Study on the pathophysiology of Haemonchus contortus (Rudolph, 1803) in calves with the use of radioisotopes

International Nuclear Information System (INIS)

Gennari, S.M.

1989-01-01

This study was conducted to investigate the pathophysiology of Haemonchus contortus infection in friesian calves. The animals were divided in two groups of four calves each. One group was infected, orally, with 500 Haemonchus contortus larvae per kg body weight and the other one as the worm-free control. Five weeks after infection they were all housed in metabolic crates and injected with 125 I-bovine albumin, 51 Cr-red cell and 59 Fe-transferrin to study the albumin metabolism, the erythokinetic and ferrokinetic. The results showed that there was a significant reduction in the mean haematocrit values and an evidence of lower weight gains over the period of experiment in the infected calves compared with the controls. There was also a change in the distribution of albumin from the extravascular to the intravascular pool and a significant increase in the plasma and blood volumes of infected calves. The study of ferrokinetic showed an increase in the iron turnover and in the rate of iron utilization in the infected group. The blood and albumin loss by the gastrointestinal tract were similar in both groups. (author)

Effectiveness of Ivermectin and Albendazole against Haemonchus contortus in Sheep in West Java, Indonesia.

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Puspitasari, Silvia; Farajallah, Achmad; Sulistiawati, Erni; Muladno

2016-02-01

Administering a half dose of an anthelmintic is a simple method for detecting resistance in parasites infesting small ruminants. When a single anthelmintic fails in native sheep from Indonesia, a combination of anthelmintics from different chemical classes with different modes of action are administered as an alternative parasite-control strategy. This study compared the anthelmintic efficacy of ivermectin (IVM) and albendazole (ABZ) given either separately as a single dose or half dose or co-administered to sheep naturally infected with Haemonchus contortus. Twelve sheep from Bogor, West Java, Indonesia were divided into the following six treatment groups: half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, combined IVM + ABZ, and control. The treatment efficacy was determined using the faecal egg count reduction test (FECRT) at day 0 (pre-treatment) and post-treatment at days 7, 14, 21, 28, 35, and 42. The efficacies of half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, and the combination treatment ranged from -1900% to 100%, 99% to 100%, -167% to 100%, -467% to 89%, and -200% to 100%, respectively. The FECRT for the half-dose IVM, half-dose ABZ, full-dose ABZ showed that H. contortus is resistant to half-dose IVM and ABZ. Full-dose IVM was effective against H. contortus. The combined treatment was more effective against H. contortus than ABZ alone.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay.

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Hördegen, P; Cabaret, J; Hertzberg, H; Langhans, W; Maurer, V

2006-11-03

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyl-tetrazolium (MTT) reduction assay. Pyrantel tartrate was used as reference anthelmintic. Bromelain, the enzyme complex of the stem of Ananas comosus (Bromeliaceae), the ethanolic extracts of seeds of Azadirachta indica (Meliaceae), Caesalpinia crista (Caesalpiniaceae) and Vernonia anthelmintica (Asteraceae), and the ethanolic extracts of the whole plant of Fumaria parviflora (Papaveraceae) and of the fruit of Embelia ribes (Myrsinaceae) showed an anthelmintic efficacy of up to 93%, relative to pyrantel tartrate. Based on these results obtained with larval Haemonchus contortus, the modified MTT reduction assay could be a possible method for testing plant products with anthelmintic properties.

Antigenic Cross-reactivity among Haemonchus contortus, Oesophagostomum columbianum and Trichuris ovis of Goat

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Ruma JAS

2016-12-01

Full Text Available Background: Cross antigenicity is the major problem in developing a reliable tool for immunodiagnosis and immunoprophylaxis of parasitic diseases. Mixed infection due to different types of gastrointestinal parasites is more common than single species infection under field condition.Methods: The present study was undertaken to detect antigenic cross-reactivity among Haemonchus contortus, Oesophagostomum columbianum and Trichuris ovis of goats by SDS-PAGE and western blot analysis using hyperimmune sera (HIS rose in rabbit separately against the antigens of the three nematode species.Results: Thirteen, 16 and 14 polypeptides in crude somatic antigen (CSAg of H. contortus (CSAg-Hc, O. columbianum (CSAg-Oc and T. ovis (CSAg-To, respectively, were resolved in SDS PAGE analyses. It was revealed that 54 kDa peptide was shared by H.contortus and O. columbianum, whereas 47 kDa peptide was shared by O. columbianum and T. ovis. Western blot analyses revealed that three immunogenic polypeptides (MW 54, 49 and 42 kDa in CSAg-Hc, five in CSAg-Oc (54, 47, 44, 38 and 35.5 kDa and CSAg-To and five polypeptides (90, 51, 47, 39.5 and 31 kDa in CSAg-To cross-reacted with the heterologous HIS. Four species-specific immunoreactive polypeptides (92, 85, 65 and 39 kDa of H. contortus and two (72 & 26 kDa in O. columbianum were also identified in the study. Conclusion: The shared polypeptides and species-specific polypeptides might be evaluated as protective antigen and subsequently exploitation for developing immunodiagnostic and for immunoprophylactic tools of for these common nematode species. 

Antigenic Cross-reactivity among Haemonchus contortus, Oesophagostomum columbianum and Trichuris ovis of Goat.

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Jas, Ruma; Ghosh, Joydeb; DAS, Kinsuk

2016-01-01

Cross antigenicity is the major problem in developing a reliable tool for immunodiagnosis and immunoprophylaxis of parasitic diseases. Mixed infection due to different types of gastrointestinal parasites is more common than single species infection under field condition. The present study was undertaken to detect antigenic cross-reactivity among Haemonchus contortus, Oesophagostomum columbianum and Trichuris ovis of goats by SDS-PAGE and western blot analysis using hyperimmune sera (HIS) rose in rabbit separately against the antigens of the three nematode species. Thirteen, 16 and 14 polypeptides in crude somatic antigen (CSAg) of H. contortus (CSAg-Hc), O. columbianum (CSAg-Oc) and T. ovis (CSAg-To), respectively, were resolved in SDS PAGE analyses. It was revealed that 54 kDa peptide was shared by H.contortus and O. columbianum , whereas 47 kDa peptide was shared by O. columbianum and T. ovis . Western blot analyses revealed that three immunogenic polypeptides (MW 54, 49 and 42 kDa) in CSAg-Hc, five in CSAg-Oc (54, 47, 44, 38 and 35.5 kDa) and CSAg-To and five polypeptides (90, 51, 47, 39.5 and 31 kDa) in CSAg-To cross-reacted with the heterologous HIS. Four species-specific immunoreactive polypeptides (92, 85, 65 and 39 kDa) of H. contortus and two (72 & 26 kDa) in O. columbianum were also identified in the study. The shared polypeptides and species-specific polypeptides might be evaluated as protective antigen and subsequently exploitation for developing immunodiagnostic and for immunoprophylactic tools of for these common nematode species.

Safety and serologic response to a Haemonchus contortus vaccine in alpacas.

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VanHoy, Grace; Carman, Michelle; Habing, Greg; Lakritz, Jeffrey; Hinds, C Austin; Niehaus, Andrew; Kaplan, Ray M; Marsh, Antoinette E

2018-03-15

Haemonchosis in camelids remains a challenging disease to treat, and prevention has become increasingly problematic due to widespread anthelmintic resistance. Barbervax ® is an adjuvanted vaccine containing natural H-11, H-gal-GP antigens obtained from Haemonchus contortus adults via a proprietary process and solubilized in Quil A. This vaccine is approved for use in Australia, after demonstrating its safety and efficacy in sheep and goats. There are no published studies evaluating Barbervax in other ruminants/pseudoruminants such as camelids which can be parasitized with H. contortus. The vaccine utilizes a mixture of the parasite gut mucosal membrane enzymes including H-gal-GP and H11, involved in digesting a blood meal from the host. This study monitored the safety profile of the Barbervax ® vaccine in a group of adolescent alpacas. Although designed into the original study of vaccine efficacy, the experimental infection with viable H. contortus third stage larvae could not be completed due to lack of detectable significant variation of infection following experimental challenge. Twelve alpacas (158 + 15 days) were randomized to vaccination with Barbervax ® or no treatment. Three doses of Barbervax ® were administered at 3 week intervals and investigators involved in animal monitoring and sample collection were blinded to the groupings. Clinical pathologic parameters were evaluated 7 days before vaccination, and 1 and 2 months post-vaccination. Daily clinical observations were made and specific observations regarding the injection site and rectal temperatures were monitored in each alpaca twice daily for 1 week following vaccination. Fecal egg counts, packed cell volume, and total protein were monitored following challenge with 1500 H. contortus larvae on days 42, 46, and 50. An increase in rectal temperature for a duration of 2 days (range 2-4 days) was observed post-vaccination. Vaccinated alpacas were lethargic for 2-3 days following vaccination

Effectiveness of Ivermectin and Albendazole against Haemonchus contortus in Sheep in West Java, Indonesia

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Puspitasari, Silvia; Farajallah, Achmad; Sulistiawati, Erni; Muladno

2016-01-01

Administering a half dose of an anthelmintic is a simple method for detecting resistance in parasites infesting small ruminants. When a single anthelmintic fails in native sheep from Indonesia, a combination of anthelmintics from different chemical classes with different modes of action are administered as an alternative parasite-control strategy. This study compared the anthelmintic efficacy of ivermectin (IVM) and albendazole (ABZ) given either separately as a single dose or half dose or co-administered to sheep naturally infected with Haemonchus contortus. Twelve sheep from Bogor, West Java, Indonesia were divided into the following six treatment groups: half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, combined IVM + ABZ, and control. The treatment efficacy was determined using the faecal egg count reduction test (FECRT) at day 0 (pre-treatment) and post-treatment at days 7, 14, 21, 28, 35, and 42. The efficacies of half-dose IVM, full-dose IVM, half-dose ABZ, full-dose ABZ, and the combination treatment ranged from −1900% to 100%, 99% to 100%, −167% to 100%, −467% to 89%, and −200% to 100%, respectively. The FECRT for the half-dose IVM, half-dose ABZ, full-dose ABZ showed that H. contortus is resistant to half-dose IVM and ABZ. Full-dose IVM was effective against H. contortus. The combined treatment was more effective against H. contortus than ABZ alone. PMID:27019686

Morphological variation between isolates of the nematode Haemonchus contortus from sheep and goat populations in Malaysia and Yemen.

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Gharamah, A A; Rahman, W A; Siti Azizah, M N

2014-03-01

Haemonchus contortus is a highly pathogenic nematode parasite of sheep and goats. This work was conducted to investigate the population and host variations of the parasitic nematode H. contortus of sheep and goats from Malaysia and Yemen. Eight morphological characters were investigated, namely the total body length, cervical papillae, right spicule, left spicule, right barb, left barb, gubernaculum and cuticular ridge (synlophe) pattern. Statistical analysis showed the presence of morphological variation between populations of H. contortus from Malaysia and Yemen, with minor variation in the synlophe pattern of these isolates. Isolates from each country were grouped together in the scatterplots with no host isolation. Body, cervical papillae and spicule lengths were the most important characters that distinguished between populations of the two countries. This variation between Malaysia and Yemen may be attributed to geographical isolation and the possible presence of a different isolate of this worm in each country.

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus.

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Sandra Alves Araújo

Full Text Available Gastrointestinal nematodes are a significant concern for animal health and well-being, and anthelmintic treatment is mainly performed through the use of chemical products. However, bioactive compounds produced by plants have shown promise for development as novel anthelmintics. The aim of this study is to assess the anthelmintic activity of protein fractions from Spigelia anthelmia on the gastrointestinal nematode Haemonchus contortus. Plant parts were separated into leaves, stems and roots, washed with distilled water, freeze-dried and ground into a fine powder. Protein extraction was performed with sodium phosphate buffer (75 mM, pH 7.0. The extract was fractionated using ammonium sulfate (0-90% and extensively dialyzed. The resulting fractions were named LPF (leaf protein fraction, SPF (stem protein fraction and RPF (root protein fraction, and the protein contents and activities of the fractions were analyzed. H. contortus egg hatching (EHA, larval exsheathment inhibition (LEIA and larval migration inhibition (LMIA assays were performed. Proteomic analysis was conducted, and high-performance liquid chromatography (HPLC chromatographic profiles of the fractions were established to identify proteins and possible secondary metabolites. S. anthelmia fractions inhibited H. contortus egg hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1. During LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg mL-1. According to LMIA, the fractions from roots, stems and leaves also reduced the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1, respectively. Protein analysis indicated the presence of plant defense proteins in the S. anthelmia fractions, including protease, protease inhibitor, chitinase and others. Conversely, secondary metabolites were absent in the S. anthemia fractions. These results suggest that S. anthelmia proteins are promising for the control of the gastrointestinal nematode H

Effect of sericea lespedeza leaf meal pellets on adult female Haemonchus contortus in goats.

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Kommuru, D S; Whitley, N C; Miller, J E; Mosjidis, J A; Burke, J M; Gujja, S; Mechineni, A; Terrill, T H

2015-01-15

Sericea lespedeza (SL; Lespedeza cuneata) is a perennial warm-season forage rich in condensed tannins (CT) that has been reported to have anthelmintic activity against small ruminant gastrointestinal nematodes (GIN), particularly Haemonchus contortus, a highly pathogenic blood-feeder, but the mechanism of action of CT against H. contortus is not clearly understood. An experiment with young goats was designed to study the effect of SL leaf meal pellets on (1) a mature H. contortus infection, and (2) the surface appearance of adult H. contortus female worms. Thirty-six female and castrated male Boer crossbred goats artificially infected with H. contortus larvae were fed 75% SL leaf meal pellets or alfalfa pellets (18 goats/treatment group) in a 28-day confinement feeding trial. Fecal and blood samples were collected weekly for fecal egg count (FEC) and packed cell volume (PCV) determination, respectively, and all goats were slaughtered at the end of the trial for adult GIN recovery and counting. Five adult female H. contortus were recovered from the abomasum of two goats from each treatment group and from a prior study in which 75% and 95% SL leaf meal pellets or a commercial feed pellet were group-fed to grazing goats (270 days old, Spanish males, 10/treatment group) at 0.91 kg/head/d for 11 weeks. Adult GIN collected were fixed and examined for evidence of surface damage using scanning electron microscopy. Feeding 75% SL pellets to young goats in confinement reduced (P<0.05) FEC compared with control animals, while total worm numbers and PCV were not influenced by treatment. Three out of the 5 adult H. contortus recovered from SL treatment goats in the confinement feeding trial had cuticular surface damage, while no damage was observed on worms from the control group. All five worms observed from both SL treatments in the grazing study showed a shrunken, disheveled cuticular surface, whereas this was not observed on worms from control animals. Overall, this work

Survival and infectivity studies of in-vitro cultivated larvae of Haemonchus contortus in Sheep and Goats in Nigeria

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A O Sonibare

Full Text Available The survival and infectivity of indigenous isolates of Haemonchus contortus and in vitro cultivated infective larvae (L3 were studied. Active motile gravid females, adult and immature parasites were morphologically and morphometrically characterized and isolated from Haemonchus infected goats. These were subsequently inoculated into the abomasum of parasite free lambs through left sided laparatomy for production of fertile eggs to be used for coproculture. Harvested faeces from these lambs were cultured in a nutritive medium and kept in the incubator at 320C and room temperature 24-280C to obtain in vitro cultivated L of H. contortus. The infectivity and survival of in vitro cultivated L3 stored in refrigerator at 4-80C and under room temperature 24-280C were evaluated. Incubation period of 3 coproculture was observed to be shorter in medium at 320C than under room temperature 24-280C. The storage of in vitro cultivated L under different temperature range showed that the quantity of active larvae in a given volume 3 decreased at p<0.05 from the 5th day under 24-280C while at 4-80C became significantly lower from 20th day. Infectivity of isolate kept at 4-80C was sustained for 18-20 days while at 24-280C it was maintained for 10-12 days. Laboratory invitro cultivated L of H. contortus had limited infectivity and larvae mortality increase with time under different 3 temperatures. [Vet. World 2011; 4(12.000: 533-536

In vitro and in vivo Nematocidal Activity of Allium sativum and Tagetes erecta Extracts Against Haemonchus contortus.

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Palacio- Landín, Josefina; Mendoza-de Gives, Pedro; Salinas-Sánchez, David Osvaldo; López-Arellano, María Eugenia; Liébano-Hernández, Enrique; Hernández-Velázquez, Victor Manuel; Valladares-Cisneros, María Guadalupe

2015-12-01

In the Mexican ethno-medicine, a number of plants have shown a successful anthelmintic activity. This fact could be crucial to identify possible green anti-parasitic strategies against nematodes affecting animal production. This research evaluated the in vitro and in vivo nematocidal effects of two single and combined plant extracts: bulbs of Allium sativum (n-hexane) and flowers of Tagetes erecta (acetone). The in vivo assay evaluated the administration of extracts either individually or combined against Haemonchus contortus in experimentally infected gerbils. The in vitro larvicidal activity percentage (LAP) of A. sativum and T. erecta extracts against H. contortus (L3) was determined by means of individual and combined usage of the extracts. Similarly, the extracts were evaluated in terms of reduction in the parasitic population in gerbils infected with H. contortus by individual and combined usage. The LAP at 40 mg/mL was 68% with A. sativum and 36.6% with T. erecta. The combination caused 83.3% mortality of parasites. The oral administration of A. sativum and T. erecta extracts at 40 mg/mL, caused 68.7% and 53.9% reduction of the parasitic burden, respectively. Meanwhile, the combined effect of both extracts shown 87.5% reduction. This study showed evidence about the effect of A. sativum and T. erecta plant extracts by means of individual and combined usage against H. contortus in in vitro and in vivo bioassays in artificially H. contortus-infected gerbils as a model.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay

OpenAIRE

Hördegen, P.; Cabaret, J.; Hertzberg, H.; Langhans, W.; Maurer, V.

2006-01-01

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyltetrazolium (MTT) reduction assay. Pyrantel tartrate was u...

Climate-driven changes to the spatio-temporal distribution of the parasitic nematode, Haemonchus contortus, in sheep in Europe.

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Rose, Hannah; Caminade, Cyril; Bolajoko, Muhammad Bashir; Phelan, Paul; van Dijk, Jan; Baylis, Matthew; Williams, Diana; Morgan, Eric R

2016-03-01

Recent climate change has resulted in changes to the phenology and distribution of invertebrates worldwide. Where invertebrates are associated with disease, climate variability and changes in climate may also affect the spatio-temporal dynamics of disease. Due to its significant impact on sheep production and welfare, the recent increase in diagnoses of ovine haemonchosis caused by the nematode Haemonchus contortus in some temperate regions is particularly concerning. This study is the first to evaluate the impact of climate change on H. contortus at a continental scale. A model of the basic reproductive quotient of macroparasites, Q0 , adapted to H. contortus and extended to incorporate environmental stochasticity and parasite behaviour, was used to simulate Pan-European spatio-temporal changes in H. contortus infection pressure under scenarios of climate change. Baseline Q0 simulations, using historic climate observations, reflected the current distribution of H. contortus in Europe. In northern Europe, the distribution of H. contortus is currently limited by temperatures falling below the development threshold during the winter months and within-host arrested development is necessary for population persistence over winter. In southern Europe, H. contortus infection pressure is limited during the summer months by increased temperature and decreased moisture. Compared with this baseline, Q0 simulations driven by a climate model ensemble predicted an increase in H. contortus infection pressure by the 2080s. In northern Europe, a temporal range expansion was predicted as the mean period of transmission increased by 2-3 months. A bimodal seasonal pattern of infection pressure, similar to that currently observed in southern Europe, emerges in northern Europe due to increasing summer temperatures and decreasing moisture. The predicted patterns of change could alter the epidemiology of H. contortus in Europe, affect the future sustainability of contemporary

Effects of commonly used chemical fertilizers on development of free-living stages of Haemonchus contortus in experimentally infected pasture

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Tapas Kumar Roul

2017-07-01

Full Text Available Aim: The effects of N-P-K fertilizers in the form of urea, single super phosphate and muriate of potash on development of free-living stages of Haemonchus contortus were studied. Materials and Methods: Five parasite free experimental plots of 1 mx1 m area, each of paddy leaves (15-day-old and an equal number of Cynodon dactylon grass were infested with about 10x104 eggs/ml phosphate buffer saline along with the application of the calculated amount of fertilizers solution. On the 10th day of posttreatment, the pasture was cut, processed, larvae recovered by Baermann method and counted, which was expressed as number of L3 per kg dry matter (DM of pasture. Results: The average recovered population of L3 of H. contortus per kg DM varied significantly (p0.05. Conclusion: This study shown that when N-P-K fertilizers administered at recommended level, significantly reduced larval translation of H. contortus minimizing pasture infectivity for the free range grazing animals.

Effect of copper oxide wire particles dosage and feed supplement level on Haemonchus contortus infection in lambs.

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Burke, J M; Miller, J E; Olcott, D D; Olcott, B M; Terrill, T H

2004-09-02

The objective of the experiment was to determine the optimal dose of copper oxide wire particles (COWPs) to reduce infection of Haemonchus contortus in male lambs. Five to six-month-old hair breed lambs were housed on concrete and fed 450 (L; n = 25) or 675 g (H; n = 25) corn/soybean meal supplement and bermudagrass hay. In July, lambs were inoculated with 10,000 L(3) larvae (97% H. contortus; Day 0). Lambs were administered 0, 2, 4, or 6 g COWP on Day 28. Concentrations of copper in the liver were determined. There were no effects of supplement level on concentrations of copper in the liver and a linear relationship existed between COWP treatment and concentrations of copper in liver (P copper in the liver of the COWP treatment groups. PCV values were more favorable for lambs fed the higher level of supplement, especially when FEC were greater than 8000 epg.

An in silico analysis of the glutamate dehydrogenases of Teladorsagia circumcincta and Haemonchus contortus

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SIMON BROWN

2014-04-01

Full Text Available Nematode glutamate dehydrogenase (GDH amino acid sequences are very highly conserved (68-99% identity and are also very similar to those of the bovine and human enzymes (54-60% identity. The residues involved in binding nucleotides or substrates are completely conserved and tend to be located in highly conserved regions of the sequence. Based on the strong homology between the bovine, Teladorsagia circumcincta and Haemonchus contortus GDH sequences, models of the structure of the T. circumcincta and H. contortus monomers were constructed. The structure of the T. circumcincta monomer obtained using SWISS-MODEL was very similar to that of the bovine enzyme monomer and the backbone of the polypetide deviated very little from that of the bovine enzyme monomer. Despite the sequence differences between the bovine and T. circumcincta enzymes, the relative positions and orientations of the residues involved in ligand binding were very similar. The reported Km for NADP+ of T. circumcincta is about 35 and times that of the bovine enzyme, whereas the Kms of the two enzymes for glutamate, -ketoglutarate and NAD(PH are much more similar. The residue corresponding to S267 of the bovine enzyme is involved in binding the 2′-phosphate of NADP+ and is replaced in the T. circumcincta and H. contortus sequences by a tryptophan. The partial occlusion of the NAD(P-binding site by the tryptophan sidechain and the loss of at least one potential H-bond provided by the serine may explain the lower affinity of the T. circumcincta for NADP+.

Genetic diversity of Haemonchus contortus isolated from sympatric wild blue sheep (Pseudois nayaur) and sheep in Helan Mountains, China.

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Shen, Dong-Dong; Wang, Ji-Fei; Zhang, Dan-Yu; Peng, Zhi-Wei; Yang, Tian-Yun; Wang, Zhao-Ding; Bowman, Dwight D; Hou, Zhi-Jun; Liu, Zhen-Sheng

2017-09-19

Haemonchus contortus is known among parasitic nematodes as one of the major veterinary pathogens of small ruminants and results in great economic losses worldwide. Human activities, such as the sympatric grazing of wild with domestic animals, may place susceptible wildlife hosts at risk of increased prevalence and infection intensity with this common small ruminant parasite. Studies on phylogenetic factors of H. contortus should assist in defining the amount of the impact of anthropogenic factors on the extent of sharing of agents such as this nematode between domestic animals and wildlife. H. contortus specimens (n = 57) were isolated from wild blue sheep (Pseudois nayaur) inhabiting Helan Mountains (HM), China and additional H. contortus specimens (n = 20) were isolated from domestic sheep that were grazed near the natural habitat of the blue sheep. Complete ITS2 (second internal transcribed spacer) sequences and partial sequences of the nad4 (nicotinamide dehydrogenase subunit 4 gene) gene were amplified to determine the sequence variations and population genetic diversities between these two populations. Also, 142 nad4 haplotype sequences of H. contortus from seven other geographical regions of China were retrieved from database to further examine the H. contortus population structure. Sequence analysis revealed 10 genotypes (ITS2) and 73 haplotypes (nad4) among the 77 specimens, with nucleotide diversities of 0.007 and 0.021, respectively, similar to previous studies in other countries, such as Pakistan, Malaysia and Yemen. Phylogenetic analyses (BI, MP, NJ) of nad4 sequences showed that there were no noticeable boundaries among H. contortus populations from different geographical origin and population genetic analyses revealed that most of the variation (94.21%) occurred within H. contortus populations. All phylogenetic analyses indicated that there was little genetic differentiation but a high degree of gene flow among the H. contortus populations among

Anthelmintic activity of acetone extracts from South African plants used on egg hatching of Haemonchus contortus

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Gerda Fouche

2016-07-01

Full Text Available The nematode, Haemonchus contortus, is responsible for major economic losses in the livestock industry. The management of parasites such as H. contortus has been through the use of synthetic parasiticides. This has resulted in the presence of residues in meat and milk, which affects food safety. The development of resistance to available anthelmintics coupled with their high cost has further complicated matters. This has led to the investigation of alternative methods to manage nematodes, including the use of plants and plant extracts as a potential source of novel anthelmintics. Acetone extracts were prepared from 15 South African plant species and their anthelmintic activity determined using the egg hatch assay (EHA. The leaf extract of Cleome gynandra had the best inhibitory activity (68% ± 3% at a concentration of 2.5 mg/mL, followed by the stem extract of Maerua angolensis (65% ± 5%. The extracts had a relatively low toxicity on Vero cells determined by the MTT (3-(4,5-dimethylthiazol-2-yl-2,5- diphenyltetrazolium bromide cellular assay.

Interactions Between Nutrition and Infections With Haemonchus contortus and Related Gastrointestinal Nematodes in Small Ruminants.

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Hoste, H; Torres-Acosta, J F J; Quijada, J; Chan-Perez, I; Dakheel, M M; Kommuru, D S; Mueller-Harvey, I; Terrill, T H

2016-01-01

Interactions between host nutrition and feeding behaviour are central to understanding the pathophysiological consequences of infections of the digestive tract with parasitic nematodes. The manipulation of host nutrition provides useful options to control gastrointestinal nematodes as a component of an integrated strategy. Focussed mainly on the Haemonchus contortus infection model in small ruminants, this chapter (1) illustrates the relationship between quantitative (macro- and micro-nutrients) and qualitative (plant secondary metabolites) aspects of host nutrition and nematode infection, and (2) shows how basic studies aimed at addressing some generic questions can help to provide solutions, despite the considerable diversity of epidemiological situations and breeding systems. Copyright © 2016 Elsevier Ltd. All rights reserved.

Chemical composition and efficacy in the egg-hatching inhibition of essential oil of Piper aduncum against Haemonchus contortus from sheep

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Gisele Lopes Oliveira

Full Text Available Piper aduncum L., Piperaceae, has been used to treat mainly inflammatory diseases and has shown several biological activities such as insecticidal and larvicidal. The aim of this study was to analyze the chemical composition of essential oil of P. aduncum and its efficacy to egg-hatching inhibition of Haemonchus contortus from sheep. The essential oil was obtained from leaves and analysed by gas chromatography coupled to flame ionization detector and gas chromatography coupled to mass spectrometry. It was possible to characterize 22 different substances, among them monoterpenes (80.6% and sesquiterpenes (13.9%. The major compound was identified as 1,8-cineole (55.8%. Eggs of the nematode were exposed to four concentrations of the essential oil. Levamisole phosphate was used as positive control. The essential oil showed to be effective in inhibiting H. contortus hatchability and the LC90 was calculated as 8.9 mg.ml-1. These results can point out the P. aduncumessential oil and its chemical components as potential alternative to control of H. contortus

Dried, ground banana plant leaves (Musa spp.) for the control of Haemonchus contortus and Trichostrongylus colubriformis infections in sheep.

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Gregory, L; Yoshihara, E; Ribeiro, B L M; Silva, L K F; Marques, E C; Meira, E B S; Rossi, R S; Sampaio, P H; Louvandini, H; Hasegawa, M Y

2015-12-01

To evaluate the anthelmintic effect of Musa spp. leaves, 12 animals were artificially infected with Haemonchus contortus, and another 12 animals were infected with Trichostrongylus colubriformis. Then, both treatment groups were offered 400 g of dried ground banana plant leaves, and the control animals were offered only 1000 g of coast cross hay. During the trials, the animals received weekly physical examinations. The methods used to evaluate the efficiency of this treatment were packed cell volume, total plasma protein and faecal egg counts, and egg hatchability tests were performed on days -2, +3, +6, +9, +13 and +15. Coproculture tests were performed on day -2 to confirm monospecific infections. In the FEC and EHT, a statistically significant difference (0.04, 0.005; p  0.05) for Haemochus contortus group in all tests. Our results confirmed previous findings suggesting that dried ground banana plant leaves possess anthelmintic activity.

Core alpha1-->3-fucose is a common modification of N-glycans in parasitic helminths and constitutes an important epitope for IgE from Haemonchus contortus infected sheep

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van Die, I.; Gomord, V.; Kooyman, F. N.; van den Berg, T. K.; Cummings, R. D.; Vervelde, L.

1999-01-01

Synthesis of parasite specific IgE plays a critical role in the defence against helminth infections. We report here that IgE from serum from Schistosoma mansoni infected mice and Haemonchus contortus infected sheep recognizes complex-type N-glycans from Arabidopsis thaliana, which contain

Comparative in vitro efficacy of kefir produced from camel, goat, ewe and cow milk on Haemonchus contortus.

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Alimi, D; Rekik, M; Akkari, H

2018-04-18

One of the great challenges of veterinary parasitology is the search for alternative methods for controlling gastrointestinal parasites in small ruminants. Milk kefir is a traditional source of probiotic, with great therapeutic potential. The objective of this study was to investigate the anthelmintic effects of kefir on the abomasal nematode Haemonchus contortus from sheep. The study used camel, goat, ewe and cow milk as a starting material, to produce camel, goat, cow and ewe milk kefir. All kefirs showed a significant concentration-dependent effect on H. contortus egg hatching at all tested concentrations. The highest inhibition (100%) of eggs was observed with camel milk kefir at a concentration 0.125 mg/ml. In relation to the effect of kefirs on the survival of adult parasites, all kefirs induced concentration-dependent mortality in adults, with variable results. The complete mortality (100%) of adults of H. contortus occurred at concentrations in the range 0.25-2 mg/ml. The highest inhibition of motility (100%) of worms was observed after 8 h post exposure with camel milk kefir at 0.25 mg/ml. These findings indicate that kefir can be considered a potential tool to control haemonchosis in sheep. Further investigations are needed to assess the active molecules in kefir responsible for its anthelmintic properties and to investigate similar in vivo effects.

Effect of six tropical tanniferous plant extracts on larval exsheathment of Haemonchus contortus Efeito de seis extratos de plantas taniníferas tropicais sobre o desembainhamento larvar de Haemochus contortus

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Lorena Mayana Beserra de Oliveira

2011-06-01

Full Text Available Tanniferous plants represent a promising alternative for controlling gastrointestinal nematodes of small ruminants. This experiment evaluated the effects of extracts from the leaf and stem of Anadenanthera colubrina, Leucaena leucocephala and Mimosa tenuiflora on larval exsheathment of Haemonchus contortus in vitro and verified the role of tannins in this process. Third-stage larvae of H. contortus were incubated with extracts for 3 hours and were exposed to sodium hypochlorite solution. The extracts were tested at 300 µg.mL-1 and accompanied by controls: phosphate buffer solution (PBS and polyvinyl polypyrrolidone (PVPP. The larval exsheathment was evaluated for 60 minutes, and the results were subjected to the Kruskal-Wallis test (p 0.05, except for L. leucocephala and M. tenuiflora leaf extracts. However, pre-incubation with PVPP of these two extracts significantly changed larval exsheathment when compared to the non-treated extracts (p Plantas taniníferas representam uma promissora alternativa de controle dos nematóides gastrintestinais de pequenos ruminantes. Esse experimento avaliou in vitro os efeitos dos extratos das folhas e caules de Anadenanthera colubrina, Leucaena leucocephala e Mimosa tenuiflora sobre o desembainhamento larvar de Haemonchus contortus e verificou o papel dos taninos nesse processo. Larvas de terceiro estádio de H. contortus foram incubadas com 300 µg.mL-1 de extrato por 3 horas e expostas a uma solução de hipoclorito de sódio. O ensaio foi acompanhado por controles: solução salina tamponada com fosfato (PBS e polivinilpolipirrolidona (PVPP. O desembainhamento larvar foi avaliado durante 60 minutos e os resultados submetidos ao teste Kruskal-Wallis (p 0,05, exceto nos extratos das folhas de L. leucocephala e M. tenuiflora. Entretanto, a pré-incubação desses dois extratos com PVPP alterou significativamente o desembainhamento quando comparado com extratos não-tratados (p < 0,05. Esses resultados sugerem

In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus.

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Zenebe, Selamawit; Feyera, Teka; Assefa, Solomon

2017-01-01

Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods . Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Higher concentrations of the extracts (10 and 5 mg/ml) had a significantly superior adulticidal activity ( p molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus .

Comparison of parasitological and productive traits of Criollo lambs native to the central Mexican Plateau and Suffolk lambs experimentally infected with Haemonchus contortus.

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Alba-Hurtado, F; Romero-Escobedo, E; Muñoz-Guzmán, M A; Torres-Hernández, G; Becerril-Pérez, C M

2010-09-20

The study compares the parasitological and productive traits of Criollo lambs native to the central Mexican Plateau (CNCMP) and Suffolk (SU) lambs experimentally infected with Haemonchus contortus. CNCMP lambs (n=20) and SU lambs (n=15) were infected with L3 of H. contortus while five lambs of each genotype were kept as controls. Fecal egg count (FEC), packed cell volume (PCV), blood eosinophil number (BEN), ocular mucous membrane color (as measured by the FAMACHA index), changes in body condition score (BCS) and cumulative live weight gain (CLWG) were measured weekly during a 20-week period. On week 20, all animals were euthanized and the number of adult worms (AW) in the abomasum was counted. Infected SU lambs had higher (p<0.05) FEC and AW mean values compared to CNCMP lambs, which had a higher mean BEN count (p<0.05). Infected lambs had lower PCV values than controls, regardless of genotype, and had a negative correlation (r=-0.84, p<0.05) with the FAMACHA index. BCS tended to decline for infected SU lambs and increased slightly for infected CNCMP lambs. CLWG differed in all groups (p<0.05); infected SU lambs gained 12.1+/-1.9kg, infected CNCMP lambs gained 18.8+/-0.7kg, control SU lambs gained 34.6+/-1.6kg, and control CNCMP lambs gained 26.9+/-0.8kg. In conclusion, CNCMP lambs had a smaller worm burden, a better ability to maintain their productive traits, and were less affected by infection with Haemonchus contortus.

Annotation of two large contiguous regions from the Haemonchus contortus genome using RNA-seq and comparative analysis with Caenorhabditis elegans.

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Roz Laing

Full Text Available The genomes of numerous parasitic nematodes are currently being sequenced, but their complexity and size, together with high levels of intra-specific sequence variation and a lack of reference genomes, makes their assembly and annotation a challenging task. Haemonchus contortus is an economically significant parasite of livestock that is widely used for basic research as well as for vaccine development and drug discovery. It is one of many medically and economically important parasites within the strongylid nematode group. This group of parasites has the closest phylogenetic relationship with the model organism Caenorhabditis elegans, making comparative analysis a potentially powerful tool for genome annotation and functional studies. To investigate this hypothesis, we sequenced two contiguous fragments from the H. contortus genome and undertook detailed annotation and comparative analysis with C. elegans. The adult H. contortus transcriptome was sequenced using an Illumina platform and RNA-seq was used to annotate a 409 kb overlapping BAC tiling path relating to the X chromosome and a 181 kb BAC insert relating to chromosome I. In total, 40 genes and 12 putative transposable elements were identified. 97.5% of the annotated genes had detectable homologues in C. elegans of which 60% had putative orthologues, significantly higher than previous analyses based on EST analysis. Gene density appears to be less in H. contortus than in C. elegans, with annotated H. contortus genes being an average of two-to-three times larger than their putative C. elegans orthologues due to a greater intron number and size. Synteny appears high but gene order is generally poorly conserved, although areas of conserved microsynteny are apparent. C. elegans operons appear to be partially conserved in H. contortus. Our findings suggest that a combination of RNA-seq and comparative analysis with C. elegans is a powerful approach for the annotation and analysis of strongylid

PREVALENCE OF HAEMONCHUS CONTORTUS IN NATURALLY INFECTED SMALL RUMINANTS GRAZING IN THE POTOHAR AREA OF PAKISTAN

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F. R. CHAUDARY, M. F. U. KHAN1 AND M. QAYYUM

2007-04-01

Full Text Available The present study was carried out to investigate the prevalence and seasonal trend of the Haemonchus contortus in sheep and goats in the Potohar areas of northern Punjab, Pakistan from December 2004 to January 2006. Faecal samples collected from 968 sheep and 961 goats of different breeds were examined by the modified McMaster technique using saturated solution of sodium chloride. Results revealed that the infection was significantly (P<0.05 higher in sheep compared to goats. The peak infection level was recorded during rainy season (July-October. On the other hand, low infection level was noted from December upto May. In sheep, highest log faecal egg counts (LFECs were recorded in Islamabad, followed by Attock, Jhelum and Chakwal. However, in goats the LFECs trend was highest in Islamabad, followed by Jhelum, Attock and Chakwal districts. A significant (P<0.05 variability in LFECs was noted between sheep and goat breeds from site-site, while no significant difference was observed between breeds at the same site. Hairy (Jattal goats and Salt-Range (Latti sheep breeds exhibited significantly reduced LFECs level along with higher packed cell volume (PCV and haemoglobin (Hb levels compared to other breeds. Moreover, FAMACHA© chart scoring in relation with worm infection (FECs was more valid in sheep than goats. High prevalence of H. contortus in Potohar areas was due to favourable agro-climatic conditions that favour the development and survival of the free-living stages of H. contortus. The findings are discussed with regard to their relevance for strategic control of haemonchosis in small ruminants.

Atividade ovicida e larvicida in vitro do óleo essencial de Eucalyptus globulus sobre Haemonchus contortus Ovicidal and larvicidal activity in vitro of Eucalyptus globulus essential oils on Haemonchus contortus

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Iara T. F. Macedo

2009-09-01

Full Text Available O objetivo deste trabalho foi avaliar o efeito do óleo essencial de Eucalyptus globulus (OEEG sobre a eclosão e desenvolvimento de larvas de Haemonchus contortus. A determinação da composição química do OEEG foi feita por meio de cromatografia gasosa acoplada à espectrometria de massa. No teste de eclosão de ovos (TEO, utilizaram-se as concentrações 21,75; 17,4; 8,7; 5,43 e 2,71 mg.mL-1; e no de desenvolvimento larvar (TDL as concentrações foram 43,5; 21,75; 10,87; 5,43 e 2,71 mg.mL-1. Cada ensaio foi acompanhado por um controle positivo, 0,02 mg.mL-1 tiabendazol, no TEO e 0,008 mg.mL-1 ivermectina no TDL, e por um controle negativo com o diluente Tween 80 (3%. A eficácia máxima obtida pelo OEEG sobre ovos foi de 99,3% na concentração de 21,75 mg.mL-1 e, sobre larvas, 98,7% na concentração de 43,5 mg.mL-1. A concentração do OEEG, que inviabilizou 50% dos ovos e das larvas do parasito, foi de 8,3 e 6,92 mg.mL-1, respectivamente. A análise química do óleo identificou como principal componente o monoterpeno 1,8-cineol. O OEEG apresentou atividade ovicida e larvicida in vitro sobre H. contortus, determinando um bom potencial para utilização no controle de nematóides gastrintestinais de ovinos e caprinos.The objective of this work was to evaluate ovicidal and larvicidal effects of Eucalyptus globulus essential oil (EGEO on Haemonchus contortus. The chemical composition determination of EGEO was through gas chromatography and mass spectrometry. Egg hatch test (EHT was performed in concentrations 21.75; 17.4; 8.7; 5.43 e 2.71 mg.mL-1. In larval development test (LDT were used the concentrations 43.5; 21.75; 10.87; 5.43 e 2.71 mg.mL-1. Each trial was conducted by negative control with Tween 80 (3% and positive control, 0.02 mg.mL -1 of thiabendazole in EHT and 0.008 mg.mL-1 of ivermectin in LDT. The maximum effectiveness of EGEO on eggs was 99.3% in concentration of 21.75 mg.mL-1 and on larvae was 98.7% in concentration 43

Metabolism of albendazole, ricobendazole and flubendazole in Haemonchus contortus adults: Sex differences, resistance-related differences and the identification of new metabolites

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Lucie Raisová Stuchlíková

2018-04-01

Full Text Available Haemonchus contortus (family Trichostrongylidae, Nematoda, a hematophagous gastrointestinal parasite found in small ruminants, has a great ability to develop resistance to anthelmintic drugs. We studied the biotransformation of the three benzimidazole anthelmintics: albendazole (ABZ, ricobendazole (albendazole S-oxide; RCB and flubendazole (FLU in females and males of H. contortus in both a susceptible ISE strain and resistant IRE strain. The ex vivo cultivation of living nematodes in culture medium with or without the anthelmintics was used. Ultrasensitive UHPLC/MS/MS analysis revealed 9, 7 and 12 metabolites of ABZ, RCB and FLU, respectively, with most of these metabolites now described in the present study for the first time in H. contortus. The structure of certain metabolites shows the presence of biotransformation reactions not previously reported in nematodes. There were significant qualitative and semi-quantitative differences in the metabolites formed by male and female worms. In most cases, females metabolized drugs more extensively than males. Adults of the IRE strain were able to form many more metabolites of all the drugs than adults of the ISE strain. Some metabolites were even found only in adults of the IRE strain. These findings suggest that increased drug metabolism may play a role in resistance to benzimidazole drugs in H. contortus. Keywords: Drug resistance, Drug metabolism, Anthelmintics, Benzimidazole, Nematode

Early detection of Haemonchus contortus infection in sheep using three different faecal occult blood tests

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A.V. Rodríguez

2015-07-01

Full Text Available Haemonchus contortus is a blood-sucking parasite causing the presence of faecal occult blood (FOB. The objective was to study three different FOB tests in order to have a new indicator of H. contortus infection in sheep that could be included in the genetic evaluation system as an alternative selection criterion to faecal worm egg count (FEC. A total of 29 Corriedale lambs were experimentally infected with 10.000 larvae of H. contortus. Stool samples were recorded for FEC and FOB tests (Hexagon, Hematest® and Multistix®, blood for packed cell volume (PCV, haemoglobin, white and red blood cell count (RBC, and FAMACHA© for scoring anaemia. At the end of the experiment lambs were slaughtered to worm burden count. Field infection was achieved in 309 Merino lambs under natural parasite challenge. FEC data were normalized through logarithmic transformation (LnFEC. Pearson correlation was estimated to examine the relationship between all traits. The three tests were able to detect the presence of FOB at day 11. FEC, PCV and RBC decreased to sub-normal values from day 18. FAMACHA© score 3 was considered to be indicative of anaemia. Most of the correlations were of high magnitude, with the exception of Multistix® test that was moderately correlated with haematological parameters, LnFEC and FEC. In field infection, most samples were negative to FOB tests and the correlations were lower than those calculated under experimental infection. In conclusion, FOB tests were able to detect haemonchosis earlier than FEC under high experimental parasite challenge. However, they were not able to detect FOB under natural mixed parasite challenge. FAMACHA© and PCV demonstrated to be good indicators of Haemonchosis, having moderate to high correlations with FEC.

Gastrointestinal parasites of captive European bison Bison bonasus (L.) with a sign of reduced efficacy of Haemonchus contortus to fenbendazole.

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Pyziel, Anna M; Björck, Sven; Wiklund, Rikard; Skarin, Moa; Demiaszkiewicz, Aleksander W; Höglund, Johan

2018-01-01

The history of European bison Bison bonasus Linnaeus, 1758 has been stormy since its extinction in the wild after the First World War. Due to the fact that the species was restored from just 12 founders, further expansion has suffered from low genetic variability, rendering the bison vulnerable to various pathogens due to inbreeding depression. Parasites are recognised as a key biological threat to bison population. Thus, parasitological examination including monitoring of the level of anthelmintic resistance in a herd should be a routine procedure involved in management and protection of European bison. This study was conducted in a group of 27 bison kept in a European bison breeding centre in Sweden. In April 2015, a faecal egg count reduction test (FECRT) was performed in animals with ≥ 100 gastrointestinal nematode (GIN) eggs per gram faeces, to determine effectiveness of fenbendazole (FBZ) treatment. Additionally, the third stage larvae were cultured for molecular examination by a conventional PCR as well as by real-time quantitative PCR (q-PCR) for detection of the blood-sucking nematode Haemonchus contortus. Faecal sampling was conducted 1 day before and 8 days after deworming each animal. Anthelmintic treatment turned to be entirely efficient toward intestinal nematodes of genera Nematodirus and Trichuris, whereas shedding of strongylid eggs from the subfamily Ostertagiinae was reduced from 81 to 30%. Polymerase chain reaction (PCR) on cultured third-stage larvae (L3) before treatment was positive for H. contortus, Ostertagia ostertagi and Cooperia oncophora, whereas post-treatment examination revealed exclusively the DNA of H. contortus. Thus, only H. contortus was involved in post-treatment faecal egg count (FEC). FECRT showed that the reduction in strongylid FEC to FBZ in the examined bison herd was 87% (95%-confidence intervals [95% CI] = 76-93), suggesting reduced efficacy of FBZ to strongylid GIN including mainly H. contortus.

The nutritional status affects the complete blood count of goats experimentally infected with Haemonchus contortus.

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Cériac, S; Jayles, C; Arquet, R; Feuillet, D; Félicité, Y; Archimède, H; Bambou, J-C

2017-11-09

Gastrointestinal nematode (GIN) remains the most important pathogenic constraint of small ruminant production worldwide. The improvement of the host immune response against GIN though breeding for improved animal resistance, vaccination and nutritional supplementation appear as very promising methods. The objective of this study was to investigate the effect of four nutritional status differing in protein and energy levels (Hay: 5.1 MJ/Kg of dry matter (DM) and 7.6% of crude protein (CP), Ban: 8.3 MJ/Kg of DM and 7.5% of CP, Soy: 7.6 MJ/Kg of DM and 17.3% of CP, BS: 12.7 MJ/Kg of DM and 7.4% of CP) on the haematological disturbances due to Haemonchus contortus infection in Creole kid goats. No significant effect of the nutritional status was observed for faecal egg count (FEC) but the experimental infection induced haematological disturbances whose intensity and lengthening were dependent on the nutritional status. A transient marked regenerative macrocytic hypochromic anaemia as revealed by a decrease of packed cell volume (PCV), red blood cells (RBC) and hemoglobin and an increase of reticulocytes was observed in all infected groups except Hay. In this latter, the anaemia settled until the end of the experiment. Furthermore, H. contortus induced a thrombocytopenia significantly more pronounced in the group under the lowest nutritional status in term of protein (Hay and Ban). A principal component analysis revealed that the variables that discriminated the nutritional status were the average daily gain (ADG) and the PCV, considered as measures of the level of resilience to H. contortus infection. Moreover, the variables that discriminated infected and non-infected animals were mostly related to the biology of RBC (i.e. size and hemoglobin content) and they were correlated with FEC. The severity and the lengthening of the regenerative anaemia and the thrombocytopenia induced by H. contortus have been affected by the nutritional status. The protein enriched

Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids.

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Bambou, J C; Archimède, H; Arquet, R; Mahieu, M; Alexandre, G; González-Garcia, E; Mandonnet, N

2011-06-10

The aim of the present study was to test the effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids. One trial with three replicates involved a total of 154 female kids that were chosen from three successive cohorts of the Creole flock of INRA-Gardel in 2007. The kids were placed into four treatments according to the amount of concentrate they received: G0 (no concentrate and a quality Dichantium spp. hay ad libitum, HAY), G1 (HAY+100g commercial concentrate d(-1)), G2 (HAY+200 g commercial concentrate d(-1)), G3 (HAY+300 g commercial concentrate d(-1)). The G0-G3 groups were infected with a single dose of 10,000 H. contortus third stage larvae (L(3)) at Day 0 (D0). Each infected group was comprised of one half resistant and one half susceptible genetically indexed kids. The average breeding values on egg excretion at 11 months of age were distant of 0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0, G1, G2 and G3, respectively. The faecal egg count (FEC), packed cell volume (PCV), eosinophilia (EOSI) and dry matter intake (DMI) indices were monitored weekly until 42 days post-infection. Enzyme-linked immunosorbent assay was carried out on serum samples to determine the level of IgA anti-H. contortus L(3) crude extracts and adult excretion/secretion products (ESP). The 10,000 L(3) dose received by the kids induced a severe infection: 8000 eggs per gram at the FEC peak, a PCV less than 15% and mortality. Interestingly, the supplemented animals in G3 showed a higher level of EOSI but a lower level of IgA anti-L3 and IgA anti-ESP than non-supplemented animals (G0). Resistant and susceptible kids had significantly different FEC variations within the groups. Susceptible kids had a 1.6 times higher egg output than resistant kids in G0. This difference was not found in the supplemented groups. The results of this study showed that supplementary feeding improved resistance of Creole kids to H. contortus

The potential to control Haemonchus contortus in indigenous South African goats with copper oxide wire particles.

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Vatta, A F; Waller, P J; Githiori, J B; Medley, G F

2009-06-10

The high prevalence of resistance of Haemonchus contortus to all major anthelmintic groups has prompted investigations into alternative control methods in South Africa, including the use of copper oxide wire particle (COWP) boluses. To assess the efficacy of COWP against H. contortus in indigenous South African goats, 18 male faecal egg-count-negative goats were each given ca.1200 infective larvae of H. contortus three times per week during weeks 1 and 2 of the experiment. These animals made up an "established" infection group (ESTGRP). At the start of week 7, six goats were each given a 2-g COWP bolus orally; six goats received a 4-g COWP bolus each and six animals were not treated. A further 20 goats constituted a "developing" infection group (DEVGRP). At the beginning of week 1, seven of the DEVGRP goats were given a 2-g COWP bolus each; seven goats were treated with a 4-g COWP bolus each and no bolus was given to a further six animals. During weeks 1-6, each of these DEVGRP goats was given ca. 400 H. contortus larvae three times per week. All 38 goats were euthanized for worm recovery from the abomasa and small intestines in week 11. In the ESTGRP, the 2-g and 4-g COWP boluses reduced the worm burdens by 95% and 93%, respectively compared to controls (mean burden+/-standard deviation, SD: 23+/-33, 30+/-56 and 442+/-518 worms, P=0.02). However, in the DEVGRP goats, both the 2-g and 4-g COWP treatments were ineffective in reducing the worm burdens relative to the controls (mean burdens+/-SD: 1102+/-841, 649+/-855, 1051+/-661 worms, P=0.16). Mean liver copper levels did not differ between the ESTGRP goats treated with 2-g COWP, 4-g COWP or no COWP (mean+/-standard error of the mean, SEM, in ppm: 93.7+/-8.3; 101.5+/-8.3; 71.8+/-8.3, P=0.07) nor did they differ between the DEVGRP goats (mean+/-SEM, in ppm: 74.1+/-9.1; 75.4+/-9.1; 74.9+/-10.0, P>0.99). The copper values were considered adequate, but not high, for goats. The COWP boluses have the potential to be used

Eco-epidemiologia de Haemonchus contortus bahienses, ecotipo presente en ovinos de zonas aridas de Venezuela Eco-epidemiologia de Haemonchus contortus bahiensis, ecotipo presente em ovinos de zonas áridas da Venezuela Ecoepidemiology of Haemonchus contortus bahiensis: ecotype present in sheep of Venezuelan arid zone

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Gustavo Morales

1987-09-01

Full Text Available Haemonchus contortus bahiensis Grisi, 1974 parasita a los ovinos de zonas áridas de Venezuela. En las hembras de dicho ecotipo la distribución de frecuencias por nosotros observada fue de un 15,32% para el morfotipo con lengüeta supravulvar, de un 51,61% para el botonado y de un 33,07% para el liso. Correspondiéndole a dicho ecotipo un índice de diversidad de Shannon-Weaver de 1,44 bits. Las medidas corporales, el tamaño de los huevos y el symlophe son similares en los tres morfotipos. La disposición espacial tanto de los machos como de las hembras en el seno de la población de hospedadores es de tipo contagiosa siendo bien descrita por el parámetro K de la distibución binominal negativa. Se evidenció la existencia de una compleja interacción entre la abundancia, la agregación y la prevalencia de dicho ecotipo y se discute su importancia sobre el equilibrio hospedador-parásito.Nas fêmesas do dito ecotipo a distribuição de freqüência observada foi de 15,32% para o morfotipo com lingueta supra-vulvar, de 51,61% para o tipo com botão vulvare de 33,07% para o tipo liso. A esse ecotipo corresponde um índice de diversidade de Shannon-Weaver igual a 1,44 bits. As medidas corporais, o tamanho dos ovos e o número de estrias cuticulares longitudinais são semelhantes nos três morfotipos. A disposição espacial, tanto dos machos quanto das fêmeas na população de hospedeiros é do tipo contagiosa, sendo bem descrita pelo parâmetro K da distribuição binomial negativa. Se verificou a existência de uma complexa interação entre a abundância, a agregação e a prevalência do citado ecotipo, discutindo-se sua importância no equilíbrio hospedeiro-parasito.The frequency distribution of female Haemonchus contortus bahiensis Grisi, 1974 in sheep from Venezuelan arid zones is 15.32% for the type with vulvar flap, 51.6% for the vulvar-knob and 33.07% for smooth type. A Shannon-Weaver diversity index corresponding to 1.44 bits was

Variability of resistance in Black Bengal goats naturally infected with Haemonchus contortus.

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Kumar, Ratnesh; Ranjan, Sanjeev; Vishnu, P Guru; Negi, Mamta; Senapati, P K; Charita, V Gnani

2015-03-01

A total 290 Black Bengal goats (6 buck, 109 doe and 175 kids born from 11 sires) were studied to evaluate the variability of resistance in Black Bengal goats naturally infected with Haemonchus contortus. The variability of resistance in Black Bengal goat was studied for both genetic and non-genetic factors like village, sex, age dam, sire, dam resistance group and offspring resistance group. Male kids have slightly higher resistance than female kids although it was not significant. Resistance of kids was increased as age increases and kid population showed significantly different resistance status among the offspring resistant groups. The doe population showed significantly different LEPG as per the resistance group in all the collections. The present study found that the resistance of kids under sire were varied significantly and observed that the kids under sire 1, 6-8 were significantly more resistant than the kids of the sire 2, 5 and 11 in 3rd collection and it is also noticed that maternal genetic effect has a very little impact on resistance of kids. Males (buck) were most resistant and the kids were least resistant and the resistance of dam was in between the male and kids population.

In Vitro Anthelmintic Activity of Crude Extracts of Aerial Parts of Cissus quadrangularis L. and Leaves of Schinus molle L. against Haemonchus contortus

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Selamawit Zenebe

2017-01-01

Full Text Available Background. Haemonchus contortus, the causative agent of Haemonchosis, is the most economically important parasite in small ruminant production. Control with chemotherapy has not been successful due to rapid emergence of drug-resistant strains. There is a continuous search for alternative leads particularly from plants. The study aimed to evaluate the anthelmintic activity of crude methanolic extracts of leaves of Schinus molle and aerial parts of Cissus quadrangularis against H. contortus. Methods. Adult motility test and egg hatching inhibition assay were employed to investigate the in vitro adulticidal and egg hatching inhibitory effects of the extracts. Results. Higher concentrations of the extracts (10 and 5 mg/ml had a significantly superior adulticidal activity (p<0.05 compared to the negative control and lower concentration levels, which was comparable to albendazole. Similarly, the relative egg hatch inhibition efficacy of S. molle and C. quadrangularis extracts indicated a maximum of 96% and 88% egg hatch inhibition, respectively, within the 48 hrs of exposure at 1 mg/ml. Conclusion. The current study evidenced that the crude methanolic extracts of the plants have promising adulticidal and egg hatching inhibitory effects against H. contortus.

Biotransformation of albendazole and activities of selected detoxification enzymes in Haemonchus contortus strains susceptible and resistant to anthelmintics.

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Vokřál, Ivan; Jirásko, Robert; Stuchlíková, Lucie; Bártíková, Hana; Szotáková, Barbora; Lamka, Jiří; Várady, Marián; Skálová, Lenka

2013-09-23

The increased activity of drug-metabolizing enzymes can protect helminths against the toxic effect of anthelmintics. The aim of this study was to compare the metabolism of the anthelmintic drug albendazole (ABZ) and the activities of selected biotransformation and antioxidant enzymes in three different strains of Haemonchus contortus: the ISE strain (susceptible to common anthelmintics), the BR strain (resistant to benzimidazole anthelmintics) and the WR strain (multi-resistant). H. contortus adults were collected from the abomasum of experimentally infected lambs. In vitro (subcellular fractions of H. contortus homogenate) as well as ex vivo (living nematodes cultivated in flasks with medium) experiments were performed. HPLC with spectrofluorimetric and mass-spectrometric detection was used in the analysis of ABZ metabolites. The in vitro activities of oxidation/antioxidation and conjugation enzymes toward model substrates were also assayed. The in vitro data showed significant differences between the susceptible (ISE) and resistant (BR, WR) strains regarding the activities of peroxidases, catalase and UDP-glucosyltransferases. S-oxidation of ABZ was significantly lower in BR than in the ISE strain. Ex vivo, four ABZ metabolites were identified: ABZ sulphoxide and three ABZ glucosides. In the resistant strains BR and WR, the ex vivo formation of all ABZ glucosides was significantly higher than in the susceptible ISE strain. The altered activities of certain detoxifying enzymes might partly protect the parasites against the toxic effect of the drugs as well as contribute to drug-resistance in these parasites. Copyright © 2013 Elsevier B.V. All rights reserved.

In vitro AND in vivo ANTHELMINTIC EFFECT OF Coffea arabica RESIDUES AGAINTS AN Haemonchus contortus ISOLATE WITH LOW SUSCEPTIBILITY TO TANNINS

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G.I. Ortiz-Ocampo

2016-05-01

Full Text Available The present study evaluated in vitro and in vivo anthelmintic (AH effect of the percolated by-product of Coffea arabica against a Haemonchus contortus isolate with low susceptibility to tannins. Firstly, the susceptibility to tannins of Paraíso H. contortus isolate was evaluated with the in vitro larval exsheathment inhibition assay (LEIA using an acetone:water (70:30 extract of Acacia pennatula. Afterwards, the in vitro AH effect of acetone:water extracts of percolated C. arabica by-products (Clean and Smooth (CS and Shade Grown (SG Starbucks® were evaluated. Then, the in vivo AH effect was determined in growing sheep (4 months old and 15.2 kg bodyweight, infected with H. contortus (Paraiso isolate and fed with a diet containing the percolated by-product of C. arabica. Three experimental groups were formed (n = 6 lambs: GC Group (control diet without C. arabica, GCA Group (feed containing 10% C. arabica and, GCA+PEG (polyethilenglycol Group (feed with 10% C. arabica + PEG as a tannin inhibitor. Fecal samples were obtained daily from the rectum of lambs (days 1 to 20 of the study. The H. contortus isolate tolerated the A. pennatula extract at concentrations of 150 and 300 μg extract/ml PBS (P > 0.05, which confirmed its low susceptibility to tannins. The C. arabica CS extract reduced the exsheatment from 150 μg of extract/ml PBS and the SG variety from 1200 μg of extract/ml PBS (P0.05. In conclusion, the C. arabica percolated by-product extracts showed in vitro AH effect against exsheathment. However, the consumption of feed containing 10% of the percolated by-product did not show an in vivo AH effect in sheep infected with the Paraiso H. contortus isolate. The latter could be due to the low susceptibility towards the secondary compounds of tannin rich plants of the H. contortus isolate used in this study.

Differences in transcription between free-living and CO2-activated third-stage larvae of Haemonchus contortus

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Zhong Weiwei

2010-04-01

Full Text Available Abstract Background The disease caused by Haemonchus contortus, a blood-feeding nematode of small ruminants, is of major economic importance worldwide. The infective third-stage larva (L3 of this gastric nematode is enclosed in a cuticle (sheath and, once ingested with herbage by the host, undergoes an exsheathment process that marks the transition from the free-living (L3 to the parasitic (xL3 stage. This study explored changes in gene transcription associated with this transition and predicted, based on comparative analysis, functional roles for key transcripts in the metabolic pathways linked to larval development. Results Totals of 101,305 (L3 and 105,553 (xL3 expressed sequence tags (ESTs were determined using 454 sequencing technology, and then assembled and annotated; the most abundant transcripts encoded transthyretin-like, calcium-binding EF-hand, NAD(P-binding and nucleotide-binding proteins as well as homologues of Ancylostoma-secreted proteins (ASPs. Using an in silico-subtractive analysis, 560 and 685 sequences were shown to be uniquely represented in the L3 and xL3 stages, respectively; the transcripts encoded ribosomal proteins, collagens and elongation factors (in L3, and mainly peptidases and other enzymes of amino acid catabolism (in xL3. Caenorhabditis elegans orthologues of transcripts that were uniquely transcribed in each L3 and xL3 were predicted to interact with a total of 535 other genes, all of which were involved in embryonic development. Conclusion The present study indicated that some key transcriptional alterations taking place during the transition from the L3 to the xL3 stage of H. contortus involve genes predicted to be linked to the development of neuronal tissue (L3 and xL3, formation of the cuticle (L3 and digestion of host haemoglobin (xL3. Future efforts using next-generation sequencing and bioinformatic technologies should provide the efficiency and depth of coverage required for the determination of the

In vitro anthelmintic activity of five tropical legumes on the exsheathment and motility of Haemonchus contortus infective larvae.

