had higher growth and survival rates compared with treatment 1 in which their differences have also been significant (P<0.05). In the case of stress tests, results indicated that the highest survival rate has been reported when specimens were offered a diet containing high levels of highly unsaturated fatty acids with vitamin C. So that in stage PL1 in the salinity stress tests 10 and 20 ppt the highest survival rate was observed in treatment 3. As for the second, treatment 2 showed a significant difference (P<0.05) with treatment 3. It is worth mentioning that treatment 3 showed a higher survival rate compared to treatment 2 due to the synergistic properties of vitamin C. The difference between these two treatments with treatment 1 and control was also significant. No significant difference was observed in formalin stress test 100 ppm in this stage between treatments 3 and 2 which shows the highest survival rate. But their difference with treatments 1 and control was significant (P<0.05). Also, in stage PL5 in the salinity stress tests 10 and 20 ppt the highest survival rate was observed in treatment 3 which showed no significant difference (P<0.05) with control treatment. While their difference in the amount of survival rate with treatment 1 and 2 was significant (P<0.05). In this stage, the highest observed survival rate in formalin stress test 100 ppm included treatments control, 3 and 2 among which there were no significant differences (P<0.05). While the difference between these three treatments with treatment 1 was significant.

Characterization of acid-sensing ion channels in dorsal horn neurons of rat spinal cord

Acid-sensing ion channels (ASICs) are ligand-gated cation channels activated by extracellular protons. In periphery, they contribute to sensory transmission, including that of nociception and pain. Here we characterized ASIC-like currents in dorsal horn neurons of the rat spinal cord and their functional modulation in pathological conditions. Reverse transcriptase-nested PCR and Western blotting showed that three ASIC isoforms, ASIC1a, ASIC2a, and ASIC2b, are expressed at a high level in dorsal horn neurons. Electrophysiological and pharmacological properties of the proton-gated currents suggest that homomeric ASIC1a and/or heteromeric ASIC1a + 2b channels are responsible for the proton-induced currents in the majority of dorsal horn neurons. Acidification-induced action potentials in these neurons were compatible in a pH-dependent manner with the pH dependence of ASIC-like current. Furthermore, peripheral complete Freund's adjuvant-induced inflammation resulted in increased expression of both ASIC1a and ASIC2a in dorsal horn. These results support the idea that the ASICs of dorsal horn neurons participate in central sensory transmission/modulation under physiological conditions and may play important roles in inflammation-related persistent pain.

Coupling between NMDA receptor and acid-sensing ion channel contributes to ischemic neuronal death

Acid-sensing ion channels (ASICs) composed of ASIC1a subunit exhibit a high Ca2+ permeability and play important roles in synaptic plasticity and acid-induced cell death. Here, we show that ischemia enhances ASIC currents through the phosphorylation at Ser478 and Ser479 of ASIC1a, leading to exacerbated ischemic cell death. The phosphorylation is catalyzed by Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity, as a result of activation of NR2B-containing N-methyl-D-aspartate subtype of glutamate receptors (NMDARs) during ischemia. Furthermore, NR2B-specific antagonist, CaMKII inhibitor, or overexpression of mutated form of ASIC1a with Ser478 or Ser479 replaced by alanine (ASICla-S478A, ASIC1a-S479A) in cultured hippocampal neurons prevented ischemia-induced enhancement of ASIC currents, cytoplasmic Ca2+ elevation, as well as neuronal death. Thus, NMDAR-CaMKII cascade is functionally coupled to ASICs and contributes to acidotoxicity during ischemia. Specific blockade of NMDAR/CaMKII-ASIC coupling may reduce neuronal death after ischemia and other pathological conditions involving excessive glutamate release and acidosis.

Upregulation of acid-sensing ion channel ASIC1a in spinal dorsal horn neurons contributes to inflammatory pain hypersensitivity

Development of chronic pain involves alterations in peripheral nociceptors as well as elevated neuronal activity in multiple regions of the CNS. Previous pharmacological and behavioral studies suggest that peripheral acid-sensing ion channels (ASICs) cont

Nutritive value of fish of Lake Tanganyika [part] I: Amino acid composition

The amino acid composition was determined of three economic important fish species of Lake Tanganyika, representing 95% of the tolal catch:Stolothrissa tanganicae Regan, Limnothrissa miodon Boulenger and Luciolates stapersii Boulenger. The analysis of the whole sun dried fish indicate that, compared with Tilapia macrochir, beef and the whole hen's egg, the fish is of a very high nutritive value. The free amino acids in the dried fish count for an average of 11 %, indicating the degree of auteolysis.

Effects of gamma-aminobutyric acid, progesterone and ionophore A23187 on acrosome reaction of tree shrew sperm in vitro: examination of acrosome reaction with an improved fluorescence microscopy

A number of acrosome reaction (AR) initiators have been found to be effective in inducing AR of human, laboratory and domestic animal sperm. Using an improved simple fluorescence microscopy, effects of gamma-aminobutyric acid (GABA), progesterone and ionophore A23187 on sperm AR of tree shrew, a useful animal model in biomedical research, have been investigated. Spontaneous AR in 4.92-7.53% of viable sperm was observed. Complete AR in 10.31-18.25% of viable tree shrew sperm was obviously induced by 5 mu M and 10 mu M calcium ionophore A23187, 1 mM GABA, and 5 mu M progesterone, and there were no significant differences between their abilities to initiate complete AR. No significant differences of AR percentages between 1- and 2-h treatments with A23187, progesterone and/or GABA were observed. These results suggested that the responses of tree shrew sperm to these AR initiators are similar to that of human and other mammalian sperm. (C) 1997 Elsevier Science B.V.

Purification, characterization and biological activities of the L-amino acid oxidase from Bungarus fasciatus snake venom

L-Amino acid oxidases (LAAOs) are widely distributed in snake venoms, which contribute to the toxicity of venoms. However, LAAO from Bungarus fasciatus (B. fasciatus) snake venom has not been isolated previously. In the present study, LAAO from B. fasciat