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von Son-de Fernex, Elke; Alonso-Díaz, Miguel Angel; Valles-de la Mora, Braulio; Capetillo-Leal, Concepción M

2012-08-01

This study investigated the in vitro anthelmintic (AH) activity of five tropical legume plants [Arachis pintoi CIAT 22160 (A.p. 22160), Gliricidia sepium, Cratylia argentea (C.a. Yacapani), C. argentea CIAT 22386 (C.a. 22386), C. argentea Veranera (C.a. Veranera)] against Haemonchus contortus infective larvae and the role of tannins/polyphenolic compounds in the AH effect. Lyophilized leaf extracts of each plant were evaluated using the Larval Exsheathment Inhibition Assay (LEIA) and the larval migration inhibition assay (LMIA). The role of tannins/polyphenolic compounds in the AH effect was evaluated in both assays using polyethylene glycol (PEG) to remove tannins from the solutions. At the highest concentration (1200μg of extract/ml), A. pintoi 22160, C.a. Yacapani, C.a. Veranera and C.a. 22386 completely inhibited the exsheathment process of H. contortus (P<0.01). At the same concentration (1200μg of extract/ml), the inhibition of larval migration for C.a. 22386, C.a. Veranera and G. sepium was 66.0%, 35.9% and 39.2% (relative to the PBS control), respectively. In both bioassays (LEIA and LMIA), the AH effect shown by each plant was blocked after the addition of polyethylene glycol (PEG), corroborating the role of tannins/polyphenolic compounds. Copyright © 2012 Elsevier Inc. All rights reserved.

The efficacy of a slow-release albendazole capsule against Haemonchus contortus with known resistance to albendazole

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Alan D. Fisher

2013-12-01

Full Text Available Controlled-release albendazole capsules (CRCs are currently registered for use in Australia and New Zealand as anthelmintic treatment in sheep. However, reports on the efficacy of such products on resistant parasite populations are sometimes controversial. This is the first study to report on the efficacy of such products under South African field conditions in sheep harbouring a population of Haemonchus contortus with known multiple anthelmintic resistance, including to albendazole. Treatment groups were comprised of CRC-treated and single dose albendazole-treated sheep, as well as negative controls. Groups were compared by using faecal egg count reduction tests, FAMACHA© anaemia scoring, conception rates and comparative weight gains over three and a half months. Based on a comparison of faecal egg counts, no advantage could be found using CRCs. Moreover, the use of the product actually decreased weight gain when compared with the control group animals.

Feed resource selection of Criollo goats artificially infected with Haemonchus contortus: nutritional wisdom and prophylactic self-medication.

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Ventura-Cordero, J; González-Pech, P G; Jaimez-Rodriguez, P R; Ortiz-Ocampo, G I; Sandoval-Castro, C A; Torres-Acosta, J F J

2018-06-01

Previous cafeteria studies suggested that a moderate natural gastrointestinal nematode (GIN) infection did not modify the resource selection of adult Criollo goats towards tannin-rich plants compared with worm-free goats. A higher infection with Haemonchus contortus could trigger a change in the resource selection behaviour towards tannin-rich foliage. Alternatively, goats might select plant species solely to meet their nutritional requirements. A cafeteria study investigated the effect of a high artificial infection with H. contortus on the feed resource selection of goats. Adult Criollo goats (37.5±4.8 kg BW) with browsing experience were distributed in two groups: the infected group (IG) with six animals artificially infected with H. contortus (6000 L3/animal); and the non-infected group (NIG) with six animals maintained worm-free. The experiment included two 5-day periods with additional 5-day adaptation period. In the first period, animals were offered foliage of five plant species with a decreasing gradient of condensed tannins (CT) (Mimosa bahamensis, Gymnopodium floribundum, Havardia albicans, Acacia pennatula, Lysiloma latisiliqum), and three plant species with negligible CT content (Leucaena leucocephala, Piscidia piscipula and Brosimum alicastrum). In the second period the foliage of B. alicastrum was withdrawn. A grain-based concentrate feed was offered daily at 1% BW in DM basis. Dry matter and nutrient intake was determined. Foliage selection of each experimental group was determined using the Chesson selection index. The H. contortus egg count per gram of faeces (EPG) was determined for infected goats twice daily. Chesson index showed a similar pattern of foliage selection on periods 1 and 2. Mean EPG of goats in IG was 2028±259 EPG during period 1 and 1 293±198 EPG during period 2 (P>0.05). During period 1, the selection pattern was highest for B. alicastrum (tannin-free), followed by a tannin-rich plant (M. bahamensis). These two plants remained

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

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Fernanda S. Fortes

2013-02-01

Full Text Available Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM and moxidectin (MOX effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT. Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL. The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50 dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL, identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001 and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA guidance.

Impact of the post-weaning parasitism history on an experimental Haemonchus contortus infection in Creole goat kids.

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Ceï, W; Mahieu, M; Philibert, L; Arquet, R; Alexandre, G; Mandonnet, N; Bambou, J C

2015-01-15

Gastrointestinal nematode (GIN) infections have an important negative impact on small ruminant production. The selection of genotypes resistant to these parasitic infections is a promising alternative control strategy. Thus, resistance against GIN is an important component of small ruminant breeding schemes, based on phenotypic measurements of resistance in immune mature infected animals. In this study we evaluated both the impact of the post-weaning parasitism history on the response to an experimental Haemonchus contortus infection of resistant and susceptible Creole kids chosen on the basis of their estimated breeding value, and the interaction with the kid's genetic status. During the post-weaning period (from 3 months until 7 months of age) Creole kids were reared at pasture according to four different levels of a mixed rotational stocking system with Creole cattle: 100% (control), 75% (GG75), 50% (GG50), and 25% (GG25) of the total stocking rate of the pasture. The level of infection of the kids decreased significantly at 50% and 25% of the total stocking rate. After the post-weaning period at pasture, at 11 months of age kids were experimentally infected with H. contortus. The faecal egg counts (FEC) were significantly lower in the groups showing the highest FEC at pasture. This result suggests that a degree of protection against an experimental H. contortus infection occurred during the post-weaning period and was dependant on the level of parasitism. Interestingly, no interaction was observed between this level of protection and the genetic status. In conclusion, the level of post-weaning natural parasitism history at pasture would not influence the genetic status evaluation. More generally our results suggest that it would be better to expose kids to a high level of gastrointestinal parasitism during the post-weaning period in order to increase the basal level of resistance thereafter. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of two in vitro methods for the detection of ivermectin resistance in Haemonchus contortus in sheep

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Urda Dolinská M.

2016-06-01

Full Text Available Gastrointestinal parasitic nematodes in sheep cause severe economic losses. Anthelmintics are the most commonly used drugs for prophylaxis and therapy against parasitic helminths. The problem of drug resistance has developed for all commercially available anthelmintics in several genera and classes of helminths. In vitro and in vivo tests are used to detect anthelmintic resistance. Two in vitro methods (larval migration inhibition test and micromotility test for the detection of ivermectin (IVM resistance were compared using IVM-resistant and IVM-susceptible isolates of Haemonchus contortus. The degree of resistance for each test was expressed as a resistance factor (RF. The micromotility test was more sensitive for quantitatively measuring the degree of resistance between susceptible and resistant isolates. The RFs for this test for IVM and eprinomectin ranged from 1.00 to 108.05 and from 3.87 to 32.32, respectively.

E-ADA activity in erythrocytes of lambs experimentally infected with Haemonchus contortus and its possible functional correlations with anemia.

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Da Silva, Aleksandro S; Schafer, Andressa S; Aires, Adelina R; Tonin, Alexandre A; Pimentel, Victor C; Oliveira, Camila B; Zanini, Daniela; Schetinger, Maria R C; Lopes, Sonia T A; Leal, Marta L R

2013-12-01

The aim of this study was to evaluate the ecto-adenosine deaminase (E-ADA) activity in erythrocytes of lambs experimentally infected with Haemonchus contortus, correlating it with the degrees of anemia of the experimental animals. A total of 14 healthy lambs, with negative fecal exam for parasites, were to carry out the present study. They were divided into two groups, composed by seven animals: Group A represented the healthy animals (uninfected), while in Group B the animals were infected with 15,000 larvae of H. contortus. Blood was drawn on the days 15, 45 and 75 post-infection (PI) in order to perform the hematological analysis, as well as the mensuration of E-ADA activity in erythrocytes. Parasitological stool exam were performed on the same days mentioned above to follow up the evolution of the infection, as well to determine the number of eggs per gram of feces (EPG). On day 15PI, the animals presented negative EPG and there was not significant (P>0.05) difference between groups in relation to E-ADA activity and hematologic parameters. Animals in Group B had positive EPG for helminths on days 45 and 75 PI, accompanied by varying degrees of anemia, when compared to Group A. At the same periods E-ADA activity was significantly (PADA in erythrocytes and hematocrit on days 45 (r = -0.76) and 75 (r = -0.85)PI. Based on these results and in the scientific literature, it is possible to conclude that the E-ADA may participate on mechanisms related with the pathogenesis and host response against anemia caused by H. contortus. Copyright © 2013 Elsevier Ltd. All rights reserved.

Ovicidal and larvicidal activity of the crude extracts from Phytolacca icosandra against Haemonchus contortus.

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Hernández-Villegas, M M; Borges-Argáez, R; Rodriguez-Vivas, R I; Torres-Acosta, J F J; Méndez-Gonzalez, M; Cáceres-Farfan, M

2011-06-30

The development of anthelmintic resistance has impacted on the success of conventional anthelmintics (AH) for the control of gastrointestinal nematodes in grazing/browsing sheep and goats. Medicinal plants from the traditional herbolary in Mexico may provide new candidates that can be explored as alternative sources of AHs for ruminants. This study evaluated the leaf extracts derived from Phytolacca icosandra against infective L(3) larvae and eggs from Haemonchus contortus collected from sheep. Three extracts of different polarities were obtained from the leaf plants using ethanol, n-hexane and dichloromethane as the solvents. The effectiveness of the in vitro AH activity of the plant extracts was evaluated using larval migration inhibition (LMI) and egg hatch (EHA) assays. For the LMI assays, the ethanolic extract of P. icosandra showed 55.4% inhibition of larval migration at 2mg/mL (p<0.05). The dichloromethane extract of P. icosandra showed 67.1% inhibition of migration at 3mg/mL (p<0.05) and a dose-dependent response with an LD(50) of 0.90 mg/mL. The n-hexane extract failed to show inhibition of larval migration at any concentration explored. In the EHA for the ethanol extract, the lowest concentration tested (0.15 mg/mL) resulted in inhibition of egg hatching greater than 72.6%. Therefore, the LD(50) could not be calculated for this extract. The LD(50) of the dichloromethane extract of P. icosandra was 0. 28 mg/mL. An egg hatch inhibition greater than 90% was observed with both the ethanolic and dichloromethane extracts when using a concentration of 0.90 mg/mL or higher. The n-hexane extract failed to show egg hatch inhibition at any concentration tested. The AH activity reported for P. icosandra could be attributable to the flavonoids, steroids, terpenoids, coumarins and/or saponins that were present in the ethanolic and dichloromethane extracts. A combination of more than one component may also explain the observed AH activity against the H. contortus life

Effectiveness of copper oxide wire particles for Haemonchus contortus control in sheep.

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Knox, M R

2002-04-01

To assess the efficacy of copper oxide wire particles (COWP) for the control of H contortus infections in grazing sheep. In experiment 1, 40 worm-free Merino hoggets (11 to 12 months of age) were divided into four equal groups and allocated to separate 0.8 ha pasture plots. Two groups then received 2.5 g COWP whereas the other two groups were untreated. From 1 week after COWP treatment all lambs received a weekly infection of 2000 H contortus larvae. At week 8, six sheep from the untreated group were then allocated to two groups and treated with either 2.5 or 5.0 g of COWP to establish therapeutic efficacy of treatment. Experiment 2 followed a similar protocol but was conducted with 40 worm-free Merino lambs (3 to 4 months of age) and no assessment of therapeutic efficacy was made. In experiment 1 no significant difference in faecal worm egg counts was observed between treatments and faecal worm egg counts remained less than 3000 epg in all animals. Total worm counts were reduced by 37% by COWP treatment (P = 0.055). Both 2.5 g and 5.0 g doses of COWP at 8 weeks of infection reduced faecal worm egg counts by > 85% with the higher dose giving an earlier response to treatment. In experiment 2, faecal worm egg counts at 4 and 6 weeks were reduced by more than 90% in the COWP treated lambs and worm numbers were 54% lower after 6 weeks when all remaining untreated lambs had to be treated for haemonchosis. Mean faecal worm egg counts in the COWP lambs remained below 3500 epg and clinical disease did not develop in the majority of lambs before the end of the experiment at 10 weeks. Treatment with COWPs appears to have the potential to reduce establishment and worm fecundity of Haemonchus spp for an extended period and may offer livestock producers a supplementary means of reducing larval contamination of pasture particularly in areas where anthelmintic resistance is a problem and copper supplementation is likely to be beneficial.

In vitro anthelmintic activity of Combretum molle (R. Br. ex G. Don) (Combretaceae) against Haemonchus contortus ova and larvae.

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Ademola, I O; Eloff, J N

2010-04-19

Parasitic nematodes, especially Haemonchus contortus (Rudolphi), are among the most common and economically important causes of disease in sheep and goats owned by pastoralists and small holder farmers in Africa. The control of these infections relies mainly on the use of anthelmintic drugs. However, herbal preparations are widely used by pastoralists and small holder farmers for the treatment of their livestock against helminth parasites. The anthelmintic effect of acetone leaf extract and fractions of Combretum molle was investigated to determine the relative efficacy of the components against gastrointestinal sheep nematodes. The fractions were obtained by solvent:solvent extraction from the acetone extract. These were evaluated for nematocidal activity by means of an egg hatch (EHA) and larval a development and viability assay (LDVA) in vitro. The effect of the test extracts on the hatchability of eggs and development of first to third stage larvae and the survival rate of the third stage larvae. H. contortus, were used to determine the relative bioactivities. Best-fit LC(50) values were computed using global model of nonlinear regression curve-fitting. The extracts inhibited egg hatching and development of the larvae of H. contortus in a concentration-dependent manner. Best-fit LC(50) values for the egg hatch test were 0.866, 0.333, 0.833, 0.747, and 0.065mg/mL for acetone extract, n-butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. The best-fit LC(50) values for the LDVA were 0.604, 0.362, 1.077, 0.131 and 0.318mg/mL for the acetone extract, butanol, hexane, chloroform, and 35% water in methanol fractions, respectively. In the EHA the 35% water in methanol fraction was significantly more active than all the other fractions (pmolle leaf could find application in anthelmintic therapy in veterinary practice.

[Ovicidal and larvicidal activity in vitro of Eucalyptus globulus essential oils on Haemonchus contortus].

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Macedo, Iara T F; Bevilaqua, Claudia M L; de Oliveira, Lorena M B; Camurça-Vasconcelos, Ana L F; Vieira, Luiz da S; Oliveira, Fabrício R; Queiroz-Junior, Eudson M; Portela, Bruno G; Barros, Renata S; Chagas, Ana C S

2009-01-01

The objective of this work was to evaluate ovicidal and larvicidal effects of Eucalyptus globulus essential oil (EGEO) on Haemonchus contortus. The chemical composition determination of EGEO was through gas chromatography and mass spectrometry. Egg hatch test (EHT) was performed in concentrations 21.75; 17.4; 8.7; 5.43 e 2.71 mg x mL(-1). In larval development test (LDT) were used the concentrations 43.5; 21.75; 10.87; 5.43 e 2.71 mg x mL(-1). Each trial was conducted by negative control with Tween 80 (3%) and positive control, 0.02 mg x mL(-1) of thiabendazole in EHT and 0.008 mg x mL(-1) of ivermectin in LDT. The maximum effectiveness of EGEO on eggs was 99.3% in concentration of 21.75 mg x mL(-1) and on larvae was 98.7% in concentration 43.5 mg x mL(-1). The concentration of EGEO that inhibits 50% of the eggs and larvae was 8.3 and 6.92 mg x mL(-1), respectively. The oil chemical analysis identified as main component the monoterpen 1,8-cineol. EGEO presented ovicidal and larvicidal activities in vitro, revealing a good potential for use in the control of sheep and goat gastrointestinal nematodes.

Site-Directed Mutagenesis Study Revealed Three Important Residues in Hc-DAF-22, a Key Enzyme Regulating Diapause of Haemonchus contortus

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Yan Huang

2017-11-01

Full Text Available Haemonchus contortus (H. contortus is one of the most important parasites of small ruminants, especially goats and sheep. The complex life cycle of this nematode is a main obstacle for the control and prevention of haemonchosis. So far, a special form of arrested development called diapause different from the dauer stage in Caenorhabditis elegans (C. elegans has been found in many parasitic nematodes. In our previous study, we have characterized a novel gene Hc-daf-22 from H. contortus sharing high homology with Ce-daf-22 and functional analysis showed this gene has similar biological function with Ce-daf-22. In this study, Hc-daf-22 mutants were constructed using site-directed mutagenesis, and carried out rescue experiments, RNA interference (RNAi experiments and in vitro enzyme activity analysis with the mutants to further explore the precise function site of Hc-DAF-22. The results showed that Hc-daf-22 mutants could be expressed in the rescued ok693 worms and the expression positions were mainly in the intestine which was identical with that of Hc-daf-22 rescued worms. Through lipid staining we found that Hc-daf-22 could rescue daf-22 mutant (ok693 from the fatty acid metabolism deficiency while Hc-daf-22 mutants failed. Brood size and body length analyses in rescue experiment along with body length and life span analyses in RNAi experiment elucidated that Hc-daf-22 resembled Ce-daf-22 in effecting the development and capacity of C. elegans and mutants impaired the function of Hc-daf-22. Together with the protease activity assay, this research revealed three important active resides 84C/299H/349H in Hc-DAF-22 by site-directed mutagenesis.

Duddingtonia flagrans chlamydospores in nutritional pellets: effect of storage time and conditions on the trapping ability against Haemonchus contortus larvae.

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Fitz-Aranda, J A; Mendoza-de-Gives, P; Torres-Acosta, J F J; Liébano-Hernández, E; López-Arellano, M E; Sandoval-Castro, C A; Quiroz-Romero, H

2015-01-01

The study evaluated the effect of storage time and conditions of nutritional pellets (NP) containing Duddingtonia flagrans chlamydospores on its in vitro trapping ability against Haemonchus contortus L3 larvae. The treated batch (200 NP) contained 4 × 106 chlamydospores of the FTH0-8 strain, whereas the control batch (200 NP) was produced without spores. Both NP batches were exposed to four experimental storage conditions: (T1) shelves (indoors); (T2) refrigeration (4°C); (T3) outdoors under a roof; and (T4) 100% outdoors. Each group comprised 48 NP with spores and 48 NP without spores (control). The ability of D. flagrans spores to trap H. contortus L3 larvae was evaluated for 8 weeks for each storage condition. For that purpose, six randomly selected NP with spores were compared to their respective control NP. Each NP was individually crushed. The crushed material (1 g) was placed on the surface of a 2% water agar plate with 200 H. contortus L3 larvae. Plates were sealed and were incubated at room temperature for 8 days. The whole content of every plate was transferred to a Baermann apparatus to recover the remaining larvae. There was a clear larval reduction in the NP with spores, compared to the respective control NP in the four storage conditions (PT2), 77 ± 6.1 (T1), 81.5 ± 3.8 (T4) and 82.1 ± 2.5 (T3). Larval reductions were similar at all times and were not affected by storage conditions or storage time (R 20.05). The long-term shelf-life of the chlamydospores in the NP suggests that this spore dosage technology is a viable option.

Site-Directed Mutagenesis Study Revealed Three Important Residues in Hc-DAF-22, a Key Enzyme Regulating Diapause of Haemonchus contortus.

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Huang, Yan; Zheng, Xiuping; Zhang, Hongli; Ding, Haojie; Guo, Xiaolu; Yang, Yi; Chen, Xueqiu; Zhou, Qianjin; Du, Aifang

2017-01-01

Haemonchus contortus ( H. contortus ) is one of the most important parasites of small ruminants, especially goats and sheep. The complex life cycle of this nematode is a main obstacle for the control and prevention of haemonchosis. So far, a special form of arrested development called diapause different from the dauer stage in Caenorhabditis elegans ( C. elegans ) has been found in many parasitic nematodes. In our previous study, we have characterized a novel gene Hc-daf-22 from H. contortus sharing high homology with Ce-daf-22 and functional analysis showed this gene has similar biological function with Ce-daf-22 . In this study, Hc-daf-22 mutants were constructed using site-directed mutagenesis, and carried out rescue experiments, RNA interference (RNAi) experiments and in vitro enzyme activity analysis with the mutants to further explore the precise function site of Hc-DAF-22. The results showed that Hc-daf-22 mutants could be expressed in the rescued ok693 worms and the expression positions were mainly in the intestine which was identical with that of Hc-daf-22 rescued worms. Through lipid staining we found that Hc-daf-22 could rescue daf-22 mutant ( ok693 ) from the fatty acid metabolism deficiency while Hc-daf-22 mutants failed. Brood size and body length analyses in rescue experiment along with body length and life span analyses in RNAi experiment elucidated that Hc-daf-22 resembled Ce-daf-22 in effecting the development and capacity of C. elegans and mutants impaired the function of Hc-daf-22 . Together with the protease activity assay, this research revealed three important active resides 84C/299H/349H in Hc-DAF-22 by site-directed mutagenesis.

Age of Haemonchus contortus third stage infective larvae is a factor influencing the in vitro assessment of anthelmintic properties of tannin containing plant extracts.

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Castañeda-Ramírez, G S; Mathieu, C; Vilarem, G; Hoste, H; Mendoza-de-Gives, P; González-Pech, P G; Torres-Acosta, J F J; Sandoval-Castro, C A

2017-08-30

The larval exsheathment inhibition assay (LEIA) of infective larvae (L 3 ) is an in vitro method used to evaluate the anthelmintic (AH) activity of tannin-containing plant extracts against different species of gastrointestinal nematodes, including Haemonchus contortus. Some conditions remain to be defined in order to standardize the LEIA, i.e. the optimal age of larvae produced from donor animals to use in the assays. Therefore, this study aimed at identifying the effect of age and age-related vitality of H. contortus infective larvae produced under tropical conditions, on the in vitro AH activity measured with the LEIA. The same acetone:water (70:30) extract from Acacia pennatula leaves was used to perform respective LEIA tests with H. contortus L 3 of different ages (1-7 weeks). Each week, the L 3 were tested against different concentrations of extract (1200, 600, 400, 200, 100, 40μg/mL of extract) plus a PBS control. Bioassays were performed with a benzimidazole (Bz) resistant H. contortus (Paraíso) strain. In order to identify changes in L 3 vitality on different weeks (1-7), two assays testing larval motility were included only with PBS: the larval migration assay (LMA) and the larval motility observation assay (LMOA). Mean effective concentrations causing 50% and 90% exsheathment inhibition (EC 50 , EC 90 ) were obtained for every week using respective Probit analyses. On the first week, the larvae had lowest EC 50 and EC 90 (39.4 and 65.6μg/mL) compared to older larvae (P0.05), while older larvae tended to show higher EC 50 and EC 90 (Page of larvae (r≥-0.83; P age. More stable efficacy results were found between two to five weeks of age. Copyright © 2017 Elsevier B.V. All rights reserved.

Study on the irradiated vaccine of the haemonchus contortus worm in lamb

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Soekardji; Arifin, M.; Murnihati, I. (National Atomic Energy Agency, Jakarta (Indonesia). Pasar Djumat Research Centre)

1983-12-01

The effect of /sup 60/Co gamma rays on the third larvae of haemonchus contortus has been studied in lambs. Inoculation of 1.000 irradiated L/sub 3/ from BPPH (Balai Penelitian Penyakit Hewan) Bogor strain (400, 500, 600 and 700 Gy) was done. The production of eggs were examined, also the effect of radiation on anaemia, and live weight gain of the lambs. The eggs production in every lamb was: control group (K)=116.0 +- 94.83; R/sub 1/ (radiation group at 400 Gy)=93.5 +- 64.05; R/sub 2/(radiation group at 500 Gy)=29.63 +- 19.67; R/sub 3/(radiation group at 600 Gy)=173.3 +- 181.34; and R/sub 4/(radiation group at 700 Gy)=125.2 +- 114.22. The specific clinical symptoms were anaemia; the data obtained were K=31.38 +- 0.49%; R/sub 1/=30.76 +- 1.21%; R/sub 2/=39.50 +- 2.27%; R/sub 3/=28.65 +- 0.92% and R/sub 4/=28.60 +- 1.44%. Average weight gain of the surviving animals was K=0.96 +- 0.49kg; R/sub 1/=3.82 +- 1.56kg; R/sub 2/=3.38 +- 0.84kg; R/sub 3/=1.57 +- 0.53kg; and R/sub 4/=1.99 +- 0.90kg. The experimental analysis of these data showed that the groups inoculated with irradiated L/sub 3/ compared with control were highly significant different, while among the 5 group were respectively highly significant.

Development of a tetra-primer ARMS-PCR for detecting the E198A SNP in the isotype-1 β-tubulin gene of Haemonchus contortus populations in China.

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Zongze, Zhang; Xin, Yang; Awais, Ali Ahmad; Weiqiang, Lei; Chunqun, Wang; Di, Wenda; Yanqin, Zhou; Junlong, Zhao; Rui, Fang; Min, Hu

2018-03-15

The tetra-primer ARMS-PCR is a rapid, simple and low cost method for single nucleotide polymorphism (SNP) genotyping and has been used to detect SNPs associated with diseases and drug resistance. E198A in the isotype-1 β-tubulin gene is one of the three SNPs associated with benzimidazole resistance in parasitic nematode Haemonchus contortus. However, up to now, only PCR-RFLP method was used to test E198A in H. contortus. In the present study, we developed a tetra-primer ARMS-PCR to detect E198A in H. contortus and the accuracy of the results was compared with that of PCR-coupled sequencing. The results showed that optimization of PCR reaction system, especially the proportion of the amount of inner and outer primers, could achieve desirable amplification effect. Three different profiles displaying three distinct genotypes could be identified clearly and intuitively on the agarose gel where the samples with amplified PCR products containing two bands of 433 bp and 200 bp in size indicated susceptible homozygous (SS), those with PCR products containing two bands of 433 bp and 284 bp in length indicated resistant homozygous (RR) and the samples with amplified PCR products containing three bands of 433 bp, 284 bp and 200 bp in size indicated heterozygous (RS). The results showed that the established method can be successfully applied to the detection of E198A in H. contortus, which has high accuracy and is easy to perform. Copyright © 2018 Elsevier B.V. All rights reserved.

Phylogenetic Study of Haemonchus Species from Iran Based On Morpho-Molecular Characterization.

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Behnam Meshgi

2015-06-01

Full Text Available Haemonchosis has a negative effect on the farming industry throughout the world, especially in the tropic and sub-tropic countries. The present study was carried out to differentiate Haemonchus species from its main hosts in Iran, including sheep, goat and camel.The identification took place based on the morphometrics of the spicules and molecular characters. Two hundred seventy adult male nematodes were collected from the abomasums of different ruminants (90 samples from each animal at the slaughterhouses from different localities in Iran. Samples were morphologically identified according to the spicules' morphometric measurements. In the section on molecular study, 10 samples of each Haemonchus isolates were genetically examined. A simple PCR-restriction fragment length polymorphism (PCR-RFLP assay of the second internal transcribed spacer of ribosomal DNA (ITS2-rDNA were described to confirm the PCR results.PCR-RFLP profile obtained from the restriction enzyme HPa1 in H. contortus and H. longistipes indicated 1 (278 bp and 2 (113 and 135 bp different fragments, respectively. The morphological parameters clearly distinguish H. contortus from H. longistipes. Moreover, regarding the ITS2-rDNA, sequences of 295 bp and 314 bp were obtained from H. contortus and H. longistipes, respectively.The genotypic results are in agreement with the phenotypic findings of both species.

Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus

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Lucas Atehmengo Ngongeh

2015-01-01

Full Text Available Response of the West African Dwarf (WAD goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS goat isolate (RSHc and the WAD goat isolate (WADHc (isolated from WAD goats, was studied by experimental infections of 4–6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3 of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP, faecal egg counts (FEC, worm burden (WB, body weight (BW, packed cell volume (PCV, and body condition score (BCS were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of 19.63±0.26 and 19.50±0.19, respectively. Goats infected with WADHc isolate had significantly higher FEC (P=0.004 and WB (P=0.001. BW were significantly higher (P=0.004 both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (P=0.001. There was a significant drop in PCV (P=0.004 of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host.

Comparative Response of the West African Dwarf Goats to Experimental Infections with Red Sokoto and West African Dwarf Goat Isolates of Haemonchus contortus.

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Ngongeh, Lucas Atehmengo; Onyeabor, Amaechi

2015-01-01

Response of the West African Dwarf (WAD) goats to two different isolates of Haemonchus contortus, the Red Sokoto (RS) goat isolate (RSHc) and the WAD goat isolate (WADHc) (isolated from WAD goats), was studied by experimental infections of 4-6-month-old male WAD goat kids. Group 1 and Group 2 goats were each infected with 4500 infective larvae (L3) of RSHc and WADHc, respectively. Group 3 animals served as uninfected control. Prepatent period (PPP), faecal egg counts (FEC), worm burden (WB), body weight (BW), packed cell volume (PCV), and body condition score (BCS) were determined. WAD goats infected with RSHc isolate and the ones infected with WADHc isolate had mean PPP of 19.63 ± 0.26 and 19.50 ± 0.19, respectively. Goats infected with WADHc isolate had significantly higher FEC (P = 0.004) and WB (P = 0.001). BW were significantly higher (P = 0.004) both in the controls and in Group 2 goats infected with WADHc isolate than in Group 1 goats infected with the RSHc isolate. BCS of animals in both infected groups dropped significantly (P = 0.001). There was a significant drop in PCV (P = 0.004) of both infected groups in comparison. Both isolates of H. contortus were pathogenic to the host.

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in grazing South African goats.

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Vatta, A F; Waller, P J; Githiori, J B; Medley, G F

2012-11-23

A study was conducted to examine the duration of anthelmintic effect of copper oxide wire particles (COWP) in grazing goats, as data for the persistence of efficacy of COWP in this host species is limited. Forty-eight indigenous male goats were infected naturally by grazing them on Haemonchus contortus-infected pasture. When the faecal egg count (FEC) in the goats was 3179 ± 540 eggs per gram of faeces (mean ± standard error), half the animals were treated with 4 g COWP (day 0; mean live weight=25.5 ± 0.8 kg). Eight treated (COWP) and eight non-treated (CONTROL) goats were removed from the pasture on each of days 7, 28 and 56, maintained for 27 or 29 days in concrete pens and then humanely slaughtered for nematode recovery. Mean liver copper levels were in the high range in the goats removed from pasture at day 7 (treated: 191 ± 19.7 ppm; untreated: 120 ± 19.7 ppm; P=0.022), but had dropped to normal levels at days 28 and 56. The mean H. contortus burdens of the treated versus the non-treated goats were, respectively, 184 ± 48 and 645 ± 152 for the goats removed from pasture at day 7 (71% reduction; P=0.004), 207 ± 42 and 331 ± 156 at day 28 (37% reduction; P=0.945) and 336 ± 89 and 225 ± 53 at day 56 (-49% reduction; P=0.665). Weekly monitoring of FECs after treatment until slaughter indicated that the COWP-treated goats had lower FECs than the controls, the treatment main effect being significant at days 7, 28 and 56 (Pcopper levels return to normal two to three months after COWP treatment. Copyright © 2012 Elsevier B.V. All rights reserved.

Anthelmintic effect of carob pods and sainfoin hay when fed to lambs after experimental trickle infections with Haemonchus contortus and Trichostrongylus colubriformis

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Arroyo-Lopez Celia

2014-01-01

Full Text Available The aim of the study was to compare the in vivo anthelmintic activity of sainfoin hay (Onobrychis viciifolia and carob pod meal (Ceratonia siliqua against gastrointestinal nematodes. Seven days before infection, 64 naive lambs were assigned to four different groups: Group S received sainfoin hay and group CAR was fed with carob pods. The remaining lambs received lucerne hay (Medicago sativa and were assigned to positive (non-treated, NT and negative (treated, T control groups (treatment with albendazole. On day 0, lambs were artificially trickle infected for 6 weeks, with a mixture of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis. Parasitological and pathophysiological parameters were measured repeatedly during the 2-month study. Compared to the NT group, decreases in egg excretion were observed in the CAR and S groups with significant differences only found for sainfoin (p < 0.05. At necropsy, group S showed decreases in the total worm numbers of both nematode species with significant differences for H. contortus. In contrast, no differences were noticed for the CAR group. Compared to the NT group, lower values for fecundity of female H. contortus were found in the S and CAR groups, however differences were non-significant. No differences in body weight gains were found between groups. Consistent results were found showing significantly higher packed cell volume (PCV values in the T and S groups compared to NT and CAR groups. Overall, these results confirm a positive effect associated with the feeding of lambs with tanniniferous resources on host resilience (PCV values and against gastrointestinal parasitic nematodes by affecting some biological traits of worm populations (e.g. eggs per gram of faeces and worm numbers. However, the anthelmintic effects differed between the two tannin-containing resources, which might be associated with the quantity and/or quality of secondary metabolites (condensed tannins and

Piper aduncum against Haemonchus contortus isolates: cross resistance and the research of natural bioactive compounds

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Yousmel Alemán Gaínza

Full Text Available Abstract The anthelminthic activity of the essential oil (EO of Piper aduncum L. was tested in vitro on eggs and larvae of resistant (Embrapa2010 and susceptible (McMaster isolates of Haemonchus contortus. The EO was obtained by steam distillation and its components identified by chromatography. EO concentrations of 12.5 to 0.02 mg/mL were used in the egg hatch test (EHT and concentrations of 3.12 to 0.01 mg/mL in the larval development test (LDT. Inhibition concentrations (IC were determined by the SAS Probit procedure, and significant differences assessed by ANOVA followed by Tukey’s test. In the EHT, the IC50 for the susceptible isolate was 5.72 mg/mL. In the LDT, the IC50 and IC90 were, respectively, 0.10 mg/mL and 0.34 mg/mL for the susceptible isolate, and 0.22 mg/mL and 0.51 mg/mL for the resistant isolate. The EO (dillapiole 76.2% was highly efficacious on phase L1. Due to the higher ICs obtained for the resistant isolate, it was raised the hypothesis that dillapiole may have a mechanism of action that resembles those of other anthelmintic compounds. We further review and discuss studies, especially those conducted in Brazil, that quantified the major constituents of P. aduncum-derived EO.

Piper aduncum against Haemonchus contortus isolates: cross resistance and the research of natural bioactive compounds.

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Gaínza, Yousmel Alemán; Fantatto, Rafaela Regina; Chaves, Francisco Celio Maia; Bizzo, Humberto Ribeiro; Esteves, Sérgio Novita; Chagas, Ana Carolina de Souza

2016-01-01

The anthelminthic activity of the essential oil (EO) of Piper aduncum L. was tested in vitro on eggs and larvae of resistant (Embrapa2010) and susceptible (McMaster) isolates of Haemonchus contortus. The EO was obtained by steam distillation and its components identified by chromatography. EO concentrations of 12.5 to 0.02 mg/mL were used in the egg hatch test (EHT) and concentrations of 3.12 to 0.01 mg/mL in the larval development test (LDT). Inhibition concentrations (IC) were determined by the SAS Probit procedure, and significant differences assessed by ANOVA followed by Tukey's test. In the EHT, the IC50 for the susceptible isolate was 5.72 mg/mL. In the LDT, the IC50 and IC90 were, respectively, 0.10 mg/mL and 0.34 mg/mL for the susceptible isolate, and 0.22 mg/mL and 0.51 mg/mL for the resistant isolate. The EO (dillapiole 76.2%) was highly efficacious on phase L1. Due to the higher ICs obtained for the resistant isolate, it was raised the hypothesis that dillapiole may have a mechanism of action that resembles those of other anthelmintic compounds. We further review and discuss studies, especially those conducted in Brazil, that quantified the major constituents of P. aduncum-derived EO.

Phenobarbital induction and chemical synergism demonstrate the role of UDP-glucuronosyltransferases in detoxification of naphthalophos by Haemonchus contortus larvae.

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Kotze, Andrew C; Ruffell, Angela P; Ingham, Aaron B

2014-12-01

We used an enzyme induction approach to study the role of detoxification enzymes in the interaction of the anthelmintic compound naphthalophos with Haemonchus contortus larvae. Larvae were treated with the barbiturate phenobarbital, which is known to induce the activity of a number of detoxification enzymes in mammals and insects, including cytochromes P450 (CYPs), UDP-glucuronosyltransferases (UDPGTs), and glutathione (GSH) S-transferases (GSTs). Cotreatment of larvae with phenobarbital and naphthalophos resulted in a significant increase in the naphthalophos 50% inhibitory concentration (IC50) compared to treatment of larvae with the anthelmintic alone (up to a 28-fold increase). The phenobarbital-induced drug tolerance was reversed by cotreatment with the UDPGT inhibitors 5-nitrouracil, 4,6-dihydroxy-5-nitropyrimidine, probenecid, and sulfinpyrazone. Isobologram analysis of the interaction of 5-nitrouracil with naphthalophos in phenobarbital-treated larvae clearly showed the presence of strong synergism. The UDPGT inhibitors 5-nitrouracil, 4,6-dihydroxy-5-nitropyrimidine, and probenecid also showed synergistic effects with non-phenobarbital-treated worms (synergism ratio up to 3.2-fold). This study indicates that H. contortus larvae possess one or more UDPGT enzymes able to detoxify naphthalophos. In highlighting the protective role of this enzyme group, this study reveals the potential for UDPGT enzymes to act as a resistance mechanism that may develop under drug selection pressure in field isolates of this species. In addition, the data indicate the potential for a chemotherapeutic approach utilizing inhibitors of UDPGT enzymes as synergists to increase the activity of naphthalophos against parasitic worms and to combat detoxification-mediated drug resistance if it arises in the field. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Conservation of a microRNA cluster in parasitic nematodes and profiling of miRNAs in excretory-secretory products and microvesicles of Haemonchus contortus.

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Henry Y Gu

2017-11-01

Full Text Available microRNAs are small non-coding RNAs that are important regulators of gene expression in a range of animals, including nematodes. We have analysed a cluster of four miRNAs from the pathogenic nematode species Haemonchus contortus that are closely linked in the genome. We find that the cluster is conserved only in clade V parasitic nematodes and in some ascarids, but not in other clade III species nor in clade V free-living nematodes. Members of the cluster are present in parasite excretory-secretory products and can be detected in the abomasum and draining lymph nodes of infected sheep, indicating their release in vitro and in vivo. As observed for other parasitic nematodes, H. contortus adult worms release extracellular vesicles (EV. Small RNA libraries were prepared from vesicle-enriched and vesicle-depleted supernatants from both adult worms and L4 stage larvae. Comparison of the miRNA species in the different fractions indicated that specific miRNAs are packaged within vesicles, while others are more abundant in vesicle-depleted supernatant. Hierarchical clustering analysis indicated that the gut is the likely source of vesicle-associated miRNAs in the L4 stage, but not in the adult worm. These findings add to the growing body of work demonstrating that miRNAs released from parasitic helminths may play an important role in host-parasite interactions.

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in sheep.

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Galindo-Barboza, A J; Torres-Acosta, J F J; Cámara-Sarmiento, R; Sandoval-Castro, C A; Aguilar-Caballero, A J; Ojeda-Robertos, N F; Reyes-Ramírez, R; España-España, E

2011-03-10

The aim was to determine the persistent efficacy of copper oxide wire particles (COWP) against Haemonchus contortus in sheep, using the harmonization guidelines protocol. Thirty-six male lambs (2 months old) reared free of gastrointestinal nematodes were used (average body weight of 10.8±3.8kg). Before and for the duration of the study, lambs were kept in raised cages with slatted floors and were offered ad libitum a complete mixed diet. Animals were divided into six groups (n=6): one non-treated control group (G0) and five groups treated with one COWP capsule (1.7g of copper oxide; Copinox(®)). Animals in each group were treated on pre-defined dates before the artificial infection was applied: days -35 (G1), -28 (G2), -21 (G3), -14 (G4) and -7 (G5). On day 0 animals were infected with 3700 H. contortus infective larvae per animal. Animals were humanely slaughtered between days 22 and 23 post-infection. The abomasums were individually washed to obtain the contents. These organs were subjected to separate artificial digestions. Adult parasites were counted from the abomasum contents and the larvae from the digested material. Worm burden geometric means were calculated for each group. A significant worm burden reduction in either of the treated groups (G1, G2, G3, G4, and G5) compared to the control (G0) was considered as persistence of the anthelmintic effect. Copper levels were determined from individual liver samples of each animal. The geometric mean worm burden of the control group (G0) was 1959. Compared to the control, worm burdens geometric means were significantly reduced in groups G1 (1108), G4 (528) and G5 (1063) (P<0.03). Efficacies in G1, G4 and G5 were 43.4%, 73.0% and 45.7% respectively. No significant reduction was found for G2 (1342) and G3 (1430). A larger quantity of Cu was found in the livers of treated animals compared to the control group (P<0.05) except for G3 (P=0.06). A negative association between Cu liver content and worm burdens was

A study of the influence of breed and strain on the susceptibility of sheep to Haemonchus controtus infection

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Castelino, J.

1976-07-01

Breed appears to influence the susceptibility of sheep to Haemonchus contortus infection. Experiments on Masai and Merino sheep indicated that the Masai appear innately capable of reacting more vigorously to throw off H. contortus infection, this reaction being enhanced by previous experience. Following challenge, tangible resistance develops consistently in Masai, compared to a partial resistance, at best, in Merino. Resistance appears to be a short-lived phenomenon in both breeds. A field trial was carried out, to determine the influence of hemoglobin type on the susceptibility of Merino sheep. Sheep of hemoglobin types AA, AB and BB were used. A lower egg production occurred in hemoglobin type AA sheep, contrary to results obtained in the laboratory where ''massive'' experimental infection (250 Haemonchus larvae/kg liveweight) was effected. Tracer techniques involving 51 Cr- labelled erythrocytes and 125 I-tagged plasma were applied

Effect of the infection with the nematode Haemonchus contortus (Strongylida: Trichostrongylidae on the haematological, biochemical, clinical and reproductive traits in rams

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Mariem Rouatbi

2016-08-01

Full Text Available This study aimed to investigate the effect of Haemonchus contortus infection on rams’ haematological, biochemical and clinical parameters and reproductive performances. A total number of 12 Barbarine rams (control and infected were included in the experiment. The infected group received 30 000 H. contortus third-stage larvae orally. Each ram’s ejaculate was immediately evaluated for volume, sperm cell concentration and mortality rate. At the end of the experiment (day 82 post-infection, which lasted 89 days, serial blood samples were collected in order to assess plasma testosterone and luteinising hormone (LH concentrations. There was an effect of time, infection and their interaction on haematological parameters (p < 0.001. In infected rams, haematocrit, red blood cell count and haemoglobin started to decrease from 21 days post-infection. There was an effect of time and infection for albumin. For total protein, only infection had a statistically significant effect. For glucose, only time had a statistically significant effect. Concentrations were significantly lower in infected rams compared to control animals. A significant effect of infection and time on sperm concentrations and sperm mortality was observed. The effect of infection appears in time for sperm concentrations at days 69 and 76 post-infection. Sperm mortality rate was significantly higher in infected animals at day 46 post-infection when compared to control group (p < 0.05. Finally, plasma testosterone traits (average concentration, cumulated levels during the sampling period and pulse frequency were depressed in infected rams when compared to control counterparts; none of these endocrine traits were affected for plasma LH.

Single vs. double dose of copper oxide wire particles (COWP) for treatment of anthelmintic resistant Haemonchus contortus in weanling lambs.

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Schweizer, Nikki M; Foster, Derek M; Knox, William B; Sylvester, Hannah J; Anderson, Kevin L

2016-10-15

Haemonchus contortus parasitism is a major disease of sheep, with these parasites frequently demonstrating multi-drug class anthelmintic resistance. Copper oxide wire particles (COWP) have shown potential as adjuncts or alternatives to anthelmintics in resistant flocks. The purpose of this study was to compare the efficacy of two different COWP treatment regimens or placebo in the control of H. contortus in weaned lambs within a flock historically shown to have multi-drug resistant H. contortus using the DrenchRite ® assay. Data from 43 lambs within 3 treatment groups in a double blind study were included in the experiment. Treatments were administered as a total of 2 boluses, each given on separate occasions (day 0 and day 42), so that each lamb received either 2 placebos, a single dose of 2g COWP followed by placebo, or two doses of 1g COWP. Strongyle-type fecal egg counts (FEC) were performed at initial treatment (day 0), on day 10, at second treatment (day 42), on day 52, and at study end (day 84). At the start of the trial, mean±standard deviation FEC were 1634.4±825.2, 2241.7±1496.8, and 2013.3±1194.2epg for the 2g, 1g×2, and control groups, respectively. At the end of the trial, FEC were 757.1±825.3, 483.4±557.2, and 1660.0±1345.3epg for the 2g, 1g×2, and control groups, respectively. Lambs given a 2g single dose of COWP or a 1g dose of COWP twice had reductions in strongyle-type FEC (p≤0.01) from trial start to trial end, whereas lambs given placebo did not. Average daily gains did not differ significantly among groups. Although copper is potentially toxic to sheep, no signs of toxicity were observed during this trial, which was consistent with similar studies at this treatment dose. The study indicated that administering COWP to lambs at weaning reduced FEC. Copyright © 2016 Elsevier B.V. All rights reserved.

Anthelmintic effect of carob pods and sainfoin hay when fed to lambs after experimental trickle infections with Haemonchus contortus and Trichostrongylus colubriformis.

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Arroyo-Lopez, Celia; Manolaraki, Foteini; Saratsis, Anastasios; Saratsi, Katerina; Stefanakis, Alexandros; Skampardonis, Vasileios; Voutzourakis, Nikolaos; Hoste, Hervé; Sotiraki, Smaragda

2014-01-01

The aim of the study was to compare the in vivo anthelmintic activity of sainfoin hay (Onobrychis viciifolia) and carob pod meal (Ceratonia siliqua) against gastrointestinal nematodes. Seven days before infection, 64 naive lambs were assigned to four different groups: Group S received sainfoin hay and group CAR was fed with carob pods. The remaining lambs received lucerne hay (Medicago sativa) and were assigned to positive (non-treated, NT) and negative (treated, T) control groups (treatment with albendazole). On day 0, lambs were artificially trickle infected for 6 weeks, with a mixture of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis. Parasitological and pathophysiological parameters were measured repeatedly during the 2-month study. Compared to the NT group, decreases in egg excretion were observed in the CAR and S groups with significant differences only found for sainfoin (p < 0.05). At necropsy, group S showed decreases in the total worm numbers of both nematode species with significant differences for H. contortus. In contrast, no differences were noticed for the CAR group. Compared to the NT group, lower values for fecundity of female H. contortus were found in the S and CAR groups, however differences were non-significant. No differences in body weight gains were found between groups. Consistent results were found showing significantly higher packed cell volume (PCV) values in the T and S groups compared to NT and CAR groups. Overall, these results confirm a positive effect associated with the feeding of lambs with tanniniferous resources on host resilience (PCV values) and against gastrointestinal parasitic nematodes by affecting some biological traits of worm populations (e.g. eggs per gram of faeces and worm numbers). However, the anthelmintic effects differed between the two tannin-containing resources, which might be associated with the quantity and/or quality of secondary metabolites (condensed tannins and/or other

Effects of single or trickle Haemonchus contortus experimental infection on digestibility and host responses of naïve Creole kids reared indoor.

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Bambou, J C; Cei, W; Camous, S; Archimède, H; Decherf, A; Philibert, L; Barbier, C; Mandonnet, N; González-García, E

2013-01-31

The objective of this study was to compare the effects of the type of Haemonchus contortus experimental infection (trickle infection, TI versus single infection, SI) on feed intake, nutrients digestibility, parasitological and haematological measures, and plasma leptin in Creole kids. The animals were infected over 2 periods (challenge 1 and challenge 2) of 6 weeks each, corresponding respectively to the primary and the secondary infection. Periods prior infection (1 week each) were considered as controls. The primary infection was realized with 35 Creole kids (18.40±3.76 kg BW) housed in individual boxes and fed a hay-based diet. The secondary infection continued with 29 kids (21.90±3.40 kg BW) from the initial 35. A total of 6 kids and 8 kids were slaughtered for measuring nematode burden at the end of the primary and the secondary infection, respectively. Measurements of nutrients digestibility were made at 0, 3 and 5 weeks post-infection for both challenges. Faecal egg count (FEC), blood eosinophilia and packed cell volume (PCV) were monitored weekly. Feed intake (dry matter intake, DMI) and nutrients digestibility were negatively affected by H. contortus infection only during the primary infection. Plasma leptin changed significantly over time (P=0.0002) but was not affected by the infection type. Effect of infection type was observed only on crude protein digestibility during the primary infection, which was lower in the TI group (P<0.01). The overall level of blood eosinophilia was significantly higher in the TI group (P<0.0001) during both challenges. The overall FEC mean was significantly higher in the SI compared with the TI groups, during both challenges (P<0.02). These results were related to the mean female length significantly higher in the SI group compared with the TI group during challenge 1 (P=0.004), and the number of adult nematode significantly lower in the TI group compared with the SI group during the challenge 2 (P=0.05). The results

Hc-daf-2 encodes an insulin-like receptor kinase in the barber's pole worm, Haemonchus contortus, and restores partial dauer regulation.

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Li, Facai; Lok, James B; Gasser, Robin B; Korhonen, Pasi K; Sandeman, Mark R; Shi, Deshi; Zhou, Rui; Li, Xiangrui; Zhou, Yanqin; Zhao, Junlong; Hu, Min

2014-06-01

Infective L3s (iL3s) of parasitic nematodes share common behavioural, morphological and developmental characteristics with the developmentally arrested (dauer) larvae of the free-living nematode Caenorhabditis elegans. It is proposed that similar molecular mechanisms regulate entry into or exit from the dauer stage in C. elegans, and the transition from free-living to parasitic forms of parasitic nematodes. In C. elegans, one of the key factors regulating the dauer transition is the insulin-like receptor (designated Ce-DAF-2) encoded by the gene Ce-daf-2. However, nothing is known about DAF-2 homologues in most parasitic nematodes. Here, using a PCR-based approach, we identified and characterised a gene (Hc-daf-2) and its inferred product (Hc-DAF-2) in Haemonchus contortus (a socioeconomically important parasitic nematode of ruminants). The sequence of Hc-DAF-2 displays significant sequence homology to insulin receptors (IR) in both vertebrates and invertebrates, and contains conserved structural domains. A sequence encoding an important proteolytic motif (RKRR) identified in the predicted peptide sequence of Hc-DAF-2 is consistent with that of the human IR, suggesting that it is involved in the formation of the IR complex. The Hc-daf-2 gene was transcribed in all life stages of H. contortus, with a significant up-regulation in the iL3 compared with other stages. To compare patterns of expression between Hc-daf-2 and Ce-daf-2, reporter constructs fusing the Ce-daf-2 or Hc-daf-2 promoter to sequence encoding GFP were microinjected into the N2 strain of C. elegans, and transgenic lines were established and examined. Both genes showed similar patterns of expression in amphidial (head) neurons, which relate to sensation and signal transduction. Further study by heterologous genetic complementation in a daf-2-deficient strain of C. elegans (CB1370) showed partial rescue of function by Hc-daf-2. Taken together, these findings provide a first insight into the roles of Hc

Structural and functional characterization of a novel gene, Hc-daf-22, from the strongylid nematode Haemonchus contortus.

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Guo, Xiaolu; Zhang, Hongli; Zheng, Xiuping; Zhou, Qianjin; Yang, Yi; Chen, Xueqiu; Du, Aifang

2016-07-29

The strongylid nematode Haemonchus contortus is a parasite of major concern for modern livestock husbandry because hostile environmental conditions may induce diapause in the early fourth-stage larvae. A new gene Hc-daf-22 was identified which is the homologue of Ce-daf-22 and human SCPx. Genome walking and RACE were performed to obtain the whole cDNA and genomic sequence of this gene. Using qRT-PCR with all developmental stages as templates to explore the transcription level and micro-injection was applied to confirm the promoter activity of the 5'-flanking region. Overexpression, rescue and RNA interference experiments were performed in N2, daf-22 mutant (ok 693) strains of C. elegans to study the gene function of Hc-daf-22. The full length gene of Hc-daf-22 (6,939 bp) contained 16 exons separated by 15 introns, and encoded a cDNA of 1,602 bp (533 amino acids, estimated at about 59.3 kDa) with a peak in L3 and L4 in transcriptional level. The Hc-DAF-22 protein was consisted of a 3-oxoacyl-CoA thiolase domain and a SCP2 domain and evolutionarily conserved. The 1,548 bp fragment upstream of the 5'-flanking region was confirmed to have promoter activity compared with 5'-flanking region of Ce-daf-22. The rescue experiment by micro-injection of daf-22 (ok693) mutant strain showed significant increase in body size and brood size in the rescued worms with significantly reduced or completely absent fat granules confirmed by Oil red O staining, indicating that Hc-daf-22 could partially rescue the function of Ce-daf-22. Furthermore, RNAi with Hc-daf-22 could partially silence the endogenous Ce-daf-22 in N2 worms and mimic the phenotype of daf-22 (ok693) mutants. The gene Hc-daf-22 was isolated and its function identified using C. elegans as a model organism. Our results indicate that Hc-daf-22 shared similar characteristics and function with Ce-daf-22 and may play an important role in peroxisomal β-oxidation and the development in H. contortus.

Introgression of ivermectin resistance genes into a susceptible Haemonchus contortus strain by multiple backcrossing.

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Elizabeth Redman

2012-02-01

Full Text Available Anthelmintic drug resistance in livestock parasites is already widespread and in recent years there has been an increasing level of anthelmintic drug selection pressure applied to parasitic nematode populations in humans leading to concerns regarding the emergence of resistance. However, most parasitic nematodes, particularly those of humans, are difficult experimental subjects making mechanistic studies of drug resistance extremely difficult. The small ruminant parasitic nematode Haemonchus contortus is a more amenable model system to study many aspects of parasite biology and investigate the basic mechanisms and genetics of anthelmintic drug resistance. Here we report the successful introgression of ivermectin resistance genes from two independent ivermectin resistant strains, MHco4(WRS and MHco10(CAVR, into the susceptible genome reference strain MHco3(ISE using a backcrossing approach. A panel of microsatellite markers were used to monitor the procedure. We demonstrated that after four rounds of backcrossing, worms that were phenotypically resistant to ivermectin had a similar genetic background to the susceptible reference strain based on the bulk genotyping with 18 microsatellite loci and individual genotyping with a sub-panel of 9 microsatellite loci. In addition, a single marker, Hcms8a20, showed evidence of genetic linkage to an ivermectin resistance-conferring locus providing a starting point for more detailed studies of this genomic region to identify the causal mutation(s. This work presents a novel genetic approach to study anthelmintic resistance and provides a "proof-of-concept" of the use of forward genetics in an important model strongylid parasite of relevance to human hookworms. The resulting strains provide valuable resources for candidate gene studies, whole genome approaches and for further genetic analysis to identify ivermectin resistance loci.

Evaluation of Copper Supplementation to Control Haemonchus contortus Infections of Sheep in Sweden

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Rydzik A

2004-09-01

Full Text Available A pen study was conducted to assess the effect of providing daily copper mineral supplement, or copper wire particle (COWP capsules, on established or incoming mixed nematode infections in young sheep. For lambs with established (6 week old infections, COWP resulted in 97% and 56% reduction of the adult and early L4 stages of H. contortus, respectively, compared with controls (p Teladorsagia circumcincta infections in the COWP lambs compared with controls (p H. contortus infections, but lack of parasites during the grazing season prevented an adequate assessment from being made. These results indicate that there is little, if any, benefit from a parasite control standpoint in recommending copper therapy, specifically to control parasites in Swedish sheep flocks.

Interaction between the effects of evaporation rate and amount of simulated rainfall on development of the free-living stages of Haemonchus contortus.

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O'Connor, Lauren J; Kahn, Lewis P; Walkden-Brown, Stephen W

2008-08-17

A factorial experiment (3 x 4 x 2 x 3) was conducted in programmable incubators to investigate interaction between the effects of rainfall amount, rainfall distribution and evaporation rate on development of Haemonchus contortus to L3. Sheep faeces containing H. contortus eggs were incubated on sterilised soil under variable temperatures typical of summer in the Northern Tablelands of NSW, Australia. Simulated rainfall was applied in 1 of 3 amounts (12, 24 or 32 mm) and 4 distributions (a single event on the day after deposition, or the same total amount split in 2, 3 or 4 equal events over 2, 3 or 4 days, respectively). Samples were incubated at either a Low or High rate of evaporation (Low: 2.1-3.4 mm/day and High: 3.8-6.1 mm/day), and faeces and soil were destructively sampled at 4, 7 and 14 days post-deposition. Recovery of L3 from the soil (extra-pellet L3) increased over time (up to 0.52% at day 14) and with each increment of rainfall (12 mm: evaporation rate (0.01%) compared with the Low evaporation rate (0.31%). All rainfall amounts yielded significantly different recoveries of L3 under Low evaporation rates but there was no difference between the 12 and 24 mm treatments under the High evaporation rate. The distribution of simulated rainfall did not significantly affect recovery of infective larvae. Faecal moisture content was positively associated with L3 recovery, as was the ratio of cumulative precipitation and cumulative evaporation (P/E), particularly when measured in the first 4 days post-deposition. The results show that evaporation rate plays a significant role in regulating the influence of rainfall amount on the success of L3 transmission.

Blood parameters of sheep with high infection of Haemonchus contortus and treated with “mushroom of the sun” (Agaricus blazei

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Gabriela Almeida Bastos

2016-04-01

Full Text Available Blood and parasitological parameters of sheep experimentally infected with Haemonchus contortus and treated with “mushroom of the sun” (Agaricus blazei were assessed. Lambs infected with the nematode were divided into three groups: treated with basidiocarp powder from the fungus, wormed with trichlorfon, and a control group that did not receive anthelmintic. Split-plot design analysis was performed where the treatments were defined as plots and four periods of collection were defined as subplots. Interaction between the treatments and the collection periods for the fecal egg counts per gram of feces (FEC was observed. Treatment with “mushroom of the sun” showed anthelmintic efficacies ranging from 28.6 to 54.2%. Similar performances to weight gain were observed among the groups. A significant interaction between treatments and evaluation periods was observed for erythrocyte counts, hematocrit value, and serum concentrations of albumin and urea. The values of the mean corpuscular volume, erythrocyte distribution width, and leukocyte, neutrophil, lymphocyte, monocyte, and eosinophil counts, as well as the mean platelet volume, varied among the collection days. The administration of “mushroom of the sun” significantly reduced the FEC, did not influence on the erythrocyte count and hematocrit value, which remained within normal limits. However it increased the serum concentrations of albumin.

Anthelmintic efficacy of five tropical native Australian plants against Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats (Capra hircus).

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Moreno, F C; Gordon, I J; Knox, M R; Summer, P M; Skerrat, L F; Benvenutti, M A; Saumell, C A

2012-06-08

The study of the anthelmintic properties of plants rich in plant secondary metabolites can provide ecologically sound methods for the treatment of parasites on grazing animals. The purpose of the present study was to evaluate the anthelmintic effect of five tropical native Australian plant species rich in plant secondary metabolites on adult Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats. Thirty young, nematode-free goats were infected with 2500 H. contortus and 5000 T. colubriformis infective larvae thrice weekly for a week (day 1-7 of the experiment). On day 27 after first infection, the goats were allocated into six groups of five animals per group. From day 28 to day 35, fresh leaves from Acacia salicina, Acacia nilotica, Eucalyptus corymbia, Casuarina cunninghamiana and Eucalyptus drepanophylla were included in the goats diet. Five groups were offered leaves from one of these plant species and one group, the untreated control, received only the basal diet formulated with 20% Medicago sativa and 80% Avena sativa. Following plant material administration, the goats were monitored daily until day 40 and then slaughtered on day 41. Total faecal worm egg output, total production of larvae recovered from faecal cultures, total post-mortem worm burdens and the per capita fecundity of female worms were estimated. The toxicity of the plant species for the goats was measured by histopathological analyses of liver and kidney samples. Results showed that goats feeding on the plant material rich in plant secondary metabolites had significantly lower egg output compared to the control goats (P0.05), the per capita fecundity was significantly reduced by E. corymbia, A. nilotica and A. salicina (Pgoats can benefit from the short-term ingestion of plant secondary metabolites, which reduce the total faecal egg output and thus decrease the potential for re-infection from the pasture. Crown Copyright © 2012. Published by Elsevier B.V. All

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in grazing South African goats

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Vatta, A.F.; Waller, P.J.; Githiori, J.B.; Medley, G.F.

2012-01-01

A study was conducted to examine the duration of anthelmintic effect of copper oxide wire particles (COWP) in grazing goats, as data for the persistence of efficacy of COWP in this host species is limited. Forty-eight indigenous male goats were infected naturally by grazing them on Haemonchus contortus-infected pasture. When the faecal egg count (FEC) in the goats was 3179 ± 540 eggs per gram of faeces (mean ± standard error), half the animals were treated with 4 g COWP (day 0; mean live weight = 25.5 ± 0.8 kg). Eight treated (COWP) and eight non-treated (CONTROL) goats were removed from the pasture on each of days 7, 28 and 56, maintained for 27 or 29 days in concrete pens and then humanely slaughtered for nematode recovery. Mean liver copper levels were in the high range in the goats removed from pasture at day 7 (treated: 191 ± 19.7 ppm; untreated: 120 ± 19.7 ppm; P = 0.022), but had dropped to normal levels at days 28 and 56. The mean H. contortus burdens of the treated versus the non-treated goats were, respectively, 184 ± 48 and 645 ± 152 for the goats removed from pasture at day 7 (71% reduction; P = 0.004), 207 ± 42 and 331 ± 156 at day 28 (37% reduction; P = 0.945) and 336 ± 89 and 225 ± 53 at day 56 (−49% reduction; P = 0.665). Weekly monitoring of FECs after treatment until slaughter indicated that the COWP-treated goats had lower FECs than the controls, the treatment main effect being significant at days 7, 28 and 56 (P goats removed from pasture at day 28 (P ≤ 0.001). Packed cell volumes increased during the course of the experiment (day, P goats removed from pasture at day 28 (CONTROL 28 d, 28.65 ± 0.52% goats, extending at most to 28 days after treatment. However, repeated COWP administration at three-month intervals may be safe, given that liver copper levels return to normal two to three months after COWP treatment. PMID:22789299

Predatory activity of Butlerius nematodes and nematophagous fungi against Haemonchus contortus infective larvae

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Manoel Eduardo da Silva

Full Text Available Abstract The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3 in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8. Two thousand H. contortus infective larvae (L3 were added to each group. Additionally, the following treatments were established: Group 1 – 2000 Butlerius spp. larvae; group 2 – A. musiformis (1x107 conidia; group 3 – T. esau (1x107 conidia; group 4 – C. rosea (1x107 conidia, group 5 – D. flagrans (1x107conidia and Group 6 – no biological controller (control group. The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

Evaluation of the phenotypic performance of a Red Maasai and Dorper double backcross resource population: indoor trickle challenge with Haemonchus contortus.

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Mugambi, J M; Audho, J O; Njomo, S; Baker, R L

2005-02-28

Six F(1) Dorper (D) x Red Maasai (R) rams were mated to both D and R ewes to produce backcross lambs. These six double backcross resource families are being analysed to identify quantitative trait loci that may be controlling resistance to gastro-intestinal (GI) nematode parasites, mainly Haemonchus contortus. After assessing the phenotypic performance of the lambs following exposure to natural infections, the surviving lambs were drenched and moved indoors for an artificial challenge with H. contortus along with straightbred D and R lambs. A total of 1317 lambs were included in the analyses consisting of 523 3/4D, 580 3/4R, 87 D and 127 R. The D lambs were significantly and consistently heavier than R lambs and 3/4D were significantly heavier than the 3/4R lambs. The difference between the backcrosses was about half of that between the straightbreds. Resistance was assessed in terms of faecal egg counts (FEC) and total worm counts (TWC) at necropsy while packed cell volume (PCV) was used to assess resilience to weekly oral doses of 2500 infective larvae of H. contortus. No significant breed differences were observed for log transformed FEC (LFEC). A significant breed difference in PCV was recorded. The backcrosses had the higher values and while no differences were observed between the straightbreds, 3/4D had significantly higher PCV than the 3/4R. Despite the absence of breed differences in FEC the R and the 3/4R had significantly fewer worms than the D and the 3/4D. The D had significantly longer worms than the R and the 3/4D had significantly longer worms than the 3/4R. Worms recovered from D had more eggs than those recovered from R. Similarly worms from 3/4D contained more eggs than those from 3/4R. Thus, on a breed basis the breed with more worms had longer worms. In contrast, when, in a small part of the experiment two doses of larvae were used to check for any breed by dose interactions, worms from the low dose (and hence fewer worms) animals were longer

The anthelmintic effect of aqueous methanol extract of Combretum molle (R. Br. x. G. Don) (Combretaceae) in lambs experimentally infected with Haemonchus contortus.

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Simon, M K; Ajanusi, O J; Abubakar, M S; Idris, A L; Suleiman, M M

2012-06-08

The aqueous methanol extract from the stem-bark of Combretum molle was evaluated for anthelmintic activity in lambs infected with Haemonchus contortus using faecal egg count (FEC) reduction assay. The extract showed a dose-dependent reduction in FEC in infected animals. At doses of 500, 1000 and 2000 mg kg(-1), the extract caused FEC reduction of 63%, 69.25% and 96.23%, respectively. Similarly, the standard anthelmintic (albendazole) at a dose of 200 mg kg(-1) produced FEC reduction of 99.24%. FEC reduction produced by the extract at doses of 500 and 1000 mg kg(-1) is below the minimum standard of 90% FEC recommended by the World Association for the Advancement of Veterinary Parasitology (WAAVP). However, there was no significant (P>0.05) difference between the means of groups treated with 1000 mg kg(-1) and 2000 mg kg(-1) compared to that of albendazole. In this study, C. molle has shown a promising anthelmintic activity against experimental haemonchosis. Nonetheless, further studies to evaluate its detailed toxicity are required for the plant extract to be developed into a useful anthelmintic drug. There is also the need to evaluate other parts of the plant (root, leaves, fruits, etc.) for the same effect. Copyright © 2012 Elsevier B.V. All rights reserved.

Impact of the post-weaning nutritional history on the response to an experimental Haemonchus contortus infection in Creole goats and Black Belly sheep.

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Ceï, W; Salah, N; Paut, C; Dumoulin, P-J; Arquet, R; Félicité, Y; Alexandre, G; Archimède, H; Bambou, J-C

2016-03-15

In small ruminants, the response against gastrointestinal nematode (GIN) infections is influenced not only by the host genotype and the physiological stage but also by environmental factors, particularly the nutritional status at the time of infection. In this study we evaluated the long-term effect and the interaction between the host species and the nutritional history on the response to GIN infection in two animal models differing in their phenotypic growth and their level of GIN resistance: Black Belly sheep and Creole goats. Lambs and kids were subjected to three distinct nutritional conditions at weaning: low dietary conditions (100% of the theoretical energy requirement for maintenance, corresponding to 548v. 484KJ/Kg BW(0.75) for lambs and kids respectively and 6% of crude protein, CP), medium dietary conditions (150% of the theoretical energy requirement for maintenance and 13% CP) and high dietary conditions (200% of the theoretical energy requirement for maintenance and 20% CP). This 3-months period was followed by a 1-month period on the medium dietary conditions for all the animals before an experimental Haemonchus contortus infection. We monitored the impact of the nutritional history (nutritional condition after weaning), on the intensity of the GIN infection by measuring individual faecal egg counts (FEC), growth rate (ADG), blood eosinophil counts and other pathophysiological parameters. The FEC, growth rate and blood eosinophil counts were significantly affected by the nutritional history in lambs but not in kids. The lowest FEC was found for lambs placed in high dietary conditions, however during the same period body weight loss was observed in this group. In low dietary conditions, kids were more resistant than lambs and the ADG was higher in lambs. However, the anaemia and the level of serum pepsinogen, marker of the abomasal mucosa integrity, were higher in kids. Our data suggest that the impact of the post-weaning nutritional history on the

Benzimidazole -Resistance in Haemonchus Contortus: New PCR-RFLP Method for the Detection of Point Mutation at Codon 167 of Isotype 1 Β-Tubulin Gene

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A Eslami

2012-12-01

Full Text Available Background: Due to the lack of a suitable and economic test for the analysis of the polymorphism at codon 167, we developed a new PCR-RFLP technique, based on a modified forward primer (UT-HC167 MF-primer, to identify simultaneously the SNPs at codons 167 and 200 of isotype 1 β-tubu­lin gene of Haemonchus contortus.Methods: There already are several safe and easy methods for identification of point mutations at codons 198 and 200. Due to the lack of a reliable and easy method for the detection of the single nucleo­tide polymorphism (SNP at codon 167, we developed an innovative PCR-RFLP technique based on a modified forward primer (UT-HC167 MF-primer, in which the nucleotide T at the posi­tion 443 was substituted through a nucleotide A creating a restriction site for restriction endonuc­lease SnaB I in the nucleotide sequences including codon 167. A total of 138 adult male H. contortus were collected from three different geo-climatic areas of Iran. The isolated genomic DNA of each single worm was amplified by PCR using primers flanking codon 167. The PCR product (527 bp was then amplified by semi-nested PCR using the UT-HC167 MF-primer and the reverse primer achiev­ing a PCR product of 451 bp in length. This PCR product was subsequently digested with the restriction endonucleases SnaB I and TaaI for analysis of the mutations at codons 167 and 200, respec­tively.Results: All worms had two alleles encoding for phenylalanine (BZss homozygote for both codons.Conclusion: Using the UT-HC167 MF-primer and a suitable reverse primer designed upstream from codon 200, it is possible to amplify a PCR product which can be used for analysis of the SNPs at all three mentioned codons using RFLP.

ANTHELMINTIC EFFECTS OF DRIED GROUND BANANA PLANT LEAVES (MUSA SPP.) FED TO SHEEP ARTIFICIALLY INFECTED WITH HAEMONCHUS CONTORTUS AND TRICHOSTRONGYLUS COLUBRIFORMIS.

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Gregory, Lilian; Yoshihara, Eidi; Silva, Leandro Kataoaka Fernandes; Marques, Eduardo Carvalho; Ribeiro, Bruno Leonardo Mendonça; de Souza Meira, Enoch Brandão; Rossi, Rodolfo Santos; do Amarante, Alessandro Francisco Talamini; Hasegawa, Marjorie Yumi

2017-01-01

Helminths is a endoparasites that cause the major losses for profitable sheep production in Brazil. The increased development of resistant strains of endoparasites have enforced the search for sustainable alternatives. The aim of this paper was to provide information about endoparasites control with banana leaves in infected sheep as alternative control strategies and see its viability. In this study, we performed two trials to investigate the anthelmintic properties of banana leaves on endoparasites in sheep. In Trial 1, twelve sheep were artificially infected with Trichostrongylus colubriformis ; in Trial 2, eleven sheep were artificially infected with Haemonchus contortus . Clinical examinations, packed cell volume, total protein, faecal egg counts (FECs) and egg hatchability tests (EHTs) were performed. At the end of the trials, the sheep were humanely slaughtered, and total worm counts were performed. In Trial 1 and 2, no significant FEC decreases were note but significant diference in EHTs were observed. Total worm counts, clinical and haematological parameters did not reveal significant changes between the treatment and control groups. These results suggest that feeding dried ground banana plant leaves to sheep may reduce the viability of Trichostrongylus colubriformis eggs, and this anthelmintic activity is potentially exploitable as part of an integrated parasite management programme. However, further investigation is needed to establish the optimal dosage, develop a convenient delivery form and confirm the economic feasibility of using banana plantation byproducts as feed for ruminant species. Abbreviations: Coproculture test (CT)., Faecal egg count (FEC)., Egg hatchability test (EHT).

Experiences with integrated concepts for the control of Haemonchus contortus in sheep and goats in the United States.

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Terrill, Thomas H; Miller, James E; Burke, Joan M; Mosjidis, Jorge A; Kaplan, Ray M

2012-05-04

The generally warm, moist environmental conditions in the southern United States (U.S.) are ideal for survival and growth of the egg and larval stages of Haemonchus contortus and other gastrointestinal nematodes (GIN) of sheep and goats. Consequently, infection with GIN is the greatest threat to economic small ruminant production in this region. With anthelmintic resistance now reaching epidemic proportions in small ruminants in the U.S., non-chemical control alternatives are critically needed. The Southern Consortium for Small Ruminant Parasite Control (SCSRPC) was formed in response to this crisis and over the last decade has successfully validated the use of several novel control technologies, including FAMACHA(©) for the implementation of targeted selective treatments (TST), copper oxide wire particles (COWP), nematode-trapping fungi, and grazing or feeding hay of the high-tannin perennial legume sericea lespedeza [Lespedeza cuneata (Dum.-Cours. G. Don)]. Producer attitudes toward GIN control in the U.S. have been shifting away from exclusive dependence upon anthelmintics toward more sustainable, integrated systems of parasite control. Some novel control technologies have been readily adopted by producers in combination with appropriate diagnostic tools, such as FAMACHA(©). Others techniques are still being developed, and will be available for producer use as they are validated. Although new drugs will likely be available to U.S. goat and sheep producers in the future, these will also be subject to development of anthelmintic resistance. Therefore, the adoption and implementation of sustainable GIN control principles will remain important. With emerging markets for grass-fed or organic livestock, there will continue to be a critical need for research and outreach on development and on-farm application of integrated GIN control systems for small ruminants in the U.S. for the foreseeable future. Copyright © 2011 Elsevier B.V. All rights reserved.

In vitro and in vivo evaluation of the activity of pineapple (Ananas comosus) on Haemonchus contortus in Santa Inês sheep.

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Domingues, Luciana Ferreira; Giglioti, Rodrigo; Feitosa, Karina Alves; Fantatto, Rafaela Regina; Rabelo, Márcio Dias; de Sena Oliveira, Márcia Cristina; Bechara, Gervásio Henrique; de Oliveira, Gilson Pereira; Barioni Junior, Waldomiro; de Souza Chagas, Ana Carolina

2013-10-18

The development of resistance to anthelmintics has prompted research into alternative methods of controlling intestinal nematodes in ruminants. This study aimed to assess the activity of Ananas comosus on Haemonchus contortus in Santa Inês sheep. The aqueous extract of pineapple skin (AEPS), bromelain from pineapple stems (B4882) and residue from pineapple processing was evaluated in in vitro and in vivo tests. The enzymatic activity of substances was analyzed by the azocasein method. The egg hatch test (EHT) and larval development test (LDT) were performed using the Embrapa2010 isolate of H. contortus. In the in vivo test, 36 sheep artificially infected with H. contortus were divided into six groups: G1: 2 g/kg BW of the aqueous extract administered for three days; G2: 2 g/kg BW of the industrial pineapple residue for 60 days; G3: 180 mg/animal of bromelain in a single dose; G4: negative control I; G5: positive control (levamisole phosphate); and G6: negative control II. The eggs per gram (EPG) in the feces were counted till 28 days after treatment. LC₅₀ and LC₉₀ were obtained by the probit procedure, while the in vivo test results were analyzed by GLM. The aqueous extract in the in vitro and in vivo test, the bromelain and industrial residue presented 0.102, 0.157, 1.864 and 0.048 enzyme units/mL, respectively. In the egg hatch test, the LC₅₀ and LC₉₀ were respectively 31 and 81 mg/mL for the aqueous extract and 0.50 and 2 mg/mL for bromelain. In the larval development test, the LC₅₀ and LC₉₀ were respectively 1.7 and 7.3 mg/mL for the aqueous extract and 0.019 and 0.086 mg/mL for bromelain. In the in vivo test, the general efficacies of the treatments in relation to the negative control were 22.6%, 42.2%, 3.65% and 89% for the aqueous extract, industrial pineapple residue, bromelain and positive control respectively. The transformed EPG values were 3.19 ± 0.59, 3.32 ± 0.25, 2.85 ± 0.66, 3.44 ± 0.50, 2.28 ± 0.93 and 2.75 ± 0.94 for

A report on the metabolism of iron in goats artificially infected with Haemonchus contortus

International Nuclear Information System (INIS)

Perpuse, W.G.; Yumul, B.Y.; Anden, A.

1976-03-01

The determination of iron metabolism in goats artificially infected with N. contortus using tracer method has been conducted. Radioferric chloride ( 59 Fe) was given orally and intravenously and the distribution and utilization in goats were determined. Results showed that anemic goats have a higher absorption rate compared to non-anemic goats. Of all the organs examined the bone marrow showed the highest activity

Evaluation of multiple low doses of copper oxide wire particles compared with levamisole for control of Haemonchus contortus in lambs.

Science.gov (United States)

Burke, J M; Miller, J E

2006-06-30

High levels of anthelmintic resistance in gastrointestinal nematodes (GIN) of small ruminants have created the need for alternative approaches to parasite control. Copper oxide wire particles (COWP; 2g) have proven effective in decreasing GIN infection in lambs. However, the risk of copper toxicity has limited the usefulness of this approach. Recently, smaller doses (0.5 and 1g) have proven effective in GIN control, reducing the risk of toxicity. The objective of this study was to examine the effectiveness and risk of toxicity using multiple small doses of COWP for GIN control in lambs between weaning and market weight. Dorper crossbred ram lambs were orally administered levamisole (Levasol, 8.0mg/kg; n=8), 0.5g (n=9), or 1g COWP (n=9) at weaning (Day 0; 118+/-2 days of age; late May 2005) and again at 6-week intervals for a total of four treatments. A pooled fecal culture determined that Haemonchus contortus was the predominant gastrointestinal parasite at weaning. Lambs grazed bermudagrass pastures and were supplemented with up to 500g corn/soybean meal and free choice trace mineralized salt. Fecal egg counts (FEC), packed cell volume (PCV), and plasma aspartate aminotransferase (AST) activity were determined every 14 days and lambs weighed every 28 days. GIN infection reached a peak at Day 42 (high FEC, low PCV). COWP effectively reduced FEC on Days 0 and 42 compared with the previous week, but did not reduce FEC on Days 84 and 126 (treatment by time interaction, Pcopper in the liver on Day 155 were greater in COWP-treated lambs (Pcopper toxicity.

Evaluación del efecto de las artemisininas provenientes del extracto etanólico de Artemisia cina sobre L3 de Haemonchus contortus en una técnica de explantes abomasales

Directory of Open Access Journals (Sweden)

Rosa Isabel Higuera-Piedrahita

2016-01-01

Full Text Available El nuevo enfoque de control integrado de parásitos obliga a buscar alternativas que consideren el medioambiente, es decir, sostenibles y sustentables. El objetivo de este trabajo fue fraccionar el extracto etanólico de la planta Artemisia cina, obtener artemisininas y conocer el efecto de estas sobre la capacidad de asociación de las L3 de Haemonchus contortus en explantes abomasales. El extracto etanólico se fraccionó por medio de la metodología establecida para Artemisia japónica, y se identificaron las artemisininas por medio de cromatografía en capa fina, teniendo como referencia artemisininas comerciales. Las artemisininas se utilizaron sobre L3 desenvainada, se realizó la técnica de explantes abomasales por triplicado y se compararon así: levamisol (7.5 mg/ml, artemisinina comercial (1 mg/ml, agua y seis diferentes fracciones de A. cina a dosis de 1 mg/ml: Ac3k, Ac3b, Ac3a, Ac3h, Ac3i, Ac4b. No se obtuvieron diferencias significativas entre las fracciones de A. cina y artemisinina comercial (p>0.05. Se concluye que no existió efecto de las artemisininas obtenidas del extracto etanólico de A. cina sobre la capacidad de asociación de las L3 de H. contortus al tejido abomasal. Es importante continuar con más estudios de la artemisinina para determinar sobre qué fase del parásito afecta su viabilidad.

Haemonchosis and haemoparasites of small ruminants reared in ...

African Journals Online (AJOL)

The abomasa were examined by Hansen and Perry method for the presence of Haemonchus contortus while blood samples were examined using the thin blood smear and Haematocrit Centrifugation Techniques (HCT). The prevalence of Haemonchus contortus in small ruminants was 80.3% with goats and sheep having ...

Effects of in vitro exposure to ivermectin and levamisole on the expression patterns of ABC transporters in Haemonchus contortus larvae

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Ali Raza

2016-08-01

Full Text Available This study investigated the interaction of ATP binding cassette (ABC transport proteins with ivermectin (IVM and levamisole (LEV in larvae of susceptible and resistant isolates of Haemonchus contortus in vitro by measuring transcription patterns following exposure to these anthelmintics. Furthermore, we studied the consequences of drug exposure by measuring the sensitivity of L3 to subsequent exposure to higher drug concentrations using larval migration assays. The most highly transcribed transporter genes in both susceptible and resistant L3 were pgp-9.3, abcf-1, mrp-5, abcf-2, pgp-3, and pgp-10. The resistant isolate showed significantly higher transcription of pgp-1, pgp-9.1 and pgp-9.2 compared to the susceptible isolate. Five P-gp genes and the haf-6 gene showed significantly higher transcription (up to 12.6-fold after 3 h exposure to IVM in the resistant isolate. Similarly, five P-gp genes, haf-6 and abcf-1 were transcribed at significantly higher levels (up to 10.3-fold following 3 h exposure to LEV in this isolate. On the other hand, there were no significant changes in transcriptional patterns of all transporter genes in the susceptible isolate following 3 and 6 h exposure to IVM or LEV. In contrast to these isolate-specific transcription changes, both isolates showed an increase in R-123 efflux following exposure to the drugs, suggesting that the drugs stimulated activity of existing transporter proteins in both isolates. Exposure of resistant larvae to IVM or LEV resulted, in some instances, in an increase in the proportion of the population able to migrate at the highest IVM concentrations in subsequent migration assays. The significant increase in transcription of some ABC transporter genes following 3 h exposure to both IVM and LEV in the resistant isolate only, suggests that an ability to rapidly upregulate protective pathways in response to drugs may be a component of the resistance displayed by this isolate.

The N- and C-terminal carbohydrate recognition domains of Haemonchus contortus galectin bind to distinct receptors of goat PBMC and contribute differently to its immunomodulatory functions in host-parasite interactions.

Science.gov (United States)

Lu, MingMin; Tian, XiaoWei; Yang, XinChao; Yuan, Cheng; Ehsan, Muhammad; Liu, XinChao; Yan, RuoFeng; Xu, LiXin; Song, XiaoKai; Li, XiangRui

2017-09-05

Hco-gal-m is a tandem-repeat galectin isolated from the adult worm of Haemonchus contortus. A growing body of studies have demonstrated that Hco-gal-m could exert its immunomodulatory effects on host peripheral blood mononuclear cells (PBMC) to facilitate the immune evasion. Our previous work revealed that C-terminal and N-terminal carbohydrate recognition domains (CRD) of Hco-gal-m had different sugar binding abilities. However, whether different domains of Hco-gal-m account differently for its multiple immunomodulatory functions in the host-parasite interaction remains to be elucidated. We found that the N-terminal CRD of Hco-gal-m (MNh) and the C-terminal CRD (MCh) could bind to goat peripheral blood mononuclear cells by distinct receptors: transmembrane protein 63A (TMEM63A) was a binding receptor of MNh, while transmembrane protein 147 (TMEM147) was a binding receptor of MCh. In addition, MCh was much more potent than MNh in inhibiting cell proliferation and inducing apoptosis, while MNh was much more effective in inhibiting NO production. Moreover, MNh could suppress the transcription of interferon-γ (IFN-γ), but MCh not. Our data suggested that these two CRDs of Hco-gal-m bind to distinct receptors and contributed differently to its ability to downregulate host immune response. These results will improve our understanding of galectins from parasitic nematodes contributing to the mechanism of parasitic immune evasion and continue to illustrate the diverse range of biological activities attributable to the galectin family.

Study on the use of sheep serum post vaccination of three phase larva (L3) Haemonchus contortus worm irradiated inoculated to rabbit

International Nuclear Information System (INIS)

Partodihardjo, Sukardji; Suryadi, Helmi; Purwanti, Endang; Adiwinata, Gatot; Irtisam

1998-01-01

Study has been done on rabbit to find out the effect of giving sheep serum post vaccination with irradiated gastric worm challenged with infective H. contortus larva. Forty eight local rabbit divided into 3 groups : K, VI and V2, each groups: consists of 4 rabbit, by using 4 times repetition. K group which only inoculated 10,000L3 H. contortus infective. V1 = group which twice serum inoculated with 21 days interval, inoculation dose were 0,3 ml serum each, and challenged with 10.000 L3 H.contortus infective. V2 = group which 3 times serum inoculation with 21 days interval, inoculation dose 0,3 ml serum each and challenged with 10.000 L3 H.contortus infective. Design model used with Completely Random Design (RAL), Duncan was used as inter treatment test. Parameter observed are total protein fraction, Larva L4 content in gaster, cumulative weight gain, and death percentage rabbits. Result of study showed that on average parameter total protein fraction on K + 2,0086 ± 0,2353 mg/dl, V1 = 3,2781 ± 0,9227 mg/dl and V2 = 5,5035 ± 1,6794 mg/d. Average parameter L4 death showed on K = 10, V1 = 8, and V2 = 4, while L4 alive on K = 3, V1 + 1 and V2 = 0. Commutative Average parameter PCV value on K = 19,91 ± 1,24%, V1= 27,31 ± 1,22%, and V2 35,71 ± 0,56 %. Average parameter weight gain per week on K = 24,46 ± 0,74 g, V1 = 26,04 ± 2.23 g, and V2 = 28,93 ± 1,67 g. Percentage average parameter death rate on K = 37,5 ± 14,46%, V1=18,75 ± 23,90% and V2 = 0± 0%. The conclusion of the study result was serum inoculation on rabbits have positive result on reducing anaemia rate, elevating weight gain, reduce death percentage, reduced L4 alive content in gaster, and able oneelevate immunity respond content that is total protein fraction on rabbits. (author)

Between-breed variations in resistance/resilience to gastrointestinal nematodes among indigenous goat breeds in Uganda

NARCIS (Netherlands)

Onzima, R.B.; Mukiibi, Robert; Ampaire, A.; Benda-Beckmann, von K.; Kanis, E.

2017-01-01

Gastrointestinal nematodes (GINs), Haemonchus contortus, are a major health problem in goat production. Resistance to H. contortus, the most prevalent GIN in Uganda, was studied among three indigenous goat breeds to assess their differences. Twelve male goats of each breed approximately 7 months old

[Distribution of nematode parasites of the digestive system in sheep (Ovis aries) and goats (Capra hircus) of the Piedmontese and Valdostano Alpine arc].

Science.gov (United States)

Balbo, T; Costantini, R; Gallo, M G; Lanfranchi, P

1977-01-01

A survey, carried out on gastro-intestinal nematodes of sheep and goats of Piemonte and of Valle d'Aosta (87 sheep and 12 goats) has shown the presence of the following species in sheep, Bunostomum trigonocephalum, Chabertia ovina, Cooperia curticei, Haemonchus contortus, Marshallagia marshalli, Nematodirus abnormalis, Nematodirus filicollis, Nematodirus helvetianus, Nematodirus spathiger, Oesophagostomum venulosum, Ostertagia circumcincta, Ostertagia lyrata, Ostertagia trifurcata, Skrjabinema ovis, Trichostrongylus axei, Trichostronglus colubriformis, Trichostronglus vitrinus, Trichuris ovis and Trichuris skrjabini; in goats, Bunostomum trigonocephalum, Chabertia ovina, Haemonchus contortus, Nematodirus filicollis, Nematodirus helvetianus, Oesophagostomum venulosum, Ostertagia circumcincta, Ostertagia ostertagi, Ostertagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis and Trichostrongylus vitrinus. The percentage of each species in the two host is given in the text table.

BKR.2014.003 (Sonibare) b

African Journals Online (AJOL)

Femi Olorunniji

2014-03-31

Mar 31, 2014 ... constituted 33.33% prevalence rate, while Haemonchus contortus and Oesophagostomum columbianum mixed infection ... However, in WAD, the eosinophil count ... Caprine parasitic gastrointestinal infection is worldwide in.

Anthelmintic-resistant strains of Haemonchus contortus and Trichostrongylus colubriformis isolated from an organic sheep and goat farm in Denmark

DEFF Research Database (Denmark)

Pena-Espinoza, Miguel Angel; Enemark, Heidi L.; Thansborg, Stig M.

2013-01-01

A suspected case of anthelmintic resistance (AR) was investigated in an organic dairy sheep and goat farm. The herd was established in 2007 by purchase of animals from a number of other farms. Selection for the study was based on history of anthelmintic-treatment failure. Forty-eight lambs and 48......) or 10 mg/kg closantel (CLO). Kids were treated with 10 mg/kg FBZ, 0.3 mg/kg MOX, 14 mg/kg LEV, 0.2 mg/kg IVM or 10 mg/kg CLO. FECs were performed at day of treatment and 14 days post treatment. In a subsequent investigation, faeces from adult goats were cultured to obtain 3rd-stage nematode larvae (L3...... %. This is the first isolation of BZ-resistant H. contortus and T. colubriformis in Denmark and highlights the need for continuous surveillance of AR in conventional and organic farms....

Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock.

Science.gov (United States)

George, Melissa M; Lopez-Soberal, Lorraine; Storey, Bob E; Howell, Sue B; Kaplan, Ray M

2018-04-01

Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3) of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM) compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin) and Haemonchus contortus (eprinomectin, ivermectin, moxidectin). Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC 50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 μM), 100% inhibition of motility was never achieved and EC 50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring resistance to AM

Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock

Directory of Open Access Journals (Sweden)

Melissa M. George

2018-04-01

Full Text Available Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3 of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin and Haemonchus contortus (eprinomectin, ivermectin, moxidectin. Concentrations tested with the Worminator ranged from 0.156 to 40 μM. Differences in EC50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 μM, 100% inhibition of motility was never achieved and EC50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring

The immunogenic effect of irradiated Hemonchus contortus larvae in goats

International Nuclear Information System (INIS)

de la Cruz, B.; Perfuse, W.

1980-01-01

The development of radiation-attenuated vaccines against parasitic disease has been reported in literature. The use of irradiated larvae as vaccines enables one to take advantage of the special immunological properties of living parasites; the radiation treatment can largely suppress the pathogenic effects of the vaccines and the reproductive capacity of the parasites involved. This report deals with our attempts to produce a radiation-attenuated vaccine against Hemonchus contortus infection in adult goats. Hemonchus contortus infection in ruminants is prevalent in the Philippines, especially among cattle and goats. The results presented indicate that a certain degree of resistance against Hemonchus contortus infection can be induced in adult goats immunized with irradiated larvae. The shelf-life of the vaccine was also determined. (author)

In vivo Efficacy of Vernonia amygdalina (Compositae Against Natural Helminth Infection in Bunaji (Bos indicus Calves

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C. B. I. Alawa ab*, A. M. Adamu, J. O. Gefub, O. J. Ajanusic, P. A. Abdud and N. P. Chiezeyb

2010-10-01

Full Text Available Fifteen Bunaji calves (Bos indicus averaging 105±12.5 Kg liveweight and approximately nine months of age with natural helminth infection were distributed into three treatment groups of five animals each. Animals were either treated orally with aqueous extract of Vernonia amygdalina at a dose concentration of 1.1g/Kg body weight, a conventional anthelmintic or left untreated. V. amygdalina treatment produced 59.5% reduction in eggs per gram (EPG of faeces which was significantly different (P<0.001 from the untreated control (-17.24%, whereas levamisol hydrochloride treatment produced 100% reduction in EPG. A total of six genera of helminths were recovered from the gastrointestinal tracts and liver of experimental animals. These were Haemonchus contortus, Trichostrongylus spp, Bunostomum spp, Oesophagostomum spp, Fasciola spp and Dicrocoelium spp. There was significant difference (P<0.001 in worm load between the different treatment groups. Except for Haemonchus spp, animals in the untreated group had significantly (P<0.001 higher worm load for all the genera of helminth recovered than those of the V. amygdalina treated group, indicating that V. amygdalina had no effect on Haemonchus contortus.

Fauna helmintológica de ovinos provenientes da microrregião de Jaboticabal, estado de São Paulo, Brasil

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Willian Giquelin Maciel

2014-03-01

Full Text Available Avaliou-se a prevalência e a contagem parasitária das diferentes espécies de helmintos de ovinos provenientes da microrregião de Jaboticabal, região Noroeste de São Paulo. Para tanto, foram utilizados 66 animais naturalmente infectados, de quatro a 36 meses de idade, criados em regime extensivo. Os resultados necroscópicos revelaram a presença de sete gêneros e 12 espécies, com a seguinte prevalência e a média de parasitismo: Haemonchus contortus: 100,0% (2947,2; Trichostrongylus colubriformis: 90,9% (3048,8; Cooperia curticei: 56,0% (256,5; Oesophagostomum columbianum: 48,4% (36,0; Cooperia punctata: 30,3% (94,5; Trichostrongylus axei: 22,7% (26,5; Strongyloides papillosusi: 19,6% (83,0; Haemonchus contortus (L4: 7,5% (17,2; Cooperia pectinatai: 10,6% (12,9; Trichuris ovis: 10,6% (0,6; Cooperia spatulata 4,5% (0,3; Capillaria bovis: 4,5% (0,1. A carga parasitária média foi de 6.524,7 helmintos por animal. Haemonchus contortus (Adultos e L4 e Trichostrongylus colubriformis corresponderam a 45,4% e 46,7% da carga parasitária média total, respectivamente. Pode-se concluir que as duas espécies de helmintos mais abundantes e importantes da microrregião de Jaboticabal/São Paulo foram Trichostrongylus colubriformis e Haemonchus contortus, sendo que essas duas espécies perfizeram 92,1% da distribuição percentual dos helmintos recolhidos de todos os animais. Tais resultados demonstram a importância em se realizar um monitoramento das contagens de ovos por grama de fezes (OPG dos rebanhos desta região, quando o método FAMACHA for empregado em uma determinada propriedade, uma vez que este método de controle, geralmente, não permite diagnosticar os danos/sinais clínicos desencadeados nos animais pelo T. colubriformis, em função de essa espécie não possuir hábito de hematofagismo sobre os hospedeiros.

Browse Title Index

African Journals Online (AJOL)

Items 51 - 100 of 238 ... Vol 3, No 1 (2013), Digit loss due to Demodex spp. infestation in a dog: ... Early detection of Haemonchus contortus infection in sheep using three ... of ensiled and dried pomegranate seeds for ruminants using in vitro gas ...

Examination of commercially available copper oxide wire particles in combination with albendazole for control of gastrointestinal nematodes in lambs

Science.gov (United States)

Alternatives to synthetic anthelmintics remain critical due to the prevalence of anthelmintic resistance. The objective of the experiment was to determine the efficacy of copper oxide wire particles (COWP) from three commercial sources to control Haemonchus contortus in lambs. Naturally infected Ka...

some nutritional aspects of haemonchosis in experimentally infested ...

African Journals Online (AJOL)

nutritional aberration has been described as anorexia in both pure (Evans, Blunt & Southcott, 1963) and mixed infestations where Haemonchus contortus was prominent. (Clark, Ortlepp, Bosman, Laurence, Groenewald & Quin,. 1951; Shumard et al. 1957). Further observations on nutritional aspects of a pure infestation of ...

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus.

Science.gov (United States)

Salles, Hévila Oliveira; Braga, Ana Carolina Linhares; Nascimento, Maria Thayana dos Santos Canuto do; Sousa, Ana Márjory Paiva; Lima, Adriano Rodrigues; Vieira, Luiz da Silva; Cavalcante, Antônio Cézar Rocha; Egito, Antonio Silvio do; Andrade, Lúcia Betânia da Silva

2014-01-01

Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus

Directory of Open Access Journals (Sweden)

Hévila Oliveira Salles

Full Text Available Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (0.05, Bonferroni test. Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.

A Biological Study of Larvae and Adult Hemonchus contortus in Goat

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Yuswandi Yuswandi

2015-11-01

experimental animal in vivo. Before the goat necropsied, the diagnosis of H. contortus egg was done every two day post infection and started two weeks after infection. The data was analyzed descriptively. The results showed that the capability of the egg development of the worm to the L3 stadium was 0,33%, the capability of the L3 development to adult was 32,42%, and the prepaten period of H. contortus was 21 days.

In vitro anthelmintic effect of two medicinal plants (Anogeissus ...

African Journals Online (AJOL)

STORAGESEVER

2009-09-15

Sep 15, 2009 ... A study was conducted to evaluate Anogeissus leiocarpus leaf and Daniellia oliveri stem barks as effective remedy for gastrointestinal parasites. The anthelmintic activity of these extracts on eggs, first stage larvae and adults of Haemonchus contortus was examined by in vitro tests. The extracts were.

Studies on the Leucocytic Response to Experimental Infection with ...

African Journals Online (AJOL)

Also, neutrophil numbers declined significantly (P < 0.05) in red fronted gazelles infected either singly with Trypanosoma brucei or concurrently with both parasites while those infected singly with Haemonchus contortus experienced a significant (P <0.05) rise in neutrophil counts which became evident from day 30 post ...

Interaction between Trypanosoma brucei and Haemonchus ...

African Journals Online (AJOL)

In order to investigate the immunomodulatory influence of concurrent T. brucei and H. contortus infection in West African Dwarf (WAD) goats, 28 infected and 7 uninfected (control) of 8-9 months old male WAD goats were studied. The infected goats were separated into resistant (Class 1) and susceptible (Class 2) Faecal ...

Epidemiological study on gastrointestinal tract hel- minthosis of ...

African Journals Online (AJOL)

zone practice a crop - livestock mixed farming and keep combination of live- ... of animals (Goat and sheep), sex of animals (male and female), age of animals .... Mean worm burden of Haemonchus contortus was significantly high in older ..... high and mid land portion of the study site where sheep were forced to graze.

IN VITRO ANTIHELMINTIC ACTIVITY OF THREE PLANT SPECIES TRADITIONALLY USED IN TABASCO, MEXICO.

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Judith Espinosa-Moreno

2016-03-01

Full Text Available Antihelmintic activity against Haemonchus contortus was searched as a new alternative for animal health improvement using three plants (Cydista aequinoctialis L., Heliotropium indicum L. and Momordica charantia L. of traditional usage in the rural area of Tabasco State, Mexico. Aqueous extracts were obtained of dried material. Each extract was diluted in distilled water to obtain a concentration of 20 mg ml-1. The antihelmintic activity was evaluated at 24, 48 y 72 h of exposition with 100 μL of each extract and 50 L3 unsheathed larvaes of H. contortus in a 96 well ELISA plaque.;/span;

Efficacy of closantel against ivermectin- and fenbendazole-resistant Haemonchus sp. in sheep in Ontario, Canada.

Science.gov (United States)

Westers, T; Jones-Bitton, A; Menzies, P; Van Leeuwen, J; Poljak, Z; Peregrine, A S

2016-09-15

In Ontario, Canada, widespread resistance to ivermectin and fenbendazole, the only readily available ovine anthelmintics, has been documented, primarily in Haemonchus sp. In other parts of the world, closantel has been used to control such infections; however, the drug was not currently licensed for use in Canada and the USA. A randomized controlled trial was conducted on six client-owned farms in Ontario in 2013 and 2014 to determine the efficacy of closantel (Flukiver ® 5% Oral Suspension, Elanco Animal Health, 10mg/kg bodyweight) against ivermectin- and fenbendazole-resistant Haemonchus sp. infections in periparturient ewes and grazing lambs. Three farms were randomly assigned to treat all ewes, and three farms were randomly assigned to selectively treat individual ewes at lambing, using predetermined criteria. Fecal samples were collected from a minimum of 15 randomly selected ewes and 13 lambs per group on each farm at the time of treatment and approximately 14days later. Trichostrongyle-type fecal egg counts (FEC) were performed using a modified McMaster technique with a lower detection limit of 8.3 eggs per gram of feces (epg). Haemonchus-specific FECs were determined by multiplying FECs by the proportion of Haemonchus sp. identified from coproculture for each farm; Haemonchus-specific FEC reductions were calculated for each farm. Twenty grazing lambs had FECs conducted monthly, and when mean monthly FECs surpassed 200 epg, all lambs were randomly allocated to either closantel, positive control (ivermectin, fenbendazole, or levamisole) or negative control groups. Pre-treatment Haemonchus-specific mean FECs ranged from 27 to 3359 epg in ewes and 0-5698 epg in lambs. Efficacy of closantel against Haemonchus sp. ranged from 99% (95% CI: 97%-99%) to 100% in recently lambed ewes on all farms in both years (total n=274 ewes), and from 99% (95% CI: 98%-99%) to 100% in grazing lambs in both years on all but one farm (total n=171 lambs). On the latter farm, a whole

1100-IJBCS-Article-Abiodoum Olounlade

African Journals Online (AJOL)

DR GATSING

(Bignoniaceae) et de Zanthoxylum zanthoxyloïdes (Rutaceae), leur effet inhibiteur a été évalué in vitro sur la migration larvaire de ... Les extraits de Newbouldia laevis et de Zanthoxylum zanthoxyloïdes inhibent in vitro la migration larvaire de. Haemonchus contortus. ... médicaments, à la malfaçon et la contrefaçon de.

Comparison of constitutive and thiabendazole-induced expression of five cytochrome P450 genes in fourth-stage larvae of Haemonchus contortus isolates with different drug susceptibility identifies one gene with high constitutive expression in a multi-resistant isolate

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Esra Yilmaz

2017-12-01

Full Text Available Benzimidazoles (BZs remain amongst the most widely used anthelmintic drug classes against gastro-intestinal nematode infections, although their efficacy is increasingly compromised by resistance. The primary underlying mechanisms for BZ resistance are single-nucleotide polymorphisms (SNPs in the isotype 1 β-tubulin gene causing the substitutions F167Y, E198A or F200Y. However, resistance is believed to be multi-genic and previous studies have shown that isolates carrying 90–100% F200Y can vary considerably in their resistance level in the egg hatch assay (EHA. Cytochrome P450 monooxygenases (CYPs are associated with drug resistance in mammals and arthropods and have been considered as mediators of anthelmintic resistance. In Caenorhabditis elegans, several members of the CYP34/35 and CYP31 families are BZ and/or xenobiotic inducible and thiabendazole (TBZ is metabolised by CYP35D1. Here, expression of all 5 CYPs closely related to the C. elegans CYP34/35 and CYP31 families was investigated in fourth-stage larvae of two susceptible and three BZ-resistant Haemonchus contortus isolates following in vitro exposure to TBZ for 3 and 6 h using real-time RT-PCR. The resistance status of all isolates was determined using EHAs and quantification of resistance-associated β-tubulin SNPs using pyrosequencing. While none of the CYPs was TBZ inducible, constitutive expression of CYP34/35 family member HCOI100383400 was significantly 2.4–3.7-fold higher in the multi-drug resistant WR isolate with the strongest BZ resistance phenotype compared to susceptible and intermediate-level BZ-resistant isolates. Although this increase is only moderate, HCOI100383400 might still be involved in high-level BZ resistance by further decreasing susceptibility in isolates already carrying 100% of a β-tubulin SNP causing BZ resistance. Lower transcript levels were observed for all CYPs in the intermediately resistant IRE isolate in comparison to the susceptible Hc

Prevalence of Ovine Haemonchosis in Wukro, Ethiopia

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Lidya Gebresilassie

2015-01-01

Full Text Available Background. Haemonchosis caused by Haemonchus contortus is a predominant, highly pathogenic, and economically important disease of sheep and goats. Objective. Assessing the prevalence of Haemonchus parasite and its associated risk factors in sheep slaughtered at different restaurants of Wukro. Methods. Cross-sectional study using random sampling from November 2013 to April 2014 in a total of 384 sheep was conducted and SPSS version 20 software using descriptive statistics was used for data analysis and P0.05. Conclusion. The current finding revealed that significant numbers of sheep were affected by the parasites. Hence strategic deworming with good husbandry practice should be implemented.

Seasonal prevalence of gastrointestinal nematode parasites of sheep in Northern region of Nile Delta, Egypt.

Science.gov (United States)

Khalafalla, Reda E; Elseify, Mahmoud A; Elbahy, Nasr M

2011-02-01

Over 1 year, from January to December 1999, a total of 173 slaughtered sheep at Al-Mahala abattoir were examined for presence of nematode parasites. Eighteen sheep (10.4%) were infected with eight different species of nematodes. The prevalence rates of detected nematode parasites were; Haemonchus contortus (3.5%), Haemonchus placei (1.7%), Trichuris ovis (5.8%), Parabronema skrjabini (2.9%), Ostertagia trifurcata (1.2%), Chabertia ovina (0.6%) and Strongyloides papillosus (0.6%), and Graphidiops species (2.9%). The seasonal prevalence of the infection with the nematode parasites was studied and the highest rate was during autumn (15.2%) followed by summer (11.1%) and winter (9.4%) while the lowest rate was during spring (5.6%).

The preparation and use of radiolabelled specific helminth antibodies

International Nuclear Information System (INIS)

Movsesijan, M.; Jovanovic, B.; Borojevic, D.; Petrovic, M.

1983-01-01

Specific antibodies from the serum of sheep infected with Haemonchus contortus were isolated by combination with a ''solid phase antigen'' (soluble antigen coupled to an activated crystalline cellulose). The antibodies were labelled with 125 I while bound to the solid phase then eluted and their potential demonstrated: (1) to determine amounts of specific antibody in unknown sera; (2) to determine amounts of soluble antigen in unknown preparations. (author)

Seasonal distribution of gastrointestinal nematode infections in sheep in a semiarid region, northeastern Brazil

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Maria de Fátima de Souza

Full Text Available The objective of this study was to determine the seasonal distribution and gastrointestinal nematode parasite load in crossbred Santa Inês tracer lambs, and to correlate the rainfall during the study period with occurrences of parasitic infections. Sixty-four male tracer lambs between the ages of four and eight months were used in the study. Two tracer lambs were inserted into the herd every 28 days to determine the pattern of infective larvae available in the environment. Variation in the fecal egg count (FEC of nematodes was observed at the study site, with many samples containing undetectable parasite loads during the dry season. The larvae identified in coprocultures wereHaemonchus sp., Trichostrongylus sp.,Cooperia sp., Strongyloides sp. andOesophagostomum sp. The nematodes recovered at necropsy were Haemonchus contortus, Trichostrongylus colubriformis, Cooperia punctata, C. pectinata, Trichuris sp.,Oesophagostomum sp. and Skrajabinema ovis. The total number of larvae and the total number of immature and adult forms recovered from the tracers showed seasonal distributions that significantly correlated with the amount of rainfall received that month (p value ≅ 0.000 in all cases . The species H. contortus was predominant in the herd and should be considered to be main pathogenic nematode species in these hosts under these conditions.

Seasonal distribution of gastrointestinal nematode infections in sheep in a semiarid region, northeastern Brazil.

Science.gov (United States)

de Souza, Maria de Fátima; Pimentel-Neto, Manoel; de Pinho, André Luís Santos; da Silva, Rízia Maria; Farias, Albeísa Cleyse Batista; Guimarães, Marcos Pezzi

2013-01-01

The objective of this study was to determine the seasonal distribution and gastrointestinal nematode parasite load in crossbred Santa Inês tracer lambs, and to correlate the rainfall during the study period with occurrences of parasitic infections. Sixty-four male tracer lambs between the ages of four and eight months were used in the study. Two tracer lambs were inserted into the herd every 28 days to determine the pattern of infective larvae available in the environment. Variation in the fecal egg count (FEC) of nematodes was observed at the study site, with many samples containing undetectable parasite loads during the dry season. The larvae identified in coprocultures were Haemonchus sp., Trichostrongylus sp., Cooperia sp., Strongyloides sp. and Oesophagostomum sp. The nematodes recovered at necropsy were Haemonchus contortus, Trichostrongylus colubriformis, Cooperia punctata, C. pectinata, Trichuris sp., Oesophagostomum sp. and Skrajabinema ovis. The total number of larvae and the total number of immature and adult forms recovered from the tracers showed seasonal distributions that significantly correlated with the amount of rainfall received that month (p value ≅ 0.000 in all cases ). The species H. contortus was predominant in the herd and should be considered to be main pathogenic nematode species in these hosts under these conditions.

Antiparasitic efficacy of ivermectin in naturally parasitized sheep.

Science.gov (United States)

Yazwinski, T A; Greenway, T; Presson, B L; Pote, L M; Featherstone, H; Williams, M

1983-11-01

Sixteen sheep harboring naturally acquired parasitisms were allocated to 1 of 2 treatment groups: (i) sheep given ivermectin in an oral solution at the dosage rate of 200 micrograms/kg of body weight, and (ii) those given the vehicle at a dosage rate of 0.25 ml/kg. All animals were necropsied at 2 weeks after treatment. Parasites and percentages of parasitic reductions, as demonstrated in this trial, were: Dictyocaulus filaria (99.4%), Oestrus ovis first stage instars (100%), Trichuris ovis (98.9%), Strongyloides papillosus (99.8%), Nematodirus spathiger (100%), arrested 4th stage Nematodirus spp (96.2%), Trichostrongylus colubriformis (100%), T axei (100%), Oster tagia circumcincta (100%), Haemonchus contortus (100%), and arrested Haemonchus spp 4th stage larvae (99.9%). The sheep showed no adverse effects due to ivermectin or vehicle administration.

Prevalence of Gastro-Intestinal Nematodes in Goats in Hyderabad and Adjoining Areas

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Nasreen Akhter*, A. G. Arijo, M. S. Phulan, Zafar Iqbal1 and K. B. Mirbahar

2011-10-01

Full Text Available The study was conducted to determine the prevalence of gastrointestinal nematodes of goats (n=1065 in and around Hyderabad using qualitative and quantitative coprological examinations. Results revealed that 43.10% (459 goats were infected with different species of nematodes including Haemonchus contortus (14.65%, Trichuris ovis (8.17%, Trichostrongylus axei (7.61%, Trichostrongylus colubriformis (6.76%, Oesphagostomum columbianum (5.35%, Ostertagia circumcincta (5.35%, Chabertia ovina (4.79% and Strongyloides papillosus (4.51%. Infections with mixed species of nematodes were recorded in 6.54% (n=30/459; T. ovis + H. contortus, 5.23% (n=24/459; C. ovina + H. contortus, 5.88% (n=27/459; S. papillosus + C. ovina, and 12.42% (n=57/459; O. circumcincta + T. ovis goats. Of the total infected (n=459; 51.4, 38.3 and 10.2% goats had light, moderate and heavy infections, respectively. The prevalence, nature and intensity of the helminthiasis in goats warrant an immediate attention to devise strategies for its control to reduce the production losses.

Control of gastrointestinal nematodes with copper oxide wire particles in a flock of lactating Polypay ewes and offspring in Iowa, USA.

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Burke, J M; Morrical, D; Miller, J E

2007-05-31

Copper oxide wire particles (COWP) have been used to reduce infection of Haemonchus contortus in hair breed lambs in southeastern USA without signs of copper toxicity. However, copper sensitivity among breeds and regions varies. The objective was to determine the effectiveness and safety of COWP in lactating Polypay ewes and their offspring grazing alfalfa/bluegrass pasture in a rotational grazing system. Mature Polypay ewes were administered 0, 0.5, 1, or 2 g (n=8 or 9/dose) COWP approximately 60 days after lambing in mid-July 2005. Their offspring were administered 0 (n=6), 0.5 or 0.75 g (n=9), 1 or 2 g (n=6) COWP 2 weeks later in late July. The primary gastrointestinal nematode was H. contortus (70%). Between Days 7 and 35, FEC were greater in 0 and 0.5 g COWP groups compared with ewes administered 2 g COWP (COWP x day, Pcopper levels, and body weight was similar among groups of ewes. FEC decreased within 7 days in COWP-treated compared with untreated lambs and remained low throughout experiment (COWP x day, Pcopper toxicity in ewes or lambs. Alternative suppression of H. contortus infections may be necessary in ewes, but COWP was effective in H. contortus management for lambs.

The effect of long-term feeding of fresh and ensiled cassava (Manihot esculenta) foliage on gastrointestinal nematode infections in goats.

Science.gov (United States)

Sokerya, S; Waller, P J; Try, P; Höglund, J

2009-02-01

The benefit of long-term feeding of fresh or ensiled cassava foliage on gastrointestinal parasite in goats was evaluated. Eighteen male goats (15.15 +/- 2.83 kg and between 4-6 months) were randomly allocated into three treatments supplemented with 200 g of wheat bran head(-1) day(-1). All groups were fed ad-libitum on either grass (CO), fresh cassava (CaF) or ensiled cassava foliage (CaS). At the beginning of the trial, each goat was inoculated with 3000 L3 containing approximately 50% Haemonchus contortus. Individual LWt, FEC and PCV were measured at weekly intervals for 10 weeks. At the termination of the experiment all goats were slaughtered for worm recovery and enumeration. The goats in CaF and CaS had similar weight gains while those in CO lost weight (p goats. PCV of all groups decreased from above 30% to around 25% at the end of the trial. The compositions of established worm burdens were mainly H. contortus (19-40%) and Trichostrongylus colubriformis (55-76%). TWB did not differ among the groups, however, CaS significantly reduced H. contortus burdens, as compared to CaF and CO (p < or = 0.005). Thus, ensiled cassava foliage reduced the H. contortus population while the fresh foliage only reduced worm fecundity.

Discordant population histories of host and its parasite: A role for ecological permeability of extreme environment?

Czech Academy of Sciences Publication Activity Database

Jirsová, Dagmar; Štefka, Jan; Jirků, Miloslav

2017-01-01

Roč. 12, č. 4 (2017), č. článku e0175286. E-ISSN 1932-6203 R&D Projects: GA ČR(CZ) GBP505/12/G112 Grant - others:GA MŠk(CZ) LM2015042 Institutional support: RVO:60077344 Keywords : fragment length polymorphism * genetic diversity * cestoda caryophyllidea * haemonchus-contortus * divergent selection * mitochondrial-dna * malaria parasites * local adaptation * faunal turnover * cichlid fishes Subject RIV: EG - Zoology OBOR OECD: Zoology Impact factor: 2.806, year: 2016

In vitro variation of glycogen content in three sheep nematodes.

Science.gov (United States)

Premvati, G; Chopra, A K

1979-06-01

In vitro variation of glycogen content under aerobic conditions was measured on fresh weight basis in 3 sheep nematodes inhabiting different niches; Haemonchus contortus, Oesophagostomum columbianum and Trichuris ovis. The parasites were saparated into species and then sexes and starved for varying periods of time up to 24 h in glucose-free physiological saline. The differences between females and males and among the species with respect to glycogen content and its rate of change with time are discussed.

In vitro larvicidal effects of ethanolic extract of Curcuma longa Linn. on Haemonchus larval stage

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Norisal Binti Nasai

2016-04-01

Full Text Available Aim: Gastrointestinal helminthosis is a global problem in small ruminant production. Most parasites have developed resistance to commonly available anthelminthic compounds, and there is currently an increasing need for new compounds with more efficacies. This study evaluated the in vitro effects of ethanolic extract of Curcuma longa (EECL as a biological nematicide against third stage Haemonchus larvae (L3 isolated from sheep. Materials and Methods: Haemonchus L3 were cultured and harvested from the feces of naturally infected sheep. EECL was prepared and three concentrations; 50, 100, and 200 mg/mL were tested for their efficacies on Haemonchus L3. Levamisole at concentration 1.5 and 3 mg/mL were used as positive controls. Results: EECL showed anthelmintic activity in a dose-dependent manner with 78% worm mortality within 24 h of exposure at the highest dose rate of 200 mg/mL. There was a 100% worm mortality rate after 2 h of levamisole (3 mg/mL admisntration. However, there was a comparable larvicidal effect between when levamisole (1.5 mg/mL and EECL (200 mg were administered. Conclusion: The study shows that EECL does exhibit good anthelmintic properties at 200 mg/mL which is comparable with levamisole at 1.5 mg/mL.

Caracterización microecologica de nemátodos parásitos presentes en caprinos de zonas áridas de Venezuela

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Gustavo Morales

1986-06-01

Full Text Available Se determinó que los caprinos de las zonas áridas de Venezuela son parasitados por lassiguientes especies de nemátodos: Haemonchus contortus, Trichostrongylus axei, T. colubriformis, Cooperia curticei, Oesophagostomun columbianum, Skrjabinema ovis y Trichuris globulosa. Los valores del índice de diversidad de Shannon-Weaver oscilaron entre 0,045 y 1,73 bits (X = 1,15 ± 0,24 bits. La diversidad máxima por mes presenta valores que van desde 1 a 2,80 bits (X = 2,49 ± 0,28 bits y la equitabilidad varió de 0,045 a 0,67 (X = 0,44 ± 0,09. Los valores obtenidos con los datos agrupados en un solo bloque son de 1,25 bits, 2,80 bits y 0,40 para el índice de diversidad de Shannon-Weaver, la diversidad máxima y la equitabilidad respectivamente. En lo referente a las asociaciones parasitarias, se encontró un primer grupo conformado por T. axei, T. colubriformis y H. contortus, un segundo grupo por T. colubriformis, H. contortus y O. columbianum, y un tercero integrado por T. colubriformis, S. ovis y T. globulosa. Los resultados obtenidos permiten además, sugerir em torno al control químico que en los meses en los cuales el valor del índice de diversidad de Shannon-Weaver se aproxime al de la diversidad máxima y el del índice de equitabilidad se aproxime a la unidad, se pueden utilizar antihelmínticos de amplio espectro.Goats from the Venezuelan northem arid zones were found infested by the following nematodes: Haemonchus contortus, Trichostrongylus axei, T. colubriformis, Cooperia curticei, Oesophagostomum columbianum, Skrjabinema ovis y Trichuris globulosa. The Shannon-Weaver index of diversity values oscillated between 0.045 and 1.73 bits (X = 1.16 ± 0.24 bits. The maximum value of montly diversity ranged from 1 to 2.80 bits (X = 2.49 ± 0.28 bits and the equitability ranged from 0.045 to 0.67 (X = 0.44 ± 0.09. Parasitic associations were found among 1 T. axei, T. colubriformis and H. contortus, 2 T. colubriformis, H. contornus and O

Unique contributions of an arginine side chain to ligand recognition in a glutamate-gated chloride channel

DEFF Research Database (Denmark)

Lynagh, Timothy; Komnatnyy, Vitaly V; Pless, Stephan A

2017-01-01

Glutamate recognition by neurotransmitter receptors often relies on arginine (Arg) residues in the binding site, leading to the assumption that charge-charge interactions underlie ligand recognition. However, assessing the precise chemical contribution of Arg side chains to protein function......-gated chloride channel from the nematode Haemonchus contortus. Our data unveil a surprisingly small contribution of charge at a conserved arginine side chain previously suggested to form a salt bridge with the ligand, glutamate. Instead, our data show that Arg contributes crucially to ligand sensitivity via...

Dinámica espacio-temporal de nematodos gastrointestinales y escarabajos estercoleros en un agroecosistema bovino de Veracruz.

OpenAIRE

Flota Bañuelos, Carolina

2012-01-01

Se analizaron los componentes del agroecosistema bovino, con la inclusión de la ivermectina sobre la abundancia y la disposición espacio-temporal de escarabajos estercoleros y de nematodos gastrointestinales. El capítulo I presenta el estudio de los nematodos gastrointestinales que afectan a los bovinos, su abundancia, su dispersión en el potrero, y el efecto de la ivermectina sobre los huevecillos en las heces. Se encontró que los parásitos que afectan al ganado son Haemonchus contortus, Tri...

Reconstruction of Cysteine Biosynthesis Using Engineered Cysteine-Free and Methionine-Free Enzymes

Science.gov (United States)

Wang, Kendrick; Fujishima, Kosuke; Abe, Nozomi; Nakahigashi, Kenji; Endy, Drew; Rothschild, Lynn J.

2016-01-01

Ten of the proteinogenic amino acids can be generated abiotically while the remaining thirteen require biology for their synthesis. Paradoxically, the biosynthesis pathways observed in nature require enzymes that are made with the amino acids they produce. For example, Escherichia coli produces cysteine from serine via two enzymes that contain cysteine. Here, we substituted alternate amino acids for cysteine and also methionine, which is biosynthesized from cysteine, in serine acetyl transferase (CysE) and O-acetylserine sulfhydrylase (CysM). CysE function was rescued by cysteine-and-methionine-free enzymes and CysM function was rescued by cysteine-free enzymes. Structural modeling suggests that methionine stabilizes CysM and is present in the active site of CysM. Cysteine is not conserved among CysE and CysM protein orthologs, suggesting that cysteine is not functionally important for its own synthesis. Engineering biosynthetic enzymes that lack the amino acids being synthesized provides insights into the evolution of amino acid biosynthesis and pathways for bioengineering.

Synthesis and Application of Aurophilic Poly(Cysteine and Poly(Cysteine-Containing Copolymers

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David Ulkoski

2017-10-01

Full Text Available The redox capacity, as well as the aurophilicity of the terminal thiol side groups, in poly(Cysteine lend a unique characteristic to this poly(amino acid or polypeptide. There are two major application fields for this polymer: (i biomedical applications in drug delivery and surface modification of biomedical devices and (ii as coating for electrodes to enhance their electrochemical sensitivity. The intended application determines the synthetic route for p(Cysteine. Polymers to be used in biomedical applications are typically polymerized from the cysteine N-carboxyanhydride by a ring-opening polymerization, where the thiol group needs to be protected during the polymerization. Advances in this methodology have led to conditions under which the polymerization progresses as living polymerization, which allows for a strict control of the molecular architecture, molecular weight and polydispersity and the formation of block copolymers, which eventually could display polyphilic properties. Poly(Cysteine used as electrode coating is typically polymerized onto the electrode by cyclic voltammetry, which actually produces a continuous, pinhole-free film on the electrode via the formation of covalent bonds between the amino group of Cysteine and the carbon of the electrode. This resulting coating is chemically very different from the well-defined poly(Cysteine obtained by ring-opening polymerizations. Based on the structure of cysteine a significant degree of cross-linking within the coating deposited by cyclic voltammetry can be assumed. This manuscript provides a detailed discussion of the ring-opening polymerization of cysteine, a brief consideration of the role of glutathione, a key cysteine-containing tripeptide, and examples for the utilization of poly(Cysteine and poly(Cysteine-containing copolymers, in both, the biomedical as well as electrochemical realm.

Evaluation the effect of albendazole against nematodes in sheep

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M. I. Al-Farwachi

2008-01-01

Full Text Available Six sheep farms in Mosul city, Iraq randomly selected, were surveyed for gastrointestinal nematodes resistant to Albendazole. On each of 6 sheep farms, 20 lambs were randomly distributed into two equal groups untreated control group, and albendazole (benzimidazole group (10 mg/kg BW. Faecal egg counts and larval cultures were done at 7, 14, and 21 days after anthelmintic treatment. Resistance was apparent for albendazole on 4 farms out of 6 (66.7%. Post-treatment larval cultures indicated: Strongyloides papillosus, Marshalligia marshalli, Nematodirus spathiger and Haemonchus contortus.

Haemonchus resistente a lactona macrocíclica em caprinos naturalmente parasitados

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Mattos Mary Jane Tweedie de

2004-01-01

Full Text Available O ivermectin, antiparasitário amplamente utilizado em caprinos jovens, foi avaliado sobre nematódeos gastrintestinais, em todas as fases parasitárias. Doze caprinos jovens, naturalmente parasitados foram utilizados no experimento, sendo um grupo de seis animais medicado com ivermectin, na dose de 200µg.kg-1 v.o. e o outro grupo utilizado como controle. Para a avaliação, utilizou-se o teste de redução na contagem de ovos (FECR, redução de larvas infectantes e carga parasitária. Amostras fecais de caprinos foram coletadas no dia da medicação (dia zero, aos sete e catorze dias. No décimo quarto dia pós-medicação, todos os caprinos foram sacrificados, para a coleta de helmintos e cálculo da carga parasitária total. A redução do número de ovos (FECR e a eficácia sobre adultos foi de 42,10 % e 32,62 %, respectivamente.O gênero Haemonchus representou 100 % da população do grupo medicado e 99,58 % do grupo controle. Estes resultados indicam que os caprinos já estavam parasitados por Haemonchus resistente ao ivermectin.

Anthelmintic resistance in a dairy cattle farm in the State of Minas Gerais Resistência anti-helmíntica em uma propriedade de bovinos leiteiros em Minas Gerais

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Maria do Socorro Veloso Leite Ferraz da Costa

2011-06-01

Full Text Available Eighty-four half-blood Gir × Holstein (F1 calves aged six months who were naturally infected by gastrointestinal helminths and maintained in rotational grazing received different anthelmintic treatments. Group A received anthelmintics according to the usual management in the property (eight treatments, seven including a macrocyclic lactone agent. Group B received strategic treatment (ivermectin 3.15% at the beginning and at the end of the rainy period. Eggs per gram of feces (EPG counts and genus of larvae from fecal cultures were determined on a monthly basis from April 2002 to December 2003. There was no significant reduction (p > 0.05 in EPG counts in any group after anthelminthic treatment, and the larvae in fecal cultures observed were Cooperia, Haemonchus, Oesophagostomum and a few Trichostrongylus. Cooperia was the most prevalent genus in the first four months of the experiment and Haemonchus in the following months. In 2003, tracer calves were introduced onto the pastures monthly and they showed high nematode burden many times throughout the year, and Cooperia punctata and Haemonchus contortus were the main species identified. The results suggest that there is anthelminthic resistance in this farm, mainly to macrocyclic lactones, and the development of immunity by crossbred animals was vital to reduce nematode burden.Oitenta e quatro bezerras meio sangue Gir × holandês (F1 com seis meses de idade, naturalmente infectadas por helmintos gastrintestinais e mantidas em pastejo rotacionado receberam diferentes tratamentos anti-helmínticos. O grupo A recebeu anti-helmínticos segundo manejo empregado na propriedade (oito tratamentos, sete com produtos à base de lactonas macrocíclicas. O grupo B recebeu tratamento estratégico (ivermectina 3,15% no inicio e final de período chuvoso. Mensalmente, no período de abril de 2002 a dezembro de 2003, foram realizadas contagens de ovos por grama de fezes (OPG e coproculturas. Não houve redu

Benzimidazole resistance of sheep nematodes in Norway confirmed through controlled efficacy test

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Domke Atle V

2012-08-01

Full Text Available Abstract Background Resistance against benzimidazoles (BZ has recently been detected in Norwegian sheep flocks through a large scale prevalence survey based on the faecal egg count reduction test (FECRT. The use of this test in combination with bulk larval culture only gives an indication of which gastrointestinal nematodes genera that are involved and these results have to be confirmed by a controlled efficacy test (CET to get accurate information about resistant nematodes populations at species level. A CET was therefore performed with larvae from two flocks where BZ resistance was previously detected through FECRT. Results The latter test confirmed the previous results in both flocks. In flock A, the BZ resistant nematode population consisted solely of Haemonchus contortus, whereas H. contortus and Teladorsagia circumcincta comprised the resistant worm population in flock B. Conclusions Some discrepancies that have been recorded between FECRT and CET results regarding time for post-treatment coproscopical examination and a temporary suppression of faecal egg excretion are discussed.

The Cysteine Protease–Cysteine Protease Inhibitor System Explored in Soybean Nodule Development

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Marian Dorcas Quain

2013-08-01

Full Text Available Almost all protease families have been associated with plant development, particularly senescence, which is the final developmental stage of every organ before cell death. Proteolysis remobilizes and recycles nitrogen from senescent organs that is required, for example, seed development. Senescence-associated expression of proteases has recently been characterized using large-scale gene expression analysis seeking to identify and characterize senescence-related genes. Increasing activities of proteolytic enzymes, particularly cysteine proteases, are observed during the senescence of legume nodules, in which a symbiotic relationship between the host plant and bacteria (Rhizobia facilitate the fixation of atmospheric nitrogen. It is generally considered that cysteine proteases are compartmentalized to prevent uncontrolled proteolysis in nitrogen-fixing nodules. In addition, the activities of cysteine proteases are regulated by endogenous cysteine protease inhibitors called cystatins. These small proteins form reversible complexes with cysteine proteases, leading to inactivation. However, very little is currently known about how the cysteine protease-cysteine protease inhibitor (cystatin system is regulated during nodule development. Moreover, our current understanding of the expression and functions of proteases and protease inhibitors in nodules is fragmented. To address this issue, we have summarized the current knowledge and techniques used for studying proteases and their inhibitors including the application of “omics” tools, with a particular focus on changes in the cysteine protease-cystatin system during nodule development.

Anthelmintic activity of Trianthema portulacastrum L. and Musa paradisiaca L. against gastrointestinal nematodes of sheep.

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Hussain, Altaf; Khan, Muhammad Nisar; Iqbal, Zafar; Sajid, Muhammad Sohail; Khan, Muhammad Kasib

2011-06-30

Evaluation of anthelmintic effects of Trianthema (T.) portulacastrum L. (Aizoaceae) whole plant and Musa (M.) paradisiaca L. (Musaceae) leaves against prevalent gastrointestinal worms of sheep was done that may justify their traditional use in veterinary clinical medicine. In vitro anthelmintic activity of the crude aqueous methanolic extract (CAME) of both the plants was determined using mature female Haemonchus (H.) contortus and their eggs in adult motility assay (AMA) and egg hatch test (EHT), respectively. In vivo anthelmintic activity of crude powder (CP) and CAME in increasing doses (1.0-8.0 g kg(-1)) was determined in sheep naturally infected with mixed species of nematodes using fecal egg count reduction test (FECRT) and larval counts. The study design also included untreated as well as treated controls. Fecal egg count reduction and larval counts from coprocultures were performed pre- and post-treatments to assess the anthelmintic activity of the plants. CAME of T. portulacastrum and M. paradisiaca showed a strong in vitro anthelmintic activity and pronounced inhibitory effects on H. contortus egg hatching as observed through AMA and EHT, respectively. Both plants exhibited dose and time dependent anthelmintic effects on live worms as well as egg hatching. M. paradisiaca (LC(50)=2.13 μg mL(-1)) was found to be more potent than T. portulacastrum (LC(50)=2.41 μg mL(-1)) in EHT. However, in vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 85.6% and 80.7% with CAME of T. portulacastrum and M. paradisiaca at 8.0 g kg(-1) on 15th day post-treatment, respectively as compared to that of Levamisole (7.5 mg kg(-1)) that caused 97.0% reduction in EPG. All the species of gastrointestinal nematodes (GINs), i.e. Haemonchus contortus, Trichostronglyus spp., Oesophagostomum columbianum and Trichuris ovis which were prevalent, found susceptible (Pparadisiaca possess strong anthelmintic activity in vitro and in vivo, thus, justifying their use in

An estimate of seasonality and intensity of infection with gastrointestinal nematodes in sheep and goats . in West Java

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Beriajaya

1998-12-01

Full Text Available Tracer Inoodle tbin-tail sheep and Kacang goats were used to measure the seasonal changes in gastrointestinal nematodes parasitism i ~'und[, .xillapek .Fodd~ 'ofis in West Java . Each 3 months for 12 months worm-free male sheep (5 and goats (5 about 5 months of age were ~ $Jri~tgldI o% ef- farmer, and managed as part of their flock for 2 months . Animals were then returned to the laboratory and maintained on "`~^taaan-ftwAiet in elevated slatted pens for 3 weeks prior to slaughter. In all trials sheep had higher faecal egg counts than goats . Egg counts were significantly lower during the late dry-early wet season due mainly to lower burdens of Oesophagostomum spp. than at other times of the year. The predominant genera recovered from faecal larval cultures were Haemonchus and Trichostrongylus . At post mortem more than 94 percent of animals were infected with Trichostrongylus colubriformis, T. axei, Haemonchus contortus, Oesophagostomum columbianwn and Strongyloides papillosus . Other species found, in descending order of occurrence, were Cooperia curticei, Trichuris ovis, Bunostornum trigonocephalum, Oesophagostomumn asperum, Capillaria bovis and Gaigena pachycelis. It was concluded that intensity of exposure of both sheep and goats to H contortus, T. axei and C. curticei was similar throughout the year, but that availability of infectioe larvae of T. colubriformis was higher during the dry than the wet season and vise versa for O. columbianum . Sheep had higher burdens of T. Colubrzformis than goats but similar numbers of other species.

Atividade anti-helmíntica de plantas nativas do continente americano: uma revisão Anthelmintic activity of native American plants: a review

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R.G. Sousa

2013-01-01

Full Text Available O presente trabalho faz uma revisão de plantas nativas do continente americano com atividade anti-helmíntica com a finalidade de subsidiar pesquisas e o desenvolvimento de novos medicamentos na área de produtos naturais. Na literatura, foram citadas 22 famílias de plantas, tais como: Moraceae, Asteraceae e Cucurbitaceae, e estudadas cerca de 35 espécies, como a Ficus sp, Cucurbita máxima, e Ruta graveolens, usadas por sua alegada atividade anti-helmíntica. Destas espécies, 26 foram ativas contra alguns helmintos: Strongyloides stercolaris, Ancylostomidae, Trichostrongylus sp, Capillaria sp, Ascaris lumbricoides, Taenia saginata, Haemonchus contortus, e Hymenolepsis diminuta. Foi verificado maior número de estudos com parasitas de animais, possivelmente devido ao desenvolvimento de resistência dos helmintos aos tratamentos convencionais disponíveis.This paper is a review of native American plants showing anthelmintic activity in order to give support to research and development of new drugs in the field of natural products. In the literature, 22 plant families were cited, such as: Moraceae, Asteraceae and Cucurbitaceae, and around 35 species were studied, including Ficus sp, Cucurbita maxima and Ruta graveolens, used for their putative anthelmintic activity. From these species, 26 were active against some helminthes: Strongyloides stercolaris, Ancylostomidae, Trichostrongylus sp, Capillaria sp, Ascaris lumbricoides, Taenia saginata, Haemonchus contortus and Hymenolepsis diminuta. Data indicated a larger number of studies with animal parasites, probably due to the resistance of these helminthes to the conventional treatments available.

Simultaneous electrochemical determination of L-cysteine and L-cysteine disulfide at carbon ionic liquid electrode.

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Safavi, Afsaneh; Ahmadi, Raheleh; Mahyari, Farzaneh Aghakhani

2014-04-01

A linear sweep voltammetric method is used for direct simultaneous determination of L-cysteine and L-cysteine disulfide (cystine) based on carbon ionic liquid electrode. With carbon ionic liquid electrode as a high performance electrode, two oxidation peaks for L-cysteine (0.62 V) and L-cysteine disulfide (1.3 V) were observed with a significant separation of about 680 mV (vs. Ag/AgCl) in phosphate buffer solution (pH 6.0). The linear ranges were obtained as 1.0-450 and 5.0-700 μM and detection limits were estimated to be 0.298 and 4.258 μM for L-cysteine and L-cysteine disulfide, respectively. This composite electrode was applied for simultaneous determination of L-cysteine and L-cysteine disulfide in two real samples, artificial urine and nutrient broth. Satisfactory results were obtained which clearly indicate the applicability of the proposed electrode for simultaneous determination of these compounds in complex matrices.

Evaluation of Eucalyptus citriodora essential oil on goat gastrointestinal nematodes Avaliação do óleo essencial de Eucalyptus citriodora sobre nematóides gastrintestinais de caprinos

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Iara Tersia Freitas Macedo

2011-09-01

Full Text Available Phytotherapy may be an alternative strategy for controlling gastrointestinal parasites. This study evaluated the anthelmintic efficacy of Eucalyptus citriodora essential oil (EcEO. The in vitro effects of EcEO were determined through testing the inhibition of egg hatching and larval development of Haemonchus contortus. EcEO was subjected to acute toxicity testing on mice, orally and intraperitoneally. The in vivo effects of EcEO were determined by the fecal egg count reduction test (FECRT in goats infected with gastrointestinal nematodes. The results showed that 5.3 mg.mL-1 EcEO inhibited egg hatching by 98.8% and 10.6 mg.mL-1 EcEO inhibited H. contortus larval development by 99.71%. The lethal doses for 50% of the mice were 4153 and 622.8 mg.kg-1, for acute toxicity orally and intraperitoneally. In the FECRT, the efficacy of EcEO and ivermectin was 66.25 and 79.16% respectively, on goat gastrointestinal nematodes eight days after treatment. EcEO showed in vitro and in vivo anthelmintic activity.Fitoterapia pode ser uma estratégia alternativa para o controle de parasitas gastrintestinais. Este estudo avaliou a eficácia anti-helmintica do óleo essencial de Eucalyptus citriodora (OeEc. Os efeitos in vitro do OeEc foram determinados através do teste de eclosão de ovos e inibição do desenvolvimento larvar de Haemonchus contortus. O OeEc foi submetido ao teste de toxicidade aguda oral e intraperitoneal, em camundongos. Os efeitos in vivo do OeEc foram avaliados através do teste de redução da contagem de ovos nas fezes (FECRT com caprinos infectados com nematóides gastrintestinais. Os resultados mostraram que 5,3 mg.mL-1 OeEc inibiram 98,8% a eclosão de ovos e 10,6 mg.mL-1 OeEc inibiram 99,71% o desenvolvimento larvar de H. contortus. As doses letais para 50% dos camundongos foram de 4153 e 622,8 mg.kg-1 pela via oral e intraperitoneal. No FECRT, a eficácia de OeEc e ivermectina foi de 66,25 e 79,16%, respectivamente, em caprinos 8 dias

Prevalence of gastrointestinal parasites in young camels in Bahrain

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M. I. Abubakr

2000-03-01

Full Text Available The prevalence of gastrointestinal parasites in young camels in Bahrain is reported for the first time. Six genera of parasites were found. The nematodes observed were Haemonchus contortus (36.47%, Nematodirus spathiger (30.59% and Trichuris sp. (10.6%; the only cestode recorded was Moniezia expansa (2.4%. The incidence of Eimeria dromedarii was 20%. Single, double, triple and quadruple parasitic infestation occurred in 41.2, 33.5, 19.4 and 5.9% of the infected animals, respectively. Balantidium coli, a protozoan parasite, was occasionally seen in young camels suffering from diarrhea at the time of sampling.

Soft Cysteine Signaling Network: The Functional Significance of Cysteine in Protein Function and the Soft Acids/Bases Thiol Chemistry That Facilitates Cysteine Modification.

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Wible, Ryan S; Sutter, Thomas R

2017-03-20

The unique biophysical and electronic properties of cysteine make this molecule one of the most biologically critical amino acids in the proteome. The defining sulfur atom in cysteine is much larger than the oxygen and nitrogen atoms more commonly found in the other amino acids. As a result of its size, the valence electrons of sulfur are highly polarizable. Unique protein microenvironments favor the polarization of sulfur, thus increasing the overt reactivity of cysteine. Here, we provide a brief overview of the endogenous generation of reactive oxygen and electrophilic species and specific examples of enzymes and transcription factors in which the oxidation or covalent modification of cysteine in those proteins modulates their function. The perspective concludes with a discussion of cysteine chemistry and biophysics, the hard and soft acids and bases model, and the proposal of the Soft Cysteine Signaling Network: a hypothesis proposing the existence of a complex signaling network governed by layered chemical reactivity and cross-talk in which the chemical modification of reactive cysteine in biological networks triggers the reorganization of intracellular biochemistry to mitigate spikes in endogenous or exogenous oxidative or electrophilic stress.

Efficacy of copper oxide wire particles against gastrointestinal nematodes in sheep and goats.

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Soli, F; Terrill, T H; Shaik, S A; Getz, W R; Miller, J E; Vanguru, M; Burke, J M

2010-02-26

Profitable sheep and goat production in the USA is severely limited by gastrointestinal nematode (GIN) parasitism, particularly by Haemonchus contortus. Copper oxide wire particles (COWP) have anti-parasitic properties in the diet of small ruminants, but efficacy of COWP may differ between sheep and goats. In a study with weaned kids (Kiko x Spanish cross, 6 months old) and lambs (Katahdin or Dorper x Blackface crosses, 5 months old), grazing the same pasture area in Central Georgia, 2g of COWP in a gel capsule was given to half the animals of each species, while the other half were given no COWP. Fecal and blood samples were taken weekly to determine GIN fecal egg counts (FEC) and blood packed cell volume (PCV). After COWP treatment, animals were grazed for 4 weeks and then slaughtered, with adult GIN recovered from the abomasum and small intestines for counting and identification to species. For both sheep and goats, COWP treatment reduced EPG (P<0.05), increased PCV (P<0.05), and lowered abomasal GIN numbers (P<0.05). For EPG, these differences were 82.5 and 90.5% for sheep and goats, respectively, 26 days after treatment, while adult H. contortus were 67.2 and 85.8% lower for COWP-treated sheep and goats, respectively. In this study, COWP treatment was equally effective against H. contortus infection in lambs and kids and appears to be an effective method of controlling H. contortus infection for up to 6 weeks in small ruminants following weaning.

Yeast genes involved in regulating cysteine uptake affect production of hydrogen sulfide from cysteine during fermentation.

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Huang, Chien-Wei; Walker, Michelle E; Fedrizzi, Bruno; Gardner, Richard C; Jiranek, Vladimir

2017-08-01

An early burst of hydrogen sulfide (H2S) produced by Saccharomyces cerevisiae during fermentation could increase varietal thiols and therefore enhance desirable tropical aromas in varieties such as Sauvignon Blanc. Here we attempted to identify genes affecting H2S formation from cysteine by screening yeast deletion libraries via a colony colour assay on media resembling grape juice. Both Δlst4 and Δlst7 formed lighter coloured colonies and produced significantly less H2S than the wild type on high concentrations of cysteine, likely because they are unable to take up cysteine efficiently. We then examined the nine known cysteine permeases and found that deletion of AGP1, GNP1 and MUP1 led to reduced production of H2S from cysteine. We further showed that deleting genes involved in the SPS-sensing pathway such as STP1 and DAL81 also reduced H2S from cysteine. Together, this study indirectly confirms that Agp1p, Gnp1p and Mup1p are the major cysteine permeases and that they are regulated by the SPS-sensing and target of rapamycin pathways under the grape juice-like, cysteine-supplemented, fermentation conditions. The findings highlight that cysteine transportation could be a limiting factor for yeast to generate H2S from cysteine, and therefore selecting wine yeasts without defects in cysteine uptake could maximise thiol production potential. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Albendazole in environment: faecal concentrations in lambs and impact on lower development stages of helminths and seed germination.

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Prchal, Lukáš; Podlipná, Radka; Lamka, Jiří; Dědková, Tereza; Skálová, Lenka; Vokřál, Ivan; Lecová, Lenka; Vaněk, Tomáš; Szotáková, Barbora

2016-07-01

Albendazole (ABZ), widely used benzimidazole anthelmintic, administered to animals enters via excrements into environment and may impact non-target organisms. Moreover, exposure of lower development stages of helminths to anthelmintics may also encourage the development of drug-resistant strains of helminths. In present project, the kinetics of ABZ (10 mg kg(-1) p.o.) and its metabolite (ABZ.SO, ABZSO2) elimination in faeces from treated Texel lambs were studied using UHPLC/MS/MS with the aim to find out their concentrations achievable in the environment. Consequently, the effect of these compounds on lower development stages of Barber's pole worm (Haemonchus contortus) and on germination of white mustard (Sinapis alba) seeds was evaluated. The results showed that ABZ concentrations in faeces excreted in 4-60 h after treatment were above the concentrations lethal for H. contortus eggs. Moreover, pre-incubation with sub-lethal doses of ABZ and ABZ.SO did not increase the resistance of H. contortus eggs and larvae to anthelmintics. On the other hand, concentrations of ABZ and ABZ.SO in faeces are so high that might have negative influence on non-target soil invertebrates. As neither ABZ nor its metabolites affect the germination of mustard seeds, phytoremediation could be considered as potential tool for detoxification of ABZ in the environment.

Crystal Structure of Mammalian Cysteine dioxygenase: A Novel Mononuclear Iron Center for Cysteine Thiol Oxidation

Energy Technology Data Exchange (ETDEWEB)

Simmons,C.; Liu, Q.; Huang, Q.; Hao, Q.; Begley, T.; Karplus, P.; Stipanuk, M.

2006-01-01

Cysteine dioxygenase is a mononuclear iron-dependent enzyme responsible for the oxidation of cysteine with molecular oxygen to form cysteinesulfinate. This reaction commits cysteine to either catabolism to sulfate and pyruvate or to the taurine biosynthetic pathway. Cysteine dioxygenase is a member of the cupin superfamily of proteins. The crystal structure of recombinant rat cysteine dioxygenase has been determined to 1.5 Angstroms resolution, and these results confirm the canonical cupin {beta}-sandwich fold and the rare cysteinyl-tyrosine intramolecular crosslink (between Cys93 and Tyr157) seen in the recently reported murine cysteine dioxygenase structure. In contrast to the catalytically inactive mononuclear Ni(II) metallocenter present in the murine structure, crystallization of a catalytically competent preparation of rat cysteine dioxygenase revealed a novel tetrahedrally coordinated mononuclear iron center involving three histidines (His86, His88, and His140) and a water molecule. Attempts to acquire a structure with bound ligand using either co-crystallization or soaks with cysteine revealed the formation of a mixed disulfide involving Cys164 near the active site, which may explain previously observed substrate inhibition. This work provides a framework for understanding the molecular mechanisms involved in thiol dioxygenation and sets the stage for exploring the chemistry of both the novel mononuclear iron center and the catalytic role of the cysteinyl-tyrosine linkage.

Metabolic profiling and in vitro assessment of anthelmintic fractions of Picria fel-terrae Lour.

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Rasika Kumarasingha

2016-12-01

Full Text Available Anthelmintic resistance is widespread in gastrointestinal nematode populations, such that there is a consistent need to search for new anthelmintics. However, the cost of screening for new compounds is high and has a very low success rate. Using the knowledge of traditional healers from Borneo Rainforests (Sarawak, Malaysia, we have previously shown that some traditional medicinal plants are a rich source of potential new anthelmintic drug candidates. In this study, Picria fel-terrae Lour. plant extract, which has previously shown promising anthelmintic activities, was fractionated via the use of a solid phase extraction cartridge and each isolated fraction was then tested on free-living nematode Caenorhabditis elegans and the parasitic nematode Haemonchus contortus. We found that a single fraction was enriched for nematocidal activity, killing ≥90% of C. elegans adults and inhibiting the motility of exsheathed L3 of H. contortus, while having minimal cytotoxic activity in mammalian cell culture. Metabolic profiling and chemometric analysis of the effective fraction indicated medium chained fatty acids and phenolic acids were highly represented.

Enhancement of thioredoxin/glutaredoxin-mediated L-cysteine synthesis from S-sulfocysteine increases L-cysteine production in Escherichia coli

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2012-01-01

Background Escherichia coli has two L-cysteine biosynthetic pathways; one is synthesized from O-acetyl L-serine (OAS) and sulfate by L-cysteine synthase (CysK), and another is produced via S-sulfocysteine (SSC) from OAS and thiosulfate by SSC synthase (CysM). SSC is converted into L-cysteine and sulfite by an uncharacterized reaction. As thioredoxins (Trx1 and Trx2) and glutaredoxins (Grx1, Grx2, Grx3, Grx4, and NrdH) are known as reductases of peptidyl disulfides, overexpression of such reductases might be a good way for improving L-cysteine production to accelerate the reduction of SSC in E. coli. Results Because the redox enzymes can reduce the disulfide that forms on proteins, we first tested whether these enzymes catalyze the reduction of SSC to L-cysteine. All His-tagged recombinant enzymes, except for Grx4, efficiently convert SSC into L-cysteine in vitro. Overexpression of Grx1 and NrdH enhanced a 15-40% increase in the E. coliL-cysteine production. On the other hand, disruption of the cysM gene cancelled the effect caused by the overexpression of Grx1 and NrdH, suggesting that its improvement was due to the efficient reduction of SSC under the fermentative conditions. Moreover, L-cysteine production in knockout mutants of the sulfite reductase genes (ΔcysI and ΔcysJ) and the L-cysteine synthase gene (ΔcysK) each decreased to about 50% of that in the wild-type strain. Interestingly, there was no significant difference in L-cysteine production between wild-type strain and gene deletion mutant of the upstream pathway of sulfite (ΔcysC or ΔcysH). These results indicate that sulfite generated from the SSC reduction is available as the sulfur source to produce additional L-cysteine molecule. It was finally found that in the E. coliL-cysteine producer that co-overexpress glutaredoxin (NrdH), sulfite reductase (CysI), and L-cysteine synthase (CysK), there was the highest amount of L-cysteine produced per cell. Conclusions In this work, we showed that Grx1 and

21 CFR 184.1271 - L-Cysteine.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true L-Cysteine. 184.1271 Section 184.1271 Food and... Substances Affirmed as GRAS § 184.1271 L-Cysteine. (a) L-Cysteine is the chemical L-2-amino-3... of total L-cysteine per 100 parts of flour in dough as a dough strengthener as defined in § 170.3(o...

The mitochondrial toxicity of cysteine-S-conjugates: Studies with pentachlorobutadienyl-L-cysteine

International Nuclear Information System (INIS)

Wallin, A.

1990-01-01

Nephrotoxic cysteine conjugates, arising from mercapturate biosynthesis, can perturb the mitochondrial membrane potential and calcium homeostasis in renal epithelial cells. Activation of these cysteine conjugates to reactive species by mitochondrial β-lyases results in covalent binding and mitochondrial damage. PCBC and related cysteine conjugates inhibit ADP-stimulated respiration in mitochondria respiring on alpha-ketoglutrate/malate and succinate indicating that both dehydrogenases may be targets. The respiratory inhibition is blocked by aminooxyacetic acid, an inhibitor of the β-lyase. Hence, metabolic activation is required implying that covalent binding of reactive intermediates may be important to the mitochondrial injury. Binding of 35 S-fragments has been found for 5 conjugates with varying degrees of mitochondrial toxicity. PCBC is more lipophilic and has a higher affinity for cellular membranes than other cysteine conjugates. PCBC rapidly depolarizes the inner membrane potential resulting in an inhibition of mitochondrial oxidative phosphorylation and calcium upon sequestration. Consequently, mitochondria and renal epithelial cells exposed to PCBC show a sudden release of calcium upon exposure to PCBC which is followed by a later increase in state 4 respiration leading to an inhibition of oxidative phosphorylation. The primary effect of other cysteine conjugates is an inhibition of the dehydrogenases, thus inhibiting state 3 respiration

Synergistic interaction of ten essential oils against Haemonchus contortus in vitro

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Anthelmintic resistance in sheep gastrointestinal nematodes is a worldwide problem. Multi-drug resistant haemonchosis is the most serious impediment for small ruminant systems, and there are no new drug candidates currently under development. Molecules from natural sources have demonstrated anthelmi...

L-Cysteine Metabolism and Fermentation in Microorganisms.

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Takagi, Hiroshi; Ohtsu, Iwao

L-Cysteine is an important amino acid both biologically and commercially. Although most amino acids are industrially produced by microbial fermentation, L-cysteine has been mainly produced by protein hydrolysis. Due to environmental and safety problems, synthetic or biotechnological products have been preferred in the market. Here, we reviewed L-cysteine metabolism, including biosynthesis, degradation, and transport, and biotechnological production (including both enzymatic and fermentation processes) of L-cysteine. The metabolic regulation of L-cysteine including novel sulfur metabolic pathways found in microorganisms is also discussed. Recent advancement in biochemical studies, genome sequencing, structural biology, and metabolome analysis has enabled us to use various approaches to achieve direct fermentation of L-cysteine from glucose. For example, worldwide companies began to supply L-cysteine and its derivatives produced by bacterial fermentation. These companies successfully optimized the original metabolism of their private strains. Basically, a combination of three factors should be required for improving L-cysteine fermentation: that is, (1) enhancing biosynthesis: overexpression of the altered cysE gene encoding feedback inhibition-insensitive L-serine O-acetyltransferase (SAT), (2) weakening degradation: knockout of the genes encoding L-cysteine desulfhydrases, and (3) exploiting export system: overexpression of the gene involved in L-cysteine transport. Moreover, we found that "thiosulfate" is much more effective sulfur source than commonly used "sulfate" for L-cysteine production in Escherichia coli, because thiosulfate is advantageous for saving consumption of NADPH and relating energy molecules.

21 CFR 184.1272 - L-Cysteine monohydrochloride.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true L-Cysteine monohydrochloride. 184.1272 Section 184... Listing of Specific Substances Affirmed as GRAS § 184.1272 L-Cysteine monohydrochloride. (a) L-Cysteine... ingredient is used to supply up to 0.009 part of total L-cysteine per 100 parts of flour in dough as a dough...

Assay of cysteine dioxygenase activity

International Nuclear Information System (INIS)

Bagley, P.J.; Stipanuk, M.H.

1990-01-01

It has been proposed that rat liver contains two cysteine dioxygenase enzymes which convert cysteine to cysteinesulfinic acid, one which is stimulated by NAD + and has a pH optimum of 6.8 and one which is not stimulated by NAD + and has a pH optimum of 9.0. This led the authors to reinvestigate assay conditions for measuring cysteine dioxygenase activity in rat liver homogenate. An HPLC method, using an anion exchange column (Dionex Amino-Pac trademark PA1 (4x250 mm)) was used to separate the [ 35 S]cysteinesulfinic acid produced from [ 35 S]cysteine in the incubation mixture. They demonstrated that inclusion of hydroxylamine prevented further metabolism of cysteinesulfinic acid. which occurred rapidly in the absence of hydroxylamine

On the Dynamical Behavior of the Cysteine Dioxygenase-l-Cysteine Complex in the Presence of Free Dioxygen and l-Cysteine.

Science.gov (United States)

Pietra, Francesco

2017-11-01

In this work, viable models of cysteine dioxygenase (CDO) and its complex with l-cysteine dianion were built for the first time, under strict adherence to the crystal structure from X-ray diffraction studies, for all atom molecular dynamics (MD). Based on the CHARMM36 FF, the active site, featuring an octahedral dummy Fe(II) model, allowed us observing water exchange, which would have escaped attention with the more popular bonded models. Free dioxygen (O 2 ) and l-cysteine, added at the active site, could be observed being expelled toward the solvating medium under Random Accelerated Molecular Dynamics (RAMD) along major and minor pathways. Correspondingly, free dioxygen (O 2 ), added to the solvating medium, could be observed to follow the same above pathways in getting to the active site under unbiased MD. For the bulky l-cysteine, 600 ns of trajectory were insufficient for protein penetration, and the molecule was stuck at the protein borders. These models pave the way to free energy studies of ligand associations, devised to better clarify how this cardinal enzyme behaves in human metabolism. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

π-Clamp-mediated cysteine conjugation

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Zhang, Chi; Welborn, Matthew; Zhu, Tianyu; Yang, Nicole J.; Santos, Michael S.; van Voorhis, Troy; Pentelute, Bradley L.

2016-02-01

Site-selective functionalization of complex molecules is one of the most significant challenges in chemistry. Typically, protecting groups or catalysts must be used to enable the selective modification of one site among many that are similarly reactive, and general strategies that selectively tune the local chemical environment around a target site are rare. Here, we show a four-amino-acid sequence (Phe-Cys-Pro-Phe), which we call the ‘π-clamp’, that tunes the reactivity of its cysteine thiol for site-selective conjugation with perfluoroaromatic reagents. We use the π-clamp to selectively modify one cysteine site in proteins containing multiple endogenous cysteine residues. These examples include antibodies and cysteine-based enzymes that would be difficult to modify selectively using standard cysteine-based methods. Antibodies modified using the π-clamp retained binding affinity to their targets, enabling the synthesis of site-specific antibody-drug conjugates for selective killing of HER2-positive breast cancer cells. The π-clamp is an unexpected approach to mediate site-selective chemistry and provides new avenues to modify biomolecules for research and therapeutics.

Rethinking Cysteine Protective Groups: S-Alkylsulfonyl-l-Cysteines for Chemoselective Disulfide Formation.

Science.gov (United States)

Schäfer, Olga; Huesmann, David; Muhl, Christian; Barz, Matthias

2016-12-12

The ability to reversibly cross-link proteins and peptides grants the amino acid cysteine its unique role in nature as well as in peptide chemistry. We report a novel class of S-alkylsulfonyl-l-cysteines and N-carboxy anhydrides (NCA) thereof for peptide synthesis. The S-alkylsulfonyl group is stable against amines and thus enables its use under Fmoc chemistry conditions and the controlled polymerization of the corresponding NCAs yielding well-defined homo- as well as block co-polymers. Yet, thiols react immediately with the S-alkylsulfonyl group forming asymmetric disulfides. Therefore, we introduce the first reactive cysteine derivative for efficient and chemoselective disulfide formation in synthetic polypeptides, thus bypassing additional protective group cleavage steps. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Evidence for cysteine sulfinate as a neurotransmitter

International Nuclear Information System (INIS)

Recasens, M.; Varga, V.; Nanopoulos, D.; Saadoun, F.; Vincendon, G.; Benavides, J.

1982-01-01

The Na + -independent binding of L-[ 3 H]cysteine sulfinate and L-[ 3 H]cysteine sulfinate uptake were investigated in rat brain membranes and vesicles. Specific binding of L-[ 3 H]cysteine sulfinate was saturable and occurred by a single high affinity process with a Ksub(b) of 100 nM +- 9 and a capacity (Bsub(max)) of 2.4 +- 0.22 pmol/mg protein. The regional distribution of the binding of L-[ 3 H]cysteine sulfinate in the brain was found to be heterogeneous. The rate of L-[ 3 H]cysteine sulfinate uptake shows a biphasic dependence on the concentration of L-cysteine sulfinate, corresponding to a high affinity (27.2 μM) and a low affinity (398 μM) transport system. The maximum L-[ 3 H]cysteine sulfinate uptake is reached at 2min and the uptake increases as a function of the sodium concentration. Chloride and potassium ions stimulate the uptake. (Auth.)

Reduction of Guanosyl Radical by Cysteine and Cysteine-Glycine Studied by Time-Resolved CIDNP

NARCIS (Netherlands)

Morozova, O.B.; Kaptein, R.; Yurkovskaya, A.V.

2012-01-01

As a model for chemical DNA repair, reduction of guanosyl radicals in the reaction with cysteine or the dipeptide cysteine-glycine has been studied by time-resolved chemically induced dynamic nuclear polarization (CIDNP). Radicals were generated photochemically by pulsed laser irradiation of a

Serine protease inhibitors of parasitic helminths.

Science.gov (United States)

Molehin, Adebayo J; Gobert, Geoffrey N; McManus, Donald P

2012-05-01

Serine protease inhibitors (serpins) are a superfamily of structurally conserved proteins that inhibit serine proteases and play key physiological roles in numerous biological systems such as blood coagulation, complement activation and inflammation. A number of serpins have now been identified in parasitic helminths with putative involvement in immune regulation and in parasite survival through interference with the host immune response. This review describes the serpins and smapins (small serine protease inhibitors) that have been identified in Ascaris spp., Brugia malayi, Ancylostoma caninum Onchocerca volvulus, Haemonchus contortus, Trichinella spiralis, Trichostrongylus vitrinus, Anisakis simplex, Trichuris suis, Schistosoma spp., Clonorchis sinensis, Paragonimus westermani and Echinococcus spp. and discusses their possible biological functions, including roles in host-parasite interplay and their evolutionary relationships.

L-Cysteine metabolism and its nutritional implications.

Science.gov (United States)

Yin, Jie; Ren, Wenkai; Yang, Guan; Duan, Jielin; Huang, Xingguo; Fang, Rejun; Li, Chongyong; Li, Tiejun; Yin, Yulong; Hou, Yongqing; Kim, Sung Woo; Wu, Guoyao

2016-01-01

L-Cysteine is a nutritionally semiessential amino acid and is present mainly in the form of L-cystine in the extracellular space. With the help of a transport system, extracellular L-cystine crosses the plasma membrane and is reduced to L-cysteine within cells by thioredoxin and reduced glutathione (GSH). Intracellular L-cysteine plays an important role in cellular homeostasis as a precursor for protein synthesis, and for production of GSH, hydrogen sulfide (H(2)S), and taurine. L-Cysteine-dependent synthesis of GSH has been investigated in many pathological conditions, while the pathway for L-cysteine metabolism to form H(2)S has received little attention with regard to prevention and treatment of disease in humans. The main objective of this review is to highlight the metabolic pathways of L-cysteine catabolism to GSH, H(2)S, and taurine, with special emphasis on therapeutic and nutritional use of L-cysteine to improve the health and well-being of animals and humans. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cysteine Protease Zymography: Brief Review.

Science.gov (United States)

Wilkesman, Jeff

2017-01-01

Cysteine proteases play multiple roles in basically all aspects of physiology and development. In plants, they are involved in growth and development and in accumulation and mobilization of storage proteins. Furthermore, they are engaged in signalling pathways and in the response to biotic and abiotic stresses. In animals and also in humans, they are responsible for senescence and apoptosis, prohormone processing, and ECM remodelling. When analyzed by zymography, the enzyme must be renaturated after SDS-PAGE. SDS must be washed out and substituted by Triton X-100. Gels are then further incubated under ideal conditions for activity detection. Cysteine proteases require an acidic pH (5.0-6.0) and a reducing agent, usually DTT. When screening biological samples, there is generally no previous clue on what peptidase class will be present, neither optimal proteolysis conditions are known. Hence, it is necessary to assess several parameters, such as incubation time, pH, temperature, influence of ions or reducing agents, and finally evaluate the inhibition profile. For detection of cysteine peptidase activity, the use of specific inhibitors, such as E-64, can be used to prevent the development of cysteine peptidase activity bands and positively confirm its presence. Here four different protocols to assess cysteine protease activity from different sources are presented.

The role of cysteine residues in the sulphate transporter, SHST1: construction of a functional cysteine-less transporter.

Science.gov (United States)

Howitt, Susan M

2005-05-20

We investigated the role of cysteine residues in the sulphate transporter, SHST1, with the aim of generating a functional cysteine-less variant. SHST1 contains five cysteine residues and none was essential for function. However, replacement of C421 resulted in a reduction in transport activity. Sulphate transport by C205 mutants was dependent on the size of the residue at this position. Alanine at position 205 resulted in a complete loss of function whereas leucine resulted in a 3-fold increase in sulphate transport relative to wild type SHST1. C205 is located in a putative intracellular loop and our results suggest that this loop may be important for sulphate transport. By replacing C205 with leucine and the other four cysteine residues with alanine, we constructed a cysteine-less variant of SHST1 that has transport characteristics indistinguishable from wild type. This construct will be useful for further structure and function studies of SHST1.

Mass spectrometric analysis of L-cysteine metabolism: physiological role and fate of L-cysteine in the enteric protozoan parasite Entamoeba histolytica.

Science.gov (United States)

Jeelani, Ghulam; Sato, Dan; Soga, Tomoyoshi; Watanabe, Haruo; Nozaki, Tomoyoshi

2014-11-04

L-cysteine is essential for virtually all living organisms, from bacteria to higher eukaryotes. Besides having a role in the synthesis of virtually all proteins and of taurine, cysteamine, glutathione, and other redox-regulating proteins, L-cysteine has important functions under anaerobic/microaerophilic conditions. In anaerobic or microaerophilic protozoan parasites, such as Entamoeba histolytica, L-cysteine has been implicated in growth, attachment, survival, and protection from oxidative stress. However, a specific role of this amino acid or related metabolic intermediates is not well understood. In this study, using stable-isotope-labeled L-cysteine and capillary electrophoresis-time of flight mass spectrometry, we investigated the metabolism of L-cysteine in E. histolytica. [U-(13)C3, (15)N]L-cysteine was rapidly metabolized into three unknown metabolites, besides L-cystine and L-alanine. These metabolites were identified as thiazolidine-4-carboxylic acid (T4C), 2-methyl thiazolidine-4-carboxylic acid (MT4C), and 2-ethyl-thiazolidine-4-carboxylic acid (ET4C), the condensation products of L-cysteine with aldehydes. We demonstrated that these 2-(R)-thiazolidine-4-carboxylic acids serve for storage of L-cysteine. Liberation of L-cysteine occurred when T4C was incubated with amebic lysates, suggesting enzymatic degradation of these L-cysteine derivatives. Furthermore, T4C and MT4C significantly enhanced trophozoite growth and reduced intracellular reactive oxygen species (ROS) levels when it was added to cultures, suggesting that 2-(R)-thiazolidine-4-carboxylic acids are involved in the defense against oxidative stress. Amebiasis is a human parasitic disease caused by the protozoan parasite Entamoeba histolytica. In this parasite, L-cysteine is the principal low-molecular-weight thiol and is assumed to play a significant role in supplying the amino acid during trophozoite invasion, particularly when the parasites move from the anaerobic intestinal lumen to highly

The IRC7 gene encodes cysteine desulphydrase activity and confers on yeast the ability to grow on cysteine as a nitrogen source.

Science.gov (United States)

Santiago, Margarita; Gardner, Richard C

2015-07-01

Although cysteine desulphydrase activity has been purified and characterized from Saccharomyces cerevisiae, the gene encoding this activity in vivo has never been defined. We show that the full-length IRC7 gene, encoded by the YFR055W open reading frame, encodes a protein with cysteine desulphydrase activity. Irc7p purified to homogeneity is able to utilize l-cysteine as a substrate, producing pyruvate and hydrogen sulphide as products of the reaction. Purified Irc7p also utilized l-cystine and some other cysteine conjugates, but not l-cystathionine or l-methionine, as substrates. We further show that, in vivo, the IRC7 gene is both necessary and sufficient for yeast to grow on l-cysteine as a nitrogen source, and that overexpression of the gene results in increased H2 S production. Strains overexpressing IRC7 are also hypersensitive to a toxic analogue, S-ethyl-l-cysteine. While IRC7 has been identified as playing a critical role in converting cysteine conjugates to volatile thiols that are important in wine aroma, its biological role in yeast cells is likely to involve regulation of cysteine and redox homeostasis. Copyright © 2015 John Wiley & Sons, Ltd.

Design and synthesis of novel insecticides based on the serotonergic ligand 1-[(4-aminophenyl)ethyl]-4-[3-(trifluoromethyl)phenyl]piperazine (PAPP).

Science.gov (United States)

Cai, Mingyi; Li, Zhong; Fan, Feng; Huang, Qingchun; Shao, Xusheng; Song, Gonghua

2010-03-10

1-[(4-Aminophenyl)ethyl]-4-[3-(trifluoromethyl)phenyl]piperazine (PAPP) is a 5-HT(1A) agonist and was reported to display high affinity for serotonin (5-HT) receptor from the parasitic nematode Haemonchus contortus . The present investigation explored the possibility of using PAPP as a lead compound of new insecticides with novel mode of action. On the basis of the PAPP scaffold, a series of 1-arylmethyl-4-[(trifluoromethyl)pyridin-2-yl]piperazine derivatives were designed, synthesized, and evaluated for biological activities against the armyworm Pseudaletia separata (Walker). Bioassays showed that most of the target compounds displayed certain growth-inhibiting activities or larvicidal activities against armyworm. The quantitative structure-activity relationship (QSAR) for growth-inhibiting activities was also analyzed and established.

Nuclear techniqes in the study of genetic resistance to gastrointestinal nematode infections of sheep

International Nuclear Information System (INIS)

Dargie, J.D.

1984-01-01

The paper reviews genetic resistance of sheep to gastrointestinal nematodes from the standpoint of resistance to the parasites themselves and of resistance to the diseases they produce. Attention is focused on infections with the abomasal parasite Haemonchus contortus and the small intestinal nematode Trichostrongylus colubriformis, and on the role of nuclear techniques both in verifying the existence of genetically based differences in resistance to these parasites and in gaining an understanding of the mechanisms involved. It is concluded that resistance to disease per se is much less important than resistance to parasite establishment and survival and that genetic studies could contribute substantially to the identification of the factors and variables responsible for the present inability to successfully vaccinate young animals against these infections. (author)

Physicochemical properties of the modeled structure of astacin metalloprotease moulting enzyme NAS-36 and mapping the druggable allosteric space of Heamonchus contortus, Brugia malayi and Ceanorhabditis elegans via molecular dynamics simulation.

Science.gov (United States)

Sharma, Om Prakash; Agrawal, Sonali; Kumar, M Suresh

2013-12-01

Nematodes represent the second largest phylum in the animal kingdom. It is the most abundant species (500,000) in the planet. It causes chronic, debilitating infections worldwide such as ascariasis, trichuriasis, hookworm, enterobiasis, strongyloidiasis, filariasis and trichinosis, among others. Molecular modeling tools can play an important role in the identification and structural investigation of molecular targets that can act as a vital candidate against filariasis. In this study, sequence analysis of NAS-36 from H. contortus (Heamonchus contortus), B. malayi (Brugia malayi) and C. elegans (Ceanorhabditis elegans) has been performed, in order to identify the conserved residues. Tertiary structure was developed for an insight into the molecular structure of the enzyme. Molecular Dynamics Simulation (MDS) studies have been carried out to analyze the stability and the physical properties of the proposed enzyme models in the H. contortus, B. malayi and C. elegans. Moreover, the drug binding sites have been mapped for inhibiting the function of NAS-36 enzyme. The molecular identity of this protease could eventually demonstrate how ex-sheathment is regulated, as well as provide a potential target of anthelmintics for the prevention of nematode infections.

Probes of the catalytic site of cysteine dioxygenase.

Science.gov (United States)

Chai, Sergio C; Bruyere, John R; Maroney, Michael J

2006-06-09

The first major step of cysteine catabolism, the oxidation of cysteine to cysteine sulfinic acid, is catalyzed by cysteine dioxygenase (CDO). In the present work, we utilize recombinant rat liver CDO and cysteine derivatives to elucidate structural parameters involved in substrate recognition and x-ray absorption spectroscopy to probe the interaction of the active site iron center with cysteine. Kinetic studies using cysteine structural analogs show that most are inhibitors and that a terminal functional group bearing a negative charge (e.g. a carboxylate) is required for binding. The substrate-binding site has no stringent restrictions with respect to the size of the amino acid. Lack of the amino or carboxyl groups at the alpha-carbon does not prevent the molecules from interacting with the active site. In fact, cysteamine is shown to be a potent activator of the enzyme without being a substrate. CDO was also rendered inactive upon complexation with the metal-binding inhibitors azide and cyanide. Unlike many non-heme iron dioxygenases that employ alpha-keto acids as cofactors, CDO was shown to be the only dioxygenase known to be inhibited by alpha-ketoglutarate.

Subcellular distribution of glutathione and cysteine in cyanobacteria.

Science.gov (United States)

Zechmann, Bernd; Tomasić, Ana; Horvat, Lucija; Fulgosi, Hrvoje

2010-10-01

Glutathione plays numerous important functions in eukaryotic and prokaryotic cells. Whereas it can be found in virtually all eukaryotic cells, its production in prokaryotes is restricted to cyanobacteria and proteobacteria and a few strains of gram-positive bacteria. In bacteria, it is involved in the protection against reactive oxygen species (ROS), osmotic shock, acidic conditions, toxic chemicals, and heavy metals. Glutathione synthesis in bacteria takes place in two steps out of cysteine, glutamate, and glycine. Cysteine is the limiting factor for glutathione biosynthesis which can be especially crucial for cyanobacteria, which rely on both the sufficient sulfur supply from the growth media and on the protection of glutathione against ROS that are produced during photosynthesis. In this study, we report a method that allows detection and visualization of the subcellular distribution of glutathione in Synechocystis sp. This method is based on immunogold cytochemistry with glutathione and cysteine antisera and computer-supported transmission electron microscopy. Labeling of glutathione and cysteine was restricted to the cytosol and interthylakoidal spaces. Glutathione and cysteine could not be detected in carboxysomes, cyanophycin granules, cell walls, intrathylakoidal spaces, periplasm, and vacuoles. The accuracy of the glutathione and cysteine labeling is supported by two observations. First, preadsorption of the antiglutathione and anticysteine antisera with glutathione and cysteine, respectively, reduced the density of the gold particles to background levels. Second, labeling of glutathione and cysteine was strongly decreased by 98.5% and 100%, respectively, in Synechocystis sp. cells grown on media without sulfur. This study indicates a strong similarity of the subcellular distribution of glutathione and cysteine in cyanobacteria and plastids of plants and provides a deeper insight into glutathione metabolism in bacteria.

A novel potentiometric biosensor for selective L-cysteine determination using L-cysteine-desulfhydrase producing Trichosporon jirovecii yeast cells coupled with sulfide electrode

International Nuclear Information System (INIS)

Hassan, Saad S.M.; El-Baz, Ashraf F.; Abd-Rabboh, Hisham S.M.

2007-01-01

Trichosporon jirovecii yeast cells are used for the first time as a source of L-cysteine desulfhydrase enzyme (EC 4.4.1.1) and incorporated in a biosensor for determining L-cysteine. The cells are grown under cadmium stress conditions to increase the expression level of the enzyme. The intact cells are immobilized on the membrane of a solid-state Ag 2 S electrode to provide a simple L-cysteine responsive biosensor. Upon immersion of the sensor in L-cysteine containing solutions, L-cysteine undergoes enzymatic hydrolysis into pyruvate, ammonia and sulfide ion. The rate of sulfide ion formation is potentiometrically measured as a function of L-cysteine concentration. Under optimized conditions (phosphate buffer pH 7, temperature 37 ± 1 deg. C and actual weight of immobilized yeast cells 100 mg), a linear relationship between L-cysteine concentration and the initial rate of sulfide liberation (dE/dt) is obtained. The sensor response covers the concentration range of 0.2-150 mg L -1 (1.7-1250 μmol L -1 ) L-cysteine. Validation of the assay method according to the quality control/quality assurance standards (precision, accuracy, between-day variability, within-day reproducibility, range of measurements and lower limit of detection) reveals remarkable performance characteristics of the proposed biosensor. The sensor is satisfactorily utilized for determination of L-cysteine in some pharmaceutical formulations. The lower limit of detection is ∼1 μmol L -1 and the accuracy and precision of the method are 97.5% and ±1.1%, respectively. Structurally similar sulfur containing compounds such as glutathione, cystine, methionine, and D-cysteine do no interfere

Protein cysteine oxidation in redox signaling

DEFF Research Database (Denmark)

Forman, Henry Jay; Davies, Michael J; Krämer, Anna C

2017-01-01

Oxidation of critical signaling protein cysteines regulated by H2O2 has been considered to involve sulfenic acid (RSOH) formation. RSOH may subsequently form either a sulfenyl amide (RSNHR') with a neighboring amide, or a mixed disulfide (RSSR') with another protein cysteine or glutathione. Previ...

Cysteine Biosynthesis Controls Serratia marcescens Phospholipase Activity.

Science.gov (United States)

Anderson, Mark T; Mitchell, Lindsay A; Mobley, Harry L T

2017-08-15

Serratia marcescens causes health care-associated opportunistic infections that can be difficult to treat due to a high incidence of antibiotic resistance. One of the many secreted proteins of S. marcescens is the PhlA phospholipase enzyme. Genes involved in the production and secretion of PhlA were identified by screening a transposon insertion library for phospholipase-deficient mutants on phosphatidylcholine-containing medium. Mutations were identified in four genes ( cyaA , crp , fliJ , and fliP ) that are involved in the flagellum-dependent PhlA secretion pathway. An additional phospholipase-deficient isolate harbored a transposon insertion in the cysE gene encoding a predicted serine O -acetyltransferase required for cysteine biosynthesis. The cysE requirement for extracellular phospholipase activity was confirmed using a fluorogenic phospholipase substrate. Phospholipase activity was restored to the cysE mutant by the addition of exogenous l-cysteine or O -acetylserine to the culture medium and by genetic complementation. Additionally, phlA transcript levels were decreased 6-fold in bacteria lacking cysE and were restored with added cysteine, indicating a role for cysteine-dependent transcriptional regulation of S. marcescens phospholipase activity. S. marcescens cysE mutants also exhibited a defect in swarming motility that was correlated with reduced levels of flhD and fliA flagellar regulator gene transcription. Together, these findings suggest a model in which cysteine is required for the regulation of both extracellular phospholipase activity and surface motility in S. marcescens IMPORTANCE Serratia marcescens is known to secrete multiple extracellular enzymes, but PhlA is unusual in that this protein is thought to be exported by the flagellar transport apparatus. In this study, we demonstrate that both extracellular phospholipase activity and flagellar function are dependent on the cysteine biosynthesis pathway. Furthermore, a disruption of cysteine

Pulse photolysis of NADH in the presence of cysteine

International Nuclear Information System (INIS)

Scheel, H.E.

1976-01-01

In the UV irradiation of NADH under anaerobic conditions, cysteine, which often acts as a radioprotective substance, has a sensitizing effect. With the aid of pulse photolysis, it was studied which reaction mechanisms in the presence or absence of cysteine are responsible for the damage to NADH in aqueous solution. In the absence of cysteine, the characteristic NADH absorption at 340 nm is reduced immediately after UV quanta have been absorbed by the adenine fraction of the molecules; in the presence of cysteine, a secondary reaction causes additional damage. The spectra of the intermediate products of NADH and cysteine have been recorded for different cysteine concentrations, and the reaction constants have been determined. These values suggest that the sensitizing effect is due to a reaction of NADH with radical anions produced by photolysis. (orig.) [de

Effect of (L)-cysteine on acetaldehyde self-administration.

Science.gov (United States)

Peana, Alessandra T; Muggironi, Giulia; Fois, Giulia R; Zinellu, Manuel; Sirca, Donatella; Diana, Marco

2012-08-01

Acetaldehyde (ACD), the first metabolite of ethanol, has been implicated in several behavioural actions of alcohol, including its reinforcing effects. Recently, we reported that l-cysteine, a sequestrating agent of ACD, reduced oral ethanol self-administration and that ACD was orally self-administered. This study examined the effects of l-cysteine pre-treatment during the acquisition and maintenance phases of ACD (0.2%) self-administration as well as on the deprivation effect after ACD extinction and on a progressive ratio (PR) schedule of reinforcement. In a separate PR schedule of reinforcement, the effect of l-cysteine was assessed on the break-point produced by ethanol (10%). Furthermore, we tested the effect of l-cysteine on saccharin (0.2%) reinforcement. Wistar rats were trained to self-administer ACD by nose poking on a fixed ratio (FR1) schedule in 30-min daily sessions. Responses on an active nose-poke caused delivery of ACD solution, whereas responses on an inactive nose-poke had no consequences. l-cysteine reduced the acquisition (40 mg/kg), the maintenance and the deprivation effect (100 mg/kg) of ACD self-administration. Furthermore, at the same dose, l-cysteine (120 mg/kg) decreased both ACD and ethanol break point. In addition, l-cysteine was unable to suppress the different responses for saccharin, suggesting that its effect did not relate to an unspecific decrease in a general motivational state. Compared to saline, l-cysteine did not modify responses on inactive nose-pokes, suggesting an absence of a non-specific behavioural activation. Taken together, these results could support the hypotheses that ACD possesses reinforcing properties and l-cysteine reduces motivation to self-administer ACD. Copyright © 2012 Elsevier Inc. All rights reserved.

CHARACTERIZATION OF DANSYLATED CYSTEINE, GLUTATHIONE DISULFIDE, CYSTEINE AND CYSTINE BY NARROW BORE LIQUID CHROMATOGRAPHY/ELECTROSPRAY IONIZATION MASS SPECTROMETRY

Science.gov (United States)

A method using reversed phase high performance liquid chromatography/electrospray ionization-mass spectrometric (RP-LC/ESI-MS) method has been developed to confirm the identity of dansylated derivatives of cysteine and glutathione, and their respective dimers. Cysteine, GSH, CSSC...

Ovine haemonchosis: Digestive pathophysiology and vaccination trials

International Nuclear Information System (INIS)

Dakkak, A.

1988-01-01

Infection with Haemonchus contortus induced an increase in abomasal permeability, as indicated by an increased sodium concentration of the abomasal contents and a hypochlorhydria as revealed by high pH values and low concentrations of K + and Cl - . Measurement of the abomasal transmural potential difference (PD) showed that the mucosal integrity was affected within 20 min after intraruminal administration of larvae. Electromyographic studies indicated a hypomotility of the rumen, reticulum and omasum, and a hypermotility of the abomasum and intestine. These disturbances were associated with an accelerated transit and an increased flow rate of digesta in the intestine. Irradiation of larvae (400 Gy) removed these effects, although there was a minor damage to the mucosal integrity, as detected by a slight increase in PD. In vaccinated sheep (two infections each of 10,000 gamma irradiated larvae given at an interval of 30 days) the adult worm populations resulting from challenge infections with H. contortus or Ostertagia circumcincta, given either separately or combined, were reduced by 86-97%, but the number of immature worms located in the mucosa was higher by 70-92%. In sheep exposed to natural multispecies infection, vaccination had a variable effect on faecal egg count. (author). 40 refs, 7 figs, 1 tab

Comparison of biotransformation and efficacy of aminoacetonitrile anthelmintics in vitro.

Science.gov (United States)

Stuchlíková, Lucie; Lecová, Lenka; Jirásko, Robert; Lamka, Jiří; Vokřál, Ivan; Szotáková, Barbora; Holčapek, Michal; Skálová, Lenka

2016-02-01

The present in vitro study was designed to test and compare anthelmintic activity, hepatotoxicity, and biotransformation of four selected aminoacetonitrile derivatives (AADs): monepantel (MOP, anthelmintic approved for the treatment), AAD-970, AAD-1154, and AAD-1336. Micro-agar larval development test, MTT test of cytotoxicity, and biotransformation study coupled with Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technique were used for this purpose. Larvae of two Haemonchus contortus strains (drug susceptible and multi-drug resistant) and primary cultures of rat and ovine hepatocytes served as model systems. All AADs (including MOP) exhibited significant larvicidal effect in H. contortus susceptible as well as multi-resistant strains, much higher than those of reference anthelmintics thiabendazole and flubendazole. AAD-1154 provides the best results for most tested parameters among all AADs in this study. The cytotoxicity test showed that all AADs can be considered as nontoxic for hepatocytes. In the biotransformation study, Phase I and Phase II metabolites of AADs were identified and schemes of possible metabolic pathways in ovine hepatocytes were proposed. Biotransformation of MOP was much more extensive than biotransformation of other AADs. Based on obtained results, AAD-1154 and AAD-1336 can be considered as promising candidates for further in vivo testing. Copyright © 2015 John Wiley & Sons, Ltd.

Efficacy of an ivermectin controlled-release capsule against nematode and arthropod endoparasites in sheep.

Science.gov (United States)

Rehbein, S; Batty, A F; Barth, D; Visser, M; Timms, B J; Barrick, R A; Eagleson, J S

1998-03-28

Five controlled trials were conducted in Germany or in the United Kingdom, using 74 female sheep of merino or Dorset horn breeds, to evaluate the efficacy of an ivermectin controlled-release capsule against naturally acquired or induced infections of gastrointestinal nematodes, lungworms and nasal bot larvae and against incoming infections with gastrointestinal and pulmonary nematodes. Half of the animals were treated with one ivermectin controlled-release capsule that delivered ivermectin at the rate of 1.6 mg per day for 100 days while the other half remained untreated. Parasites were counted 21, 28, 35 or 56 days after administration of the capsule. The treatment was highly effective (> or = 99 per cent) against established parasites of the following species: Haemonchus contortus (adults and fourth-stage larvae), Ostertagia circumcincta, O pinnata, O trifurcata, Ostertagia species fourth-stage larvae, Trichostrongylus axei, T colubriformis, T vitrinus, Cooperia curticei, Nematodirus battus, N filicollis, Strongyloides papillosus, Chabertia ovina, Oesophagostomum venulosum, Trichuris ovis, Tr skrjabini, Dictyocaulus filaria, Protostrongylus rufescens and Oestrus ovis (larvae). The treatment prevented the establishment of the gastrointestinal nematodes H contortus, O circumcincta, T axei, T colubriformis, C curticei, N battus, N filicollis, Ch ovina, Oe vennulosum and the establishment of the lungworm D filaria by > 99 per cent compared with untreated controls (P < or = 0.01).

Effect of feeding Mucuna pruriens on helminth parasite infestation in lambs.

Science.gov (United States)

Huisden, C M; Adesogan, A T; Gaskin, J M; Courtney, C H; Raji, A M; Kang, T

2010-02-17

Mucuna pruriens is a tropical legume anecdotally reputed to have anthelmintic properties. This study was conducted to examine the validity of such claims. The aim of this study was to determine if ingestion of Mucuna seeds reduces helminth parasite infestation in lambs. Thirty-six Dorper x Katahdin ram lambs were assigned to three treatments, a cottonseed meal based control diet, a diet in which Mucuna replaced cottonseed meal and the control diet with levamisole (7.5mg/kg body weight) administration. All diets were isonitrogenous and isocaloric. The 12 lambs in each treatment were assigned randomly to 4 pens, each containing 3 lambs. Lambs were trickle infected three times per week by gavage with infectious Haemonchus contortus larvae (2000 larvae/lamb) for 3 weeks. Levamisole treatment decreased fecal egg counts by 87% and abomasal worm counts by 83%. Mucuna intake did not statistically affect fecal egg counts or abomasal worm counts, though numerical (P>0.10) reductions of 7.4% and 18.1%, respectively were evident. Anemia indicators, feed intake, and lamb growth were unaffected by treatment. Levamisole reduced the Haemonchus parasite burden in lambs significantly but feeding Mucuna reduced the burden by levels unlikely to eliminate the clinical effects of parasitism. Copyright 2009 Elsevier Ireland Ltd. All rights reserved.

Epidemiologia das helmintoses gastrintestinais de ovinos no Planalto Catarinense

Directory of Open Access Journals (Sweden)

Ramos César Itaqui

2004-01-01

Full Text Available Este trabalho foi desenvolvido em três propriedades rurais nos municípios de Lages, São Joaquim e Campos Novos, estado de Santa Catarina, com os objetivos de determinar a prevalência, a intensidade e a variação sazonal de helmintos gastrintestinais e pulmonares em ovinos no Planalto Catarinense. Com base nos resultados aintenção é propor um esquema estratégico de controle. Para isso, foram utilizados mensalmente três cordeiros traçadores por propriedade, os quais, antes de serem conduzidos às mesmas, foram estabulados por 30 dias e executados tratamentos supressivos com anti-helmínticos de diferentes princípios ativos, com exames parasitológicos semanais para verificar a total eliminação de infecção parasitária. A seguir, foram encaminhados às três propriedades onde permaneceram em pastejo por 28 dias, sendo posteriormente recolhidos ao estábulo por mais 20 dias. Após foram sacrificados e realizada a coleta de alíquotas de 10% dos conteúdos do abomaso e intestino delgado, todos os helmintos do intestino grosso e pulmão. As maiores infecções por Haemonchus contortus ocorreram durante o período de outubro a março. O parasitismo por Trichostrongylus axei e Trichostrongylus colubriformis teve índices crescentes ao longo dos anos experimentais, apresentando maior pico de maio a outubro de 1999. Predominaram as espécies, no abomaso: Haemonchus contortus (100%; Trichostrongylus axei (98,7%; Trichostrongylus colubriformis (1,3%; Teladorsagia circumcincta (100%; Ostertagia ostertagi (100%. No intestino delgado: Trichostrongylus colubriformis (100%; Cooperia punctata (69,1%; Cooperia pectinata (18,4%; Cooperia curticei (6,9%; Cooperia oncophora (4,8%; Cooperia spatulata (0,8% e Nematodirus spathiger (100%. No intestino grosso: Oesophagostomum venulosum (100% e Trichuris ovis (100%. No pulmão, não foram encontrados parasitos.

Draft genome of neurotropic nematode parasite Angiostrongylus cantonensis, causative agent of human eosinophilic meningitis.

Science.gov (United States)

Yong, Hoi-Sen; Eamsobhana, Praphathip; Lim, Phaik-Eem; Razali, Rozaimi; Aziz, Farhanah Abdul; Rosli, Nurul Shielawati Mohamed; Poole-Johnson, Johan; Anwar, Arif

2015-08-01

Angiostrongylus cantonensis is a bursate nematode parasite that causes eosinophilic meningitis (or meningoencephalitis) in humans in many parts of the world. The genomic data from A. cantonensis will form a useful resource for comparative genomic and chemogenomic studies to aid the development of diagnostics and therapeutics. We have sequenced, assembled and annotated the genome of A. cantonensis. The genome size is estimated to be ∼260 Mb, with 17,280 genomic scaffolds, 91X coverage, 81.45% for complete and 93.95% for partial score based on CEGMA analysis of genome completeness. The number of predicted genes of ≥300 bp was 17,482. A total of 7737 predicted protein-coding genes of ≥50 amino acids were identified in the assembled genome. Among the proteins of known function, kinases are the most abundant followed by transferases. The draft genome contains 34 excretory-secretory proteins (ES), a minimum of 44 Nematode Astacin (NAS) metalloproteases, 12 Homeobox (HOX) genes, and 30 neurotransmitters. The assembled genome size (260 Mb) is larger than those of Pristionchus pacificus, Caenorhabditis elegans, Necator americanus, Caenorhabditis briggsae, Trichinella spiralis, Brugia malayi and Loa loa, but smaller than Haemonchus contortus and Ascaris suum. The repeat content (25%) is similar to H. contortus. The GC content (41.17%) is lower compared to P. pacificus (42.7%) and H. contortus (43.1%) but higher compared to C. briggsae (37.69%), A. suum (37.9%) and N. americanus (40.2%) while the scaffold N50 is 42,191. This draft genome will facilitate the understanding of many unresolved issues on the parasite and the disorder it causes. Copyright © 2015 Elsevier B.V. All rights reserved.

Incidence of Haemonchus spp. and effect on haematocrit and eye colour in goats farmed under resource-poor conditions in South Africa.

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Vatta, A F; Krecek, R C; Letty, B A; van der Linde, M J; Grimbeek, R J; de Villiers, J F; Motswatswe, P W; Molebiemang, G S; Boshoff, H M; Hansen, J W

2002-01-03

The diversity and predominance of nematode genera in goats of resource-poor farmers at Rust de Winter, Gauteng Province, Impendle, KwaZulu-Natal Province, and Kraaipan, North-West Province, South Africa, was determined by means of a longitudinal study of the nematode faecal egg counts (FECs) and differential third-stage nematode larvae. The animals were bled for haematocrit determination and scored for pallor of ocular mucous membranes using the FAMACHA( Copyright) method, an assay for clinical evaluation of anaemia caused by Haemonchus spp. Animals considered to be in danger of dying from anaemia caused by haemonchosis were selectively treated with an anthelmintic. Lower haematocrit values were registered during periods of heavier Haemonchus infection, which occurred from December/January to March for Rust de Winter; from December to March/April for Impendle; and from November/December to February or April for Kraaipan. There was agreement too between the lower haematocrits and paler mucous membranes scored according to the FAMACHA( Copyright) method. The use of this system may be recommended as part of an integrated approach to worm control in goats kept in the resource-poor areas studied.

Heterologous expression of Hordeum vulgare cysteine protease in yeast

DEFF Research Database (Denmark)

Rosenkilde, Anne Lind; Dionisio, Giuseppe; Holm, Preben B

Cysteine Proteases accounts for more than 90 % of the total proteolytic activity in the degradation of barley seed storage proteins during germination. Several Cysteine proteases have been identified in barley. One of the key enzymes, Hordeum vulgare endoprotease B2 (HvEPB2) was cloned with and w......Cysteine Proteases accounts for more than 90 % of the total proteolytic activity in the degradation of barley seed storage proteins during germination. Several Cysteine proteases have been identified in barley. One of the key enzymes, Hordeum vulgare endoprotease B2 (HvEPB2) was cloned...

Modulation of ion transport across rat distal colon by cysteine

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Martin eDiener

2012-03-01

Full Text Available The aim of this study was to identify the actions of stimulation of endogenous production of H2S by cysteine, the substrate for the two H2S-producing enzymes, cystathionin-beta-synthase and cystathionin-gamma-lyase, on ion transport across rat distal colon. Changes in short-circuit current (Isc induced by cysteine were measured in Ussing chambers. Free cysteine caused a concentration-dependent, transient fall in Isc, which was sensitive to amino-oxyacetate and beta-cyano-L-alanine, i.e. inhibitors of H2S-producing enzymes. In contrast, Na cysteinate evoked a biphasic change in Isc, i.e. an initial fall followed by a secondary increase, which was also reduced by these enzyme inhibitors. All responses were dependent on the presence of Cl- and inhibited by bumetanide, suggesting that free cysteine induces an inhibition of transcellular Cl- secretion, whereas Na cysteinate – after a transient inhibitory phase – activates anion secretion. The assumed reason for this discrepancy is a fall in the cytosolic pH induced by free cysteine, but not by Na cysteinate, as observed in isolated colonic crypts loaded with the pH-sensitive dye, BCECF. Intracellular acidification is known to inhibit epithelial K+ channels. Indeed, after preinhibition of basolateral K+ channels with tetrapentylammonium or Ba2+, the negative Isc induced by free cysteine was reduced significantly. In consequence, stimulation of endogenous H2S production by Na cysteinate causes, after a short inhibitory response, a delayed activation of anion secretion, which is missing in the case of free cysteine, probably due to the cytosolic acidification. In contrast, diallyl trisulfide, which is intracellularly converted to H2S, only evoked a monophasic increase in Isc without the initial fall observed with Na cysteinate. Consequently, time course and amount of produced H2S seem to strongly influence the functional response of the colonic epithelium evoked by this gasotransmitter.

Prevalence of Helminth Infections in Dairy Animals of Nestle Milk Collection Areas of Punjab (Pakistan

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M.K. Khan

2010-02-01

Full Text Available The objective of the current research project was to document the prevalent helminths of dairy animals of Nestle milk collection areas of Punjab (Pakistan. For this purpose, seven high milk-producing areas of Punjab province including Farooqa, Kot Adu, Dunya Pur, Layyah, Mor Mandi, Shorkot and Jalapur were selected. The animals were randomly selected and screened for parasitic eggs through standard coprological examination procedures. The helminth species found prevalent in the study areas included; Ascaris vitulorum, Fasciola hepatica, Haemonchus contortus, Bunostomum phlebotomum, Ostertagia circumcinta, Oesophagostomum radiatum, and Trichostrongylus spp. The possible determinants associated with the prevalence of these parasites were also studied in this project. The results of this study provided a basic epidemiological data for planning a wide scaled helminth control program in the above-mentioned high producing areas of Pakistan.

Synthesis, antioxidative and whitening effects of novel cysteine derivatives

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Ha, Ji Hoon; Kim, Kyoung Mi; Jeong, Yoon Ju; Park, Young Min; Lee, Jae Young; Park, Soo Nam [Dept. of Fine Chemistry, Cosmetic R and D Center, Cosmetic Industry Coupled Collaboration Center, Seoul National University of Science and Technology, Seoul (Korea, Republic of); Park, Jino [Daebong LS. Ltd, Incheon (Korea, Republic of)

2017-01-15

Recently, development of biocompatibility functional cosmetic agents as antioxidant or whitening agent has increased. In this study, synthetic cysteine derivatives (DBLS-21, -24, and -33) were developed containing syringic acid and cysteine moieties (l-cysteine ethyl ester, N-acetyl cysteine methyl ester, and N-acetyl cysteine ethyl ester), and their antioxidative and whitening activities were evaluated. The cellular protective effect (τ{sub 50}) of DBLS-21 was 51.1 min at 50 μM on {sup 1}O{sub 2} -induced hemolysis of erythrocytes. This activity was slightly higher than that of α-tocopherol (43.6 min) as a lipophilic antioxidant. In the melanogenesis inhibitory effect, DBLS-21, -24, and -33 was 1.6-, 1.8-, and 2.5-fold higher than arbutin, respectively. In particular, DBLS-21 and -33 was 112.8- and 6.1-fold higher than arbutin, respectively (293.4 μM) on tyrosinase inhibition activity (IC{sub 50} ). But DBLS-24 had no tyrosinase inhibitory activity. These results suggest that cysteine derivatives possess potential for use as an antioxidant agent (DBLS-21) and whitening agents (all derivatives) in cosmetics.

Synthesis, antioxidative and whitening effects of novel cysteine derivatives

International Nuclear Information System (INIS)

Ha, Ji Hoon; Kim, Kyoung Mi; Jeong, Yoon Ju; Park, Young Min; Lee, Jae Young; Park, Soo Nam; Park, Jino

2017-01-01

Recently, development of biocompatibility functional cosmetic agents as antioxidant or whitening agent has increased. In this study, synthetic cysteine derivatives (DBLS-21, -24, and -33) were developed containing syringic acid and cysteine moieties (l-cysteine ethyl ester, N-acetyl cysteine methyl ester, and N-acetyl cysteine ethyl ester), and their antioxidative and whitening activities were evaluated. The cellular protective effect (τ_5_0) of DBLS-21 was 51.1 min at 50 μM on "1O_2 -induced hemolysis of erythrocytes. This activity was slightly higher than that of α-tocopherol (43.6 min) as a lipophilic antioxidant. In the melanogenesis inhibitory effect, DBLS-21, -24, and -33 was 1.6-, 1.8-, and 2.5-fold higher than arbutin, respectively. In particular, DBLS-21 and -33 was 112.8- and 6.1-fold higher than arbutin, respectively (293.4 μM) on tyrosinase inhibition activity (IC_5_0 ). But DBLS-24 had no tyrosinase inhibitory activity. These results suggest that cysteine derivatives possess potential for use as an antioxidant agent (DBLS-21) and whitening agents (all derivatives) in cosmetics

Electrostatic influence of local cysteine environments on disulfide exchange kinetics.

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Snyder, G H; Cennerazzo, M J; Karalis, A J; Field, D

1981-11-10

The ionic strength dependence of the bimolecular rate constant for reaction of the negative disulfide 5,5'-dithiobis (2-nitrobenzoic acid) with cysteines in fragments of naturally occurring proteins was determined by stopped-flow spectroscopy. The Debye-Hückel relationship was applied to determine the effective charge at the cysteine and thereby determine the extent to which nearby neighbors in the primary sequence influence the kinetics. Corrections for the secondary salt effect on cysteine pKs were determined by direct spectrometric pH titration of sulfhydryl groups or by observation of the ionic strength dependence of kinetics of cysteine reaction with the neutral disulfide 2,2'-dithiodipyridine. Quantitative expressions was verified by model studies with N-acetyl-cystein. At ionic strengths equal to or greater than 20 mM, the net charge at the polypeptide cysteine site is the sum of the single negative charge of the thiolate anion and the charges of the amino acids immediately preceding and following the cysteine in the primary sequence. At lower ionic strengths, more distant residues influence kinetics. At pH 7.0, 23 degree C, and an ionic strength of 20 mM, rate constants for reaction of the negative disulfide with a cysteine having two positive neighbors, one positive and one neutral neighbor, or two neutral neighbors are 132000, 3350, and 367 s-1 M-1, respectively. This corresponds to a contribution to the activation energy of 0.65- 1.1 kcal/mol per ion pair involved in collision between the cysteine and disulfide regions. The results permit the estimation that cysteine local environments may provide a means of achieving a 10(6)-fold range in rate constants in disulfide exchange reactions in random-coil proteins. This range may prove useful in developing strategies for directing disulfide pairing in synthetic proteins.

Fluoresence quenching of riboflavin in aqueous solution by methionin and cystein

International Nuclear Information System (INIS)

Droessler, P.; Holzer, W.; Penzkofer, A.; Hegemann, P.

2003-01-01

The fluorescence quantum distributions, fluorescence quantum yields, and fluorescence lifetimes of riboflavin in methanol, DMSO, water, and aqueous solutions of the sulphur atom containing amino acids methionin and cystein have been determined. In methanol, DMSO, and water (pH=4-8) only dynamic fluorescence reduction due to intersystem crossing and internal conversion is observed. In aqueous methionin solutions of pH=5.25-9 a pH independent static and dynamic fluorescence quenching occurs probably due to riboflavin anion-methionin cation pair formation. In aqueous cystein solutions (pH range from 4.15 to 9) the fluorescence quenching increases with rising pH due to cystein thiolate formation. The cystein thiol form present at low pH does not react with neutral riboflavin. Cystein thiolate present at high pH seems to react with neutral riboflavin causing riboflavin deprotonation (anion formation) by cystein thiolate reduction to the cystein thiol form

Field efficacy of four anthelmintics and confirmation of drug-resistant nematodes by controlled efficacy test and pyrosequencing on a sheep and goat farm in Denmark.

Science.gov (United States)

Peña-Espinoza, Miguel; Thamsborg, Stig M; Demeler, Janina; Enemark, Heidi L

2014-12-15

We describe a case of anthelmintic resistance on one of the largest organic small ruminant farms in Denmark. The flock was established in 2007 by purchase of animals from other Danish farms and had history of clinical parasitism, high mortality of young stock and anthelmintic treatment failure. In October 2011, 40 lambs and 40 kids were selected for a faecal egg count reduction test (FECRT) with fenbendazole (FBZ), ivermectin (IVM), moxidectin (MOX) and levamisole (LEV). Lambs were treated with the recommended sheep dose of each product while kids received the sheep dose of IVM, 1.5× sheep dose of MOX and 2× sheep dose of FBZ and LEV. Untreated lambs and kids were also included and three methods for calculating faecal egg count (FEC) reduction were compared. In a subsequent investigation, a controlled efficacy test (CET) with FBZ and IVM was performed in lambs infected with Haemonchus contortus and Trichostrongylus colubriformis isolated from adult goats on the farm. Recovered specimens of H. contortus were subjected to pyrosequencing for detection of single nucleotide polymorphisms (SNPs) related to benzimidazole (BZ) resistance. During the FECRT, FECs in untreated lambs dropped significantly by 47%. No FEC reduction was detected in untreated kids. After FBZ treatments, FEC reductions in lambs and kids ranged from 15 to 54% and 49-56%, respectively, according to the different calculation methods. Post IVM treatments, FEC reductions in lambs and kids varied between 71-90% and 81-83%, correspondingly. LEV and MOX reduced FECs by 98-100% in both species. In the CET, FBZ reduced H. contortus worm counts by 52-56% and no reduction in T. colubriformis counts were detected after treatment. IVM eliminated 100% of H. contortus and reduced T. colubriformis counts by 84-92%, according to different calculation methods. Pyrosequencing of isolated H. contortus revealed increased frequencies of the BZ resistance-related SNP in codon 200 of the β-tubulin isotype 1 gene

Cysteine Supplementation May be Beneficial in a Subgroup of Mitochondrial Translation Deficiencies.

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Bartsakoulia, Marina; Mϋller, Juliane S; Gomez-Duran, Aurora; Yu-Wai-Man, Patrick; Boczonadi, Veronika; Horvath, Rita

2016-08-30

Mitochondrial encephalomyopathies are severe, relentlessly progressive conditions and there are very few effective therapies available to date. We have previously suggested that in two rare forms of reversible mitochondrial disease (reversible infantile respiratory chain deficiency and reversible infantile hepatopathy) supplementation with L-cysteine can improve mitochondrial protein synthesis, since cysteine is required for the 2-thiomodification of mitochondrial tRNAs. We studied whether supplementation with L-cysteine or N-acetyl-cysteine (NAC) results in any improvement of the mitochondrial function in vitro in fibroblasts of patients with different genetic forms of abnormal mitochondrial translation. We studied in vitro in fibroblasts of patients carrying the common m.3243A>G and m.8344A>G mutations or autosomal recessive mutations in genes affecting mitochondrial translation, whether L-cysteine or N-acetyl-cysteine supplementation have an effect on mitochondrial respiratory chain function. Here we show that supplementation with L-cysteine, but not with N-acetyl-cysteine partially rescues the mitochondrial translation defect in vitro in fibroblasts of patients carrying the m.3243A>G and m.8344A>G mutations. In contrast, N-acetyl-cysteine had a beneficial effect on mitochondrial translation in TRMU and MTO1 deficient fibroblasts. Our results suggest that L-cysteine or N-acetyl-cysteine supplementation may be a potential treatment for selected subgroups of patients with mitochondrial translation deficiencies. Further studies are needed to explore the full potential of cysteine supplementation as a treatment for patients with mitochondrial disease.

Barley (Hordeum vulgare L.) cysteine proteases: heterologous expression, purification and characterization

DEFF Research Database (Denmark)

Rosenkilde, Anne Lind; Dionisio, Giuseppe; Holm, Preben Bach

2011-01-01

During germination of barley seeds, mobilization of protein is essential and cysteine proteases accounts for more than 90 % of the total proteolytic activity in the degradation of barley seed storage proteins. Cysteine proteases exist as pro-enzyme and is activated through reduction of the active...... site cysteines and by removal of the pro-domain. The complement of cysteine proteases is comprehensive and for detailed studies of the individual components of this complement, a fast and efficient eukaryotic expression platform is highly desirable. A cDNA clone of the barley key cysteine endoprotease...

Cysteine homeostasis plays an essential role in plant immunity.

Science.gov (United States)

Álvarez, Consolación; Bermúdez, M Ángeles; Romero, Luis C; Gotor, Cecilia; García, Irene

2012-01-01

• Cysteine is the metabolic precursor of essential biomolecules such as vitamins, cofactors, antioxidants and many defense compounds. The last step of cysteine metabolism is catalysed by O-acetylserine(thiol)lyase (OASTL), which incorporates reduced sulfur into O-acetylserine to produce cysteine. In Arabidopsis thaliana, the main OASTL isoform OAS-A1 and the cytosolic desulfhydrase DES1, which degrades cysteine, contribute to the cytosolic cysteine homeostasis. • Meta-analysis of the transcriptomes of knockout plants for OAS-A1 and for DES1 show a high correlation with the biotic stress series in both cases. • The study of the response of knockout mutants to plant pathogens shows that des1 mutants behave as constitutive systemic acquired resistance mutants, with high resistance to biotrophic and necrotrophic pathogens, salicylic acid accumulation and WRKY54 and PR1 induction, while oas-a1 knockout mutants are more sensitive to biotrophic and necrotrophic pathogens. However, oas-a1 knockout mutants lack the hypersensitive response associated with the effector-triggered immunity elicited by Pseudomonas syringae pv. tomato DC3000 avrRpm1. • Our results highlight the role of cysteine as a crucial metabolite in the plant immune response. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

Influence du feu et du pâturage sur l'évolution de la phytomasse d'une savane à Heteropogon contortus de la région de Sakaraha (sud-ouest de Madagascar

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Rakotoarimanana, V.

2008-01-01

Full Text Available Effects of Fire and Grazing on Grass Phytomass Evolution in a Heteropogon contortus Savanna (Sakaraha Region, South West Madagascar. An experimental study of the effects of fire and grazing was carried out in the Sakaraha area (S. W. Madagascar. The objectives were to evaluate the combined or separate effects of fire and grazing on the green biomass and total phytomass of a Heteropogon contortus-Poupartia caffra dominated savanna. Three burning treatments (no burning, early burning and late burning and two grazing treatments (grazed, ungrazed were compared. A three-year study showed that (1 both burning treatments led to a delayed vegetation phenophase but a higher standing phytomass at the end of the growth cycle, (2 grazing treatment led to a significant decrease of the standing phytomass and of the contribution of Poaceae.

L-cysteine suppresses ghrelin and reduces appetite in rodents and humans.

Science.gov (United States)

McGavigan, A K; O'Hara, H C; Amin, A; Kinsey-Jones, J; Spreckley, E; Alamshah, A; Agahi, A; Banks, K; France, R; Hyberg, G; Wong, C; Bewick, G A; Gardiner, J V; Lehmann, A; Martin, N M; Ghatei, M A; Bloom, S R; Murphy, K G

2015-03-01

High-protein diets promote weight loss and subsequent weight maintenance, but are difficult to adhere to. The mechanisms by which protein exerts these effects remain unclear. However, the amino acids produced by protein digestion may have a role in driving protein-induced satiety. We tested the effects of a range of amino acids on food intake in rodents and identified l-cysteine as the most anorexigenic. Using rodents we further studied the effect of l-cysteine on food intake, behaviour and energy expenditure. We proceeded to investigate its effect on neuronal activation in the hypothalamus and brainstem before investigating its effect on gastric emptying and gut hormone release. The effect of l-cysteine on appetite scores and gut hormone release was then investigated in humans. l-Cysteine dose-dependently decreased food intake in both rats and mice following oral gavage and intraperitoneal administration. This effect did not appear to be secondary to behavioural or aversive side effects. l-Cysteine increased neuronal activation in the area postrema and delayed gastric emptying. It suppressed plasma acyl ghrelin levels and did not reduce food intake in transgenic ghrelin-overexpressing mice. Repeated l-cysteine administration decreased food intake in rats and obese mice. l-Cysteine reduced hunger and plasma acyl ghrelin levels in humans. Further work is required to determine the chronic effect of l-cysteine in rodents and humans on appetite and body weight, and whether l-cysteine contributes towards protein-induced satiety.

Cysteine proteases as potential antigens in antiparasitic DNA vaccines

DEFF Research Database (Denmark)

Jørgensen, Louise von Gersdorff; Buchmann, Kurt

2011-01-01

En litteraturgennemgang af muligheder for at bruge cystein proteaser som antigener i antiparasitære vacciner.......En litteraturgennemgang af muligheder for at bruge cystein proteaser som antigener i antiparasitære vacciner....

Immunological unresponsiveness of the neonatal ruminant to gastrointestinal helminths

International Nuclear Information System (INIS)

Soulsby, E.J.L.

1981-01-01

Parasitic gastro-enteritis of domestic ruminants is a disease syndrome which is most usually seen in young animals in their first grazing season. Although this may be due, in part, to greater susceptibility of young animals to the pathogenic effects of parasitic infection, there is also good evidence that young animals are less able to mount a satisfactory protective immune response or a response which will reject an existing infection. This phenomenon is exemplified by Haemonchus contortus and Trichostrongylus spp. infection in sheep, but the phenomenon is recognized in other species including neonatal rodents (e.g. rats infected with Nippostrongylus brasiliensis) and has been demonstrated in neonatal cattle infected with Taenia saginata. The present consideration will deal mainly with the failure of lambs to mount an effective immune response to gastrointestinal nematodes during the neonatal period. (author)

Elimination of hydrogen sulphide and β substitution in cystein, catalyzed by the cysteine-lyase of hens yolk-sac and yolk (1961)

International Nuclear Information System (INIS)

Chapeville, F.; Fromageot, P.

1961-01-01

The yolk of incubated hen's eggs contains a pyridoxal phosphate activated enzyme, free of iron, copper, magnesium and calcium. This enzyme activates the β-carbon atom of cysteine. Its reactivity is demonstrated by the ease with which this β-carbon fixes various sulfur containing substances in which the sulfur has reducing properties: inorganic sulfide, sulfide or cysteine itself. In the absence of substances able to react with the β-carbon atom, the active complex, consisting of the enzyme and the aminated tri-carbon chain, is hydrolysed to pyruvic acid and ammonia. The liberation of hydrogen sulfide thus appears to be the consequence either of the substitution of the β-carbon atom of cysteine or of the decomposition of the complex which this aminoacid forms with the enzyme studied. The latter seems therefore to possess an activity which differs from the activity of the desulfhydrases as yet known. We suggest to call this enzyme cystein-lyase. (authors) [fr

Acetaldehyde Removal from Indoor Air through Chemical Absorption Using L-Cysteine

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Miyuki Noguchi

2010-09-01

Full Text Available The irreversible removal of acetaldehyde from indoor air via a chemical reaction with amino acids was investigated. To compare effectiveness, five types of amino acid (glycine, L-lysine, L-methionine, L-cysteine, and L-cystine were used as the reactants. First, acetaldehyde-laden air was introduced into aqueous solutions of each amino acid and the removal abilities were compared. Among the five amino acids, L-cysteine solution showed much higher removal efficiency, while the other amino acids solutions didn’t show any significant differences from the removal efficiency of water used as a control. Next, as a test of the removal abilities of acetaldehyde by semi-solid L-cysteine, a gel containing L-cysteine solution was put in a fluororesin bag filled with acetaldehyde gas, and the change of acetaldehyde concentration was measured. The L-cysteine-containing gel removed 80% of the acetaldehyde in the air within 24 hours. The removal ability likely depended on the unique reaction whereby acetaldehyde and L-cysteine rapidly produce 2-methylthiazolidine-4-carboxylic acid. These results suggested that the reaction between acetaldehyde and L-cysteine has possibilities for irreversibly removing toxic acetaldehyde from indoor air.

Measuring site occupancy: a new perspective on cysteine oxidation.

Science.gov (United States)

Rogowska-Wrzesinska, Adelina; Wojdyla, Katarzyna; Williamson, James; Roepstorff, Peter

2014-10-01

Site occupancy is an extremely important aspect of quantification of protein modifications. Knowing the degree of modification of each oxidised cysteine residue is critical to understanding the biological role of these modifications. Yet modification site occupancy is very often overlooked, in part because there are very few analytical tools that allow such measurements. Here we present a new strategy, which provides quantitative analysis of cysteine S-nitrosylation (SNO) and S-sulfenylation (SOH) simultaneously at the resolution of single cysteine and allows for determination of relative oxidation occupancy of the modification site. We show that, on one hand, heavily modified cysteines are not necessarily involved in the response to oxidative stress. On the other hand residues with low modification level can be dramatically affected by mild oxidative imbalance. We make use of high resolution mass spectrometry. The method relies on differential reduction of "total" cysteines, SNO cysteines and SOH cysteines with TCEP, sodium ascorbate and sodium arsenite respectively followed by iodoTMT(TM) alkylation. Enrichment of iodoTMT(TM)-containing peptides is performed using anti-TMT antibody. In vivo model of mild oxidative stress in Escherichia coli is used. To induce endogenous SNO bacteria were grown anaerobically in minimal media supplemented with fumarate or nitrate. Short-term treatment with submilimolar levels of hydrogen peroxide were used to induce SOH. We have quantified 114 SNO/SOH modified peptides corresponding to 90 proteins. Only 6 modified peptides changed significantly under mild oxidative stress. Quantitative information allowed us to determine relative modification site occupancy of each identified modified residue and pin point heavily modified ones. The method proved to be precise and sensitive enough to detect and quantify endogenous levels of oxidative stress on proteome-wide scale and brings a new perspective on the role of the modification site

Emission of hydrogen sulfide by leaf tissue in response to L-cysteine

International Nuclear Information System (INIS)

Sekiya, J.; Schmidt, A.; Wilson, L.G.; Filner, P.

1982-01-01

Leaf discs and detached leaves exposed to L-cysteine emitted a volatile sulfur compound which was proven by gas chromatography to be H 2 S. This phenomenon was demonstrated in all nine species tested (Cucumis sativus, Cucurbita pepo, Nicotiana tabacum, Coleus blumei, Beta vulgaris, Phaseolus vulgaris, Medicago sativa, Hordeum vulgare, and Gossypium hirsutum). The emission of volatile sulfur by cucumber leaves occurred in the dark at a similar rate to that in the light. The emission of leaf discs reached the maximal rate, more than 40 picomoles per minute per square centimeter, 2 to 4 hours after starting exposure to L-cysteine; then it decreased. In the case of detached leaves, the maximum occurred 5 to 10 h after starting exposure. The average emission rate of H 2 S during the first 4 hours from leaf discs of cucurbits in response to 10 millimolar L-cysteine, was usually more than 40 picomoles per minute per square centimeter, i.e. 0.24 micromoles per hour per square decimeter. Leaf discs exposed to 1 millimolar L-cysteine emitted only 2% as much as did the discs exposed to 10 millimolar L-cysteine. The emission from leaf discs and from detached leaves lasted for at least 5 and 15 hours, respectively. However, several hours after the maximal emission, injury of the leaves, manifested as chlorosis, was evident. H 2 S emission was a specific consequence of exposure to L-cysteine; neither D-cysteine nor L-cysteine elicited H 2 S emission. Aminooxyacetic acid, an inhibitor of pyridoxal phosphate dependent enzymes, inhibited the emission. In a cell free system from cucumber leaves, H 2 S formation and its release occurred in response to L-cysteine. Feeding experiments with [ 35 S]t-cysteine showed that most of the sulfur in H 2 S was derived from sulfur in the L-cysteine supplied

Lysosomal cysteine peptidases - Molecules signaling tumor cell death and survival.

Science.gov (United States)

Pišlar, Anja; Perišić Nanut, Milica; Kos, Janko

2015-12-01

Lysosomal cysteine peptidases - cysteine cathepsins - are general intracellular protein-degrading enzymes that control also a variety of specific physiological processes. They can trigger irreversible events leading to signal transduction and activation of signaling pathways, resulting in cell survival and proliferation or cell death. In cancer cells, lysosomal cysteine peptidases are involved in multiple processes during malignant progression. Their translocation from the endosomal/lysosomal pathway to nucleus, cytoplasm, plasma membrane and extracellular space enables the activation and remodeling of a variety of tumor promoting proteins. Thus, lysosomal cysteine peptidases interfere with cytokine/chemokine signaling, regulate cell adhesion and migration and endocytosis, are involved in the antitumor immune response and apoptosis, and promote cell invasion, angiogenesis and metastasis. Further, lysosomal cysteine peptidases modify growth factors and receptors involved in tyrosine kinase dependent pathways such as MAPK, Akt and JNK, thus representing key signaling tools for the activation of tumor cell growth and proliferation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Electrostatics of cysteine residues in proteins: Parameterization and validation of a simple model

Science.gov (United States)

Salsbury, Freddie R.; Poole, Leslie B.; Fetrow, Jacquelyn S.

2013-01-01

One of the most popular and simple models for the calculation of pKas from a protein structure is the semi-macroscopic electrostatic model MEAD. This model requires empirical parameters for each residue to calculate pKas. Analysis of current, widely used empirical parameters for cysteine residues showed that they did not reproduce expected cysteine pKas; thus, we set out to identify parameters consistent with the CHARMM27 force field that capture both the behavior of typical cysteines in proteins and the behavior of cysteines which have perturbed pKas. The new parameters were validated in three ways: (1) calculation across a large set of typical cysteines in proteins (where the calculations are expected to reproduce expected ensemble behavior); (2) calculation across a set of perturbed cysteines in proteins (where the calculations are expected to reproduce the shifted ensemble behavior); and (3) comparison to experimentally determined pKa values (where the calculation should reproduce the pKa within experimental error). Both the general behavior of cysteines in proteins and the perturbed pKa in some proteins can be predicted reasonably well using the newly determined empirical parameters within the MEAD model for protein electrostatics. This study provides the first general analysis of the electrostatics of cysteines in proteins, with specific attention paid to capturing both the behavior of typical cysteines in a protein and the behavior of cysteines whose pKa should be shifted, and validation of force field parameters for cysteine residues. PMID:22777874

Protein modification by acrolein: Formation and stability of cysteine adducts

OpenAIRE

Cai, Jian; Bhatnagar, Aruni; Pierce, William M.

2009-01-01

The toxicity of the ubiquitous pollutant and endogenous metabolite, acrolein, is due in part to covalent protein modifications. Acrolein reacts readily with protein nucleophiles via Michael addition and Schiff base formation. Potential acrolein targets in protein include the nucleophilic side chains of cysteine, histidine, and lysine residues as well as the free amino terminus of proteins. Although cysteine is the most acrolein-reactive residue, cysteine-acrolein adducts are difficult to iden...

Targeting cysteine proteases in trypanosomatid disease drug discovery.

Science.gov (United States)

Ferreira, Leonardo G; Andricopulo, Adriano D

2017-12-01

Chagas disease and human African trypanosomiasis are endemic conditions in Latin America and Africa, respectively, for which no effective and safe therapy is available. Efforts in drug discovery have focused on several enzymes from these protozoans, among which cysteine proteases have been validated as molecular targets for pharmacological intervention. These enzymes are expressed during the entire life cycle of trypanosomatid parasites and are essential to many biological processes, including infectivity to the human host. As a result of advances in the knowledge of the structural aspects of cysteine proteases and their role in disease physiopathology, inhibition of these enzymes by small molecules has been demonstrated to be a worthwhile approach to trypanosomatid drug research. This review provides an update on drug discovery strategies targeting the cysteine peptidases cruzain from Trypanosoma cruzi and rhodesain and cathepsin B from Trypanosoma brucei. Given that current chemotherapy for Chagas disease and human African trypanosomiasis has several drawbacks, cysteine proteases will continue to be actively pursued as valuable molecular targets in trypanosomatid disease drug discovery efforts. Copyright © 2017. Published by Elsevier Inc.

Chiral supramolecular gold-cysteine nanoparticles: Chiroptical and nonlinear optical properties

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Isabelle Russier-Antoine

2016-10-01

Full Text Available Cysteine is a sulfur-containing amino acid that easily coordinates to soft metal ions and grafts to noble metal surfaces. We report a simple synthetic approach for the production of chiral gold-cysteine polymeric nanoparticles soluble in water. Conjugation of cysteine with gold in a polymeric way, leading to ~50 nm diameter nanoparticles, resulted in the generation of new characteristic circular dichroism (CD signals in the region of 250–400 nm, whereas no CD signal changes were found with cysteine alone. We also investigate their nonlinear optical properties after two-photon absorption. Two-photon emission spectra and first hyper-polarizabilities, as obtained by the hyper-Rayleigh scattering technique, of these particles are presented.

Cysteine peroxidase activity in rat blood plasma | Razygraev ...

African Journals Online (AJOL)

The rat plasma found to be able to accelerate greatly the H2O2-dependent oxidation of cysteine. The activity was a characteristic of a protein fraction precipitated at 30—44% ammonium sulfate saturation, and the specific activity in protein fraction was significantly higher than in plasma. Cysteine:H2O2 oxidoreductase ...

Characterization of an acetylcholine receptor gene of Haemonchus contortus in relation to levamisole resistance

NARCIS (Netherlands)

Hoekstra, R.; Visser, A.; Wiley, L. J.; Weiss, A. S.; Sangster, N. C.; Roos, M. H.

1997-01-01

The anthelminitic drug levamisole is thought to bind to nicotinic acetylcholine receptors of nematodes. It is possible that resistance to this drug is associated with either a change in binding characteristics or a reduction in the number of nicotinic acetylcholine receptors. Therefore, the

Characterisation of an acetylcholine receptor gene of Haemonchus contortus in relation to levamisole resistance

NARCIS (Netherlands)

Hoekstra, R.; Visser, A.; Wiley, L.; Weiss, A.S.; Sangster, N.C.; Roos, M.H.

1997-01-01

The anthelmintic drug levamisole is thought to bind to nicotinic acetylcholine receptors of nematodes. It is possible that resistance to this drug is associated with either a change in binding characteristics or a reduction in the number of nicotinic acetylcholine receptors. Therefore, the molecular

Synergistic inhibition of Haemonchus contortus exsheathment by flavonoid monomers and condensed tannins

DEFF Research Database (Denmark)

Klongsiriwet, Chaweewan; Quijada, Jessica; Williams, Andrew Richard

2015-01-01

This study investigated the separate and combined anthelmintic (AH) effects of different phenolic compounds, including condensed tannins and flavonoids, all of which are known to occur in willow leaves, a potentially valuable dry season feed. A range of contrasting model tannins, which span the w...

Potential Test of Irradiated Vaccine for Haemonchus Contortus Worm and Food Supplement on Sheep

International Nuclear Information System (INIS)

Sukardji Partodiharjo; Arifin, M.; Endang Yuliawati; Enuh Rahardjo

2004-01-01

An experiment of vaccines was carried out is one of Nematoda worm present in abomasums or stomach of sheep or goat. The case in the field was high enough, may teach 60% the aim of this study is on observe the effect of gastric worm vaccination irradiated 5000 L 3 post challenge wild strain 5000 L 3 , The parameter of observe were clinic, blood twice description number of adult worms, with 3 treatments as follows; control (K), V 2 = twice vaccinations without challenge V 1 twice vaccinations with challenge. The result of the study the average of gain (gram); V 1 =97, V 2 =91 and K=31.20 (P 6 ), V 1 =6.50, V 2 =5.90 and K=6.10 (P 1 =29.30, V 2 =35.30 and K=27.50 (P 3 ), V 1 =9.30, V 2 =9.00 and K=7.40 (P 1 =8.30, V 2 =9.50 and K=6.20 (P 1 immunity respond of weight gain, erythrocyte and leucocyte to have immunity which is better than the respond of other treatments. (author)

Facile and green synthesis of highly stable L-cysteine functionalized copper nanoparticles

International Nuclear Information System (INIS)

Kumar, Nikhil; Upadhyay, Lata Sheo Bachan

2016-01-01

Highlights: • A facile and eco-friendly method for the synthesis of L-cysteine functionalized copper nanoparticles is reported. • Synthesis of Highly stable L-cysteine functionalized copper nanoparticles (∼40 nm) was done in an aqueous medium. • FTIR analysis shows that L-cysteine bound to the nanoparticle surface via thiol group. - Abstract: A simple eco-friendly method for L-cysteine capped copper nanoparticles (CCNPs) synthesis in aqueous solution has been developed. Glucose and L-cysteine were used as reducing agent and capping/functionalizing agent, respectively. Different parameters such as capping agent concentration, pH, reaction temperature, and reducing agent concentration were optimized during the synthesis. The L-cysteine capped copper nanoparticle were characterized by ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, Particle size and zeta potential analyser, and high resolution transmission electron microscopy. Spherical shaped cysteine functionalized/capped copper nanoparticles with an average size of 40 nm were found to be highly stable at room temperature (RT) for a period of 1 month

Facile and green synthesis of highly stable L-cysteine functionalized copper nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Kumar, Nikhil, E-mail: nkumar.phd2011.bt@nitrr.ac.in; Upadhyay, Lata Sheo Bachan, E-mail: contactlataupadhyay@gmail.com

2016-11-01

Highlights: • A facile and eco-friendly method for the synthesis of L-cysteine functionalized copper nanoparticles is reported. • Synthesis of Highly stable L-cysteine functionalized copper nanoparticles (∼40 nm) was done in an aqueous medium. • FTIR analysis shows that L-cysteine bound to the nanoparticle surface via thiol group. - Abstract: A simple eco-friendly method for L-cysteine capped copper nanoparticles (CCNPs) synthesis in aqueous solution has been developed. Glucose and L-cysteine were used as reducing agent and capping/functionalizing agent, respectively. Different parameters such as capping agent concentration, pH, reaction temperature, and reducing agent concentration were optimized during the synthesis. The L-cysteine capped copper nanoparticle were characterized by ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, Particle size and zeta potential analyser, and high resolution transmission electron microscopy. Spherical shaped cysteine functionalized/capped copper nanoparticles with an average size of 40 nm were found to be highly stable at room temperature (RT) for a period of 1 month.

Overexpression of Catalase Diminishes Oxidative Cysteine Modifications of Cardiac Proteins.

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Chunxiang Yao

Full Text Available Reactive protein cysteine thiolates are instrumental in redox regulation. Oxidants, such as hydrogen peroxide (H2O2, react with thiolates to form oxidative post-translational modifications, enabling physiological redox signaling. Cardiac disease and aging are associated with oxidative stress which can impair redox signaling by altering essential cysteine thiolates. We previously found that cardiac-specific overexpression of catalase (Cat, an enzyme that detoxifies excess H2O2, protected from oxidative stress and delayed cardiac aging in mice. Using redox proteomics and systems biology, we sought to identify the cysteines that could play a key role in cardiac disease and aging. With a 'Tandem Mass Tag' (TMT labeling strategy and mass spectrometry, we investigated differential reversible cysteine oxidation in the cardiac proteome of wild type and Cat transgenic (Tg mice. Reversible cysteine oxidation was measured as thiol occupancy, the ratio of total available versus reversibly oxidized cysteine thiols. Catalase overexpression globally decreased thiol occupancy by ≥1.3 fold in 82 proteins, including numerous mitochondrial and contractile proteins. Systems biology analysis assigned the majority of proteins with differentially modified thiols in Cat Tg mice to pathways of aging and cardiac disease, including cellular stress response, proteostasis, and apoptosis. In addition, Cat Tg mice exhibited diminished protein glutathione adducts and decreased H2O2 production from mitochondrial complex I and II, suggesting improved function of cardiac mitochondria. In conclusion, our data suggest that catalase may alleviate cardiac disease and aging by moderating global protein cysteine thiol oxidation.

Electrostatics of cysteine residues in proteins: parameterization and validation of a simple model.

Science.gov (United States)

Salsbury, Freddie R; Poole, Leslie B; Fetrow, Jacquelyn S

2012-11-01

One of the most popular and simple models for the calculation of pK(a) s from a protein structure is the semi-macroscopic electrostatic model MEAD. This model requires empirical parameters for each residue to calculate pK(a) s. Analysis of current, widely used empirical parameters for cysteine residues showed that they did not reproduce expected cysteine pK(a) s; thus, we set out to identify parameters consistent with the CHARMM27 force field that capture both the behavior of typical cysteines in proteins and the behavior of cysteines which have perturbed pK(a) s. The new parameters were validated in three ways: (1) calculation across a large set of typical cysteines in proteins (where the calculations are expected to reproduce expected ensemble behavior); (2) calculation across a set of perturbed cysteines in proteins (where the calculations are expected to reproduce the shifted ensemble behavior); and (3) comparison to experimentally determined pK(a) values (where the calculation should reproduce the pK(a) within experimental error). Both the general behavior of cysteines in proteins and the perturbed pK(a) in some proteins can be predicted reasonably well using the newly determined empirical parameters within the MEAD model for protein electrostatics. This study provides the first general analysis of the electrostatics of cysteines in proteins, with specific attention paid to capturing both the behavior of typical cysteines in a protein and the behavior of cysteines whose pK(a) should be shifted, and validation of force field parameters for cysteine residues. Copyright © 2012 Wiley Periodicals, Inc.

Knockout of the murine cysteine dioxygenase gene results in severe impairment in ability to synthesize taurine and an increased catabolism of cysteine to hydrogen sulfide

Science.gov (United States)

Ueki, Iori; Roman, Heather B.; Valli, Alessandro; Fieselmann, Krista; Lam, Jimmy; Peters, Rachel; Hirschberger, Lawrence L.

2011-01-01

Cysteine homeostasis is dependent on the regulation of cysteine dioxygenase (CDO) in response to changes in sulfur amino acid intake. CDO oxidizes cysteine to cysteinesulfinate, which is further metabolized to either taurine or to pyruvate plus sulfate. To gain insight into the physiological function of CDO and the consequence of a loss of CDO activity, mice carrying a null CDO allele (CDO+/− mice) were crossed to generate CDO−/−, CDO+/−, and CDO+/+ mice. CDO−/− mice exhibited postnatal mortality, growth deficit, and connective tissue pathology. CDO−/− mice had extremely low taurine levels and somewhat elevated cysteine levels, consistent with the lack of flux through CDO-dependent catabolic pathways. However, plasma sulfate levels were slightly higher in CDO−/− mice than in CDO+/− or CDO+/+ mice, and tissue levels of acid-labile sulfide were elevated, indicating an increase in cysteine catabolism by cysteine desulfhydration pathways. Null mice had lower hepatic cytochrome c oxidase levels, suggesting impaired electron transport capacity. Supplementation of mice with taurine improved survival of male pups but otherwise had little effect on the phenotype of the CDO−/− mice. H2S has been identified as an important gaseous signaling molecule as well as a toxicant, and pathology may be due to dysregulation of H2S production. Control of cysteine levels by regulation of CDO may be necessary to maintain low H2S/sulfane sulfur levels and facilitate the use of H2S as a signaling molecule. PMID:21693692

Biotin Switch Assays for Quantitation of Reversible Cysteine Oxidation.

Science.gov (United States)

Li, R; Kast, J

2017-01-01

Thiol groups in protein cysteine residues can be subjected to different oxidative modifications by reactive oxygen/nitrogen species. Reversible cysteine oxidation, including S-nitrosylation, S-sulfenylation, S-glutathionylation, and disulfide formation, modulate multiple biological functions, such as enzyme catalysis, antioxidant, and other signaling pathways. However, the biological relevance of reversible cysteine oxidation is typically underestimated, in part due to the low abundance and high reactivity of some of these modifications, and the lack of methods to enrich and quantify them. To facilitate future research efforts, this chapter describes detailed procedures to target the different modifications using mass spectrometry-based biotin switch assays. By switching the modification of interest to a biotin moiety, these assays leverage the high affinity between biotin and avidin to enrich the modification. The use of stable isotope labeling and a range of selective reducing agents facilitate the quantitation of individual as well as total reversible cysteine oxidation. The biotin switch assay has been widely applied to the quantitative analysis of S-nitrosylation in different disease models and is now also emerging as a valuable research tool for other oxidative cysteine modifications, highlighting its relevance as a versatile, robust strategy for carrying out in-depth studies in redox proteomics. © 2017 Elsevier Inc. All rights reserved.

Evaluation of the Effects of S-Allyl-L-cysteine, S-Methyl-L-cysteine, trans-S-1-Propenyl-L-cysteine, and Their N-Acetylated and S-Oxidized Metabolites on Human CYP Activities.

Science.gov (United States)

Amano, Hirotaka; Kazamori, Daichi; Itoh, Kenji

2016-01-01

Three major organosulfur compounds of aged garlic extract, S-allyl-L-cysteine (SAC), S-methyl-L-cysteine (SMC), and trans-S-1-propenyl-L-cysteine (S1PC), were examined for their effects on the activities of five major isoforms of human CYP enzymes: CYP1A2, 2C9, 2C19, 2D6, and 3A4. The metabolite formation from probe substrates for the CYP isoforms was examined in human liver microsomes in the presence of organosulfur compounds at 0.01-1 mM by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Allicin, a major component of garlic, inhibited CYP1A2 and CYP3A4 activity by 21-45% at 0.03 mM. In contrast, a CYP2C9-catalyzed reaction was enhanced by up to 1.9 times in the presence of allicin at 0.003-0.3 mM. SAC, SMC, and S1PC had no effect on the activities of the five isoforms, except that S1PC inhibited CYP3A4-catalyzed midazolam 1'-hydroxylation by 31% at 1 mM. The N-acetylated metabolites of the three compounds inhibited the activities of several isoforms to a varying degree at 1 mM. N-Acetyl-S-allyl-L-cysteine and N-acetyl-S-methyl-L-cysteine inhibited the reactions catalyzed by CYP2D6 and CYP1A2, by 19 and 26%, respectively, whereas trans-N-acetyl-S-1-propenyl-L-cysteine showed weak to moderate inhibition (19-49%) of CYP1A2, 2C19, 2D6, and 3A4 activities. On the other hand, both the N-acetylated and S-oxidized metabolites of SAC, SMC, and S1PC had little effect on the reactions catalyzed by the five isoforms. These results indicated that SAC, SMC, and S1PC have little potential to cause drug-drug interaction due to CYP inhibition or activation in vivo, as judged by their minimal effects (IC 50 >1 mM) on the activities of five major isoforms of human CYP in vitro.

Negative modulation of the GABAA ρ1 receptor function by l-cysteine.

Science.gov (United States)

Beltrán González, Andrea N; Vicentini, Florencia; Calvo, Daniel J

2018-01-01

l-Cysteine is an endogenous sulfur-containing amino acid with multiple and varied roles in the central nervous system, including neuroprotection and the maintenance of the redox balance. However, it was also suggested as an excitotoxic agent implicated in the pathogenesis of neurological disorders such as Parkinson's and Alzheimer's disease. l-Cysteine can modulate the activity of ionic channels, including voltage-gated calcium channels and glutamatergic NMDA receptors, whereas its effects on GABAergic neurotransmission had not been studied before. In the present work, we analyzed the effects of l-cysteine on responses mediated by homomeric GABA A ρ1 receptors, which are known for mediating tonic γ-aminobutyric acid (GABA) responses in retinal neurons. GABA A ρ1 receptors were expressed in Xenopus laevis oocytes and GABA-evoked chloride currents recorded by two-electrode voltage-clamp in the presence or absence of l-cysteine. l-Cysteine antagonized GABA A ρ1 receptor-mediated responses; inhibition was dose-dependent, reversible, voltage independent, and susceptible to GABA concentration. Concentration-response curves for GABA were shifted to the right in the presence of l-cysteine without a substantial change in the maximal response. l-Cysteine inhibition was insensitive to chemical protection of the sulfhydryl groups of the ρ1 subunits by the irreversible alkylating agent N-ethyl maleimide. Our results suggest that redox modulation is not involved during l-cysteine actions and that l-cysteine might be acting as a competitive antagonist of the GABA A ρ1 receptors. © 2017 International Society for Neurochemistry.

Preparation and application of L-cysteine-doped Keggin polyoxometalate microtubes

International Nuclear Information System (INIS)

Shen Yan; Peng Jun; Zhang Huanqiu; Meng Cuili; Zhang Fang

2012-01-01

L-cysteine-doped tungstosilicate (Lcys-SiW 12 ) microtubes are prepared, and the amount of L-cysteine doped in the microtubes can be tuned to some extent. The as-prepared Lcys-SiW 12 microtubes are sensitive to ammonia gas exhibited through the distinct color change of the microtubes from light purple to dark blue after exposing to ammonia gas. A possible mechanism of the coloration is that the adsorbed ammonia molecules increase the basicity of the Lcys-SiW 12 microtubes and promote the redox reaction between L-cysteine and polyoxometalate. This is a pH-dependent solid–solid redox reaction, which is triggered by proton capture agent. The Lcys-SiW 12 microtubes show application in chemical sensors for alkaline gases. - Graphical abstract: The Lcys-SiW 12 microtubes were formed during transformation of the monolacunary Keggin-type [α-SiW 11 O 39 ] 8− to the saturated Keggin-type [α-SiW 12 O 40 ] 4− , meanwhile L-cysteine molecules were doped during the growth of the microtubes. Highlights: ► L-cysteine-doped polyoxometalate microtubes are prepared. ► Amount of L-cysteine doped in the microtubes can be tuned to some extent. ► Lcys-SiW 12 microtubes can be applied as a sensor for detecting alkaline gases. ► This is a proton capture agent-triggered solid–solid redox reaction.

Gastrointestinal parasites of sheep, municipality of Lajes, Rio Grande do Norte, Brazil Parasitos do trato gastrintestinal de ovinos, município de Lajes, Rio Grande do Norte, Brasil

Directory of Open Access Journals (Sweden)

Maria de Fátima de Souza

2012-03-01

Full Text Available This study investigated the gastrointestinal parasitism by helminths and protozoa in sheep (Ovis aries Santa Inês breed, municipality of Lajes, Rio Grande do Norte. Monthly, from April 2005 to August 2007, stool samples were collected from two tracer lambs in the first day of the experiment and performed a necropsy of these animals in 44th day. A total of 64 lambs were sampled, but only 62 lambs were slaughtered. The fecal samples were examined by sedimentation in water. The contents of the abomasum, small intestine and large intestine were examined for the recovery of helminths. The parasitological examination revealed eggs of the following groups of helminths: Strongyloidea, Strongyloides sp., Trichuris sp., and Moniezia sp. Also were found oocysts of Eimeria spp., cysts of Entamoeba ovis and Giardia duodenalis. The helminths identified from examining the contents were: Haemonchus contortus, Cooperia pectinata, Cooperia punctata, Trichostrongylus colubriformis, Moniezia expansa, Oesophagostomum sp. Skrjabinema ovis and Trichuris sp.O presente estudo investigou o parasitismo gastrintestinal por helmintos e protozoários em ovinos (Ovis aries da raça Santa Inês, no município de Lajes, Rio Grande do Norte. Mensalmente, entre abril de 2005 e agosto de 2007, foram coletadas amostras fecais de dois cordeiros traçadores no primeiro dia do experimento e realizada a necropsia desses animais no 44º dia. O total de cordeiros amostrados foi 64, mas apenas 62 foram necropsiados. As amostras fecais foram examinadas pela técnica de sedimentação espontânea em água. Os conteúdos do abomaso, intestino delgado e intestino grosso dos cordeiros necropsiados foram examinados para a recuperação dos helmintos. Os exames parasitológicos evidenciaram ovos dos seguintes grupos de helmintos: Strongyloidea, Strongyloides sp. , Trichuris sp., e Moniezia sp. Também foram encontrados oocistos de Eimeria spp., cistos de Entamoeba ovis e de Giardia duodenalis

Screening of the ‘Open Scaffolds’ collection from Compounds Australia identifies a new chemical entity with anthelmintic activities against different developmental stages of the barber's pole worm and other parasitic nematodes

Directory of Open Access Journals (Sweden)

Sarah Preston

2017-12-01

Full Text Available The discovery and development of novel anthelmintic classes is essential to sustain the control of socioeconomically important parasitic worms of humans and animals. With the aim of offering novel, lead-like scaffolds for drug discovery, Compounds Australia released the ‘Open Scaffolds’ collection containing 33,999 compounds, with extensive information available on the physicochemical properties of these chemicals. In the present study, we screened 14,464 prioritised compounds from the ‘Open Scaffolds’ collection against the exsheathed third-stage larvae (xL3s of Haemonchus contortus using recently developed whole-organism screening assays. We identified a hit compound, called SN00797439, which was shown to reproducibly reduce xL3 motility by ≥ 70%; this compound induced a characteristic, “coiled” xL3 phenotype (IC50 = 3.46–5.93 μM, inhibited motility of fourth-stage larvae (L4s; IC50 = 0.31–12.5 μM and caused considerable cuticular damage to L4s in vitro. When tested on other parasitic nematodes in vitro, SN00797439 was shown to inhibit (IC50 = 3–50 μM adults of Ancylostoma ceylanicum (hookworm and first-stage larvae of Trichuris muris (whipworm and eventually kill (>90% these stages. Furthermore, this compound completely inhibited the motility of female and male adults of Brugia malayi (50–100 μM as well as microfilariae of both B. malayi and Dirofilaria immitis (heartworm. Overall, these results show that SN00797439 acts against genetically (evolutionarily distant parasitic nematodes i.e. H. contortus and A. ceylanicum [strongyloids] vs. B. malayi and D. immitis [filarioids] vs. T. muris [enoplid], and, thus, might offer a novel, lead-like scaffold for the development of a relatively broad-spectrum anthelmintic. Our future work will focus on assessing the activity of SN00797439 against other pathogens that cause neglected tropical diseases, optimising analogs with improved biological activities and

Capability of the nematode-trapping fungus Duddingtonia flagrans to reduce infective larvae of gastrointestinal nematodes in goat feces in the southeastern United States: dose titration and dose time interval studies.

Science.gov (United States)

Terrill, T H; Larsen, M; Samples, O; Husted, S; Miller, J E; Kaplan, R M; Gelaye, S

2004-04-15

Infection with gastrointestinal nematodes, particularly Haemonchus contortus, is a major constraint to goat production in the southeastern United States. Non-anthelmintic control alternatives are needed due to increasing resistance of these nematodes to available anthelmintics. Two studies were completed in Central Georgia in August 1999, and April-May 2000, using Spanish does naturally infected with Haemonchus contortus, Trichostongylus colubriformis, and Cooperia spp. to evaluate effectiveness of nematode-trapping fungi as a biological control agent. In the first experiment, five levels of Duddingtonia flagrans spores were mixed with a complete diet and fed once daily to the does (three per treatment) in metabolism crates. The treatment concentrations were (1) 5 x 10(5), (2) 2.5 x 10(5), (3) 10(5), and (4) 5 x 10(4) spores per kilogram body weight (BW), and (5) no spores. Fungal spores were fed for the first 7 days of the 14-day trial, and fecal samples were collected daily from individual animals for analysis of fecal egg count and establishment of fecal cultures. Efficacy of the fungus at reducing development of infective larvae (L3) in the fecal cultures was evaluated. The mean reduction in L3 from day 2 of the treatment period until the day after treatment stopped (days 2-8) was 93.6, 80.2, 84.1, and 60.8% for animals given the highest to lowest spore doses, respectively. Within 3-6 days after termination of fungal spore feedings, reduction in L3 development was no longer apparent in any of the treated animals. In a second experiment, effectiveness of 2.5 x 10(5) spores of D. flagrans per kilogram BW fed to does every day, every second day, and every third day was evaluated. Reduction in L3 development by daily feeding was less in the second experiment than in the first experiment. Daily fungal spore feeding provided more consistent larval reduction than intermittant feeding (every second or third day). When fed daily under controlled conditions, D. flagrans

Retaining in-gel zymographic activity of cysteine proteases via a cysteine-supplemented running buffer.

Science.gov (United States)

Vootukuri Reddy, Sreekanth; Philpott, Mike P; Trigiante, Giuseppe

2016-10-01

Zymography is a powerful technique to separate and identify different enzymatic activities on a standard acrylamide gel. For oxidation prone enzymes such as cysteine proteases however, the oxidizing species generated by electrolysis of the gel running buffer may result in partial or complete inactivation, thus compromising the final readout. This can be only partially remedied by subsequent treatment of the gel with reducing agents. We demonstrate the generation of reactive oxidizing species during electrophoresis and discovered that supplementation of the gel running buffer with a minimum of 5 mM cysteine prevents enzyme inactivation and allows retention of proteolytic activity as measured by zymography on model substrate N α-benzoyl-l-arginine p-nitroanilide, without at the same time altering the mobilities of the gel proteins. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Antimalarial effects of vinyl sulfone cysteine proteinase inhibitors.

OpenAIRE

Rosenthal, P J; Olson, J E; Lee, G K; Palmer, J T; Klaus, J L; Rasnick, D

1996-01-01

We evaluated the antimalarial effects of vinyl sulfone cysteine proteinase inhibitors. A number of vinyl sulfones strongly inhibited falcipain, a Plasmodium falciparum cysteine proteinase that is a critical hemoglobinase. In studies of cultured parasites, nanomolar concentrations of three vinyl sulfones inhibited parasite hemoglobin degradation, metabolic activity, and development. The antimalarial effects correlated with the inhibition of falcipain. Our results suggest that vinyl sulfones or...

Transcription factor DecR (YbaO) controls detoxification of L-cysteine in Escherichia coli.

Science.gov (United States)

Shimada, Tomohiro; Tanaka, Kan; Ishihama, Akira

2016-09-01

YbaO is an uncharacterized AsnC-family transcription factor of Escherichia coli. In both Salmonella enterica and Pantoea ananatis, YbaO homologues were identified to regulate the adjacent gene encoding cysteine desulfhydrase for detoxification of cysteine. Using the genomic SELEX (systematic evolution of ligands by exponential enrichment) screening system, we identified the yhaOM operon, located far from the ybaO gene on the E. coli genome, as a single regulatory target of YbaO. In both gel shift assay in vitro and reporter and Northern blot assays in vivo, YbaO was found to regulate the yhaOM promoter. The growth of mutants lacking either ybaO or its targets yhaOM was delayed in the presence of cysteine, indicating involvement of these genes in cysteine detoxification. In the major pathway of cysteine degradation, hydrogen sulfide is produced in wild-type E. coli, but its production was not observed in each of the ybaO, yhaO and yhaM mutants. The yhaOM promoter was activated in the presence of cysteine, implying the role of cysteine in activation of YbaO. Taken together, we propose that YbaO is the cysteine-sensing transcriptional activator of the yhaOM operon, which is involved in the detoxification of cysteine. We then propose the naming of ybaO as decR (regulator of detoxification of cysteine).

Structure and mechanism leading to formation of the cysteine sulfinate product complex of a biomimetic cysteine dioxygenase model.

Science.gov (United States)

Sallmann, Madleen; Kumar, Suresh; Chernev, Petko; Nehrkorn, Joscha; Schnegg, Alexander; Kumar, Devesh; Dau, Holger; Limberg, Christian; de Visser, Sam P

2015-05-11

Cysteine dioxygenase is a unique nonheme iron enzyme that is involved in the metabolism of cysteine in the body. It contains an iron active site with an unusual 3-His ligation to the protein, which contrasts with the structural features of common nonheme iron dioxygenases. Recently, some of us reported a truly biomimetic model for this enzyme, namely a trispyrazolylborato iron(II) cysteinato complex, which not only has a structure very similar to the enzyme-substrate complex but also represents a functional model: Treatment of the model with dioxygen leads to cysteine dioxygenation, as shown by isolating the cysteine part of the product in the course of the work-up. However, little is known on the conversion mechanism and, so far, not even the structure of the actual product complex had been characterised, which is also unknown in case of the enzyme. In a multidisciplinary approach including density functional theory calculations and X-ray absorption spectroscopy, we have now determined the structure of the actual sulfinato complex for the first time. The Cys-SO2 (-) functional group was found to be bound in an η(2) -O,O-coordination mode, which, based on the excellent resemblance between model and enzyme, also provides the first support for a corresponding binding mode within the enzymatic product complex. Indeed, this is again confirmed by theory, which had predicted a η(2) -O,O-binding mode for synthetic as well as the natural enzyme. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Adsorption Dynamics and Self-Assembled L-cysteine on Au(100)

DEFF Research Database (Denmark)

Engelbrekt, Christian; Nazmutdinov, Renat R.; Yan, Jiawei

As the only amino acid with a functional thiol group, L - cysteine offers a strong perspective both for binding to gold and other metals, and for gentle immobilization of biomolecules. Binding to single - crystal, atomically planar surfaces offers the additional perspective that bound L - cysteine...... can be structurally mapped at the single - molecule level . In this work, we have followed the adsorption of L - cysteine on single - crystal Au(100) by measuring the electrode potential dynamics during the adsorption process. In situ STM revealed the structure of the self - assembled ordered layers...

Determination of free and total cyst(e)ine in plasma of dogs and cats.

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Tôrres, Cristina L; Miller, Joshua W; Rogers, Quinton R

2004-01-01

In human blood, the amino acid cysteine forms disulfide bonds with itself and with other sulfhydryl compounds in their free form and with sulfhydryls in protein. Protein-bound cysteine is lost when plasma proteins are removed before amino acid analysis. The purpose of this study was to assess the time course and extent of cyst(e)ine (cysteine + half-cystine) loss in dog and cat plasma. An equal volume of 6% sulfosalicylic acid was added to plasma aliquots at 0, 2, 4, 10, 16, 24, 36, 48, 60, and 72 hours after separation of blood cells. Tris-2-carboxyethyl-phosphine hydrochloride (TCEP - HCl), a reducing agent, was used to regenerate total plasma cyst(e)ine after 3 months of sample storage (-20 degrees C). Initial free cyst(e)ine concentrations (mean +/- SEM) were higher in canine plasma (77 +/- 4 micromol/L) than in feline plasma (37 +/- 3 micromol/L). Free plasma cyst(e)ine concentrations in dogs and cats decreased after first-order kinetics, with a half-life of 23 and 69 hours, respectively. Total plasma cysteine after TCEP - HCl treatment was similar for dogs (290 micromol/L) and cats (296 micromol/L), but the percentage of free cysteine was higher (P = .02) in dogs (27%) than in cats (13%). Over half of the cyst(e)ine, homocysteine, cysteinylglycine, and glutathione were bound in vivo to plasma proteins. These results emphasize the importance of removing plasma proteins within 1 hour after blood collection for reliable assay of free plasma cyst(e)ine.

Functional Characterization of a Novel Class of Morantel-Sensitive Acetylcholine Receptors in Nematodes.

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Elise Courtot

2015-12-01

Full Text Available Acetylcholine receptors are pentameric ligand-gated channels involved in excitatory neuro-transmission in both vertebrates and invertebrates. In nematodes, they represent major targets for cholinergic agonist or antagonist anthelmintic drugs. Despite the large diversity of acetylcholine-receptor subunit genes present in nematodes, only a few receptor subtypes have been characterized so far. Interestingly, parasitic nematodes affecting human or animal health possess two closely related members of this gene family, acr-26 and acr-27 that are essentially absent in free-living or plant parasitic species. Using the pathogenic parasitic nematode of ruminants, Haemonchus contortus, as a model, we found that Hco-ACR-26 and Hco-ACR-27 are co-expressed in body muscle cells. We demonstrated that co-expression of Hco-ACR-26 and Hco-ACR-27 in Xenopus laevis oocytes led to the functional expression of an acetylcholine-receptor highly sensitive to the anthelmintics morantel and pyrantel. Importantly we also reported that ACR-26 and ACR-27, from the distantly related parasitic nematode of horses, Parascaris equorum, also formed a functional acetylcholine-receptor highly sensitive to these two drugs. In Caenorhabditis elegans, a free-living model nematode, we demonstrated that heterologous expression of the H. contortus and P. equorum receptors drastically increased its sensitivity to morantel and pyrantel, mirroring the pharmacological properties observed in Xenopus oocytes. Our results are the first to describe significant molecular determinants of a novel class of nematode body wall muscle AChR.

Functional Characterization of a Novel Class of Morantel-Sensitive Acetylcholine Receptors in Nematodes

Science.gov (United States)

Courtot, Elise; Charvet, Claude L.; Beech, Robin N.; Harmache, Abdallah; Wolstenholme, Adrian J.; Holden-Dye, Lindy; O’Connor, Vincent; Peineau, Nicolas; Woods, Debra J.; Neveu, Cedric

2015-01-01

Acetylcholine receptors are pentameric ligand–gated channels involved in excitatory neuro-transmission in both vertebrates and invertebrates. In nematodes, they represent major targets for cholinergic agonist or antagonist anthelmintic drugs. Despite the large diversity of acetylcholine-receptor subunit genes present in nematodes, only a few receptor subtypes have been characterized so far. Interestingly, parasitic nematodes affecting human or animal health possess two closely related members of this gene family, acr-26 and acr-27 that are essentially absent in free-living or plant parasitic species. Using the pathogenic parasitic nematode of ruminants, Haemonchus contortus, as a model, we found that Hco-ACR-26 and Hco-ACR-27 are co-expressed in body muscle cells. We demonstrated that co-expression of Hco-ACR-26 and Hco-ACR-27 in Xenopus laevis oocytes led to the functional expression of an acetylcholine-receptor highly sensitive to the anthelmintics morantel and pyrantel. Importantly we also reported that ACR-26 and ACR-27, from the distantly related parasitic nematode of horses, Parascaris equorum, also formed a functional acetylcholine-receptor highly sensitive to these two drugs. In Caenorhabditis elegans, a free-living model nematode, we demonstrated that heterologous expression of the H. contortus and P. equorum receptors drastically increased its sensitivity to morantel and pyrantel, mirroring the pharmacological properties observed in Xenopus oocytes. Our results are the first to describe significant molecular determinants of a novel class of nematode body wall muscle AChR. PMID:26625142

Efficacy of the nematode-trapping fungus Duddingtonia flagrans against three species of gastro-intestinal nematodes in laboratory faecal cultures from sheep and goats.

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Waghorn, T S; Leathwick, D M; Chen, L-Y; Skipp, R A

2003-12-30

The ability of the nematode-killing fungus Duddingtonia flagrans to reduce number of infective larvae of three species of gastro-intestinal parasitic nematodes developing in dung was investigated in both goats and sheep. Groups of lambs and kids (12-20 weeks old) were given mono-specific infections of Haemonchus contortus, Ostertagia (Teladorsagia) circumcincta or Trichostrongylus colubriformis. Following patency of the infections (t1) faecal samples were collected for determination of faecal nematode egg count (FEC) and culture of parasite larvae. Groups of animals were then dosed on 2 consecutive days with one of the two dose rates of the fungus (250,000 or 500,000 spores/kg liveweight). One (t2) and 5 (t3) days after the second dose of fungus samples were again collected for FEC and culture. The number of larvae recovered from the faecal cultures at t1 and t3 were used as controls to assess the efficacy of the experimental treatment at t2. Average efficacy was 78% with group means ranging from 40 to 93%. Dose rate of fungus appeared to influence efficacy against O. circumcincta but not against H. contortus or T. colubriformis. Overall, there were no differences in the efficacy of the fungus against any of the parasite species or in either host animal. The results of this trial indicate the potential use of this fungus as a broad spectrum anti-parasite agent for use in both goats and sheep.

Effect of L-cysteine on the oxidation of cyclohexane catalyzed by manganeseporphyrin.

Science.gov (United States)

Zhou, Wei-You; Tian, Peng; Chen, Yong; He, Ming-Yang; Chen, Qun; Chen, Zai Xin

2015-06-01

Effect of L-cysteine as the cocatalyst on the oxidation of cyclohexane by tert-butylhydroperoxide (TBHP) catalyzed by manganese tetraphenylporphyrin (MnTPP) has been investigated. The results showed that L-cysteine could moderately improve the catalytic activity of MnTPP and significantly increase the selectivity of cyclohexanol. Different from imidazole and pyridine, the L-cysteine may perform dual roles in the catalytic oxidation of cyclohexane. Besides as the axial ligand for MnTPP, the L-cysteine could also react with cyclohexyl peroxide formed as the intermediate to produce alcohol as the main product. Copyright © 2015 Elsevier Ltd. All rights reserved.

Activated human CD4 T cells express transporters for both cysteine and cystine

DEFF Research Database (Denmark)

Levring, Trine Bøegh; Hansen, Ann Kathrine; Nielsen, Bodil Lisbeth

2012-01-01

Because naïve T cells are unable to import cystine due to the absence of cystine transporters, it has been suggested that T cell activation is dependent on cysteine generated by antigen presenting cells. The aim of this study was to determine at which phases during T cell activation exogenous...... cystine/cysteine is required and how T cells meet this requirement. We found that early activation of T cells is independent of exogenous cystine/cysteine, whereas T cell proliferation is strictly dependent of uptake of exogenous cystine/cysteine. Naïve T cells express no or very low levels of both...... cystine and cysteine transporters. However, we found that these transporters become strongly up-regulated during T cell activation and provide activated T cells with the required amount of cystine/cysteine needed for T cell proliferation. Thus, T cells are equipped with mechanisms that allow T cell...

THE INCIDENCE OF PARASITIC DISEASES IN LIVESTOCK IN BALI

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N. A. Suratma

2012-09-01

Full Text Available The population of livestock in Bali has continuously increased from year to year. However, some problems are encountered with parasitic infections in livestock. Parasitic infections may be caused by worms, protozoa or ectoparasites. In cattle, the most common infections are those caused by Oesophagostomum sp, Ostertagia sp, Haemonchus sp, Mecistocirrus sp, and Cooperia sp which is the most dominant. Neoascaris vitulorum was reported to be as high as 29.1% in calves. Fascioliasis in cattle was found highly prevalent, between 34.9 to 56.7% and was caused by Fasciola gigantica. Also Paramphistomum infection was reported to be highly prevalent (50.1%. In addition, Boophilus microplus was recorded as high as 36.9%. In goat and sheep, the incidence of Haemonchus contortus was 27.7% and 53.6% respectively. Infestation of Paramphistomum sp in goat was 9.27%. Concerning ectoparasites, Sarcoptes scabiei was reported to be the cause of death of 67% of young goats and up to 11% of older gats in Br. Penginuman, Gilimanuk Negara. Parasitic infections in pigs were caused by Cysticercus tenuicollis (11% and Ascaris suum (24.2% and 21.1% showed Metastrongylus apri and also Sarcoptes scabiei was reported to be the cause of skin disease in pigs. In poultry, parasitic infection were caused by Raillietina (96%, Heterakis gallinae (66.7%, Capillaria sp (6.6%, Ascardia galli (56.7%, Oxyspirura mansoni (50%, Acuaria spiralis (13.3% and Syngamus trachea (3.3%. Multiple infections are common.

Cysteine Protease Inhibitors as Chemotherapy: Lessons from a Parasite Target

Science.gov (United States)

Selzer, Paul M.; Pingel, Sabine; Hsieh, Ivy; Ugele, Bernhard; Chan, Victor J.; Engel, Juan C.; Bogyo, Matthew; Russell, David G.; Sakanari, Judy A.; McKerrow, James H.

1999-09-01

Papain family cysteine proteases are key factors in the pathogenesis of cancer invasion, arthritis, osteoporosis, and microbial infections. Targeting this enzyme family is therefore one strategy in the development of new chemotherapy for a number of diseases. Little is known, however, about the efficacy, selectivity, and safety of cysteine protease inhibitors in cell culture or in vivo. We now report that specific cysteine protease inhibitors kill Leishmania parasites in vitro, at concentrations that do not overtly affect mammalian host cells. Inhibition of Leishmania cysteine protease activity was accompanied by defects in the parasite's lysosome/endosome compartment resembling those seen in lysosomal storage diseases. Colocalization of anti-protease antibodies with biotinylated surface proteins and accumulation of undigested debris and protease in the flagellar pocket of treated parasites were consistent with a pathway of protease trafficking from flagellar pocket to the lysosome/endosome compartment. The inhibitors were sufficiently absorbed and stable in vivo to ameliorate the pathology associated with a mouse model of Leishmania infection.

Cysteine 138 mutation in HIV-1 Nef from patients with delayed disease progression

DEFF Research Database (Denmark)

Tolstrup, Martin; Laursen, Alex Lund; Gerstoft, J.

2006-01-01

on the delayed disease status. However, the results demonstrate a high incidence of a single amino acid polymorphism (cysteine 138) in HIV-1 Nef. The allelic frequency of cysteine 138 between the delayed disease progression group and the progressor group was found to be statistically significant (P = 0.......0139). The phylogeny of isolates was investigated and the variants harbouring the cysteine 138 mutation clustered independently. CONCLUSION: The present study describes a viral genetic polymorphism related to AIDS disease progression. The polymorphism (cysteine 138) has previously been reported to confer decreased...... viral replication (Premkumar DR, et al. AIDS Res Hum Retroviruses 1996; 12(4): 337-45). A sequence database search for comparative mutations revealed a high frequency of cysteine 138 in patients with reported SP AIDS...

Effects of cysteine on growth, protease production, and catalase activity of Pseudomonas fluorescens.

OpenAIRE

Himelbloom, B H; Hassan, H M

1986-01-01

Cysteine inhibits growth of and protease production by Pseudomonas fluorescens NC3. Catalase activity in P. fluorescens NC3 was increased by cysteine. The addition of exogenous hydrogen peroxide did not increase catalase activity, thus suggesting a role for the endogenous generation of hydrogen peroxide via the autoxidation of cysteine.

Cysteine and hydrogen sulfide in the regulation of metabolism:Insights from genetics and pharmacology

OpenAIRE

Carter, Roderick N; Morton, Nicholas M

2016-01-01

Abstract Obesity and diabetes represent a significant and escalating worldwide health burden. These conditions are characterized by abnormal nutrient homeostasis. One such perturbation is altered metabolism of the sulphur?containing amino acid cysteine. Obesity is associated with elevated plasma cysteine, whereas diabetes is associated with reduced cysteine levels. One mechanism by which cysteine may act is through its enzymatic breakdown to produce hydrogen sulphide (H2S), a gasotransmitter ...

Effects of L-cysteine on lead acetate induced neurotoxicity in albino mice.

Science.gov (United States)

Mahmoud, Y I; Sayed, S S

2016-07-01

Lead is a toxic heavy metal that adversely affects nervous tissues; it often occurs as an environmental pollutant. We investigated histological changes in the cerebral cortex, hippocampus and cerebellum of adult albino mice following exposure to lead acetate. We also studied the possible ameliorative effect of the chelating agent, L-cysteine, on lead-induced neurotoxicity. We divided albino mice into six groups: 1) vehicle-only control, 2) L-cysteine control, 3 and 4) treated for 7 days with 20 and 40 mg/kg lead acetate, respectively, and 5 and 6) treated for 7 days with 20 and 40 mg/kg lead acetate, respectively, followed by 50 mg/kg L-cysteine for 7 days. Lead acetate administration caused disorganization of cell layers, neuronal loss and degeneration, and neuropil vacuolization. Brain sections from lead-intoxicated mice treated with L-cysteine showed fewer pathological changes; the neuropil showed less vacuolization and the neurons appeared less damaged. L-cysteine at the dose we used only marginally alleviated lead-induced toxicity.

Depletion of circulating cyst(e)ine by oral and intravenous mesna.

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Stofer-Vogel, B.; Cerny, T.; Küpfer, A.; Junker, E.; Lauterburg, B. H.

1993-01-01

The sulfhydryl status of normal and tumour cells is critically important in determining their susceptibility to various cytostatic agents. As a sulfhydryl compound, mesna (sodium 2-mercaptoethane-sulfonate) which is used in large doses to prevent haemorrhagic cystitis associated with certain chemotherapeutic regimens might derange cellular thiol homeostasis. In order to investigate the effects of mesna on the concentrations of thiols in plasma, cysteine, glutathione and their disulfides were measured by HPLC following the oral and intravenous administration of mesna to healthy volunteers. After 7.3 mmol mesna i.v. free cysteine rose from 8.2 (95% CI 7.0-9.4) nmol ml-1 to 53.6 (47.4-59.8) nmol ml-1 at 5 min, most likely due to reduction of circulating cystine by the sulfhydryl drug. This initial rise was followed by a marked decrease of total cyst(e)ine in plasma from 276 (215-337) nmol ml-1 to a nadir of 102 (89-115) nmol ml-1 between 30-120 min after infusion, most likely due to an increased uptake of cysteine into cells and an increased urinary excretion of cyst(e)ine. Qualitatively similar changes were seen after oral mesna. The present data indicate that mesna depletes circulating cyst(e)ine and may thereby markedly alter the sulfhydryl status of cells in vivo although the drug itself is not taken up by most cells. PMID:8353049

Facile and green synthesis of highly stable L-cysteine functionalized copper nanoparticles

Science.gov (United States)

Kumar, Nikhil; Upadhyay, Lata Sheo Bachan

2016-11-01

A simple eco-friendly method for L-cysteine capped copper nanoparticles (CCNPs) synthesis in aqueous solution has been developed. Glucose and L-cysteine were used as reducing agent and capping/functionalizing agent, respectively. Different parameters such as capping agent concentration, pH, reaction temperature, and reducing agent concentration were optimized during the synthesis. The L-cysteine capped copper nanoparticle were characterized by ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, Particle size and zeta potential analyser, and high resolution transmission electron microscopy. Spherical shaped cysteine functionalized/capped copper nanoparticles with an average size of 40 nm were found to be highly stable at room temperature (RT) for a period of 1 month

Cysteine regulation of protein function--as exemplified by NMDA-receptor modulation.

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Lipton, Stuart A; Choi, Yun-Beom; Takahashi, Hiroto; Zhang, Dongxian; Li, Weizhong; Godzik, Adam; Bankston, Laurie A

2002-09-01

Until recently cysteine residues, especially those located extracellularly, were thought to be important for metal coordination, catalysis and protein structure by forming disulfide bonds - but they were not thought to regulate protein function. However, this is not the case. Crucial cysteine residues can be involved in modulation of protein activity and signaling events via other reactions of their thiol (sulfhydryl; -SH) groups. These reactions can take several forms, such as redox events (chemical reduction or oxidation), chelation of transition metals (chiefly Zn(2+), Mn(2+) and Cu(2+)) or S-nitrosylation [the catalyzed transfer of a nitric oxide (NO) group to a thiol group]. In several cases, these disparate reactions can compete with one another for the same thiol group on a single cysteine residue, forming a molecular switch composed of a latticework of possible redox, NO or Zn(2+) modifications to control protein function. Thiol-mediated regulation of protein function can also involve reactions of cysteine residues that affect ligand binding allosterically. This article reviews the basis for these molecular cysteine switches, drawing on the NMDA receptor as an exemplary protein, and proposes a molecular model for the action of S-nitrosylation based on recently derived crystal structures.

Characterization of the gene encoding serine acetyltransferase, a regulated enzyme of cysteine biosynthesis from the protist parasites Entamoeba histolytica and Entamoeba dispar. Regulation and possible function of the cysteine biosynthetic pathway in Entamoeba.

Science.gov (United States)

Nozaki, T; Asai, T; Sanchez, L B; Kobayashi, S; Nakazawa, M; Takeuchi, T

1999-11-05

The enteric protist parasites Entamoeba histolytica and Entamoeba dispar possess a cysteine biosynthetic pathway, unlike their mammalian host, and are capable of de novo production of L-cysteine. We cloned and characterized cDNAs that encode the regulated enzyme serine acetyltransferase (SAT) in this pathway from these amoebae by genetic complementation of a cysteine-auxotrophic Escherichia coli strain with the amoebic cDNA libraries. The deduced amino acid sequences of the amoebic SATs exhibited, within the most conserved region, 36-52% identities with the bacterial and plant SATs. The amoebic SATs contain a unique insertion of eight amino acids, also found in the corresponding region of a plasmid-encoded SAT from Synechococcus sp., which showed the highest overall identities to the amoebic SATs. Phylogenetic reconstruction also revealed a close kinship of the amoebic SATs with cyanobacterial SATs. Biochemical characterization of the recombinant E. histolytica SAT revealed several enzymatic features that distinguished the amoebic enzyme from the bacterial and plant enzymes: 1) inhibition by L-cysteine in a competitive manner with L-serine; 2) inhibition by L-cystine; and 3) no association with cysteine synthase. Genetically engineered amoeba strains that overproduced cysteine synthase and SAT were created. The cysteine synthase-overproducing amoebae had a higher level of cysteine synthase activity and total thiol content and revealed increased resistance to hydrogen peroxide. These results indicate that the cysteine biosynthetic pathway plays an important role in antioxidative defense of these enteric parasites.

Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation

Energy Technology Data Exchange (ETDEWEB)

Simmons, Chad R. [Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853-8001 (United States); Hao, Quan [MacCHESS at the Cornell High Energy Synchrotron Source, Cornell University, Ithaca, NY 14853-8001 (United States); Stipanuk, Martha H., E-mail: mhs6@cornell.edu [Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853-8001 (United States)

2005-11-01

Recombinant rat cysteine dioxygenase (CDO) has been expressed, purified and crystallized and X-ray diffraction data have been collected to 1.5 Å resolution. Cysteine dioxygenase (CDO; EC 1.13.11.20) is an ∼23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O{sub 2}, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Å resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Å, α = β = γ = 90°. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative.

Exercise induced upregulation of glutamate-cysteine ligase catalytic subunit and glutamate-cysteine ligase modifier subunit gene expression in Thoroughbred horses

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Jeong-Woong Park

2017-05-01

Full Text Available Objective This study was performed to reveal the molecular structure and expression patterns of horse glutamate-cysteine ligase catalytic subunit (GCLC and glutamate-cysteine ligase modifier subunit (GCLM genes whose products form glutamate cysteine ligase, which were identified as differentially expressed genes in the previous study. Methods We performed bioinformatics analyses, and gene expression assay with quantitative polymerase chain reaction (qPCR for horse GCLC and GCLM genes in muscle and blood leukocytes of Thoroughbred horses Results Expression of GCLC showed the same pattern in both blood and muscle tissues after exercise. Expression of GCLC increased in the muscle and blood of Thoroughbreds, suggesting a tissue-specific regulatory mechanism for the expression of GCLC. In addition, expression of the GCLM gene increased after exercise in both the blood and muscle of Thoroughbreds. Conclusion We established the expression patterns of GCLC and GCLM in the skeletal muscle and blood of Thoroughbred horses in response to exercise. Further study is now warranted to uncover the functional importance of these genes in exercise and recovery in racehorses.

Aspartic acid-promoted highly selective and sensitive colorimetric sensing of cysteine in rat brain.

Science.gov (United States)

Qian, Qin; Deng, Jingjing; Wang, Dalei; Yang, Lifen; Yu, Ping; Mao, Lanqun

2012-11-06

Direct selective determination of cysteine in the cerebral system is of great importance because of the crucial roles of cysteine in physiological and pathological processes. In this study, we report a sensitive and selective colorimetric assay for cysteine in the rat brain with gold nanoparticles (Au-NPs) as the signal readout. Initially, Au-NPs synthesized with citrate as the stabilizer are red in color and exhibit absorption at 520 nm. The addition of an aqueous solution (20 μL) of cysteine or aspartic acid alone to a 200 μL Au-NP dispersion causes no aggregation, while the addition of an aqueous solution of cysteine into a Au-NP dispersion containing aspartic acid (1.8 mM) causes the aggregation of Au-NPs and thus results in the color change of the colloid from wine red to blue. These changes are ascribed to the ion pair interaction between aspartic acid and cysteine on the interface between Au-NPs and solution. The concentration of cysteine can be visualized with the naked eye and determined by UV-vis spectroscopy. The signal output shows a linear relationship for cysteine within the concentration range from 0.166 to 1.67 μM with a detection limit of 100 nM. The assay demonstrated here is highly selective and is free from the interference of other natural amino acids and other thiol-containing species as well as the species commonly existing in the brain such as lactate, ascorbic acid, and glucose. The basal dialysate level of cysteine in the microdialysate from the striatum of adult male Sprague-Dawley rats is determined to be around 9.6 ± 2.1 μM. The method demonstrated here is facile but reliable and durable and is envisaged to be applicable to understanding the chemical essence involved in physiological and pathological events associated with cysteine.

Dose titration of sericea lespedeza leaf meal on Haemonchus contortus infection in lambs and kids

Science.gov (United States)

The objective of three experiments was to determine the impact of supplementing sericea lespedeza (Lespedeza cuneata; SL) in three concentrations in a loose or pelleted diet on gastrointestinal nematodes (GIN) in small ruminants. Experiments on lambs were conducted at the USDA, Agricultural Research...

35S cystein chlorhydrate preparation

International Nuclear Information System (INIS)

Emiliozzi, R.; Pichat, P.; Herbert, M.

1960-01-01

35 S cystein chlorhydrate has been prepared with a quantitative yield by electrolytic reduction of 35 S cystin in hydrochloric medium on a vibrating mercury cathode. Reprint of a paper published in Bulletin de la Societe chimique de France, no. 2653, 4. quarter 1959, p. 1544-1545 [fr

Neuronal growth on L- and D-cysteine self-assembled monolayers reveals neuronal chiral sensitivity.

Science.gov (United States)

Baranes, Koby; Moshe, Hagay; Alon, Noa; Schwartz, Shmulik; Shefi, Orit

2014-05-21

Studying the interaction between neuronal cells and chiral molecules is fundamental for the design of novel biomaterials and drugs. Chirality influences all biological processes that involve intermolecular interaction. One common method used to study cellular interactions with different enantiomeric targets is the use of chiral surfaces. Based on previous studies that demonstrated the importance of cysteine in the nervous system, we studied the effect of L- and D-cysteine on single neuronal growth. L-Cysteine, which normally functions as a neuromodulator or a neuroprotective antioxidant, causes damage at elevated levels, which may occur post trauma. In this study, we grew adult neurons in culture enriched with L- and D-cysteine as free compounds or as self-assembled monolayers of chiral surfaces and examined the effect on the neuronal morphology and adhesion. Notably, we have found that exposure to the L-cysteine enantiomer inhibited, and even prevented, neuronal attachment more severely than exposure to the D-cysteine enantiomer. Atop the L-cysteine surfaces, neuronal growth was reduced and degenerated. Since the cysteine molecules were attached to the surface via the thiol groups, the neuronal membrane was exposed to the molecular chiral site. Thus, our results have demonstrated high neuronal chiral sensitivity, revealing chiral surfaces as indirect regulators of neuronal cells and providing a reference for studying chiral drugs.

Gastrointestinal parasites of sheep, municipality of Lajes, Rio Grande do Norte, Brazil.

Science.gov (United States)

de Souza, Maria de Fátima; Pimentel-Neto, Manoel; da Silva, Rízia Maria; Farias, Albeísa Cleyse Batista; Guimarães, Marcos Pezzi

2012-01-01

This study investigated the gastrointestinal parasitism by helminths and protozoa in sheep (Ovis aries) Santa Inês breed, municipality of Lajes, Rio Grande do Norte. Monthly, from April 2005 to August 2007, stool samples were collected from two tracer lambs in the first day of the experiment and performed a necropsy of these animals in 44th day. A total of 64 lambs were sampled, but only 62 lambs were slaughtered. The fecal samples were examined by sedimentation in water. The contents of the abomasum, small intestine and large intestine were examined for the recovery of helminths. The parasitological examination revealed eggs of the following groups of helminths: Strongyloidea, Strongyloides sp., Trichuris sp., and Moniezia sp. Also were found oocysts of Eimeria spp., cysts of Entamoeba ovis and Giardia duodenalis. The helminths identified from examining the contents were: Haemonchus contortus, Cooperia pectinata, Cooperia punctata, Trichostrongylus colubriformis, Moniezia expansa, Oesophagostomum sp. Skrjabinema ovis and Trichuris sp.

A survey of anthelmintic resistance on ten sheep farms in Mashonaland East Province, Zimbabwe : research communication

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S. Mukaratirwa

1997-07-01

Full Text Available A survey to detect anthelmintic resistance in nematode parasites of sheep was conducted on 10 randomly-distributed farms in the Chivhu District, Mashonaland East Province, Zimbabwe. Before the survey, a questionnaire was circulated to the farmers concerning nematode parasite control. Results showed that parasite control using anthelmintic treatment was the only method practised and that the benzimidazoles were the most frequently used anthelmintic drugs. The faecal egg count reduction test was used to detect resistance. The anthelmintic groups tested were benzimidazoles, levamisole and ivermectin. Resistance to benzimidazoles was detected on 6 of 10 farms and levamisole resistance on 2 of 3 farms. Ivermectin resistance was not observed on the farms surveyed. Post-treatment larval cultures indicated that Haemonchus contortus survived administration of fenbendazole, albendazole, oxfendazole and levamisole. A Cooperia sp. strain resistant to albendazole was detected and this is the first report in Zimbabwe of a resistant parasite in this genus.

Mutagenicity and cytotoxicity of two regioisomeric mercapturic acids and cysteine S-conjugates of trichloroethylene.

NARCIS (Netherlands)

Commandeur, J.N.M.; Boogaard, P.J.; Mulder, G.J.; Vermeulen, N.P.E.

1991-01-01

The mutagenicity, cytotoxicity and metabolism of two regioisomic l-cysteine- and N-acetyl-l-cysteine-S-conjugates of trichloroethylene were studied. The 1,2-dichlorovinyl(1,2-DCV) isomers of both the cysteine conjugate and the mercapturate were much stronger mutagens in the Ames test with Salmonella

Analysis of S-nitrosothiols via Copper Cysteine (2C) and Copper Cysteine - Carbon Monoxide (3C) Methods

Science.gov (United States)

Rogers, Stephen C.; Gibbons, Lindsey B.; Griffin, Sherraine; Doctor, Allan

2012-01-01

This chapter summarizes the principles of RSNO measurement in the gas phase, utilizing ozone-based chemiluminescence and the copper cysteine (2C) ± carbon monoxide (3C) reagent. Although an indirect method for quantifying RSNOs, this assay represents one of the most robust methodologies available. It exploits the NO• detection sensitivity of ozone based chemiluminscence, which is within the range required to detect physiological concentrations of RSNO metabolites. Additionally, the specificity of the copper cysteine (2C and 3C) reagent for RSNOs negates the need for sample pretreatment, thereby minimizing the likelihood of sample contamination (false positive results), NO species inter-conversion, or the loss of certain highly labile RSNO species. Herein, we outline the principles of this methodology, summarizing key issues, potential pitfalls and corresponding solutions. PMID:23116707

Loss of second and sixth conserved cysteine residues from trypsin inhibitor-like cysteine-rich domain-type protease inhibitors in Bombyx mori may induce activity against microbial proteases.

Science.gov (United States)

Li, Youshan; Liu, Huawei; Zhu, Rui; Xia, Qingyou; Zhao, Ping

2016-12-01

Previous studies have indicated that most trypsin inhibitor-like cysteine-rich domain (TIL)-type protease inhibitors, which contain a single TIL domain with ten conserved cysteines, inhibit cathepsin, trypsin, chymotrypsin, or elastase. Our recent findings suggest that Cys 2nd and Cys 6th were lost from the TIL domain of the fungal-resistance factors in Bombyx mori, BmSPI38 and BmSPI39, which inhibit microbial proteases and the germination of Beauveria bassiana conidia. To reveal the significance of these two missing cysteines in relation to the structure and function of TIL-type protease inhibitors in B. mori, cysteines were introduced at these two positions (D36 and L56 in BmSPI38, D38 and L58 in BmSPI39) by site-directed mutagenesis. The homology structure model of TIL domain of the wild-type and mutated form of BmSPI39 showed that two cysteine mutations may cause incorrect disulfide bond formation of B. mori TIL-type protease inhibitors. The results of Far-UV circular dichroism (CD) spectra indicated that both the wild-type and mutated form of BmSPI39 harbored predominantly random coil structures, and had slightly different secondary structure compositions. SDS-PAGE and Western blotting analysis showed that cysteine mutations affected the multimerization states and electrophoretic mobility of BmSPI38 and BmSPI39. Activity staining and protease inhibition assays showed that the introduction of cysteine mutations dramaticly reduced the activity of inhibitors against microbial proteases, such as subtilisin A from Bacillus licheniformis, protease K from Engyodontium album, protease from Aspergillus melleus. We also systematically analyzed the key residue sites, which may greatly influence the specificity and potency of TIL-type protease inhibitors. We found that the two missing cysteines in B. mori TIL-type protease inhibitors might be crucial for their inhibitory activities against microbial proteases. The genetic engineering of TIL-type protease inhibitors may be

Electronic structures of the L-cysteine film on dental alloys

International Nuclear Information System (INIS)

Ogawa, K.; Tsujibayashi, T.; Takahashi, K.; Azuma, J.; Kakimoto, K.; Kamada, M.

2011-01-01

Research highlights: → The electronic structures of dental alloys and L-cysteine film were studied by PES. → The density of states in the dental alloy originates from Au and Cu as constituents. → The Cu-3d states contribute dominantly to the occupied states near the Fermi level. → The electronic structure of L-cysteine thin film is different from the thick film. → The bonding between Cu-3d and S-3sp states are formed at the interface. - Abstract: Metal-organic interfaces have been attracting continuous attention in many fields including basic biosciences. The surface of dental alloys could be one of such interfaces since they are used in a circumstance full of organic compounds such as proteins and bacteria. In this work, electronic structures of Au-dominant dental alloys, which have Ag and Cu besides Au, and those of L-cysteine on the dental alloys have been studied by photoelectron spectroscopy with synchrotron radiation. It was found that the density of states in the dental alloy originate from gold and copper as constituents, and the Cu-3d states contribute dominantly to the occupied states near the Fermi level. It was also found that the electronic structure of the L-cysteine thin film on the dental alloy is different from that of the L-cysteine thick film. The result indicates the formation of the orbital bonding between Cu-3d and S-3sp states in the thin film on the dental alloy.

Electronic structures of the L-cysteine film on dental alloys

Energy Technology Data Exchange (ETDEWEB)

Ogawa, K., E-mail: e7141@cc.saga-u.ac.jp [Synchrotron Light Application Center, Saga University, Saga 840-8502 (Japan); Tsujibayashi, T. [Department of Physics, Osaka Dental University, Osaka 573-1121 (Japan); Takahashi, K.; Azuma, J. [Synchrotron Light Application Center, Saga University, Saga 840-8502 (Japan); Kakimoto, K. [Department of Geriatric Dentistry, Osaka Dental University, Osaka 573-1121 (Japan); Kamada, M. [Synchrotron Light Application Center, Saga University, Saga 840-8502 (Japan)

2011-04-15

Research highlights: {yields} The electronic structures of dental alloys and L-cysteine film were studied by PES. {yields} The density of states in the dental alloy originates from Au and Cu as constituents. {yields} The Cu-3d states contribute dominantly to the occupied states near the Fermi level. {yields} The electronic structure of L-cysteine thin film is different from the thick film. {yields} The bonding between Cu-3d and S-3sp states are formed at the interface. - Abstract: Metal-organic interfaces have been attracting continuous attention in many fields including basic biosciences. The surface of dental alloys could be one of such interfaces since they are used in a circumstance full of organic compounds such as proteins and bacteria. In this work, electronic structures of Au-dominant dental alloys, which have Ag and Cu besides Au, and those of L-cysteine on the dental alloys have been studied by photoelectron spectroscopy with synchrotron radiation. It was found that the density of states in the dental alloy originate from gold and copper as constituents, and the Cu-3d states contribute dominantly to the occupied states near the Fermi level. It was also found that the electronic structure of the L-cysteine thin film on the dental alloy is different from that of the L-cysteine thick film. The result indicates the formation of the orbital bonding between Cu-3d and S-3sp states in the thin film on the dental alloy.

Oral Administration of (S)-Allyl-l-Cysteine and Aged Garlic Extract to Rats: Determination of Metabolites and Their Pharmacokinetics.

Science.gov (United States)

Park, Taehoon; Oh, Ju-Hee; Lee, Joo Hyun; Park, Sang Cheol; Jang, Young Pyo; Lee, Young-Joo

2017-11-01

( S )-Allyl-l-cysteine is the major bioactive compound in garlic. ( S )-Allyl-l-cysteine is metabolized to ( S )-allyl-l-cysteine sulfoxide, N -acetyl-( S )-allyl-l-cysteine, and N -acetyl-( S )-allyl-l-cysteine sulfoxide after oral administration. An accurate LC-MS/MS method was developed and validated for the simultaneous quantification of ( S )-allyl-l-cysteine and its metabolites in rat plasma, and the feasibility of using it in pharmacokinetic studies was tested. The analytes were quantified by multiple reaction monitoring using an atmospheric pressure ionization mass spectrometer. Because significant quantitative interference was observed between ( S )-allyl-l-cysteine and N -acetyl-( S )-allyl-l-cysteine as a result of the decomposition of N -acetyl-( S )-allyl-l-cysteine at the detector source, chromatographic separation was required to discriminate ( S )-allyl-l-cysteine and its metabolites on a reversed-phase C 18 analytical column with a gradient mobile phase consisting of 0.1% formic acid and acetonitrile. The calibration curves of ( S )-allyl-l-cysteine, ( S )-allyl-l-cysteine sulfoxide, N -acetyl-( S )-allyl-l-cysteine, and N -acetyl-( S )-allyl-l-cysteine sulfoxide were linear over each concentration range, and the lower limits of quantification were 0.1 µg/mL [( S )-allyl-l-cysteine and N -acetyl-( S )-allyl-l-cysteine] and 0.25 µg/mL [( S )-allyl-l-cysteine sulfoxide and N -acetyl-( S )-allyl-l-cysteine sulfoxide]. Acceptable intraday and inter-day precisions and accuracies were obtained at three concentration levels. The method satisfied the regulatory requirements for matrix effects, recovery, and stability. The validated LC-MS/MS method was successfully used to determine the concentration of ( S )-allyl-l-cysteine and its metabolites in rat plasma samples after the administration of ( S )-allyl-l-cysteine or aged garlic extract. Georg Thieme Verlag KG Stuttgart · New York.

Fasting, but Not Aging, Dramatically Alters the Redox Status of Cysteine Residues on Proteins in Drosophila melanogaster

Directory of Open Access Journals (Sweden)

Katja E. Menger

2015-06-01

Full Text Available Altering the redox state of cysteine residues on protein surfaces is an important response to environmental challenges. Although aging and fasting alter many redox processes, the role of cysteine residues is uncertain. To address this, we used a redox proteomic technique, oxidative isotope-coded affinity tags (OxICAT, to assess cysteine-residue redox changes in Drosophila melanogaster during aging and fasting. This approach enabled us to simultaneously identify and quantify the redox state of several hundred cysteine residues in vivo. Cysteine residues within young flies had a bimodal distribution with peaks at ∼10% and ∼85% reversibly oxidized. Surprisingly, these cysteine residues did not become more oxidized with age. In contrast, 24 hr of fasting dramatically oxidized cysteine residues that were reduced under fed conditions while also reducing cysteine residues that were initially oxidized. We conclude that fasting, but not aging, dramatically alters cysteine-residue redox status in D. melanogaster.

[Protective Effect of S-isopentenyl-L-cysteine against DNA Damage in Irradiated Mice].

Science.gov (United States)

Zheng, Qi-sheng; Yu, Guang-yun; He, Xin; Jiang, Ming; Chu, Xiao-fei; Zhao, Shu-yi; Fan, Sai-jun; Liu, Pei-xun

2015-10-01

To evaluate the protective effect of S-isopentenyl-L-cysteine,a new cysteine derivative,on DNA damage induced by radiation by using acute radiation injury animal models. Forty ICR mice were randomly divided into five groups:the control group,1.0Gy gamma irradiation group,1.0Gy gamma irradiation combined with S-isopentenyl-L-cysteine group,7.2Gy gamma irradiation group,and 7.2Gy gamma irradiation combined with S-isopentenyl-L-cysteine group,with 8 mice in each group.The comet assay and bone marrow polychromatic micronucleus experiments were performed to evaluate the double-strand DNA breaks in ICR mice exposed to 1.0 and 7.2Gy gamma-ray, respectively. The tail DNA percentage,tail length,tail moment,and olive tail moment of peripheral blood lymphocytes in 7.2Gy gamma irradiation group were significantly higher than that of the control group (PL-cysteine group was significantly less than that of 7.2Gy gamma irradiation group (PL-cysteine before irradiation,the micronucleus rate of ICR mice exposed to 1.0 and 7.2Gy gamma-ray decreased from (39.5000 ± 3.3141)‰ to (28.1667±4.1345)‰ (P=0.033) and from (76.5000 ± 4.6242)‰ to (22.8333 ± 3.6553)‰(P=0.000),respectively. The bone marrow polychromatic micronucleus experiment indicated that the value of polychromatic erythrocyte (PCE)/normochromatic erythrocyte(NCE) of ICR mice exposed to 1.0 and 7.2Gy gamma-ray was less than the control group(PL-cysteine before irradiation was significantly higher than the corresponding groups (PL-cysteine has a good protective effect against DNA damage induced by radiation.

Validation of the FAMACHA© system in South American camelids.

Science.gov (United States)

Storey, Bobby E; Williamson, Lisa H; Howell, Sue B; Terrill, Thomas H; Berghaus, Roy; Vidyashankar, Anand N; Kaplan, Ray M

2017-08-30

Haemonchus contortus resistant to multiple anthelmintics threaten the viability of the small ruminant industry in areas where this parasite is prevalent. In response to this situation, the FAMACHA© system was developed and validated for use with small ruminants as a way to detect clinical anemia associated with haemonchosis. Given that H. contortus and multiple anthelmintic resistance is a similar problem in camelids, the FAMACHA© system might also provide the same benefits. To address this need, a validation study of the FAMACHA© system was conducted on 21 alpaca and llama farms over a 2-year period. H. contortus was the predominant nematode parasite on 17 of the 21 farms (10 alpaca and 7 llama farms) enrolled in the study, based on fecal culture results. The FAMACHA© card was used to score the color of the lower palpebral (lower eye lid) conjunctiva on a 1-5 scale. Packed cell volume (PCV) values were measured and compared to FAMACHA© scores using FAMACHA© score cutoffs of ≥3 or ≥4 and with anemia defined as a PCV ≤15%, ≤17%, or≤20%. PCV was significantly associated with FAMACHA© score, fecal egg count (FEC), and body condition score (BCS), regardless of the FAMACHA© cutoff score or the PCV% chosen to define clinical anemia (p<0.01 in all cases). The use of FAMACHA© scores ≥3 and PCV ≥ 15% indicating anemia provided the best sensitivity (96.4% vs 92.9% for FAMACHA© ≥4), whereas FAMACHA scores ≥ 4 and PCV ≤20% provided the best specificity (94.2% vs 69.1% for FAMACHA© ≥3). The data from this study support the FAMACHA© system as a useful tool for detecting clinical anemia in camelids suffering from haemonchosis. Parameters for making treatment decisions based on FAMACHA© score in camelids should mirror those established for small ruminants. Copyright © 2017 Elsevier B.V. All rights reserved.

Formation of Hg(II) Tetrathiolate Complexes with Cysteine at Neutral pH.

Science.gov (United States)

Warner, Thomas; Jalilehvand, Farideh

2016-04-01

Mercury(II) ions precipitate from aqueous cysteine (H 2 Cys) solutions containing H 2 Cys/Hg(II) mole ratio ≥ 2.0 as Hg( S -HCys) 2 . In absence of additional cysteine, the precipitate dissolves at pH ~12 with the [Hg( S,N -Cys) 2 ] 2- complex dominating. With excess cysteine (H 2 Cys/Hg(II) mole ratio ≥ 4.0), higher complexes form and the precipitate dissolves at lower pH values. Previously, we found that tetrathiolate [Hg( S -Cys) 4 ] 6- complexes form at pH = 11.0; in this work we extend the investigation to pH values of physiological interest. We examined two series of Hg(II)-cysteine solutions in which C Hg(II) varied between 8 - 9 mM and 80 - 100 mM, respectively, with H 2 Cys/Hg(II) mole ratios from 4 to ~20. The solutions were prepared in the pH range 7.1 - 8.8, at the pH at which the initial Hg( S -HCys) 2 precipitate dissolved. The variations in the Hg(II) speciation were followed by 199 Hg NMR, X-ray absorption and Raman spectroscopic techniques. Our results show that in the dilute solutions ( C Hg(II) = 8 - 9 mM), mixtures of di-, tri- (major) and tetrathiolate complexes exist at moderate cysteine excess ( C H2Cys ~ 0.16 M) at pH 7.1. In the more concentrated solutions ( C Hg(II) = 80 - 100 mM) with high cysteine excess ( C H2Cys > 0.9 M), tetrathiolate [Hg( S -cysteinate) 4 ] m -6 ( m = 0 - 4) complexes dominate in the pH range 7.3 - 7.8, with lower charge than for the [Hg( S -Cys) 4 ] 6- complex due to protonation of some ( m ) of the amino groups of the coordinated cysteine ligands. The results of this investigation could provide a key to the mechanism of biosorption and accumulation of Hg(II) ions in biological / environmental systems.

Transamination of cysteine-sulfinic acid by extracts of oat leaves

International Nuclear Information System (INIS)

Perez-Milan, H.; Schuack, J.; Fromageot, P.

1960-01-01

An aqueous extract of oat leaves catalyses a transamination between cysteine-sulfinic acid and α-ketoglutaric acid. Under the conditions utilized pyruvic acid is not an acceptor of the amino group. Neither cysteic nor aspartic acid are a substrate for the transaminase of cysteine-sulfinic acid. Reprint of a paper published in Biochimica et Biophysica Acta, Vol. 36, 1959, p. 73-83 [fr

Reversible targeting of noncatalytic cysteines with chemically tuned electrophiles

DEFF Research Database (Denmark)

Serafimova, Iana M; Pufall, Miles A; Krishnan, Shyam

2012-01-01

Targeting noncatalytic cysteine residues with irreversible acrylamide-based inhibitors is a powerful approach for enhancing pharmacological potency and selectivity. Nevertheless, concerns about off-target modification motivate the development of reversible cysteine-targeting strategies. Here we...... of these electrophiles into a noncovalent kinase-recognition scaffold produced slowly dissociating, covalent inhibitors of the p90 ribosomal protein S6 kinase RSK2. A cocrystal structure revealed specific noncovalent interactions that stabilize the complex by positioning the electrophilic carbon near the targeted...

L-Cysteine inhibits root elongation through auxin/PLETHORA and SCR/SHR pathway in Arabidopsis thaliana.

Science.gov (United States)

Wang, Zhen; Mao, Jie-Li; Zhao, Ying-Jun; Li, Chuan-You; Xiang, Cheng-Bin

2015-02-01

L-Cysteine plays a prominent role in sulfur metabolism of plants. However, its role in root development is largely unknown. Here, we report that L-cysteine reduces primary root growth in a dosage-dependent manner. Elevating cellular L-cysteine level by exposing Arabidopsis thaliana seedlings to high L-cysteine, buthionine sulphoximine, or O-acetylserine leads to altered auxin maximum in root tips, the expression of quiescent center cell marker as well as the decrease of the auxin carriers PIN1, PIN2, PIN3, and PIN7 of primary roots. We also show that high L-cysteine significantly reduces the protein level of two sets of stem cell specific transcription factors PLETHORA1/2 and SCR/SHR. However, L-cysteine does not downregulate the transcript level of PINs, PLTs, or SCR/SHR, suggesting that an uncharacterized post-transcriptional mechanism may regulate the accumulation of PIN, PLT, and SCR/SHR proteins and auxin transport in the root tips. These results suggest that endogenous L-cysteine level acts to maintain root stem cell niche by regulating basal- and auxin-induced expression of PLT1/2 and SCR/SHR. L-Cysteine may serve as a link between sulfate assimilation and auxin in regulating root growth. © 2014 Institute of Botany, Chinese Academy of Sciences.

The mechanism of cysteine detection in biological media by means of vanadium oxide nanoparticles

International Nuclear Information System (INIS)

Bezerra, A. G.; Barison, A.; Oliveira, V. S.; Foti, L.; Krieger, M. A.; Dhalia, R.; Viana, I. F. T.; Schreiner, W. H.

2012-01-01

We report on the interaction of vanadate nanoparticles, produced using the laser ablation in liquids synthesis, with cysteine in biological molecules. Cysteine is a very important amino acid present in most proteins, but also because cysteine and the tripeptide glutathione are the main antioxidant molecules in our body system. Detailed UV–Vis absorption spectra and dynamic light scattering measurements were done to investigate the detection of cysteine in large biological molecules. The intervalence band of the optical absorption spectra shows capability for quantitative cysteine sensing in the μM range in biological macromolecules. Tests included cytoplasmic repetitive antigen and flagellar repetitive antigen proteins of the Trypanosoma cruzi protozoa, as well as the capsid p24 proteins from Human Immunodeficiency Virus type 1 and type 2. Detailed NMR measurements for hydrogen, carbon, and vanadium nuclei show that cysteine in contact with the vanadate looses hydrogen of the sulphydryl side chain, while the vanadate is reduced. The subsequent detachment of two deprotonated molecules to form cystine and the slow return to the vanadate complete the oxidation–reduction cycle. Therefore, the vanadate acts as a charge exchanging catalyst on cysteine to form cystine. The NMR results also indicate that the nanoparticles are not formed by the common orthorhombic V 2 O 5 form.

The mechanism of cysteine detection in biological media by means of vanadium oxide nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Bezerra, A. G. [Universidade Tecnologica Federal do Parana, Departamento Academico de Fisica (Brazil); Barison, A. [Universidade Federal do Parana, Departamento de Quimica (Brazil); Oliveira, V. S. [Universidade Federal do Parana, Departamento de Fisica (Brazil); Foti, L.; Krieger, M. A. [Fundacao Oswaldo Cruz, Instituto de Biologia Molecular do Parana (Brazil); Dhalia, R.; Viana, I. F. T. [Fundacao Oswaldo Cruz, Centro de Pesquisas Aggeu Magalhaes (Brazil); Schreiner, W. H., E-mail: wido@fisica.ufpr.br [Universidade Federal do Parana, Departamento de Fisica (Brazil)

2012-09-15

We report on the interaction of vanadate nanoparticles, produced using the laser ablation in liquids synthesis, with cysteine in biological molecules. Cysteine is a very important amino acid present in most proteins, but also because cysteine and the tripeptide glutathione are the main antioxidant molecules in our body system. Detailed UV-Vis absorption spectra and dynamic light scattering measurements were done to investigate the detection of cysteine in large biological molecules. The intervalence band of the optical absorption spectra shows capability for quantitative cysteine sensing in the {mu}M range in biological macromolecules. Tests included cytoplasmic repetitive antigen and flagellar repetitive antigen proteins of the Trypanosoma cruzi protozoa, as well as the capsid p24 proteins from Human Immunodeficiency Virus type 1 and type 2. Detailed NMR measurements for hydrogen, carbon, and vanadium nuclei show that cysteine in contact with the vanadate looses hydrogen of the sulphydryl side chain, while the vanadate is reduced. The subsequent detachment of two deprotonated molecules to form cystine and the slow return to the vanadate complete the oxidation-reduction cycle. Therefore, the vanadate acts as a charge exchanging catalyst on cysteine to form cystine. The NMR results also indicate that the nanoparticles are not formed by the common orthorhombic V{sub 2}O{sub 5} form.

Replication of murine coronavirus requires multiple cysteines in the endodomain of spike protein

International Nuclear Information System (INIS)

Yang, Jinhua; Lv, Jun; Wang, Yuyan; Gao, Shuang; Yao, Qianqian; Qu, Di; Ye, Rong

2012-01-01

A conserved cysteine-rich motif located between the transmembrane domain and the endodomain is essential for membrane fusion and assembly of coronavirus spike (S) protein. Here, we proved that three cysteines within the motif, but not dependent on position, are minimally required for the survival of the recombinant mouse hepatitis virus. When the carboxy termini with these mutated motifs of S proteins were respectively introduced into a heterogeneous protein, both incorporation into lipid rafts and S-palmitoylation of these recombinant proteins showed a similar quantity requirement to cysteine residues. Meanwhile, the redistribution of these proteins on cellular surface indicated that the absence of the positively charged rather than cysteine residues in the motif might lead the dramatic reduction in syncytial formation of some mutants with the deleted motifs. These results suggest that multiple cysteine as well as charged residues concurrently improves the membrane-associated functions of S protein in viral replication and cytopathogenesis.

Replication of murine coronavirus requires multiple cysteines in the endodomain of spike protein

Energy Technology Data Exchange (ETDEWEB)

Yang, Jinhua; Lv, Jun; Wang, Yuyan; Gao, Shuang; Yao, Qianqian; Qu, Di; Ye, Rong, E-mail: yerong24@fudan.edu.cn

2012-06-05

A conserved cysteine-rich motif located between the transmembrane domain and the endodomain is essential for membrane fusion and assembly of coronavirus spike (S) protein. Here, we proved that three cysteines within the motif, but not dependent on position, are minimally required for the survival of the recombinant mouse hepatitis virus. When the carboxy termini with these mutated motifs of S proteins were respectively introduced into a heterogeneous protein, both incorporation into lipid rafts and S-palmitoylation of these recombinant proteins showed a similar quantity requirement to cysteine residues. Meanwhile, the redistribution of these proteins on cellular surface indicated that the absence of the positively charged rather than cysteine residues in the motif might lead the dramatic reduction in syncytial formation of some mutants with the deleted motifs. These results suggest that multiple cysteine as well as charged residues concurrently improves the membrane-associated functions of S protein in viral replication and cytopathogenesis.

Examination of commercially available copper oxide wire particles in combination with albendazole for control of gastrointestinal nematodes in lambs.

Science.gov (United States)

Burke, J M; Miller, J E; Terrill, T H; Smyth, E; Acharya, M

2016-01-15

Control of gastrointestinal nematodes (GIN) remains a critical issue due to the prevalence of anthelmintic resistance. The objective of the experiment was to determine the efficacy of copper oxide wire particles (COWP) from three commercial sources and a combination of COWP and albendazole to control GIN and/or Haemonchus contortus in lambs. Naturally infected Katahdin lambs in early June 2014 and 2015 were randomly assigned to receive no COWP (CON; n=9 and 12) or 2g COWP in a gel capsule as Copasure(®) (COP; n=4 and 17; Animax Ltd.), copper oxide-wire form (AUS; n=7 in 2014 only; Pharmplex), Ultracruz™ (ULT; n=8 and 15; Santa Cruz Animal Health™), no COWP and albendazole (CON+alb; n=10 in 2015 only; 15mg/kg BW; Valbazen(®); Zoetis Animal Health), or COWP+alb (n=7 and 11; in 2014, lambs were administered alb on day 3). Lambs grazed grass pastures as a group and were supplemented with 227g/lamb daily of a commercial grain mix (15% crude protein) and the same amount of alfalfa pellets. Feces were collected on days 0 (day of COWP treatment), 7, and 14 for determination of fecal egg counts (FEC). Pooled (2014) or pooled treatment group feces were cultured on days 0, 7, and 14 (2015 only) to determine GIN genera. Data were analyzed using repeated measures in a mixed model, and FEC were log transformed. The predominant GIN on day 0 was H. contortus (87%) in 2014, and there was a mixed population in 2015. The mean FEC was reduced by day 7 in AUS and ULT lambs (treatment×day, P=0.001), and all of the COWP products were similar. By day 14, the AUS FEC were lower than the CON and COP groups. When examining the combination of COWP and synthetic anthelmintic, the FEC of COWP+alb were reduced to nearly 0eggs/g (back-transformed) and lower than the other groups (treatment×day, P=0.001). The percentage of H. contortus in cultured feces was reduced to a greater extent in the COWP than CON or CON+alb groups of lambs. In a mixed GIN population, the COWP products appeared to

Dealing with the sulfur part of cysteine: four enzymatic steps degrade l-cysteine to pyruvate and thiosulfate in Arabidopsis mitochondria.

Science.gov (United States)

Höfler, Saskia; Lorenz, Christin; Busch, Tjorven; Brinkkötter, Mascha; Tohge, Takayuki; Fernie, Alisdair R; Braun, Hans-Peter; Hildebrandt, Tatjana M

2016-07-01

Amino acid catabolism is essential for adjusting pool sizes of free amino acids and takes part in energy production as well as nutrient remobilization. The carbon skeletons are generally converted to precursors or intermediates of the tricarboxylic acid cycle. In the case of cysteine, the reduced sulfur derived from the thiol group also has to be oxidized in order to prevent accumulation to toxic concentrations. Here we present a mitochondrial sulfur catabolic pathway catalyzing the complete oxidation of l-cysteine to pyruvate and thiosulfate. After transamination to 3-mercaptopyruvate, the sulfhydryl group from l-cysteine is transferred to glutathione by sulfurtransferase 1 and oxidized to sulfite by the sulfur dioxygenase ETHE1. Sulfite is then converted to thiosulfate by addition of a second persulfide group by sulfurtransferase 1. This pathway is most relevant during early embryo development and for vegetative growth under light-limiting conditions. Characterization of a double mutant produced from Arabidopsis thaliana T-DNA insertion lines for ETHE1 and sulfurtransferase 1 revealed that an intermediate of the ETHE1 dependent pathway, most likely a persulfide, interferes with amino acid catabolism and induces early senescence. © 2016 Scandinavian Plant Physiology Society.

Preparation, Crystallization and X-ray Diffraction Analysis to 1.5 A Resolution of Rat Cysteine Dioxygenase, a Mononuclear Iron Enzyme Responsible for Cysteine Thiol Oxidation

Energy Technology Data Exchange (ETDEWEB)

Simmons,C.; Hao, Q.; Stipanuk, M.

2005-01-01

Cysteine dioxygenase (CDO; EC 1.13.11.20) is an {approx}23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O2, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Angstroms resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Angstrom, {alpha} = {beta} = {gamma} = 90. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative.

Dietary L-cysteine improves the antioxidative potential and lipid metabolism in rats fed a normal diet.

Science.gov (United States)

Lee, Seulki; Han, Kyu-Ho; Nakamura, Yumi; Kawakami, Sakura; Shimada, Ken-ichiro; Hayakawa, Touru; Onoue, Hirotake; Fukushima, Michihiro

2013-01-01

L-cysteine works as a precursor of the antioxidant, glutathione. We investigated the effects of L-cysteine (1% and 2%) on lipid metabolism and the antioxidative system in rats fed a normal diet. Administering L-cysteine dependently decreased the food intake, fat mass weight and body weight dose. Dietary L-cysteine also decreased the triglyceride levels in the serum and liver. However, there were no significant differences in the hepatic TBARS and glutathione (GSH) levels among the groups. The activities of catalase and glutathione reductase in the rats receiving 2% L-cysteine were significantly higher (pL-cysteine dose-dependently affected the antioxidative enzyme activities, and the lipid levels in the serum and liver which might be related to the reduced food intake.

Enhancement of L-cysteine production by disruption of yciW in Escherichia coli.

Science.gov (United States)

Kawano, Yusuke; Ohtsu, Iwao; Takumi, Kazuhiro; Tamakoshi, Ai; Nonaka, Gen; Funahashi, Eri; Ihara, Masaki; Takagi, Hiroshi

2015-02-01

Using in silico analysis, the yciW gene of Escherichia coli was identified as a novel L-cysteine regulon that may be regulated by the transcriptional activator CysB for sulfur metabolic genes. We found that overexpression of yciW conferred tolerance to L-cysteine, but disruption of yciW increased L-cysteine production in E. coli. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Activation of human acid sphingomyelinase through modification or deletion of C-terminal cysteine.

Science.gov (United States)

Qiu, Huawei; Edmunds, Tim; Baker-Malcolm, Jennifer; Karey, Kenneth P; Estes, Scott; Schwarz, Cordula; Hughes, Heather; Van Patten, Scott M

2003-08-29

One form of Niemann-Pick disease is caused by a deficiency in the enzymatic activity of acid sphingomyelinase. During efforts to develop an enzyme replacement therapy based on a recombinant form of human acid sphingomyelinase (rhASM), purified preparations of the recombinant enzyme were found to have substantially increased specific activity if cell harvest media were stored for several weeks at -20 degrees C prior to purification. This increase in activity was found to correlate with the loss of the single free thiol on rhASM, suggesting the involvement of a cysteine residue. It was demonstrated that a variety of chemical modifications of the free cysteine on rhASM all result in substantial activation of the enzyme, and the modified cysteine responsible for this activation was shown to be the C-terminal residue (Cys629). Activation was also achieved by copper-promoted dimerization of rhASM (via cysteine) and by C-terminal truncation using carboxypeptidase Y. The role of the C-terminal cysteine in activation was confirmed by creating mutant forms of rhASM in which this residue was either deleted or replaced by a serine, with both forms having substantially higher specific activity than wild-type rhASM. These results indicate that purified rhASM can be activated in vitro by loss of the free thiol on the C-terminal cysteine via chemical modification, dimerization, or deletion of this amino acid residue. This method of activation is similar to the cysteine switch mechanism described previously for matrix metalloproteinases and could represent a means of posttranslational regulation of ASM activity in vivo.

[Plant signaling peptides. Cysteine-rich peptides].

Science.gov (United States)

Ostrowski, Maciej; Kowalczyk, Stanisław

2015-01-01

Recent bioinformatic and genetic analyses of several model plant genomes have revealed the existence of a highly abundant group of signaling peptides that are defined as cysteine-rich peptides (CRPs). CRPs are usually in size between 50 and 90 amino acid residues, they are positively charged, and they contain 4-16 cysteine residues that are important for the correct conformational folding. Despite the structural differences among CRP classes, members from each class have striking similarities in their molecular properties and function. The present review presents the recent progress in research on signaling peptides from several families including: EPF/EPFL, SP11/SCR, PrsS, RALF, LURE, and some other peptides belonging to CRP group. There is convincing evidence indicating multiple roles for these CRPs as signaling molecules during the plant life cycle, ranging from stomata development and patterning, self-incompatibility, pollen tube growth and guidance, reproductive processes, and nodule formation.

Interaction between copper oxide wire particles and Duddingtonia flagrans in lambs.

Science.gov (United States)

Burke, J M; Miller, J E; Larsen, M; Terrill, T H

2005-11-25

An experiment was completed to determine if copper oxide wire particles (COWP) had any effect on the activity of the nematode-trapping fungus Duddingtonia flagrans in growing lambs. COWP has been used recently as a dewormer in small ruminants because of nematode resistance to anthelmintics. D. flagrans has been used to control free-living stages of parasitic nematodes in livestock. Katahdin and Dorper lambs, 4 months of age, were administered no or 4 g COWP (n=24/dose) in early October 2003. Haemonchus contortus was the predominant gastrointestinal parasite during the trial, which was acquired naturally from pasture. Half the lambs from each COWP group were supplemented with corn/soybean meal with or without D. flagrans for 35 days. Fecal egg counts (FEC) and packed cell volume (PCV) were determined weekly between days 0 (day of COWP administration) and 35. Feces from lambs in each treatment group were pooled and three replicates per group were cultured for 14 days at room temperature. Larvae (L3) were identified and counted per gram of feces cultured. Treatment with COWP was effective in decreasing FEC, which remained low compared with FEC from lambs not treated with COWP. This led to an increase in PCV in these lambs (COWP x day, Pcopper on H. contortus, and the additional larval reducing effect exerted by the nematode destroying fungus D. flagrans, the expected result would be a much lower larval challenge on pasture when these two tools are used together in a sustainable control strategy.

Administration of copper oxide wire particles in a capsule or feed for gastrointestinal nematode control in goats.

Science.gov (United States)

Burke, J M; Soli, F; Miller, J E; Terrill, T H; Wildeus, S; Shaik, S A; Getz, W R; Vanguru, M

2010-03-25

Widespread anthelmintic resistance in small ruminants has necessitated alternative means of gastrointestinal nematode (GIN) control. The objective was to determine the effectiveness of copper oxide wire particles (COWP) administered as a gelatin capsule or in a feed supplement to control GIN in goats. In four separate experiments, peri-parturient does (n=36), yearling does (n=25), weaned kids (n=72), and yearling bucks (n=16) were randomly assigned to remain untreated or administered 2g COWP in a capsule (in Experiments 1, 2, and 3) or feed supplement (all experiments). Feces and blood were collected every 7 days between Days 0 and 21 (older goats) or Day 42 (kids) for fecal egg counts (FEC) and blood packed cell volume (PCV) analyses. A peri-parturient rise in FEC was evident in the untreated does, but not the COWP-treated does (COWP x date, P<0.02). In yearling does, FEC of the COWP-treated does tended to be lower than the untreated (COWP, P<0.02). FEC of COWP-treated kids were reduced compared with untreated kids (COWP x date, P<0.001). FEC of treated and untreated bucks were similar, but Haemonchus contortus was not the predominant nematode in these goats. However, total worms were reduced in COWP-fed bucks (P<0.03). In summary, it appeared that COWP in the feed was as effective as COWP in a gelatin capsule to reduce FEC in goats. COWP administration may have a limited effect where H. contortus is not the predominant nematode.

Decline in faecal worm egg counts in lambs suckling ewes treated with lipophilic anthelmintics: implications for hastening development of anthelmintic resistance.

Science.gov (United States)

Dever, M L; Kahn, L P

2015-04-30

The aim for this experiment was to look for evidence of milk transfer of anthelmintic actives from ewes to their suckling lambs by reference to lambs' faecal worm egg count (WEC). The hypothesis was that WEC will decline in lambs suckling ewes treated with anthelmintics known to be lipophilic. One group of lactating Border Leicester×Merino ewes were treated (TX) with a combination of short (2.5mg/kg monepantel) and long-acting (1mg/kg moxidectin long-acting injection and a sustained release of 4.62g albendazole over 100 days) anthelmintics to remove gastrointestinal nematode (GIN) burden on day 0. The other group of lactating ewes (UTX) and all lambs (White Suffolk sires) were not treated. Ewes and lambs grazed as a single group and were exposed to GIN (predominately Haemonchus contortus) infection from pasture. Measurements were taken on days 0 and 7. WEC of lambs suckling UTX ewes increased from 6441 to 10,341 eggs per gram (epg) between days 0 and 7, while there was a 51% reduction in WEC for lambs suckling TX ewes. Packed cell volume (PCV) was significantly higher for lambs suckling TX ewes on day 7 compared to lambs suckling UTX ewes (28.5% vs. 24.9%, p=0.039). These results suggest that lambs suckling ewes treated with lipophilic anthelmintics received a sub-therapeutic dose via milk which would increase selection within the GIN (H. contortus) population for anthelmintic resistance. Copyright © 2015 Elsevier B.V. All rights reserved.

L-Cysteine and L-AP4 microinjections in the rat caudal ventrolateral medulla decrease arterial blood pressure.

Science.gov (United States)

Takemoto, Yumi

2014-12-01

The thiol amino acid L-cysteine increases arterial blood pressure (ABP) when injected into the cerebrospinal fluid space in conscious rats, indicating a pressor response to centrally acting L-cysteine. A prior synaptic membrane binding assay suggests that L-cysteine has a strong affinity for the L-2-amino-4-phosphonobutyric acid (L-AP4) binding site. The central action of L-cysteine may be vial-AP4 sensitive receptors. The present study investigated cardiovascular responses to L-cysteine and L-ap4 microinjected into the autonomic area of the caudal ventrolateral medulla (CVLM) where inhibitory neurons regulate ABP via pre-sympathetic vasomotor neurons. Both the injection of L-cysteine and L-AP4 in the CVLM sites identified with L-glutamate produced the same depressor and bradycardic responses in urethane-anesthetized rats. Neither a prior antagonist microinjection of MK801 for the N-methyl-D-aspartate (NMDA) receptor nor CNQX for the non-NMDA receptor attenuated the responses to L-cysteine, but the combination of the two receptor blocking with an additional prior injection abolished the response. In contrast, either receptor blockade alone abolished the response to L-AP4, indicating distinct mechanisms between responses to L-cysteine and L-AP4 in the CVLM. The results indicate that the CVLM is a central active site for L-cysteine's cardiovascular response. Central L-cysteine's action could be independent of the L-AP4 sensitive receptors. Cardiovascular regulation may involve endogenous L-cysteine in the CVLM. Further multidisciplinary examinations are required to elaborate on L-cysteine's functional roles in the CVLM. Copyright © 2014 Elsevier B.V. All rights reserved.

Lifespan extension and increased resistance to environmental stressors by N-Acetyl-L-Cysteine in Caenorhabditis elegans

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Seung-Il Oh

2015-05-01

Full Text Available OBJECTIVE: This study was performed to determine the effect of N-acetyl-L-cysteine, a modified sulfur-containing amino acid that acts as a strong cellular antioxidant, on the response to environmental stressors and on aging in C. elegans. METHOD: The survival of worms under oxidative stress conditions induced by paraquat was evaluated with and without in vivo N-acetyl-L-cysteine treatment. The effect of N-acetyl-L-cysteine on the response to other environmental stressors, including heat stress and ultraviolet irradiation (UV, was also monitored. To investigate the effect on aging, we examined changes in lifespan, fertility, and expression of age-related biomarkers in C. elegans after N-acetyl-L-cysteine treatment. RESULTS: Dietary N-acetyl-L-cysteine supplementation significantly increased resistance to oxidative stress, heat stress, and UV irradiation in C. elegans. In addition, N-acetyl-L-cysteine supplementation significantly extended both the mean and maximum lifespan of C. elegans. The mean lifespan was extended by up to 30.5% with 5 mM N-acetyl-L-cysteine treatment, and the maximum lifespan was increased by 8 days. N-acetyl-L-cysteine supplementation also increased the total number of progeny produced and extended the gravid period of C. elegans. The green fluorescent protein reporter assay revealed that expression of the stress-responsive genes, sod-3 and hsp-16.2, increased significantly following N-acetyl-L-cysteine treatment. CONCLUSION: N-acetyl-L-cysteine supplementation confers a longevity phenotype in C. elegans, possibly through increased resistance to environmental stressors.

Elimination of hydrogen sulphide and {beta} substitution in cystein, catalyzed by the cysteine-lyase of hens yolk-sac and yolk (1961); Desulfhydration et {beta} substitution de la cysteine catalysees par la cysteinelyase du sac vitellin et du jaune de l'oeuf de poule (1961)

Energy Technology Data Exchange (ETDEWEB)

Chapeville, F; Fromageot, P [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

1961-07-01

The yolk of incubated hen's eggs contains a pyridoxal phosphate activated enzyme, free of iron, copper, magnesium and calcium. This enzyme activates the {beta}-carbon atom of cysteine. Its reactivity is demonstrated by the ease with which this {beta}-carbon fixes various sulfur containing substances in which the sulfur has reducing properties: inorganic sulfide, sulfide or cysteine itself. In the absence of substances able to react with the {beta}-carbon atom, the active complex, consisting of the enzyme and the aminated tri-carbon chain, is hydrolysed to pyruvic acid and ammonia. The liberation of hydrogen sulfide thus appears to be the consequence either of the substitution of the {beta}-carbon atom of cysteine or of the decomposition of the complex which this aminoacid forms with the enzyme studied. The latter seems therefore to possess an activity which differs from the activity of the desulfhydrases as yet known. We suggest to call this enzyme cystein-lyase. (authors) [French] Le sac vitellin et le jaune d'oeufs embryonnes de poule renferment une enzyme activee par le phosphate de pyridoxal, qui ne contient pas de fer, de magnesium, de cuivre ce de calcium et qui confere une reactivite particuliere au carbone {beta} de la cysteine. Cette reactivite se manifeste par l'aptitude que possede le carbone {beta} a fixer diverses molecules soufrees dont le soufre est reducteur, telles que le sulfure, le sulfite ou la cysteine elle-meme. En l'absence de reactifs capables de reagir avec le carbone {beta}, le complexe actif enzyme-chaine tricarbonee et aminee s'hydrolyse en acide pyruvique et en ammoniaque. La liberation d'hydrogene sulfure apparait ainsi comme une consequence soit de la substitution du carbone {beta} de la cysteine, soit de la decomposition du complexe qu'elle forme avec l'enzyme etudiee. Cette derniere semble donc posseder une activite distincte de celle des desulfhydrases connues jusqu'a present. Nous proposons de l'appeler cysteinelyase. (auteurs)

Interaction of cysteine and copper ions on the surface of iron: EIS, polarization and XPS study

International Nuclear Information System (INIS)

El-Deab, Mohamed S.

2011-01-01

Highlights: → The current study demonstrates a comprehensive study for Cysteine + Cu(II) ions as an efficient inhibitor as demonstrated by EIS, XPS and potentiodynamic polarization measurements, in addition to traditional weight loss measurements. → The novelty of the current work originates from the combined use of an eco-friendly compound (i.e., cysteine) with a minute amount of copper ions (in the micro molar range) as a corrosion inhibitor for low carbon steel in acidic medium. To this end, cysteine shows only moderate inhibition ca. 60% for iron which jumps up to more than 95% in the presence of micro molar range of Cu(II) ions. → Cysteine-Cu(II) blends are found superior to benzotriazole (BTAH)-Cu(II) blends in terms of their long-term stability in addition to the avoidance of the use of the well-reported highly toxic BTAH. - Abstract: This study addresses the enhancing effect of copper ions on the inhibition efficiency (IE) of cysteine (an eco-friendly compound) against the corrosion of iron in 0.5 M sulphuric acid. Electrochemical impedance spectroscopy (EIS) data revealed a significant increase in the polarization resistance (R p ) of the iron/solution interface in the presence of cysteine and Cu(II) ions instead of cysteine alone. That is, IE of 95% is obtained in the presence of 5 mM cysteine and 25 μM Cu(II) ions, compared to 66% in absence of Cu(II) ions. Moreover, electrochemical polarization measurements indicate that cysteine and Cu(II) ions blends act as mixed-type inhibitors for the corrosion of iron. The formation of Cu(I)-cysteinate complex and/or cysteine SAM at Cu atop the iron surface (as evident from X-ray photoelectron spectroscopy (XPS)) blocks the underlying iron surface and imparts a pronounced protection against its corrosion. IE of cysteine-Cu(II) blend remains effectively unchanged with immersion time indicating its high stability in the used acidic medium.

7-cysteine-pyrrole conjugate: A new potential DNA reactive metabolite of pyrrolizidine alkaloids.

Science.gov (United States)

He, Xiaobo; Xia, Qingsu; Ma, Liang; Fu, Peter P

2016-01-01

Pyrrolizidine alkaloids (PAs) require metabolic activation to exert cytotoxicity, genotoxicity, and tumorigenicity. We previously reported that (±)-6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP)-derived DNA adducts are responsible for PA-induced liver tumor formation in rats. In this study, we determined that metabolism of riddelliine and monocrotaline by human or rat liver microsomes produced 7-cysteine-DHP and DHP. The metabolism of 7-glutathionyl-DHP by human and rat liver microsomes also generated 7-cysteine-DHP. Further, reaction of 7-cysteine-DHP with calf thymus DNA in aqueous solution yielded the described DHP-derived DNA adducts. This study represents the first report that 7-cysteine-DHP is a new PA metabolite that can lead to DNA adduct formation.

Synthesis and characterization of a cysteine xyloglucan conjugate as mucoadhesive polymer

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Mangesh Bhalekar

2013-06-01

Full Text Available The aim of this study was to improve the mucoadhesive potential of xyloglucan polymer by the covalent attachment of cysteine as thiol moiety. The parent polymer xyloglucan was chemically modified by introducing sulphydryl bearing compound L-cysteine HCl. Different batches of xyloglucan-cysteine conjugates were prepared at varying reaction pH (2-6 and evaluated for optimum thiol incorporation, disulphide group content, swelling behavior, rheological properties and mucoadhesive properties. The obtained conjugates characterized in vitro by quantification of immobilized thiol groups; showed maximum thiol incorporation on xyloglucan (7.67 ± 0.14 % at pH 5. The disulphide group content was found maximum (2.83 ± 0.12 at pH 6. The water uptake at end of 4 h was 5.0 for xyloglucan and was found to decrease in thiolated derivatives with increase in thiolation. Mucoadhesion studies revealed that mucoadhesion of xyloglucan-cysteine conjugate increased more than twice compared to the unmodified polymer. The viscosity of thiomer was more than that of xyloglucan because of formation of disulphide bonds.

Development of a cysteine-deprived and C-terminally truncated GLP-1 receptor

DEFF Research Database (Denmark)

Underwood, Christina Rye; Knudsen, Lotte Bjerre; Garibay, Patrick W.

2013-01-01

The glucagon-like peptide-1 receptor (GLP-1R) belongs to family B of the G-protein coupled receptors (GPCRs), and has become a promising target for the treatment of type 2 diabetes. Here we describe the development and characterization of a fully functional cysteine-deprived and C......-terminally truncated GLP-1R. Single cysteines were initially substituted with alanine, and functionally redundant cysteines were subsequently changed simultaneously. Our results indicate that Cys174, Cys226, Cys296 and Cys403 are important for the GLP-1-mediated response, whereas Cys236, Cys329, Cys341, Cys347, Cys438...... that the membrane proximal part of the C-terminal is involved in receptor expression at the cell surface. The results show that seven cysteines and more than half of the C-terminal tail can be removed from GLP-1R without compromising GLP-1 binding or function....

Transport and activation of S-(1,2-dichlorovinyl)-L-cysteine and N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine in rat kidney proximal tubules

International Nuclear Information System (INIS)

Zhang, G.H.; Stevens, J.L.

1989-01-01

An important step in understanding the mechanism underlying the tubular specificity of the nephrotoxicity of toxic cysteine conjugates is to identify the rate-limiting steps in their activation. The rate-limiting steps in the activation of toxic cysteine conjugates were characterized using isolated proximal tubules from the rat and 35S-labeled S-(1,2-dichlorovinyl)-L-cysteine (DCVC) and N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine (NAC-DCVC) as model compounds. The accumulation by tubules of 35S radiolabel from both DCVC and NAC-DCVC was time and temperature dependent and was mediated by both Na+-dependent and independent processes. Kinetic studies with DCVC in the presence of sodium revealed the presence of two components with apparent Km and Vmax values of (1) 46 microM and 0.21 nmol/mg min and (2) 2080 microM and 7.3 nmol/mg.min. NAC-DVVC uptake was via a single system with apparent Km and Vmax values of 157 microM and 0.65 nmol/mg.min, respectively. Probenecid, an inhibitor of the renal organic anion transport system, inhibited accumulation of radiolabel from NAC-DCVC, but not from DCVC. The covalent binding of 35S label to cellular macromolecules was much greater from [35S]DCVC than from NAC-[35S]DCVC. Analysis of metabolites showed that a substantial amount of the cellular NAC-[35S]DCVC was unmetabolized while [35S]DCVC was rapidly metabolized to bound 35S-labeled material and unidentified products. The data suggest that DCVC is rapidly metabolized following transport, but that activation of NAC-DCVC depends on a slower rate of deacetylation. The results are discussed with regard to the segment specificity of cysteine conjugate toxicity and the role of disposition in vivo in the nephrotoxicity of glutathione conjugates

Cysteine Cathepsins as Regulators of the Cytotoxicity of NK and T Cells

Science.gov (United States)

Perišić Nanut, Milica; Sabotič, Jerica; Jewett, Anahid; Kos, Janko

2014-01-01

Cysteine cathepsins are lysosomal peptidases involved at different levels in the processes of the innate and adaptive immune responses. Some, such as cathepsins B, L, and H are expressed constitutively in most immune cells. In cells of innate immunity they play a role in cell adhesion and phagocytosis. Other cysteine cathepsins are expressed more specifically. Cathepsin X promotes dendritic cell maturation, adhesion of macrophages, and migration of T cells. Cathepsin S is implicated in major histocompatibility complex class II antigen presentation, whereas cathepsin C, expressed in cytotoxic T lymphocytes and natural killer (NK) cells, is involved in processing pro-granzymes into proteolytically active forms, which trigger cell death in their target cells. The activity of cysteine cathepsins is controlled by endogenous cystatins, cysteine protease inhibitors. Of these, cystatin F is the only cystatin that is localized in endosomal/lysosomal vesicles. After proteolytic removal of its N-terminal peptide, cystatin F becomes a potent inhibitor of cathepsin C with the potential to regulate pro-granzyme processing and cell cytotoxicity. This review is focused on the role of cysteine cathepsins and their inhibitors in the molecular mechanisms leading to the cytotoxic activity of T lymphocytes and NK cells in order to address new possibilities for regulation of their function in pathological processes. PMID:25520721

Cysteine proteases: Modes of activation and future prospects as pharmacological targets

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Sonia eVerma

2016-04-01

Full Text Available Proteolytic enzymes are crucial for a variety of biological processes in organisms ranging from lower (virus, bacteria and parasite to the higher organisms (mammals. Proteases cleave proteins into smaller fragments by catalyzing peptide bonds hydrolysis. Proteases are classified according to their catalytic site, and distributed into four major classes: cysteine proteases, serine proteases, aspartic proteases and metallo-proteases. This review will cover only cysteine proteases, papain family enzymes which are involved in multiple functions such as extracellular matrix turnover, antigen presentation, processing events, digestion, immune invasion, hemoglobin hydrolysis, parasite invasion, parasite egress and processing surface proteins. Therefore, they are promising drug targets for various diseases. For preventing unwanted digestion, cysteine proteases are synthesized as zymogens, and contain a pro-domain (regulatory and a mature domain (catalytic. The prodomain acts as an endogenous inhibitor of the mature enzyme. For activation of the mature enzyme, removal of the prodomain is necessary and achieved by different modes. The pro-mature domain interaction can be categorized as protein-protein interactions (PPIs and may be targeted in a range of diseases. Cysteine protease inhibitors are available that can block the active site but no such inhibitor available yet that can be targeted to block the pro-mature domain interactions and prevent it activation. This review specifically highlights the modes of activation (processing of papain family enzymes, which involve auto-activation, trans-activation and also clarifies the future aspects of targeting PPIs to prevent the activation of cysteine proteases.

Detection of l-Cysteine in wheat flour by Raman microspectroscopy combined chemometrics of HCA and PCA.

Science.gov (United States)

Cebi, Nur; Dogan, Canan Ekinci; Develioglu, Ayşen; Yayla, Mediha Esra Altuntop; Sagdic, Osman

2017-08-01

l-Cysteine is deliberately added to various flour types since l-Cysteine has enabled favorable baking conditions such as low viscosity, increased elasticity and rise during baking. In Turkey, usage of l-Cysteine as a food additive isn't allowed in wheat flour according to the Turkish Food Codex Regulation on food additives. There is an urgent need for effective methods to detect l-Cysteine in wheat flour. In this study, for the first time, a new, rapid, effective, non-destructive and cost-effective method was developed for detection of l-Cysteine in wheat flour using Raman microscopy. Detection of l-Cysteine in wheat flour was accomplished successfully using Raman microscopy combined chemometrics of PCA (Principal Component Analysis) and HCA (Hierarchical Cluster Analysis). In this work, 500-2000cm -1 spectral range (fingerprint region) was determined to perform PCA and HCA analysis. l-Cysteine and l-Cystine were determined with detection limit of 0.125% (w/w) in different wheat flour samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of novel malarial cysteine protease inhibitors using structure-based virtual screening of a focused cysteine protease inhibitor library.

Science.gov (United States)

Shah, Falgun; Mukherjee, Prasenjit; Gut, Jiri; Legac, Jennifer; Rosenthal, Philip J; Tekwani, Babu L; Avery, Mitchell A

2011-04-25

Malaria, in particular that caused by Plasmodium falciparum , is prevalent across the tropics, and its medicinal control is limited by widespread drug resistance. Cysteine proteases of P. falciparum , falcipain-2 (FP-2) and falcipain-3 (FP-3), are major hemoglobinases, validated as potential antimalarial drug targets. Structure-based virtual screening of a focused cysteine protease inhibitor library built with soft rather than hard electrophiles was performed against an X-ray crystal structure of FP-2 using the Glide docking program. An enrichment study was performed to select a suitable scoring function and to retrieve potential candidates against FP-2 from a large chemical database. Biological evaluation of 50 selected compounds identified 21 diverse nonpeptidic inhibitors of FP-2 with a hit rate of 42%. Atomic Fukui indices were used to predict the most electrophilic center and its electrophilicity in the identified hits. Comparison of predicted electrophilicity of electrophiles in identified hits with those in known irreversible inhibitors suggested the soft-nature of electrophiles in the selected target compounds. The present study highlights the importance of focused libraries and enrichment studies in structure-based virtual screening. In addition, few compounds were screened against homologous human cysteine proteases for selectivity analysis. Further evaluation of structure-activity relationships around these nonpeptidic scaffolds could help in the development of selective leads for antimalarial chemotherapy.

Cysteine as a non toxic corrosion inhibitor for copper alloys in conservation

DEFF Research Database (Denmark)

Gravgaard, Mari; van Lanschot, Jettie

2012-01-01

studies of colour changes in the corrosion products. The results obtained in this article demonstrate that cysteine could be a non-toxic alternative to BTA. Cysteine performed as well as BTA on pre-corroded coupons with bronze disease in high humidity and showed acceptable results during testing...

Identification, classification and expression pattern analysis of sugarcane cysteine proteinases

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Gustavo Coelho Correa

2001-12-01

Full Text Available Cysteine proteases are peptidyl hydrolyses dependent on a cysteine residue at the active center. The physical and chemical properties of cysteine proteases have been extensively characterized, but their precise biological functions have not yet been completely understood, although it is known that they are involved in a number of events such as protein turnover, cancer, germination, programmed cell death and senescence. Protein sequences from different cysteine proteinases, classified as members of the E.C.3.4.22 sub-sub-class, were used to perform a T-BLAST-n search on the Brazilian Sugarcane Expressed Sequence Tags project (SUCEST data bank. Sequence homology was found with 76 cluster sequences that corresponded to possible cysteine proteinases. The alignments of these SUCEST clusters with the sequence of cysteine proteinases of known origins provided important information about the classification and possible function of these sugarcane enzymes. Inferences about the expression pattern of each gene were made by direct correlation with the SUCEST cDNA libraries from which each cluster was derived. Since no previous reports of sugarcane cysteine proteinases genes exists, this study represents a first step in the study of new biochemical, physiological and biotechnological aspects of sugarcane cysteine proteases.Proteinases cisteínicas são peptidil-hidrolases dependentes de um resíduo de cisteína em seu sítio ativo. As propriedades físico-químicas destas proteinases têm sido amplamente caracterizadas, entretanto suas funções biológicas ainda não foram completamente elucidadas. Elas estão envolvidas em um grande número de eventos, tais como: processamento e degradação protéica, câncer, germinação, morte celular programada e processos de senescência. Diferentes proteinases cisteínicas, classificadas pelo Comitê de Nomenclatura da União Internacional de Bioquímica e Biologia Molecular (IUBMB como pertencentes à sub

Electrochemical behavior of cysteine at a CuGeO3 nanowires modified glassy carbon electrode

International Nuclear Information System (INIS)

Dong Yongping; Pei Lizhai; Chu Xiangfeng; Zhang Wangbing; Zhang Qianfeng

2010-01-01

A CuGeO 3 nanowire modified glassy carbon electrode was fabricated and characterized by scanning electron microscopy. The results of electrochemical impedance spectroscopy reveal that electron transfer through nanowire film is facile compared with that of bare glassy carbon electrode. The modified electrode exhibited a novel electrocatalytic behavior to the electrochemical reactions of L-cysteine in neutral solution, which was not reported previously. Two pairs of semi-reversible electrochemical peaks were observed and assigned to the processes of oxidation/reduction and adsorption/desorption of cysteine at the modified electrode, respectively. The electrochemical response of cysteine is poor in alkaline condition and is enhanced greatly in acidic solution, suggesting that hydrogen ions participate in the electrochemical oxidation process of cysteine. The intensities of two anodic peaks varied linearly with the concentration of cysteine in the range of 1 x 10 -6 to 1 x 10 -3 mol L -1 , which make it possible to sensitive detection of cysteine with the CuGeO 3 nanowire modified electrode. Furthermore, the modified electrode exhibited good reproducibility and stability.

Electrochemical behaviour of dopamine at covalent modified glassy carbon electrode with l-cysteine: preliminary results

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Carlos Alberto Martínez-Huitle

2009-01-01

Full Text Available The surface of glassy carbon (GC electrode has been modified by oxidation of L-cysteine. The covalent modified GC electrode with L-Cysteine has been studied, according the supporting electrolyte used. Favourable interactions between the L-cysteine film and DA enhance the current response compared to that at the Nafion GC and bare GC electrodes, achieving better performances than those other electrodes. This behaviour was as result of the adsorption of the cysteine layer film, compact and uniform formation; depending on L-cysteine solution (phosphate buffer or chloridric acid supporting electrolyte used for modifying GC surface. In cyclic voltammetric measurements, modified electrodes can successfully separate the oxidation/reduction DA peaks in different buffer solutions, but an evident dependence in the response was obtained as function of pH and modified electrode. The modified electrode prepared with L-cysteine/HCl solution was used to obtain the calibration curve and it exhibited a stable and sensitive response to DA. The results are described and discussed in the light of the existing literature.

Alleviative effects of s-allyl cysteine and s-ethyl cysteine on MCD diet-induced hepatotoxicity in mice.

Science.gov (United States)

Lin, Chun-che; Yin, Mei-chin; Liu, Wen-hu

2008-11-01

Alleviative effects of s-allyl cysteine (SAC) and s-ethyl cysteine (SEC) upon methionine and choline deficient (MCD) diet-induced hepatotoxicity in mice were examined. SAC or SEC at 1g/L was added into drinking water for 7 weeks with MCD diet. MCD feeding significantly increased hepatic triglyceride and cholesterol levels, and elevated the activity of glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme, fatty acid synthase (FAS) and 3-hydroxy-3-methylglutaryl coenzyme A reductase (P MCD feeding significantly lowered serum and hepatic glutathione (GSH) levels, increased malondialdehyde (MDA) and oxidized glutathione (GSSG) formation, and suppressed the activity and mRNA expression of glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (P MCD feeding significantly enhanced the mRNA expression of interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, matrix metalloproteinases-9 (MMP-9) and collagen-alpha1 (P MCD-induced hepatotoxicity.

Cysteine: a conditionally essential amino acid in low-birth-weight preterm infants?

NARCIS (Netherlands)

Riedijk, Maaike A.; van Beek, Ron H. T.; Voortman, Gardi; de Bie, Henrica M. A.; Dassel, Anne C. M.; van Goudoever, Johannes B.

2007-01-01

Cyst(e)ine can be synthesized de novo from methionine and serine and is, therefore, a nonessential amino acid in human adults. Several studies have suggested that cyst(e)ine might be a conditionally essential amino acid in preterm infants because of biochemical immaturity. No data are available on

Synthesis of l-cysteine derivatives containing stable sulfur isotopes and application of this synthesis to reactive sulfur metabolome.

Science.gov (United States)

Ono, Katsuhiko; Jung, Minkyung; Zhang, Tianli; Tsutsuki, Hiroyasu; Sezaki, Hiroshi; Ihara, Hideshi; Wei, Fan-Yan; Tomizawa, Kazuhito; Akaike, Takaaki; Sawa, Tomohiro

2017-05-01

Cysteine persulfide is an L-cysteine derivative having one additional sulfur atom bound to a cysteinyl thiol group, and it serves as a reactive sulfur species that regulates redox homeostasis in cells. Here, we describe a rapid and efficient method of synthesis of L-cysteine derivatives containing isotopic sulfur atoms and application of this method to a reactive sulfur metabolome. We used bacterial cysteine syntheses to incorporate isotopic sulfur atoms into the sulfhydryl moiety of L-cysteine. We cloned three cysteine synthases-CysE, CysK, and CysM-from the Gram-negative bacterium Salmonella enterica serovar Typhimurium LT2, and we generated their recombinant enzymes. We synthesized 34 S-labeled L-cysteine from O-acetyl-L-serine and 34 S-labeled sodium sulfide as substrates for the CysK or CysM reactions. Isotopic labeling of L-cysteine at both sulfur ( 34 S) and nitrogen ( 15 N) atoms was also achieved by performing enzyme reactions with 15 N-labeled L-serine, acetyl-CoA, and 34 S-labeled sodium sulfide in the presence of CysE and CysK. The present enzyme systems can be applied to syntheses of a series of L-cysteine derivatives including L-cystine, L-cystine persulfide, S-sulfo-L-cysteine, L-cysteine sulfonate, and L-selenocystine. We also prepared 34 S-labeled N-acetyl-L-cysteine (NAC) by incubating 34 S-labeled L-cysteine with acetyl coenzyme A in test tubes. Tandem mass spectrometric identification of low-molecular-weight thiols after monobromobimane derivatization revealed the endogenous occurrence of NAC in the cultured mammalian cells such as HeLa cells and J774.1 cells. Furthermore, we successfully demonstrated, by using 34 S-labeled NAC, metabolic conversion of NAC to glutathione and its persulfide, via intermediate formation of L-cysteine, in the cells. The approach using isotopic sulfur labeling combined with mass spectrometry may thus contribute to greater understanding of reactive sulfur metabolome and redox biology. Copyright © 2017 Elsevier Inc

Transcriptome Analysis of Maize Immature Embryos Reveals the Roles of Cysteine in Improving Agrobacterium Infection Efficiency

Science.gov (United States)

Liu, Yan; Zhang, Zhiqiang; Fu, Junjie; Wang, Guoying; Wang, Jianhua; Liu, Yunjun

2017-01-01

Maize Agrobacterium-mediated transformation efficiency has been greatly improved in recent years. Antioxidants, such as, cysteine, can significantly improve maize transformation frequency through improving the Agrobacterium infection efficiency. However, the mechanism underlying the transformation improvement after cysteine exposure has not been elucidated. In this study, we showed that the addition of cysteine to the co-cultivation medium significantly increased the Agrobacterium infection efficiency of hybrid HiII and inbred line Z31 maize embryos. Reactive oxygen species contents were higher in embryos treated with cysteine than that without cysteine. We further investigated the mechanism behind cysteine-related infection efficiency increase using transcriptome analysis. The results showed that the cysteine treatment up-regulated 939 genes and down-regulated 549 genes in both Z31 and HiII. Additionally, more differentially expressed genes were found in HiII embryos than those in Z31 embryos, suggesting that HiII was more sensitive to the cysteine treatment than Z31. GO analysis showed that the up-regulated genes were mainly involved in the oxidation reduction process. The up-regulation of these genes could help maize embryos to cope with the oxidative stress stimulated by Agrobacterium infection. The down-regulated genes were mainly involved in the cell wall and membrane metabolism, such as, aquaporin and expansin genes. Decreased expression of these cell wall integrity genes could loosen the cell wall, thereby improving the entry of Agrobacterium into plant cells. This study offers insight into the role of cysteine in improving Agrobacterium-mediated transformation of maize immature embryos. PMID:29089955

Mechanism of S-oxygenation by a cysteine dioxygenase model complex

OpenAIRE

Kumar, Devesh; Sastry, G. Narahari; Goldberg, David P.; de Visser, Sam P.

2011-01-01

In this work we present the first computational study on a biomimetic cysteine dioxygenase model complex, [FeII(LN3S)]+ where LN3S is a tetradentate ligand with a bis(imino)pyridyl scaffold and a pendant arylthiolate group. The reaction mechanism of sulfur dioxygenation with O2 was examined by density functional theory (DFT) methods, and compared to results obtained for cysteine dioxygenase. The reaction proceeds via multistate reactivity patterns on competing singlet, triplet and quintet spi...

The effect of cysteine on electrodeposition of gold nanoparticle

International Nuclear Information System (INIS)

Dolati, A.; Imanieh, I.; Salehi, F.; Farahani, M.

2011-01-01

Highlights: → Cysteine was found as an appropriate additive for electrodeposition of gold nanoparticles. → The deposition mechanism of gold nanoparticle was determined as instantaneous nucleation. → Oxygen reduction on the gold nanoparticle surface was eight times greater than that on the conventional gold deposits. - Abstract: The most applications of gold nanoparticles are in the photo-electronical accessories and bio-chemical sensors. Chloride solution with cysteine additive was used as electrolyte in gold nanoparticles electrodeposition. The nucleation and growing mechanism were studied by electrochemical techniques such as cyclic voltammetry and chronoamperometry, in order to obtain a suitable nano structure. The deposition mechanism was determined as instantaneous nucleation and the dimension of particles was controlled in nanometric particle size range. Atomic Force Microscope was used to evaluate the effect of cysteine on the morphology and topography of gold nanoparticles. Finally the catalytic property of gold nanoparticle electrodeposited was studied in KOH solution, where oxygen reduction on the gold nanoparticle surface was eight times greater than that on the conventional gold deposits.

Optical Absorption and Electric Resistivity of an l-Cysteine Film

Science.gov (United States)

Kamada, Masao; Hideshima, Takuya; Azuma, Junpei; Yamamoto, Isamu; Imamura, Masaki; Takahashi, Kazutoshi

2016-12-01

The optical and electric properties of an l-cysteine film have been investigated to understand its applicability to bioelectronics. The fundamental absorption is the allowed transition having the threshold at 5.8 eV and the absorption is due to the charge-transfer type transition from sulfur-3sp to oxygen-2p and/or carbon-2p states, while absorptions more than 9 eV can be explained with intra-atomic transitions in the functional groups. The electric resistivity is 2.0 × 104 Ω m at room temperature and increases as the sample temperature decreases. The results indicate that the l-cysteine film is a p-type semiconductor showing the hole conduction caused by the sulfur-3sp occupied states and unknown impurity or defect states as acceptors. The electron affinity of the l-cysteine film is derived as ≦-0.3 eV.

Cyst(e)ine imbalance and its effect on methionine precursor utilization in chicks.

Science.gov (United States)

Dilger, R N; Baker, D H

2008-08-01

Five 9- or 12-d chick growth bioassays were done in batteries using 2 Met-deficient diets: a purified AA-based diet containing (by analysis, as-fed) 20.3% CP, 0.12% Met, and 0.05% cyst(e)ine; and an AA-fortified corn-peanut meal diet containing (by analysis, as-fed) 19.0% CP, 0.22% Met, and 0.23% cyst(e) ine. Feed-grade DL-Met (dl-M; 99%) was compared with feed-grade DL-OH-Met, Ca (OH-M; 84%). When the purified diet was modified to contain 0.12% Met and 0.20% or greater cyst(e)ine, slope-ratio assays involving graded dosing of DL-M (0, 404, 808, and 1,212 mg of DL-M/kg) or isosulfurous levels of OH-M resulted in linear (P ine [i.e., 0.12% Met, 0.12% cyst(e)ine]. When this diet was supplemented with either 404 mg of DL-M/kg or 476 mg of OH-M/kg, BW gain and G:F responded (P 0.10). Assays 4 and 5 used the corn-peanut meal basal diet containing 0.22% total Met and 0.23% total cyst(e)ine. In both assays, addition of either 465 mg of DL-M/kg or 554 mg of OH-M/kg resulted in increased (P ine concentration. In the absence of excess cyst(e)ine, BW gain responses to DL-M and OH-M were similar, but when 0.10% excess cyst(e)ine was provided as L-cystine or feather meal, DL-M responses tended to exceed those of OH-M. Moreover, this small excess of dietary cyst(e)ine, regardless of source, depressed (P ine, when included in Met-deficient diets, has the potential to be both anorexigenic and pernicious to OH-M utilization.

Tuning the carbon nanotube photoluminescence enhancement at addition of cysteine through the change of external conditions

Energy Technology Data Exchange (ETDEWEB)

Kurnosov, N.V.; Karachevtsev, M.V.; Leontiev, V.S.; Karachevtsev, V.A., E-mail: karachevtsev@ilt.kharkov.ua

2017-01-15

The enhancement of the photoluminescence (PL) from the semiconducting single-walled carbon nanotubes suspended with single-stranded DNA (ssDNA) in water observed after amino acids doping is the largest at cysteine addition. The PL intensity increased through the passivation of p-defects on the carbon nanotube sidewall by the cysteine molecules due to thiol group. The effect of several external factors on the cysteine-induced enhancement of PL from carbon nanotubes covered with ssDNA was studied: UV irradiation, tip or bath sonication treatment of the suspension, the ionic strength and pH of aqueous suspension. It turned out that all these factors have an essential influence on the dependence of the PL enhancement on the cysteine concentration through inducing of additional defects on nanotube as well as a change of the nanotube surface coverage with polymer. The obtained experimental results demonstrated that PL from carbon nanotubes can be exploited successfully for the monitoring of cysteine concentration in aqueous solution. - Highlights: • Cysteine doping enhances carbon nanotube emission more than other amino acids do. • SWNT emission dependence on cysteine concentration is tuned by UV irradiation and pH. • Type of sonication treatment influences SWNT PL dependence on cysteine concentration. • Polymer coverage and defectiveness of nanotubes effect on nanotube emission. • Graphic abstract.

Experimental and theoretical investigation on corrosion inhibition of AA5052 aluminium alloy by L-cysteine in alkaline solution

International Nuclear Information System (INIS)

Wang, Dapeng; Gao, Lixin; Zhang, Daquan; Yang, Dong; Wang, Hongxia; Lin, Tong

2016-01-01

The corrosion inhibition of L-cysteine on AA5052 aluminium alloy in 4 mol/L NaOH solution was investigated by hydrogen gas evolution experiment, polarisation curve, galvanostatic discharge, electrochemical impedance spectroscopy measurements and quantum chemical calculations. The adsorption of L-cysteine on aluminium alloy surface obeyed the amended Langmuir's adsorption isotherm. The polarisation curves indicated that L-cysteine acted as a cathodic inhibitor to inhibit cathodic reaction. The inhibition mechanism was dominated by the geometric covering effect. The galvanostatic discharge shows that the additives restrain the hydrogen evolution and increase the anodic utilization rate. Quantum chemical calculations indicated that L-cysteine molecules mainly interacted with on the carboxyl groups on the aluminium alloy surface. A strong hybridization occurred between the s-orbital and p-orbital of reactive sites in the L-cysteine molecule and the sp-orbital of Aluminium. - Highlights: • L-cysteine was used as corrosion inhibitor for Al alloy in alkaline solution. • Adsorption of L-cysteine on Al alloy surface obeyed the amended Langmuir's isotherm. • L-cysteine molecules interacted with the carboxyl groups on the Al alloy surface. • A strong orbital hybridization occurred between the reactive sites in L-cysteine and Al.

The effect of L-cysteine on the portion-selective uptake of cadmium in the renal proximal tubule

International Nuclear Information System (INIS)

Murakami, Masataka; Sano, Kenichi; Webb, M.

1987-01-01

Cadmium (Cd), co-administered with an excess of L-cysteine, accumulates rapidly in the kidneys of the rat. After subcutaneous (s.c.) injection of 3 μmol CdCl 2 /kg body wt the concentrations of Cd in the blood and kidneys increase with the dose of cysteine over the range 0.06-5.0 mmol/kg body wt. At cysteine doses of less than 1.5 mmol/kg body wt the ratio of the concentrations of Cd in the outer medulla and cortex of the kidney remains the same as that after the injection of Cd alone. This ratio, however, is more than doubled at dose levels of 5-10 mmol cysteine/kg body wt. Hepatic uptake of Cd is unaffected by doses of cysteine below 1.5 mmol/kg body wt but decreases markedly at higher doses. In animals that are dosed simultaneously with 5 mmol cysteine/kg body wt, renal uptake of 109 Cd is known to occur in the straight segments of the proximal tubules. At a dose level of less than 1.5 mmol cysteine/kg body wt the present autoradiographical studies show that 109 Cd is taken up predominantly by the proximal convoluted tubules of the kidney cortex. At the critical dose level (1.5 mmol/kg body wt), cysteine decreases the retention of Cd at the s.c. injection site, but probably has little effect on the distribution of Cd between protein and other carrier molecules in the blood. This distribution, however, is altered at higher cysteine dose levels. It is suggested that, under the latter conditions, stable Cd-cysteine complexes are formed in the blood and are filtered readily through the glomeruli. These complexes are taken up in the kidney at the sites of cysteine reabsorption which, by studies with L-[ 35 S]-cysteine, are identified as the straight segments of the proximal tubules. (orig.)

L-Cysteine Production in Escherichia coli Based on Rational Metabolic Engineering and Modular Strategy.

Science.gov (United States)

Liu, Han; Fang, Guochen; Wu, Hui; Li, Zhimin; Ye, Qin

2018-05-01

L-cysteine is an amino acid with important physiological functions and has a wide range of applications in medicine, food, animal feed, and cosmetics industry. In this study, the L-cysteine synthesis in Escherichia coliEscherichia coli is divided into four modules: the transport module, sulfur module, precursor module, and degradation module. The engineered strain LH03 (overexpression of the feedback-insensitive cysE and the exporter ydeD in JM109) accumulated 45.8 mg L -1 of L-cysteine in 48 hr with yield of 0.4% g/g glucose. Further modifications of strains and culture conditions which based on the rational metabolic engineering and modular strategy improved the L-cysteine biosynthesis significantly. The engineered strain LH06 (with additional overexpression of serA, serC, and serB and double mutant of tnaA and sdaA in LH03) produced 620.9 mg L -1 of L-cysteine with yield of 6.0% g/g glucose, which increased the production by 12 times and the yield by 14 times more than those of LH03 in the original condition. In fed-batch fermentation performed in a 5-L reactor, the concentration of L-cysteine achieved 5.1 g L -1 in 32 hr. This work demonstrates that the combination of rational metabolic engineering and module strategy is a promising approach for increasing the L-cysteine production in E. coli. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Double blind test of L-cysteine for protection against radiation-induced side effects in man

International Nuclear Information System (INIS)

Ohshima, Toshimi; Tsukiyama, Iwao; Mio, Akihiko; Ito, Otomasa; Sugawara, Masatoshi.

1977-01-01

L-Cysteine (80 mg/capsule of active ingredient) or placebo (lactose) was administered to a total of 127 patients with breast cancer (postoperative irradiation) or uterine cervical cancer (post-operative and intracavitary irradiation). L-Cysteine was effective in 49.3% of all patients and in 52.0% of patients with breast cancer, the difference from the placebo group being statistically significant. Decrease in the white blood cell count was less in the group given L-cysteine than that given placebo, and this difference was significant especially in the 3rd week for all cases. Significant difference was also noted in the 2nd week for postoperative irradiation and in the 2nd and 3rd weeks for postoperative and intracavitary irradiation for uterine cervical cancer. Decrease of white blood cell count to less than 3,000 was significantly small in the group given L-cysteine than in the placebo group. The values of hematocrit and platelets remained within normal limits, but the values in the group treated with L-cysteine was considerably different (0.05< Po<0.10) from those in the placebo group during the 2nd, 4th, and 6th week. The blood sedimentation rate was more stable in the group given L-cysteine than in the placebo group, and considerably different (0.05< Po<0.10) in the 2nd week and significantly different in the 6th week compared to the control. Anorexia was significantly less in the group given L-cysteine, especially in the 3rd week. These results suggest that L-cysteine can serve as a protective agent against the side effects of radiotherapy. (J.P.N.)

Double blind test of L-cysteine for protection against radiation-induced side effects in man

Energy Technology Data Exchange (ETDEWEB)

Ohshima, T; Tsukiyama, I; Mio, A [Tokyo Teishin Hospital (Japan); Ito, O; Sugawara, M

1977-05-01

L-Cysteine (80 mg/capsule of active ingredient) or placebo (lactose) was administered to a total of 127 patients with breast cancer (postoperative irradiation) or uterine cervical cancer (post-operative and intracavitary irradiation). L-Cysteine was effective in 49.3% of all patients and in 52.0% of patients with breast cancer, the difference from the placebo group being statistically significant. Decrease in the white blood cell count was less in the group given L-cysteine than that given placebo, and this difference was significant especially in the 3rd week for all cases. Significant difference was also noted in the 2nd week for postoperative irradiation and in the 2nd and 3rd weeks for postoperative and intracavitary irradiation for uterine cervical cancer. Decrease of white blood cell count to less than 3,000 was significantly small in the group given L-cysteine than in the placebo group. The values of hematocrit and platelets remained within normal limits, but the values in the group treated with L-cysteine was considerably different (0.05cysteine than in the placebo group, and considerably different (0.05cysteine, especially in the 3rd week. These results suggest that L-cysteine can serve as a protective agent against the side effects of radiotherapy.

Experimental and theoretical investigation on corrosion inhibition of AA5052 aluminium alloy by L-cysteine in alkaline solution

Energy Technology Data Exchange (ETDEWEB)

Wang, Dapeng; Gao, Lixin [School of Environmental and Chemical Engineering, Shanghai University of Electric Power, Shanghai 200090 (China); Zhang, Daquan, E-mail: zhangdaquan@shiep.edu.cn [School of Environmental and Chemical Engineering, Shanghai University of Electric Power, Shanghai 200090 (China); Yang, Dong [School of Environmental and Chemical Engineering, Shanghai University of Electric Power, Shanghai 200090 (China); Wang, Hongxia; Lin, Tong [Institute for Frontier Materials, Deakin University, Geelong, VIC 3216 (Australia)

2016-02-01

The corrosion inhibition of L-cysteine on AA5052 aluminium alloy in 4 mol/L NaOH solution was investigated by hydrogen gas evolution experiment, polarisation curve, galvanostatic discharge, electrochemical impedance spectroscopy measurements and quantum chemical calculations. The adsorption of L-cysteine on aluminium alloy surface obeyed the amended Langmuir's adsorption isotherm. The polarisation curves indicated that L-cysteine acted as a cathodic inhibitor to inhibit cathodic reaction. The inhibition mechanism was dominated by the geometric covering effect. The galvanostatic discharge shows that the additives restrain the hydrogen evolution and increase the anodic utilization rate. Quantum chemical calculations indicated that L-cysteine molecules mainly interacted with on the carboxyl groups on the aluminium alloy surface. A strong hybridization occurred between the s-orbital and p-orbital of reactive sites in the L-cysteine molecule and the sp-orbital of Aluminium. - Highlights: • L-cysteine was used as corrosion inhibitor for Al alloy in alkaline solution. • Adsorption of L-cysteine on Al alloy surface obeyed the amended Langmuir's isotherm. • L-cysteine molecules interacted with the carboxyl groups on the Al alloy surface. • A strong orbital hybridization occurred between the reactive sites in L-cysteine and Al.

Effect of free cysteine on the denaturation and aggregation of holo α-lactalbumin

DEFF Research Database (Denmark)

Nielsen, Line R.; Lund, Marianne N.; Davies, Michael J.

2018-01-01

α-Lactalbumin (α-LA) is a key commercial whey protein for nutritional purposes. The holo protein (calcium saturated) is considered the most heat stable whey protein, capable of refolding from unfolded states under many conditions. This is due to the absence of free thiols (cysteine residues......) that are typically involved in thermal aggregation and thiol–disulphide exchange reactions of other whey proteins. Heating (0–120 min at 90 °C, pH 7.0) holo α-LA generates free thiols through thermal cleavage of disulphide bonds, resulting in aggregates comprising unfolded α-LA species. The addition of free cysteine...... promotes the formation of soluble aggregates, effectively decreasing the holding time required to reach a particular aggregate size in a dose-dependent manner (0.35–1.4 mM cysteine). Excess cysteine (≥14 mM) causes a destabilisation of α-LA, shown by decreased denaturation temperature and gel formation...

Enzymatic synthesis of S-phenyl-L-cysteine from keratin hydrolysis industries wastewater with tryptophan synthase.

Science.gov (United States)

Xu, Lisheng; Wang, Zhiyuan; Mao, Pingting; Liu, Junzhong; Zhang, Hongjuan; Liu, Qian; Jiao, Qing-Cai

2013-04-01

An economical method for production of S-phenyl-L-cysteine from keratin acid hydrolysis wastewater (KHW) containing L-serine was developed by recombinant tryptophan synthase. This study provides us with an alternative KHW utilization strategy to synthesize S-phenyl-L-cysteine. Tryptophan synthase could efficiently convert L-serine contained in KHW to S-phenyl-L-cysteine at pH 9.0, 40°C and Trion X-100 of 0.02%. In a scale up study, L-serine conversion rate reach 97.1% with a final S-phenyl-L-cysteine concentration of 38.6 g l(-1). Copyright © 2013 Elsevier Ltd. All rights reserved.

S-Substituted cysteine derivatives and thiosulfinate formation in Petiveria alliacea-part II.

Science.gov (United States)

Kubec, Roman; Kim, Seokwon; Musah, Rabi A

2002-11-01

Three cysteine derivatives, (R)-S-(2-hydroxyethyl)cysteine, together with (R(S)R(C))- and (S(S)R(C))-S-(2-hydroxyethyl)cysteine sulfoxides, have been isolated from the roots of Petiveria alliacea. Furthermore, three additional amino acids, S-methyl-, S-ethyl-, and S-propylcysteine derivatives, were detected. They were present only in trace amounts (<3 microg g(-1) fr. wt), precluding determination of their absolute configurations and oxidation states. In addition, four thiosulfinates, S-(2-hydroxyethyl) (2-hydroxyethane)-, S-(2-hydroxyethyl) phenylmethane-, S-benzyl (2-hydroxyethane)- and S-benzyl phenylmethanethiosulfinates, have been found in a homogenate of the roots. The formation pathways of various benzyl/phenyl-containing compounds previously found in the plant were also discussed.

A novel electrochemical sensor based on metal-organic framework for electro-catalytic oxidation of L-cysteine.

Science.gov (United States)

Hosseini, Hadi; Ahmar, Hamid; Dehghani, Ali; Bagheri, Akbar; Tadjarodi, Azadeh; Fakhari, Ali Reza

2013-04-15

A novel electrochemical sensor based on Au-SH-SiO₂ nanoparticles supported on metal-organic framework (Au-SH-SiO₂@Cu-MOF) has been developed for electrocatalytic oxidation and determination of L-cysteine. The Au-SH-SiO₂@Cu-MOF was characterized by scanning electron microscopy, transmission electron microscopy, x-ray diffraction and cyclic voltammetry. The electrochemical behavior of L-cysteine at the Au-SH-SiO₂@Cu-MOF was investigated by cyclic voltammetry. The Au-SH-SiO₂@Cu-MOF showed a very efficient electrocatalytic activity for the oxidation of L-cysteine in 0.1 M phosphate buffer solution (pH 5.0). The oxidation overpotentials of L-cysteine decreased significantly and their oxidation peak currents increased dramatically at Au-SH-SiO₂@Cu-MOF. The potential utility of the sensor was demonstrated by applying it to the analytical determination of L-cysteine concentration. The results showed that the electrocatalytic current increased linearly with the L-cysteine concentration in the range of 0.02-300 μM and the detection limit was 0.008 μM. Finally, the sensor was applied to determine L-cysteine in water and biological samples. Copyright © 2012 Elsevier B.V. All rights reserved.

Aggregation mechanism of Pd nanoparticles in L-cysteine aqueous solution studied by NEXAFS and AFM

International Nuclear Information System (INIS)

Tsukada, C.; Ogawa, S.; Mizutani, T.; Kutluk, G.; Namatame, H.; Taniguchi, M.; Yagi, S.

2012-01-01

Highlight: ► We focus on the biocompatibility of Pd nanoparticles (NPs) for L-cysteine under water environment. ► The Pd NPs have been fabricated and deposited on Si wafer by gas evaporation method. ► When the Pd NPs/Si has been dipped into L-cysteine aqueous solution, the L-cysteine has selectively adsorbed on Pd NPs surface and existed as the L-cysteine thiolate, atomic S and L-cystine. ► Moreover, the aggregation of Pd NPs occurs by the migration of Pd NPs on Si and the cross-linked reaction between L-cysteine thiolate molecules adsorbed on Pd NPs. - Abstract: We focus on the biocompatibility of Pd nanoparticles (NPs) from the point of microscopic view. Thus, as the basic research for the biocompatibility, we have investigated the adsorbates on the Pd NPs surface and the aggregation mechanism for the Pd NPs on Si substrate after dipping into L-cysteine aqueous solution by means of NEXAFS measurement and AFM observation. The Pd NPs have been fabricated and deposited on the Si wafer by the gas evaporation method. Judging from the results of NEXAFS measurement, it is clear that the L-cysteine thiolate and atomic S exist on the Pd NPs surface. The results of AFM observation show that the Pd NPs aggregate. It is thought that the aggregation of the Pd NPs occurs by both the migration of the Pd NPs on Si wafer and the cross-linked reaction.

Synthesis, characterization, mucoadhesion and biocompatibility of thiolated carboxymethyl dextran-cysteine conjugate.

Science.gov (United States)

Shahnaz, G; Perera, G; Sakloetsakun, D; Rahmat, D; Bernkop-Schnürch, A

2010-05-21

This study was aimed at improving the mucoadhesive properties of carboxymethyl dextran by the covalent attachment of cysteine. Mediated by a carbodiimide, l-cysteine was covalently attached to the polymer. The resulting CMD-cysteine conjugate (CMD-(273) conjugate) displayed 273+/-20 micromol thiol groups per gram of polymer (mean+/-S.D.; n=3). Within 2h the viscosity of an aqueous mucus/CMD-(273) conjugate mixture pH 7.4 increased at 37 degrees C by more than 85% compared to a mucus/carboxymethyl dextran mixture indicating enlarged interactions between the mucus and the thiolated polymer. Due to the immobilization of cysteine, the swelling velocity of the polymer was significantly accelerated (ppolymer disintegrated within 15 min, whereas tablets of the CMD-(273) conjugate remained stable for 160 min (means+/-S.D.; n=3). Results from LDH and MTT assays on Caco-2 cells revealed 4.96+/-0.98% cytotoxicity and 94.1+/-0.9% cell viability for the CMD-(273) conjugate, respectively. Controlled release of model compound from CMD-(273) conjugate tablets was observed over 6h. These findings suggest that CMD-(273) conjugate is a promising novel polymer for drug delivery systems providing improved mucoadhesive and cohesive properties, greater stability and biocompatibility. Copyright 2010 Elsevier B.V. All rights reserved.

Role of conserved cysteine residues in Herbaspirillum seropedicae NifA activity.

Science.gov (United States)

Oliveira, Marco A S; Baura, Valter A; Aquino, Bruno; Huergo, Luciano F; Kadowaki, Marco A S; Chubatsu, Leda S; Souza, Emanuel M; Dixon, Ray; Pedrosa, Fábio O; Wassem, Roseli; Monteiro, Rose A

2009-01-01

Herbaspirillum seropedicae is an endophytic diazotrophic bacterium that associates with economically important crops. NifA protein, the transcriptional activator of nif genes in H. seropedicae, binds to nif promoters and, together with RNA polymerase-sigma(54) holoenzyme, catalyzes the formation of open complexes to allow transcription initiation. The activity of H. seropedicae NifA is controlled by ammonium and oxygen levels, but the mechanisms of such control are unknown. Oxygen sensitivity is attributed to a conserved motif of cysteine residues in NifA that spans the central AAA+ domain and the interdomain linker that connects the AAA+ domain to the C-terminal DNA binding domain. Here we mutagenized this conserved motif of cysteines and assayed the activity of mutant proteins in vivo. We also purified the mutant variants of NifA and tested their capacity to bind to the nifB promoter region. Chimeric proteins between H. seropedicae NifA, an oxygen-sensitive protein, and Azotobacter vinelandii NifA, an oxygen-tolerant protein, were constructed and showed that the oxygen response is conferred by the central AAA+ and C-terminal DNA binding domains of H. seropedicae NifA. We conclude that the conserved cysteine motif is essential for NifA activity, although single cysteine-to-serine mutants are still competent at binding DNA.

Surface-modified CdS nanoparticles as a fluorescent probe for the selective detection of cysteine

International Nuclear Information System (INIS)

Negi, Devendra P S; Chanu, T Inakhunbi

2008-01-01

We present a novel method for the selective detection of cysteine, a sulfur-containing amino acid, which plays a crucial role in many important biological functions such as protein folding. Surface-modified colloidal CdS nanoparticles have been used as a fluorescent probe to selectively detect cysteine in the presence of other amino acids in the micromolar concentration range. Cysteine quenches the emission of CdS in the 0.5-10 μM concentration range, whereas the other amino acids do not affect its emission. Among the other amino acids, histidine is most efficient in quenching the emission of the CdS nanoparticles. The sulfur atom of cysteine plays a crucial role in the quenching process in the 0.5-10 μM concentration range. Cysteine is believed to quench the emission of the CdS nanoparticles by binding to their surface via its negatively charged sulfur atom. This method can potentially be applied for its detection in biological samples.

Biological roles of cysteine proteinases in the pathogenesis of Trichomonas vaginalis

Science.gov (United States)

Hernández, Hilda M.; Marcet, Ricardo; Sarracent, Jorge

2014-01-01

Human trichomonosis, infection with Trichomonas vaginalis, is the most common non-viral sexually transmitted disease in the world. The host-parasite interaction and pathophysiological processes of trichomonosis remain incompletely understood. This review focuses on the advancements reached in the area of the pathogenesis of T. vaginalis, especially in the role of the cysteine proteinases. It highlights various approaches made in this field and lists a group of trichomonad cysteine proteinases involved in diverse processes such as invasion of the mucous layer, cytoadherence, cytotoxicity, cytoskeleton disruption of red blood cells, hemolysis, and evasion of the host immune response. A better understanding of the biological roles of cysteine proteinases in the pathogenesis of this parasite could be used in the identification of new chemotherapeutic targets. An additional advantage could be the development of a vaccine in order to reduce transmission of T. vaginalis. PMID:25348828

Protein Topology Determines Cysteine Oxidation Fate: The Case of Sulfenyl Amide Formation among Protein Families

Science.gov (United States)

Defelipe, Lucas A.; Lanzarotti, Esteban; Gauto, Diego; Marti, Marcelo A.; Turjanski, Adrián G.

2015-01-01

Cysteine residues have a rich chemistry and play a critical role in the catalytic activity of a plethora of enzymes. However, cysteines are susceptible to oxidation by Reactive Oxygen and Nitrogen Species, leading to a loss of their catalytic function. Therefore, cysteine oxidation is emerging as a relevant physiological regulatory mechanism. Formation of a cyclic sulfenyl amide residue at the active site of redox-regulated proteins has been proposed as a protection mechanism against irreversible oxidation as the sulfenyl amide intermediate has been identified in several proteins. However, how and why only some specific cysteine residues in particular proteins react to form this intermediate is still unknown. In the present work using in-silico based tools, we have identified a constrained conformation that accelerates sulfenyl amide formation. By means of combined MD and QM/MM calculation we show that this conformation positions the NH backbone towards the sulfenic acid and promotes the reaction to yield the sulfenyl amide intermediate, in one step with the concomitant release of a water molecule. Moreover, in a large subset of the proteins we found a conserved beta sheet-loop-helix motif, which is present across different protein folds, that is key for sulfenyl amide production as it promotes the previous formation of sulfenic acid. For catalytic activity, in several cases, proteins need the Cysteine to be in the cysteinate form, i.e. a low pKa Cys. We found that the conserved motif stabilizes the cysteinate by hydrogen bonding to several NH backbone moieties. As cysteinate is also more reactive toward ROS we propose that the sheet-loop-helix motif and the constraint conformation have been selected by evolution for proteins that need a reactive Cys protected from irreversible oxidation. Our results also highlight how fold conservation can be correlated to redox chemistry regulation of protein function. PMID:25741692

Pressor response to L-cysteine injected into the cisterna magna of conscious rats involves recruitment of hypothalamic vasopressinergic neurons.

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Takemoto, Yumi

2013-03-01

The sulfur-containing non-essential amino acid L-cysteine injected into the cisterna magna of adult conscious rats produces an increase in blood pressure. The present study examined if the pressor response to L-cysteine is stereospecific and involves recruitment of hypothalamic vasopressinergic neurons and medullary noradrenergic A1 neurons. Intracisternally injected D-cysteine produced no cardiovascular changes, while L-cysteine produced hypertension and tachycardia in freely moving rats, indicating the stereospecific hemodynamic actions of L-cysteine via the brain. The double labeling immunohistochemistry combined with c-Fos detection as a marker of neuronal activation revealed significantly higher numbers of c-Fos-positive vasopressinergic neurons both in the supraoptic and paraventricular nuclei and tyrosine hydroxylase containing medullary A1 neurons, of L-cysteine-injected rats than those injected with D-cysteine as iso-osmotic control. The results indicate that the cardiovascular responses to intracisternal injection of L-cysteine in the conscious rat are stereospecific and include recruitment of hypothalamic vasopressinergic neurons both in the supraoptic and paraventricular nuclei, as well as of medullary A1 neurons. The findings may suggest a potential function of L-cysteine as an extracellular signal such as neuromodulators in central regulation of blood pressure.

Mutations at the cysteine codons of the recA gene of Escherichia coli

International Nuclear Information System (INIS)

Weisemann, J.M.; Weinstock, G.M.

1988-01-01

Each of the three cysteine residues in the Escherichia coli RecA protein was replaced with a number of other amino acids. To do this, each cysteine codon was first converted to a chain-terminating amber codon by oligonucleotide-directed mutagenesis. These amber mutants were then either assayed for function in different suppressor strains or reverted by a second round of mutagenesis with oligonucleotides that had random sequences at the amber codon. Thirty-three different amino acid substitutions were obtained. Mutants were tested for three functions of RecA: survival following UV irradiation, homologous recombination, and induction of the SOS response. It was found that although none of the cysteines is essential for activity, mutations at each of these positions can affect one or more of the activities of RecA, depending on the particular amino acid substitution. In addition, the cysteine at position 116 appears to be involved in the RecA-promoted cleavage of the LexA protein

L-cysteine reversibly inhibits glucose-induced biphasic insulin secretion and ATP production by inactivating PKM2.

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Nakatsu, Daiki; Horiuchi, Yuta; Kano, Fumi; Noguchi, Yoshiyuki; Sugawara, Taichi; Takamoto, Iseki; Kubota, Naoto; Kadowaki, Takashi; Murata, Masayuki

2015-03-10

Increase in the concentration of plasma L-cysteine is closely associated with defective insulin secretion from pancreatic β-cells, which results in type 2 diabetes (T2D). In this study, we investigated the effects of prolonged L-cysteine treatment on glucose-stimulated insulin secretion (GSIS) from mouse insulinoma 6 (MIN6) cells and from mouse pancreatic islets, and found that the treatment reversibly inhibited glucose-induced ATP production and resulting GSIS without affecting proinsulin and insulin synthesis. Comprehensive metabolic analyses using capillary electrophoresis time-of-flight mass spectrometry showed that prolonged L-cysteine treatment decreased the levels of pyruvate and its downstream metabolites. In addition, methyl pyruvate, a membrane-permeable form of pyruvate, rescued L-cysteine-induced inhibition of GSIS. Based on these results, we found that both in vitro and in MIN6 cells, L-cysteine specifically inhibited the activity of pyruvate kinase muscle isoform 2 (PKM2), an isoform of pyruvate kinases that catalyze the conversion of phosphoenolpyruvate to pyruvate. L-cysteine also induced PKM2 subunit dissociation (tetramers to dimers/monomers) in cells, which resulted in impaired glucose-induced ATP production for GSIS. DASA-10 (NCGC00181061, a substituted N,N'-diarylsulfonamide), a specific activator for PKM2, restored the tetramer formation and the activity of PKM2, glucose-induced ATP production, and biphasic insulin secretion in L-cysteine-treated cells. Collectively, our results demonstrate that impaired insulin secretion due to exposure to L-cysteine resulted from its direct binding and inactivation of PKM2 and suggest that PKM2 is a potential therapeutic target for T2D.

A simple synthesis of L-[35S]cysteine sulfinic acid

International Nuclear Information System (INIS)

Spears, R.M.; Martin, D.L.

1981-01-01

The synthesis of L-[ 35 S]cysteine sulfinic acid (L-2-amino-3-[ 35 S]sulfino-propanoic acid) in 65% yield from S-[ 35 S]cystine is described. The procedure was designed for use with milligram quantities of starting material and requires no purification of intermediates. L-[ 35 S]Cystine was converted first to its thiosulfonate. Subsequent reaction of the thiosulfonate with ammonium hydroxide generated L-[ 35 S]cysteine sulfinic acid and L-[ 35 S]cystine as the major products. The L-[ 35 S]cystine was recovered and reprocessed thereby increasing the yield. (author)

Labeling of human serum albumin with 105Rh-cysteine complexes

International Nuclear Information System (INIS)

Lo, J.M.; Pillai, M.R.A.; John, C.S.; Troutner, D.E.

1990-01-01

The conjugation of a complex formed by reacting RhCl 3 with cysteine to human serum albumin has been investigated. Approximately 50% of the rhodium (labelled with 105 Rh) was converted to the complex. Conjugation of the complex to HSA via the ECDI method resulted in yields of ∼ 40% of the total rhodium or ∼ 80% of the Rh-cysteine complex. No conjugation was observed in the absence of the ECDI. At approximately equal molar concentrations of rhodium and HSA, an average of ∼ 0.4 rhodium atoms per HSA molecule was achieved. (author)

High-performance liquid chromatography-fluorescence assay of pyruvic acid to determine cysteine conjugate beta-lyase activity : application to S-1,2-dichlorovinyl-L-cysteine and S-2-benzothiazolyl-L-cysteine

NARCIS (Netherlands)

Stijntjes, G.J.; te Koppele, J.M.; Vermeulen, N P

1992-01-01

An HPLC-fluorescence assay has been developed for the determination of the activity of rat renal cytosolic cysteine conjugate beta-lyase. The method is based on isocratic HPLC separation and fluorescence detection of pyruvic acid, derivatized with o-phenylenediamine (OPD), and is shown to be rapid,

The proteins of Fusobacterium spp. involved in hydrogen sulfide production from L-cysteine.

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Basic, Amina; Blomqvist, Madeleine; Dahlén, Gunnar; Svensäter, Gunnel

2017-03-14

Hydrogen sulfide (H 2 S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. We studied the H 2 S-producing protein expression of bacterial strains associated with periodontal disease. Further, we examined the effect of a cysteine-rich growth environment on the synthesis of intracellular enzymes in F. nucleatum polymorphum ATCC 10953. The proteins were subjected to one-dimensional (1DE) and two-dimensional (2DE) gel electrophoresis An in-gel activity assay was used to detect the H 2 S-producing enzymes; Sulfide from H 2 S, produced by the enzymes in the gel, reacted with bismuth forming bismuth sulfide, illustrated as brown bands (1D) or spots (2D) in the gel. The discovered proteins were identified with liquid chromatography - tandem mass spectrometry (LC-MS/MS). Cysteine synthase and proteins involved in the production of the coenzyme pyridoxal 5'phosphate (that catalyzes the production of H 2 S) were frequently found among the discovered enzymes. Interestingly, a higher expression of H 2 S-producing enzymes was detected from bacteria incubated without cysteine prior to the experiment. Numerous enzymes, identified as cysteine synthase, were involved in the production of H 2 S from cysteine and the expression varied among Fusobacterium spp. and strains. No enzymes were detected with the in-gel activity assay among the other periodontitis-associated bacteria tested. The expression of the H 2 S-producing enzymes was dependent on environmental conditions such as cysteine concentration and pH but less dependent on the presence of serum and hemin.

Cysteine-mediated gene expression and characterization of the CmbR regulon in Streptococcus pneumoniae

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Muhammad Afzal

2016-12-01

Full Text Available In this study, we investigated the transcriptomic response of Streptococcus pneumoniae D39 to cysteine. Transcriptome comparison of the D39 wild-type strain grown at a restricted concentration of cysteine (0.03 mM to one grown at a high concentration of cysteine (50 mM in chemically-define medium (CDM revealed elevated expression of various genes/operons, i.e. spd-0150, metQ, spd-0431, metEF, gshT, spd-0618, fhs, tcyB, metB-csd, metA, spd-1898, yvdE, and cysK, likely to be involved in the transport and utilization of cysteine and/or methionine. Microarray-based data were further confirmed by quantitative RT-PCR. Promoter lacZ-fusion studies and quantitative RT-PCR data showed that the transcriptional regulator CmbR acts as a transcriptional repressor of spd-0150, metEF, gshT, spd-0618, tcyB, metA, and yvdE, putatively involved in cysteine uptake and utilization. The operator site of CmbR in the promoter regions of CmbR-regulated genes is predicted and confirmed by mutating or deleting CmbR operator sites from the promoter regions of these genes.

Controllable synthesis of TiO2 nanomaterials by assisting with l-cysteine and ethylenediamine

KAUST Repository

Tao, Yugui

2013-11-21

This paper reports a facile l-cysteine-assisted solvothermal synthesis of TiO2 nanomaterials using ethylenediamine (En) and distilled water as solvent. The influence of reaction time, temperature, l-cysteine and solvent was initially investigated. Results demonstrated the reaction temperature, l-cysteine and En significantly imposed impact on the phase and morphology of the particles. Amorphous nanosheets, mixed-crystal nanorods and pure anatase nanoparticles were controllably synthesized by varying reaction temperature. The formation of the amorphous nanosheets and mixed-crystal nanorods were directly affected by the presence of l-cysteine and En. And the presence of En distinctly affected the crystal phase of the products, which was rarely mentioned in other studies. Moreover, the photocatalytic activities of three typical samples were excellent. The possible formation mechanism of the sample was also discussed. © 2013 Springer Science+Business Media New York.

Controllable synthesis of TiO2 nanomaterials by assisting with l-cysteine and ethylenediamine

KAUST Repository

Tao, Yugui; Cao, Ning; Pan, Jun; Sun, Yichen; Jin, Cheng; Song, Yang

2013-01-01

This paper reports a facile l-cysteine-assisted solvothermal synthesis of TiO2 nanomaterials using ethylenediamine (En) and distilled water as solvent. The influence of reaction time, temperature, l-cysteine and solvent was initially investigated. Results demonstrated the reaction temperature, l-cysteine and En significantly imposed impact on the phase and morphology of the particles. Amorphous nanosheets, mixed-crystal nanorods and pure anatase nanoparticles were controllably synthesized by varying reaction temperature. The formation of the amorphous nanosheets and mixed-crystal nanorods were directly affected by the presence of l-cysteine and En. And the presence of En distinctly affected the crystal phase of the products, which was rarely mentioned in other studies. Moreover, the photocatalytic activities of three typical samples were excellent. The possible formation mechanism of the sample was also discussed. © 2013 Springer Science+Business Media New York.

Photo-fermentative bacteria aggregation triggered by L-cysteine during hydrogen production.

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Xie, Guo-Jun; Liu, Bing-Feng; Xing, De-Feng; Nan, Jun; Ding, Jie; Ren, Nan-Qi

2013-05-03

Hydrogen recovered from organic wastes and solar energy by photo-fermentative bacteria (PFB) has been suggested as a promising bioenergy strategy. However, the use of PFB for hydrogen production generally suffers from a serious biomass washout from photobioreactor, due to poor flocculation of PFB. In the continuous operation, PFB cells cannot be efficiently separated from supernatant and rush out with effluent from reactor continuously, which increased the effluent turbidity, meanwhile led to increases in pollutants. Moreover, to replenish the biomass washout, substrate was continuously utilized for cell growth rather than hydrogen production. Consequently, the poor flocculability not only deteriorated the effluent quality, but also decreased the potential yield of hydrogen from substrate. Therefore, enhancing the flocculability of PFB is urgent necessary to further develop photo-fermentative process. Here, we demonstrated that L-cysteine could improve hydrogen production of Rhodopseudomonas faecalis RLD-53, and more importantly, simultaneously trigger remarkable aggregation of PFB. Experiments showed that L-cysteine greatly promoted the production of extracellular polymeric substances, especially secretion of protein containing more disulfide bonds, and help for enhancement stability of floc of PFB. Through formation of disulfide bonds, L-cysteine not only promoted production of EPS, in particular the secretion of protein, but also stabilized the final confirmation of protein in EPS. In addition, the cell surface elements and functional groups, especially surface charged groups, have also been changed by L-cysteine. Consequently, absolute zeta potential reached a minimum value at 1.0 g/l of L-cysteine, which obviously decreased electrostatic repulsion interaction energy based on DLVO theory. Total interaction energy barrier decreased from 389.77 KT at 0.0 g/l of L-cysteine to 127.21 kT at 1.0 g/l. Thus, the strain RLD-53 overcame the total energy barrier and

L-Cysteine Capped CdSe Quantum Dots Synthesized by Photochemical Route.

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Singh, Avinash; Kunwar, Amit; Rath, M C

2018-05-01

L-cysteine capped CdSe quantum dots were synthesized via photochemical route in aqueous solution under UV photo-irradiation. The as grown CdSe quantum dots exhibit broad fluorescence at room temperature. The CdSe quantum dots were found to be formed only through the reactions of the precursors, i.e., Cd(NH3)2+4 and SeSO2-3 with the photochemically generated 1-hydroxy-2-propyl radicals, (CH3)2COH radicals, which are formed through the process of H atom abstraction by the photoexcited acetone from 2-propanol. L-Cysteine was found to act as a suitable capping agent for the CdSe quantum dots and increases their biocompatability. Cytotoxicty effects of these quantum dots were evaluated in Chinese Hamster Ovary (CHO) epithelial cells, indicated a significant lower level for the L-cysteine capped CdSe quantum dots as compare to the bare ones.

Anti-trypanosomal activity of non-peptidic nitrile-based cysteine protease inhibitors.

Science.gov (United States)

Burtoloso, Antonio C B; de Albuquerque, Sérgio; Furber, Mark; Gomes, Juliana C; Gonçalez, Cristiana; Kenny, Peter W; Leitão, Andrei; Montanari, Carlos A; Quilles, José Carlos; Ribeiro, Jean F R; Rocha, Josmar R

2017-02-01

The cysteine protease cruzipain is considered to be a validated target for therapeutic intervention in the treatment of Chagas disease. Anti-trypanosomal activity against the CL Brener strain of T. cruzi was observed in the 0.1 μM to 1 μM range for three nitrile-based cysteine protease inhibitors based on two scaffolds known to be associated with cathepsin K inhibition. The two compounds showing the greatest potency against the trypanosome were characterized by EC50 values (0.12 μM and 0.25 μM) that were an order of magnitude lower than the corresponding Ki values measured against cruzain, a recombinant form of cruzipain, in an enzyme inhibition assay. This implies that the anti-trypanosomal activity of these two compounds may not be explained only by the inhibition of the cruzain enzyme, thereby triggering a putative polypharmacological profile towards cysteine proteases.

A S-cysteine conjugate, precursor of aroma of White Sauvignon

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Takatoshi Tominaga

1995-12-01

Full Text Available 4-mercapto-4-methylpentan-2-one (4-MMP, a strongly odorant compound responsible for the « boxtree » or « broom plant » odour of the Sauvignon wines, can be enzymaticaly released in vitro from an odourless must extract. The enzyme source used is a cell-free extract of the gastrointestinal bacterium Eubacterium limosum. This crude preparation exhibits a cysteine β-lyase activity which requires the presence of pyridoxal phosphate. The release of 4-MMP is inhibited when the substrate is previously treated with N-hydroxysuccimide acetate which reacts with a primary amine. The same bacterial extract is also able to release 4-MMP, pyruvic acid and ammonium, from S-(4-méthylpentan-2-one-L-cysteine. On the other hand, the cleavage of S-(4-méthylpentan-2-oneD,L-homocysteine and S-(4-méthylpentan-2-one- glutathione is very limited. These results suggest that the precursor of 4-MMP in Sauvignon must is a S-cysteine conjugate. Such an aroma precursor in grapes or in other fruits has never been round berore.

Selective electrochemical determination of homocysteine in the presence of cysteine and glutathione

International Nuclear Information System (INIS)

Salehzadeh, Hamid; Mokhtari, Banafsheh; Nematollahi, Davood

2014-01-01

Graphical abstract: 3,5-Di-tert-buthylcatechol was used for the selective electrochemical determination of homocysteine in the presence of cysteine and glutathione at the glassy carbon and carbon nanotube modified glassy carbon electrode. - Highlights: • Selective electrochemical determination of homocysteine. • Catalytic electron transfer of 3,5-di-tert-buthylcatechol in the presence of homocysteine. • Michael type addition reaction of electrochemically generated 3,5-di-tert-buthyl-o-benzoquinone with glutathione. - Abstract: The electrochemical oxidation of 3,5-di-tert-buthylcatechol in the presence of homocysteine was used for the selective electrochemical determination of homocysteine in the presence of cysteine and glutathione at a glassy carbon and a glassy carbon electrode modified with carbon nanotube. The results revealed that the electrochemically generated 3,5-di-tert-butylcyclohexa-3,5-diene-1,2-dione exhibits high catalytic activity toward homocysteine oxidation at reduced over-potential and low catalytic activity for oxidation of cysteine. The catalytic activity 3,5-di-tert-butylcyclohexa-3,5-diene-1,2-dione toward cysteine was suppressed in the presence of 4-N,N-dimethylaminocinnamaldehyde. Contrary to homocysteine and cysteine, the reaction of glutathione with 3,5-di-tert-butylcyclohexa-3,5-diene-1,2-dione is a substituation reaction. This method exhibits three dynamic linear ranges of 2.5 to 10 μmol L −1 , 10 to 100 μmol L −1 and 100 to 1000 μmol L −1 , and a lower detection limit (3σ) of 0.89 ± 3.53% μmol L −1 for homocysteine

Cysteine proteinases and cystatins

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Adeliana S. Oliveira

2003-01-01

Full Text Available This review describeds the definition, localization, functions and examples of cysteine proteinases and their protein inhibitors in vertebrate, non-vertebrate animals and plants. These inhibitors are related with defense mechanisms of plant against pests. It also describes the factors involved in the specific cysteine proteinase-cystatin interaction and high degree of affinity and large specificity in this interaction which are not only represented by the compatibility between amino acid residues of the active site involved in catalysis, but also of all amino acid residues that participante in the enzyme-inhibitor interaction.Nesta revisão foram descritas definições, localizações, funções e exemplos de proteinases cisteínicas e suas proteinas inibidoras em animais vertebrados e invertebrados e plantas. Tratamos principalmente com aqueles inibidores que são relatados com o mecanismo de defesa da planta contra pestes. Em adição, comentamos sobre recentes trabalhos que contribuíram para uma melhor compreenção dos fatores envolvidos na interação específica proteinase cisteínica-cistatina. Por outro lado, chamamos atenção para o alto grau de afinidade e grande especificidade na interação que não são apenas representadas pela compatibilidade entre os residuos de aminoácidos do sítio ativo envolvidos na catalise, mas também de todos os resíduos de aminoácidos que participam da interação enzima-inibidor.

Cysteine Addition Promotes Sulfide Production and 4-Fold Hg(II)-S Coordination in Actively Metabolizing Escherichia coli.

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Thomas, Sara A; Gaillard, Jean-François

2017-04-18

The bacterial uptake of mercury(II), Hg(II), is believed to be energy-dependent and is enhanced by cysteine in diverse species of bacteria under aerobic and anaerobic conditions. To gain insight into this Hg(II) biouptake pathway, we have employed X-ray absorption spectroscopy (XAS) to investigate the relationship between exogenous cysteine, cellular metabolism, cellular localization, and Hg(II) coordination in aerobically respiring Escherichia coli (E. coli). We show that cells harvested in exponential growth phase consistently display mixtures of 2-fold and 4-fold Hg(II) coordination to sulfur (Hg-S 2 and Hg-S 4 ), with added cysteine enhancing Hg-S 4 formation. In contrast, cells in stationary growth phase or cells treated with a protonophore causing a decrease in cellular ATP predominantly contain Hg-S 2 , regardless of cysteine addition. Our XAS results favor metacinnabar (β-HgS) as the Hg-S 4 species, which we show is associated with both the cell envelope and cytoplasm. Additionally, we observe that added cysteine abiotically oxidizes to cystine and exponentially growing E. coli degrade high cysteine concentrations (100-1000 μM) into sulfide. Thermodynamic calculations confirm that cysteine-induced sulfide biosynthesis can promote the formation of dissolved and particulate Hg(II)-sulfide species. This report reveals new complexities arising in Hg(II) bioassays with cysteine and emphasizes the need for considering changes in chemical speciation as well as growth stage.

Intramolecular synergistic effect of glutamic acid, cysteine and glycine against copper corrosion in hydrochloric acid solution

International Nuclear Information System (INIS)

Zhang Daquan; Xie Bin; Gao Lixin; Cai Qirui; Joo, Hyung Goun; Lee, Kang Yong

2011-01-01

The corrosion protection of copper by glutamic acid, cysteine, glycine and their derivative (glutathione) in 0.5 M hydrochloric acid solution has been studied by the electrochemical impedance spectroscopy and cyclic voltammetry. The inhibition efficiency of the organic inhibitors on copper corrosion increases in the order: glutathione > cysteine > cysteine + glutamic acid + glycine > glutamic acid > glycine. Maximum inhibition efficiency for cysteine reaches about 92.9% at 15 mM concentration level. The glutathione can give 96.4% inhibition efficiency at a concentration of 10 mM. The molecular structure parameters were obtained by PM3 (Parametric Method 3) semi-empirical calculation. The intramolecular synergistic effect of glutamic acid, cysteine and glycine moieties in glutathione is attributed to the lower energy of the lowest unoccupied molecular orbital (E LUMO ) level and to the excess hetero-atom adsorption centers and the bigger coverage on the copper surface.

Managing anthelmintic resistance in small ruminant livestock of resource-poor farmers in South Africa.

Science.gov (United States)

Vattaa, A F; Lindberg, A L E

2006-03-01

Gastrointestinal parasitism is one of the most important disease complexes of sheep and goats impacting on the resource-poor livestock farmer. Of the responsible nematodes, Haemonchus contortus, a blood-sucking worm of the abomasum, poses possibly the greatest threat. Over the past several decades, the worm has been controlled through the use of anthelmintics, but the emergence of anthelmintic resistance has threatened this chemotherapeutic approach. In Africa, the overall prevalence of anthelmintic resistance has not been extensively investigated, particularly within the resource-poor farming sector, but resistance has been reported from at least 14 countries with most of the reports emanating from Kenya and South Africa and the majority concerning H. contortus. While levels of resistance under commercial sheep farming systems in South Africa is considered to be amongst the worst in the world, resistance has also been reported from the resource-poor farming sector. Increases in productivity and reproduction of livestock and the development of markets for sale of animals are seen by international funding bodies as a way out of poverty for communities that keep livestock. This must lead to the greater need for parasite control. At such times, the risk of levels of anthelmintic resistance escalating is much greater and there is therefore a need to look at alternatives to their use. Proposed strategies include the appropriate, but judicious use of anthelmintics by application of the FAMACHA system and the use of alternatives to anthelmintics such as strategic nutrient supplementation. It is also very clear that there is a strong demand for knowledge about animal diseases, including helminthosis, and their effective management in the resource-poor livestock farming communities. This is an important challenge to meet.

Efficacy of albendazole against nematode parasites isolated from a goat farm in Ethiopia: relationship between dose and efficacy in goats.

Science.gov (United States)

Eguale, Tadesse; Chaka, Hassen; Gizaw, Daniel

2009-10-01

A suspected case of albendazole resistance in a goat farm of Hawassa University was examined using faecal egg count reduction test (FECRT), controlled anthelmintic efficacy test and egg hatch assay (EHA) to verify the development of resistance and/or the need for higher doses of the drug in goats than in sheep. The experiment was conducted in 12 sheep (2 groups: treatment versus control) and 24 goats (4 groups: 3 treatments versus control, n = 6; per group) following artificial infection with infective larvae of Haemonchus contortus and Oesophagostomum columbianum. The first group of sheep and goats were treated orally with albendazole at the dose rate of 3.8 mg/kg body weight (i.e. manufacturer's recommended dose for sheep) while the second group of sheep and the fourth group of goats were left untreated. The second and the third group of goats were treated with albendazole at 5.7 and 7.6 mg/kg respectively. The FECRT showed an efficacy of albendazole in goats to be 65.5, 81.4 and 84.1% at the dose rate of 3.8, 5.7 and 7.6 mg/kg body weight respectively while in sheep it was 62% at the dose rate of 3.8 mg/kg. Increasing the dose to 1.5 the sheep recommended dose induced minor improvement of efficacy in goats; however the efficacy was almost the same at 1.5 and twice the dose recommended for sheep. Worm counts at day 15 post-treatment revealed that H. contortus has developed resistance to albendazole. EHA results also supported these findings. On the other hand, O. columbianum was 100% susceptible at all dose levels tested.

A novel high throughput assay for anthelmintic drug screening and resistance diagnosis by real-time monitoring of parasite motility.

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Michael J Smout

Full Text Available BACKGROUND: Helminth parasites cause untold morbidity and mortality to billions of people and livestock. Anthelmintic drugs are available but resistance is a problem in livestock parasites, and is a looming threat for human helminths. Testing the efficacy of available anthelmintic drugs and development of new drugs is hindered by the lack of objective high-throughput screening methods. Currently, drug effect is assessed by observing motility or development of parasites using laborious, subjective, low-throughput methods. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a novel application for a real-time cell monitoring device (xCELLigence that can simply and objectively assess anthelmintic effects by measuring parasite motility in real time in a fully automated high-throughput fashion. We quantitatively assessed motility and determined real time IC(50 values of different anthelmintic drugs against several developmental stages of major helminth pathogens of humans and livestock, including larval Haemonchus contortus and Strongyloides ratti, and adult hookworms and blood flukes. The assay enabled quantification of the onset of egg hatching in real time, and the impact of drugs on hatch rate, as well as discriminating between the effects of drugs on motility of drug-susceptible and -resistant isolates of H. contortus. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that this technique will be suitable for discovery and development of new anthelmintic drugs as well as for detection of phenotypic resistance to existing drugs for the majority of helminths and other pathogens where motility is a measure of pathogen viability. The method is also amenable to use for other purposes where motility is assessed, such as gene silencing or antibody-mediated killing.

Gastrointestinal nematodes and anthelmintic resistance in Danish goat herds☆

Science.gov (United States)

Holm, Signe A.; Sörensen, Camilla R. L.; Thamsborg, Stig M.; Enemark, Heidi L.

2014-01-01

The prevalence of gastrointestinal parasites in Danish goats and the presence of anthelmintic resistance (AR) in 10 selected herds were investigated during April–September 2012. All Danish herds (n = 137) with 10 or more adult goats were invited to participate, and of these 27 herds met the inclusion criterion of more than 10 young kids never treated with anthelmintics. Questionnaire data on management were collected, and faecal samples from 252 kids were analysed by the McMaster technique. From all herds with a mean faecal egg count (FEC) above 300 eggs per g of faeces, pooled samples were stained with peanut agglutinin (PNA) for specific detection of Haemonchus contortus. Strongyle eggs were detected with an individual prevalence of 69%, including Nematodirus battus (3.6%) and other Nematodirus species (15.0%). Eimeria spp. were observed in 99.6% of the kids. H. contortus was found in 11 of 12 (92%) tested herds. Anthelmintics were used in 89% of the herds with mean treatment frequencies of 0.96 and 0.89 treatments per year for kids and adults, respectively. In 2011, new animals were introduced into 44% of the herds of which 25% practised quarantine anthelmintic treatments. In 10 herds the presence of AR was analysed by egg hatch assay and FEC reduction tests using ivermectin (0.3 mg/kg) or fenbendazole (10.0 mg/kg). AR against both fenbendazole and ivermectin was detected in seven herds; AR against fenbendazole in one herd, and AR against ivermectin in another herd. In conclusion, resistance to the most commonly used anthelmintics is widespread in larger goat herds throughout Denmark. PMID:25076056

Gastrointestinal nematodes and anthelmintic resistance in Danish goat herds☆

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Holm Signe A.

2014-01-01

Full Text Available The prevalence of gastrointestinal parasites in Danish goats and the presence of anthelmintic resistance (AR in 10 selected herds were investigated during April–September 2012. All Danish herds (n = 137 with 10 or more adult goats were invited to participate, and of these 27 herds met the inclusion criterion of more than 10 young kids never treated with anthelmintics. Questionnaire data on management were collected, and faecal samples from 252 kids were analysed by the McMaster technique. From all herds with a mean faecal egg count (FEC above 300 eggs per g of faeces, pooled samples were stained with peanut agglutinin (PNA for specific detection of Haemonchus contortus. Strongyle eggs were detected with an individual prevalence of 69%, including Nematodirus battus (3.6% and other Nematodirus species (15.0%. Eimeria spp. were observed in 99.6% of the kids. H. contortus was found in 11 of 12 (92% tested herds. Anthelmintics were used in 89% of the herds with mean treatment frequencies of 0.96 and 0.89 treatments per year for kids and adults, respectively. In 2011, new animals were introduced into 44% of the herds of which 25% practised quarantine anthelmintic treatments. In 10 herds the presence of AR was analysed by egg hatch assay and FEC reduction tests using ivermectin (0.3 mg/kg or fenbendazole (10.0 mg/kg. AR against both fenbendazole and ivermectin was detected in seven herds; AR against fenbendazole in one herd, and AR against ivermectin in another herd. In conclusion, resistance to the most commonly used anthelmintics is widespread in larger goat herds throughout Denmark.

Development of a colloidal gold immunochromatographic strip based on HSP70 for the rapid detection of Echinococcus granulosus in sheep.

Science.gov (United States)

Zhuo, Xunhui; Yu, Yingchao; Chen, Xueqiu; Zhang, Zhuangzhi; Yang, Yi; Du, Aifang

2017-06-15

Echinococcus granulosus is the causative pathogen of cystic echinococcosis, a serious disease endangering human and animal health. In this study, an immunochromatographic strip was developed based on the recombinant protein Heat shock protein 70 (HSP70) for the serological detection of E. granulosus. The protocol completes within 20min requiring no specialized equipment or chemical reagents, while specificity tests confirmed no cross-reactivity with positive serum of Fasciola hepatica, Haemonchus contortus, Peste des petits ruminants virus (PPRV) and Foot-and-mouth disease virus (FMDV). The strips remained stable after storage at 4°C for up to 8 months. Both immunochromatographic strip and ELISA tests were applied to detect E. granulosus antibody in a total of 728 serum samples obtained from slaughter houses in Zhejiang Province. Our data revealed positive rates of 2.61 and 1.65% by immunochromatographic strip and ELISA methods, respectively. The immunochromatographic strip test developed in this study provides a simple, specific and rapid method of E. granulosus antibody detection and infected sheep monitoring. Copyright © 2017. Published by Elsevier B.V.

Seasonal dynamics of endoparasitic infections at an organic goat farm and the impact of detected infections on milk production.

Science.gov (United States)

Kyriánová, Iveta A; Vadlejch, Jaroslav; Kopecký, Oldřich; Langrová, Iva

2017-11-01

This study evaluated patterns and species composition of parasitic infections detected over a 1-year period at an organic goat farm. As a result of coprological examination, the overall prevalence of observed strongylids (99%), coccidia of the genus Eimeria (98%), and Muellerius capillaris lungworms (93%) was calculated. The most prevalent strongylids recovered from incubated fecal samples were Haemonchus contortus (42%), genera Trichostrongylus (23%), Oesophagostomum columbianum (13%), and Teladorsagia circumcincta (11%). A maximum intensity of coccidia infection 5150 oocysts per gram, strongylids infection 9900 eggs per gram and lungworm infection 867.26 larvae per gram were detected. The various effects (including environment, host, and parasites) on milk yield, lactose, protein, and fat were evaluated using generalized linear mixed models. Milk yield (P goat. With the intensity of infection detected in our study, only protein content was affected (P goat itself can substantially decrease protein content but has much less of an effect on fat, milk yield, and lactose. Based on our results, we can conclude that a low intensity of parasitic infections does not significantly affect milk yield and the qualitative parameters of milk.

Epidemiology and effects of gastrointestinal nematode infection on milk productions of dairy ewes

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Suarez V.H.

2009-06-01

Full Text Available 66 Pampinta breed ewes were studied during milking to evaluate the infection and the effect of gastrointestinal nematode on milk production sheep system. Naturally infected ewes on pasture were randomly allocated to two groups: TG, suppressively treated group every four weeks with levamisole and UG, untreated group. Faecal nematode egg counts and larval differentiation were conducted monthly. Successive groups of worm free tracer lambs were grazed with ewes and then slaughtered for worm counts. Test-day milk yield of individual ewes was recorded and ewe machine-milking period length (MPL were estimated. Faecal egg counts and tracer nematode numbers increased towards midsummer and declined sharply toward the end of the study. TG (188.0 ± 60 liters produced more (p < 0.066 milk liters than UG (171.9 ± 52.2 and TG had significantly more extended (p < 0.041 MPL than those of UG. The present study showed that dairy sheep were negatively affected by worms, even when exposed to short periods of high acute nematode (mainly Haemonchus contortus infection.

Identification of semicarbazones, thiosemicarbazones and triazine nitriles as inhibitors of Leishmania mexicana cysteine protease CPB.

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Jörg Schröder

Full Text Available Cysteine proteases of the papain superfamily are present in nearly all eukaryotes. They play pivotal roles in the biology of parasites and inhibition of cysteine proteases is emerging as an important strategy to combat parasitic diseases such as sleeping sickness, Chagas' disease and leishmaniasis. Homology modeling of the mature Leishmania mexicana cysteine protease CPB2.8 suggested that it differs significantly from bovine cathepsin B and thus could be a good drug target. High throughput screening of a compound library against this enzyme and bovine cathepsin B in a counter assay identified four novel inhibitors, containing the warhead-types semicarbazone, thiosemicarbazone and triazine nitrile, that can be used as leads for antiparasite drug design. Covalent docking experiments confirmed the SARs of these lead compounds in an effort to understand the structural elements required for specific inhibition of CPB2.8. This study has provided starting points for the design of selective and highly potent inhibitors of L. mexicana cysteine protease CPB that may also have useful efficacy against other important cysteine proteases.

The effect of cysteine on the corrosion of 304L stainless steel in sulphuric acid

International Nuclear Information System (INIS)

Silva, A.B.; Agostinho, S.M.L.; Barcia, O.E.; Cordeiro, G.G.O.; D'Elia, E.

2006-01-01

The effect of cysteine on the corrosion of 304L stainless steel in 1 mol l -1 H 2 SO 4 was studied using open-circuit potential measurements, anodic polarization curves, electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). All the electrochemical measurements obtained in the presence of low cysteine concentration (10 -6 -10 -5 mol l -1 ) presented the same behaviour as those obtained in the absence of cysteine, a passivated steel surface. However, for higher cysteine concentrations (10 -4 -10 -2 mol l -1 ), a different behaviour was observed: the corrosion potential stabilized at a more negative value; an active region was observed in the anodic polarization curves and the electrochemical impedance diagrams showed an inductive loop at lower frequencies and a much lower polarization resistance. These results show that the presence of cysteine at high concentration turns the surface of 304L stainless steel electrochemically active, probably dissolving the passivation layer and promoting the stainless steel anodic dissolution. SEM experiments performed after immersion experiments at corrosion potential were in good agreement with the electrochemical results

The Cysteine S-Alkylation Reaction as a Synthetic Method to Covalently Modify Peptide Sequences.

Science.gov (United States)

Calce, Enrica; De Luca, Stefania

2017-01-05

Synthetic methodologies to chemically modify peptide molecules have long been investigated for their impact in the field of chemical biology. They allow the introduction of biochemical probes useful for studying protein functions, for manipulating peptides with therapeutic potential, and for structure-activity relationship investigations. The commonly used approach was the derivatization of an amino acid side chain. In this regard, the cysteine, for its unique reactivity, has been widely employed as the substrate for such modifications. Herein, we report on methodologies developed to modify the cysteine thiol group through the S-alkylation reaction. Some procedures perform the alkylation of cysteine derivatives, in order to prepare building blocks to be used during the peptide synthesis, whilst some others selectively modify peptide sequences containing a cysteine residue with a free thiol group, both in solution and in the solid phase. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Glutathione provides a source of cysteine essential for intracellular multiplication of Francisella tularensis.

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Khaled Alkhuder

2009-01-01

Full Text Available Francisella tularensis is a highly infectious bacterium causing the zoonotic disease tularemia. Its ability to multiply and survive in macrophages is critical for its virulence. By screening a bank of HimarFT transposon mutants of the F. tularensis live vaccine strain (LVS to isolate intracellular growth-deficient mutants, we selected one mutant in a gene encoding a putative gamma-glutamyl transpeptidase (GGT. This gene (FTL_0766 was hence designated ggt. The mutant strain showed impaired intracellular multiplication and was strongly attenuated for virulence in mice. Here we present evidence that the GGT activity of F. tularensis allows utilization of glutathione (GSH, gamma-glutamyl-cysteinyl-glycine and gamma-glutamyl-cysteine dipeptide as cysteine sources to ensure intracellular growth. This is the first demonstration of the essential role of a nutrient acquisition system in the intracellular multiplication of F. tularensis. GSH is the most abundant source of cysteine in the host cytosol. Thus, the capacity this intracellular bacterial pathogen has evolved to utilize the available GSH, as a source of cysteine in the host cytosol, constitutes a paradigm of bacteria-host adaptation.

E.S.R. studies of mechanisms of radiation protection effect by cysteine and cystine

International Nuclear Information System (INIS)

Xue-Peng, L.; Tie-Cheng, T.; Nian-Yun, L.

1981-01-01

By means of E.S.R. the repair mechanism of radiation induced spin transfer from dTMP to cysteine in binary system dTMP-cysteine has been confirmed. Furthermore, a new marked radiation protection effect, exerted by cysteine or cystine on thymine irradiated and observed at low temperature, has been detected. Another sort of fast protection mechanism, including electron transfer and excitation transfer, has been proposed, based on recent advances of primary radiation process of pyrimidine bases and analysed by molecular orbital theory. This fast radiation protection mechanism provides the possibility to utilize electrophilic sulfhydryl protectors for realizing excellent protection effect. (author)

CMD: A Database to Store the Bonding States of Cysteine Motifs with Secondary Structures

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Hamed Bostan

2012-01-01

Full Text Available Computational approaches to the disulphide bonding state and its connectivity pattern prediction are based on various descriptors. One descriptor is the amino acid sequence motifs flanking the cysteine residue motifs. Despite the existence of disulphide bonding information in many databases and applications, there is no complete reference and motif query available at the moment. Cysteine motif database (CMD is the first online resource that stores all cysteine residues, their flanking motifs with their secondary structure, and propensity values assignment derived from the laboratory data. We extracted more than 3 million cysteine motifs from PDB and UniProt data, annotated with secondary structure assignment, propensity value assignment, and frequency of occurrence and coefficiency of their bonding status. Removal of redundancies generated 15875 unique flanking motifs that are always bonded and 41577 unique patterns that are always nonbonded. Queries are based on the protein ID, FASTA sequence, sequence motif, and secondary structure individually or in batch format using the provided APIs that allow remote users to query our database via third party software and/or high throughput screening/querying. The CMD offers extensive information about the bonded, free cysteine residues, and their motifs that allows in-depth characterization of the sequence motif composition.

Acid-Mediated Tumor Proteolysis: Contribution of Cysteine Cathepsins

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Jennifer M Rothberg

2013-10-01

Full Text Available One of the noncellular microenvironmental factors that contribute to malignancy of solid tumors is acidic peritumoral pH. We have previously demonstrated that extracellular acidosis leads to localization of the cysteine pro-tease cathepsin B on the tumor cell membrane and its secretion. The objective of the present study was to determine if an acidic extracellular pH such as that observed in vivo (i.e., pHe 6.8 affects the activity of proteases, e.g., cathepsin B, that contribute to degradation of collagen IV by tumor cells when grown in biologically relevant three-dimensional (3D cultures. For these studies, we used 1 3D reconstituted basement membrane overlay cultures of human carcinomas, 2 live cell imaging assays to assess proteolysis, and 3 in vivo imaging of active tumor proteases. At pHe 6.8, there were increases in pericellular active cysteine cathepsins and in degradation of dye-quenched collagen IV, which was partially blocked by a cathepsin B inhibitor. Imaging probes for active cysteine cathepsins localized to tumors in vivo. The amount of bound probe decreased in tumors in bicarbonate-treated mice, a treatment previously shown to increase peritumoral pHe and reduce local invasion of the tumors. Our results are consistent with the acid-mediated invasion hypothesis and with a role for cathepsin B in promoting degradation of a basement membrane protein substrate, i.e., type IV collagen, in an acidic peritumoral environment.

Indirect flow injection determination of N-acetyl-L-cysteine using cerium(IV) and ferroin

International Nuclear Information System (INIS)

Vieira, Heberth Juliano; Fatibello-Filho, Orlando

2005-01-01

An indirect flow injection spectrophotometric procedure is proposed for the determination of N-acetyl-L-cysteine in pharmaceutical formulations. In this system, ferroin ([Fe(II)-(fen) 2 ] 2+ ) in excess, with a strong absorption at 500 nm, is oxidized by cerium(IV) yielding cerium(III) and [Fe(III)-(fen) 2 ] 3+ (colorless), thus producing a baseline. When N-acetyl-L-cysteine solution is introduced into the flow injection system, it reacts with cerium(IV) increasing the analytical signal in proportion to the drug concentration. Under optimal experimental conditions, the linearity of the analytical curve for N-acetyl-L-cysteine ranged from 6.5x10 -6 to 1.3x10 -4 mol L -1 . The detection limit was 5.0x10 -6 mol L -1 and recoveries between 98.0 and 106% were obtained. The sampling frequency was 60 determinations per hour and the RSD was smaller than 1.4% for 2.2x10 -5 mol L -1 N-acetyl-L-cysteine. (author)

Solid state radiolysis of sulphur-containing amino acids. Cysteine, cystine and methionine

International Nuclear Information System (INIS)

Franco Cataldo; Pietro Ragni; Susana Iglesias-Groth; Arturo Manchado

2011-01-01

The sulphur-containing proteinaceous amino acids l-cysteine, l-cystine and l-methionine were irradiated in the solid state to a dose of 3.2 MGy. This dose corresponds to that delivered by radionuclide decay in a timescale of 1.05 x 10 9 years to the organic matter buried at a depth >20 m in comets and asteroids. The purity of the sulphur-containing amino acids was studied by differential scanning calorimetry (DSC) before and after the solid state radiolysis and the preservation of the chirality after the radiolysis was studied by chirooptical methods (optical rotatory dispersion, ORD) and by FT-IR spectroscopy. Although the high radiation dose of 3.2 MGy delivered, all the amino acids studied show a high radiation resistance. The best radiation resistance was offered by l-cysteine. The radiolysis of l-cysteine leads to the formation of l-cystine. The radiation resistance of l-methionine is not at the level of l-cysteine but also l-methionine is able to survive the dose of 3.2 MGy. Furthermore in all cases examined the preservation of chirality after radiolysis was clearly observed by the ORD spectroscopy although a certain level of radioracemization was measured in all cases. The radioracemization is minimal in the case of l-cysteine and is more pronounced in the case of l-methionine. In conclusion, the study shows that the sulphur-containing amino acids can survive for 1.05 x 10 9 years and, after extrapolation of the data, even to the age of the Solar System i.e. to 4.6 x 10 9 years. (author)

Interactions between N-acetyl-L-cysteine protected CdTe quantum dots and doxorubicin through spectroscopic method

Energy Technology Data Exchange (ETDEWEB)

Yang, Xiupei, E-mail: xiupeiyang@163.com [Chemical Synthesis and Pollution Control Key Laboratory of Sichuan Province, Nanchong 637000 (China); College of Chemistry and Chemical Engineering, China West Normal University, Nanchong 637000 (China); Lin, Jia; Liao, Xiulin; Zong, Yingying; Gao, Huanhuan [College of Chemistry and Chemical Engineering, China West Normal University, Nanchong 637000 (China)

2015-06-15

Highlights: • CdTe quantum dots with the diameter of 3–5 nm were synthesized in aqueous solution. • The modified CdTe quantum dots showed well fluorescence properties. • The interaction between the CdTe quantum dots and doxorubicin (DR) was investigated. - Abstract: N-acetyl-L-cysteine protected cadmium telluride quantum dots with a diameter of 3–5 nm were synthesized in aqueous solution. The interaction between N-acetyl-L-cysteine/cadmium telluride quantum dots and doxorubicin was investigated by ultraviolet–visible absorption and fluorescence spectroscopy at physiological conditions (pH 7.2, 37 °C). The results indicate that electron transfer has occurred between N-acetyl-L-cysteine/cadmium telluride quantum dots and doxorubicin under light illumination. The quantum dots react readily with doxorubicin to form a N-acetyl-L-cysteine/cadmium telluride-quantum dots/doxorubicin complex via electrostatic attraction between the −NH{sub 3}{sup +} moiety of doxorubicin and the −COO{sup −} moiety of N-acetyl-L-cysteine/cadmium telluride quantum dots. The interaction of N-acetyl-L-cysteine/cadmium telluride-quantum dots/doxorubicin complex with bovine serum albumin was studied as well, showing that the complex might induce the conformation change of bovine serum due to changes in microenvironment of bovine serum.

Interactions between N-acetyl-L-cysteine protected CdTe quantum dots and doxorubicin through spectroscopic method

International Nuclear Information System (INIS)

Yang, Xiupei; Lin, Jia; Liao, Xiulin; Zong, Yingying; Gao, Huanhuan

2015-01-01

Highlights: • CdTe quantum dots with the diameter of 3–5 nm were synthesized in aqueous solution. • The modified CdTe quantum dots showed well fluorescence properties. • The interaction between the CdTe quantum dots and doxorubicin (DR) was investigated. - Abstract: N-acetyl-L-cysteine protected cadmium telluride quantum dots with a diameter of 3–5 nm were synthesized in aqueous solution. The interaction between N-acetyl-L-cysteine/cadmium telluride quantum dots and doxorubicin was investigated by ultraviolet–visible absorption and fluorescence spectroscopy at physiological conditions (pH 7.2, 37 °C). The results indicate that electron transfer has occurred between N-acetyl-L-cysteine/cadmium telluride quantum dots and doxorubicin under light illumination. The quantum dots react readily with doxorubicin to form a N-acetyl-L-cysteine/cadmium telluride-quantum dots/doxorubicin complex via electrostatic attraction between the −NH 3 + moiety of doxorubicin and the −COO − moiety of N-acetyl-L-cysteine/cadmium telluride quantum dots. The interaction of N-acetyl-L-cysteine/cadmium telluride-quantum dots/doxorubicin complex with bovine serum albumin was studied as well, showing that the complex might induce the conformation change of bovine serum due to changes in microenvironment of bovine serum

Direct detection of cysteine using functionalized BaTiO3 nanoparticles film based self-powered biosensor.

Science.gov (United States)

Selvarajan, Sophia; Alluri, Nagamalleswara Rao; Chandrasekhar, Arunkumar; Kim, Sang-Jae

2017-05-15

Simple, novel, and direct detection of clinically important biomolecules have continuous demand among scientific community as well as in market. Here, we report the first direct detection and facile fabrication of a cysteine-responsive, film-based, self-powered device. NH 2 functionalized BaTiO 3 nanoparticles (BT-NH 2 NPs) suspended in a three-dimensional matrix of an agarose (Ag) film, were used for cysteine detection. BaTiO 3 nanoparticles (BT NPs) semiconducting as well as piezoelectric properties were harnessed in this study. The changes in surface charge properties of the film with respect to cysteine concentrations were determined using a current-voltage (I-V) technique. The current response increased with cysteine concentration (linear concentration range=10µM-1mM). Based on the properties of the composite (BT/Ag), we created a self-powered cysteine sensor in which the output voltage from a piezoelectric nanogenerator was used to drive the sensor. The potential drop across the sensor was measured as a function of cysteine concentrations. Real-time analysis of sensor performance was carried out on urine samples by non-invasive method. This novel sensor demonstrated good selectivity, linear concentration range and detection limit of 10µM; acceptable for routine analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

Key Role of Cysteine Residues in Catalysis and Subcellular Localization of Sulfur Oxygenase-Reductase of Acidianus tengchongensis

DEFF Research Database (Denmark)

Chen, Z. W.; Jiang, C. Y.; She, Qunxin

2005-01-01

). The thio-modifying reagent N-ethylmaleimide and Zn2+ strongly inhibited the activities of the SORs of A. tengchongensis, suggesting that cysteine residues are important. Site-directed mutagenesis was used to construct four mutant SORs with cysteines replaced by serine or alanine. The purified mutant......Analysis of known sulfur oxygenase-reductases (SORs) and the SOR-like sequences identified from public databases indicated that they all possess three cysteine residues within two conserved motifs (V-G-P-K-V-C31 and C101-X-X-C104; numbering according to the Acidianus tengchongensis numbering system...... proteins were investigated in parallel with the wild-type SOR. Replacement of any cysteine reduced SOR activity by 98.4 to 100%, indicating that all the cysteine residues are crucial to SOR activities. Circular-dichroism and fluorescence spectrum analyses revealed that the wild-type and mutant SORs have...

Carbon Nanotubes Facilitate Oxidation of Cysteine Residues of Proteins.

Science.gov (United States)

Hirano, Atsushi; Kameda, Tomoshi; Wada, Momoyo; Tanaka, Takeshi; Kataura, Hiromichi

2017-10-19

The adsorption of proteins onto nanoparticles such as carbon nanotubes (CNTs) governs the early stages of nanoparticle uptake into biological systems. Previous studies regarding these adsorption processes have primarily focused on the physical interactions between proteins and nanoparticles. In this study, using reduced lysozyme and intact human serum albumin in aqueous solutions, we demonstrated that CNTs interact chemically with proteins. The CNTs induce the oxidation of cysteine residues of the proteins, which is accounted for by charge transfer from the sulfhydryl groups of the cysteine residues to the CNTs. The redox reaction simultaneously suppresses the intermolecular association of proteins via disulfide bonds. These results suggest that CNTs can affect the folding and oxidation degree of proteins in biological systems such as blood and cytosol.

Synthesis and characterization of arsenic-doped cysteine-capped thoria-based nanoparticles

International Nuclear Information System (INIS)

Pereira, F. J.; Díez, M. T.; Aller, A. J.

2013-01-01

Thoria materials have been largely used in the nuclear industry. Nonetheless, fluorescent thoria-based nanoparticles provide additional properties to be applied in other fields. Thoria-based nanoparticles, with and without arsenic and cysteine, were prepared in 1,2-ethanediol aqueous solutions by a simple precipitation procedure. The synthesized thoria-based nanoparticles were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (ED-XRS), Raman spectroscopy, Fourier transform infrared (FT-IR) spectroscopy and fluorescence microscopy. The presence of arsenic and cysteine, as well as the use of a thermal treatment facilitated fluorescence emission of the thoria-based nanoparticles. Arsenic-doped and cysteine-capped thoria-based nanoparticles prepared in 2.5 M 1,2-ethanediol solutions and treated at 348 K showed small crystallite sizes and strong fluorescence. However, thoria nanoparticles subjected to a thermal treatment at 873 K also produced strong fluorescence with a very narrow size distribution and much smaller crystallite sizes, 5 nm being the average size as shown by XRD and TEM. The XRD data indicated that, even after doping of arsenic in the crystal lattice of ThO 2 , the samples treated at 873 K were phase pure with the fluorite cubic structure. The Raman and FT-IR spectra shown the most characteristics vibrational peaks of cysteine together with other peaks related to the bonds of this molecule to thoria and arsenic when present

Cysteine-rich intestinal protein binds zinc during transmucosal zinc transport

International Nuclear Information System (INIS)

Hempe, J.M.; Cousins, R.J.

1991-01-01

The mechanism of zinc absorption has not been delineated, but kinetic studies show that both passive and carrier-mediated processes are involved. The authors have identified a low molecular mass zinc-binding protein in the soluble fraction of rat intestinal mucosa that could function as an intracellular zinc carrier. The protein was not detected in liver or pancreas, suggesting a role specific to the intestine. The protein binds zinc during transmucosal zinc transport and shows signs of saturation at higher luminal zinc concentrations, characteristics consistent with a role in carrier-mediated zinc absorption. Microsequence analysis of the protein purified by gel-filtration HPCL and SDS/PAGE showed complete identity within the first 41 N-terminal amino acids with the deduced protein sequence of cysteine-rich intestinal protein. These investigators showed that the gene for this protein is developmentally regulated in neonates during the suckling period, conserved in many vertebrate species, and predominantly expressed in the small intestine. Cysteine-rich intestinal protein contains a recently identified conserved sequence of histidine and cysteine residues, the LIM motif, which our results suggest confers metal-binding properties that are important for zinc transport and/or functions of this micronutrient

Cobre no controle da verminose gastrintestinal em ovinos Copper on the control of gastrintestinal nematodes in sheep

Directory of Open Access Journals (Sweden)

Isabel Gonçalves de Gonçalves

2004-02-01

Full Text Available Este estudo teve o objetivo de verificar o tempo de proteção do cobre contra reinfecções por helmintos gastrintestinais de ovinos em pastejo. Foram utilizados 32 ovinos da raça Corriedale, com aproximadamente 18 meses de idade, mantidos em campo nativo naturalmente contaminado por trichostrongilídeos. Os animais foram pesados e dosificados. Após, foram alocados a quatro grupos experimentais: T I (tratado com cobre e necropsiado no 28ºdia; T II (sem cobre e necropsiado no 28º dia; T III (tratado com cobre e necropsiado no 56ºdia; T IV (sem cobre e necropsiado no 56º dia. Os grupos T I e T III receberam, via oral, uma cápsula gelatinosa de quatro gramas, contendo 3,4 gramas de óxido de cobre. Os ovinos foram avaliados semanalmente através de contagem de ovos por grama de fezes (OPG, volume globular (VG, aspartato aminotransferase (AST e concentrações de cobre no plasma. Ao abate, foram determinados: carga parasitária, concentrações de cobre no fígado e peso dos fígados. Os resultados demostraram que os níveis de cobre no plasma não tiveram alterações significativas (P>0,05 entre os diversos tratamentos. Quanto aos níveis de cobre detectados no fígado dos animais abatidos observou-se que os lotes medicados com cobre, apresentaram níveis mais elevados que os não medicados (Pe TII. Quanto à avaliação do OPG, não foram observadas diferenças significativas entre os grupos (P>0,05. Com relação aos parasitos gastrintestinais, detectou-se que a administração de cobre contribuiu significativamente para uma menor reinfecção apenas pelo Haemonchus contortus e somente nos animais abatidos na quarta semana após o tratamento, quando a redução foi de 60% nos medicados (P0,05. Com base nesses resultados, podemos concluir que 3,4 gramas de óxido de cobre podem ser efetivas na redução das reinfecções por H. contortus, durante quatro semanas sem causar toxicidade para ovinos criados extensivamente.The aim of this work

Effects of pH, temperature, and chemical structure on the stability of S-(purin-6-yl)-L-cysteine: evidence for a novel molecular rearrangement mechanism to yield N-(purin-6-yl)-L-cysteine.

Science.gov (United States)

Elfarra, A A; Hwang, I Y

1996-01-01

The stability of S-(purin-6-yl)-L-cysteine (SPC), a kidney-selective prodrug of 6-mercaptopurine and a putative metabolite of 6-chloropurine, was investigated under various pH and temperature conditions. At room temperature, the half-life (t 1/2) of SPC at either highly acidic (pH 3.6) or basic conditions (pH 9.6) was longer than at neutral or slightly acidic or basic conditions (pH 5.7-8.75). The primary degradation product, N-(purin-6-yl)-L-cysteine (NPC), was isolated using Sephadex LH-20 chromatography and characterized by 1H NMR and FAB/MS after derivatization with 2-iodoacetic acid. These results reveal novel stability requirements and implicate the cysteinyl amino group and the purinyl N-1 nitrogen in the mechanism of SPC rearrangement to NPC. Further evidence for this hypothesis was provided by the findings that the stability of SPC in phosphate buffer (pH 7.4) at 37 degrees C was similar to that of S-(guanin-6-yl)-L-cysteine, whereas S-(purin-6-yl)-N-acetyl-L-cysteine and S-(purin-6-yl)glutathione which have their cysteine amino groups blocked were much more stable than SPC. S-(Purin-6-yl)-L-homocysteine (SPHC) was also more stable than SPC, possibly because the formation of a 6-membered ring transition state as would be expected with SPHC is kinetically less favored than the formation of a 5-membered ring transition state as would be expected with SPC. These results may explain previous in vivo metabolism results of SPC and its analogs and may contribute to a better understanding of stability of structurally related cysteine S-conjugates.

Cysteine peptidases and their inhibitors in breast and genital cancer.

Directory of Open Access Journals (Sweden)

Magdalena Milan

2010-11-01

Full Text Available Cysteine proteinases and their inhibitors probably play the main role in carcinogenesis and metastasis. The metastasis process need external proteolytic activities that pass several barriers which are membranous structures of the connective tissue which includes, the basement membrane of blood vessels. Activities of the proteinases are regulated by endogenous inhibitors and activators. The imbalance between cysteine proteinases and cystatins seems to be associated with an increase in metastatic potential in some tumors. It has also been reported that proteinase inhibitors, specific antibodies for these enzymes and inhibition of the urokinase receptor may prevent cancer cell invasion. Some proteinase inhibitor could serve as agents for cancer treatment.

Cysteine-free peptides in scorpion venom: geographical distribution ...

African Journals Online (AJOL)

GRACE

2006-12-29

Dec 29, 2006 ... In 1993, the first cysteine-free peptide was isolated from scorpion venom. ..... Venom is produced by 2 venom glands in the tail and stored in 2 ... The resistance of a variety of bacterial micro-organisms .... Biopolymers 55: 4-30.

Formation of elemental sulfur by Chlorella fusca during growth on L-cysteine ethylester

Energy Technology Data Exchange (ETDEWEB)

Krauss, F.; Schafer, W.; Schmidt, A.

1984-01-01

During growth on L-cysteine ethylester, Chlorella fusca (211-8b) accumulated a substance which contained bound sulfide, which could be liberated by reduction with dithioerythritol (DTE) as inorganic sulfide. This substance was extracted with hot methanol and purified by thin layer chromatography. This substance liberated free sulfide when incubated with mono- and dithiols, and thiocyanate was formed after heating with KCN. The isolated substance cochromatographed with authentic sulfur flower using different solvent systems for thin layer chromatography, high pressure liquid chromatography, and the identical spectrum with a relative ..beta..max at 263 nm was found. The chemical structure was confirmed by mass spectrometry showing a molecular weight of 256 m/e for the S/sub 8/ configuration. No labeled elemental sulfur was detected when the cells were grown on (/sup 35/S)sulfate and L-cysteine ethylester. C. fusca seems to have enzymes for the metabolism of elemental sulfur, since it disappeared after prolonged growth into the stationary phase. Cysteine was formed from O-acetyl-L-serine and elemental sulfur in the presence of thiol groups and purified cysteine synthase from spinach or Chlorella.

Correlation of secreted protein acidic and rich in cysteine with diabetic nephropathy

OpenAIRE

Li, Lei; Song, Hai-Yan; Liu, Kai; An, Meng-Meng

2015-01-01

To detect the serum concentrations of secreted protein acidic and rich in cysteine (SPARC) in patients with diabetic nephropathy and SPARC mRNA and protein expressions in renal tissue of db/db mice (C57BL/KsJ, diabetic nephropathy mice), thus preliminary exploration on the role of secreted protein acidic riches in cysteine in the development of diabetic nephropathy were carried out. Serum SPARC levels in normal subjects, patients with type 2 diabetes mellitus (without diabetic nephropathy), c...

Thermo-Kinetic Investigation of Comparative Ligand Effect on Cysteine Iron Redox Reaction

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Masood Ahmad Rizvi

2015-03-01

Full Text Available Transition metal ions in their free state bring unwanted biological oxidations generating oxidative stress. The ligand modulated redox potential can be indispensable in prevention of such oxidative stress by blocking the redundant bio-redox reactions. In this study we investigated the comparative ligand effect on the thermo-kinetic aspects of biologically important cysteine iron (III redox reaction using spectrophotometric and potentiometric methods. The results were corroborated with the complexation effect on redox potential of iron(III-iron(II redox couple. The selected ligands were found to increase the rate of cysteine iron (III redox reaction in proportion to their stability of iron (II complex (EDTA < terpy < bipy < phen. A kinetic profile and the catalytic role of copper (II ions by means of redox shuttle mechanism for the cysteine iron (III redox reaction in presence of 1,10-phenanthroline (phen ligand is also reported.

Resistência anti-helmíntica de nematóides gastrintestinais em ovinos, Mato Grosso do Sul Anthelmintic resistance of gastrointestinal nematodes in sheep, Mato Grosso do Sul, Brazil

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Eurico A. Sczesny-Moraes

2010-03-01

Full Text Available Entre os métodos de controle da verminose gastrintestinal em ovinos, a utilização de produtos químicos é o mais empregado. Porém, o uso indiscriminado e continuado desses produtos tem selecionado populações de helmintos resistentes aos anti-helmínticos, fenômeno relatado no mundo todo. Este trabalho teve como objetivo identificar as espécies de parasitos gastrintestinais e diagnosticar a situação da resistência anti-helmíntica em ovinos no Estado de Mato Grosso do Sul. Foram realizados testes de redução na contagem de ovos por grama de fezes (OPG em rebanhos de dezesseis propriedades rurais; as sete formulações utilizadas continham as seguintes bases farmacológicas: Albendazol, Ivermectina, Levamisole, Triclorfon, Moxidectina, Closantel e uma contendo as três primeiras associadas. As espécies identificadas nas necropsias, em ovinos adultos, foram: Haemonchus contortus, Trichostrongylus colubriformis, Cooperia curticei, C. punctata, C. pectinata e Oesophagostomum columbianum; em ordem de prevalência. As formulações contendo Albendazol e Ivermectina não apresentaram eficácia na redução de OPG nos rebanhos testados, com médias de redução de 0,7 e -19,6%, respectivamente. Closantel apresentou eficácia média de 6,7%; Levamisole, Moxidectina e Triclorfon de 28,7, 26,8 e 65%, respectivamente; a associação das três bases (Albendazol, Ivermectina e Levamisole, uma média de eficácia de 55,8%. As percentagens médias de larvas infectantes recuperadas nas coproculturas, tanto no pré como no pós-tratamento, foram semelhantes; indicando que a resistência às bases testadas está presente em todas as espécies citadas, em maior ou menor intensidade. Os dois gêneros predominantemente resistentes são Haemonchus sp., com 86,9%; seguido por Trichostrongylus sp., com média de 47,5%; Strongyloides sp. 33,6%; Oesophagostomum, sp. 21,4% e Cooperia sp. 19,7%.Among the methods of control of gastrointestinal worms in sheep

Structural influence in the interaction of cysteine with five coordinated copper complexes: Theoretical and experimental studies

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Huerta-Aguilar, Carlos Alberto; Thangarasu, Pandiyan; Mora, Jesús Gracia

2018-04-01

Copper complexes of N,N,N‧,N‧-tetrakis(pyridyl-2-ylmethyl)-1,2-diaminoethane (L1) and N,N,N‧,N‧-tetrakis(pyridyl-2-ylmethyl)-1,3-diaminopropane (L2) prepared were characterized completely by different analytical methods. The X-structure of the complexes shows that Cu(II) presents in trigonal bi-pyramidal (TBP) geometry, consisting with the electronic spectra where two visible bands corresponding to five coordinated structure were observed. Thus TD-DFT was used to analyze the orbital contribution to the electronic transitions for the visible bands. Furthermore, the interaction of cysteine with the complexes was spectrally studied, and the results were explained through DFT analysis, observing that the geometrical parameters and oxidation state of metal ions play a vital role in the binding of cysteine with copper ion. It appears that the TBP structure is being changed into octahedral geometry during the addition of cysteine to the complexes as two bands (from complex) is turned to a broad band in visible region, signifying the occupation of cysteine molecule at sixth position of octahedral geometry. In the molecular orbital analysis, the existence of a strong overlapping of HOMOs (from cysteine) with LUMOs of Cu ion was observed. The total energy of the systems calculated by DFT shows that cysteine binds favorably with copper (I) than that with Cu(II).

Cross-talk between malarial cysteine proteases and falstatin: the BC loop as a hot-spot target.

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Srinivasan Sundararaj

Full Text Available Cysteine proteases play a crucial role in the development of the human malaria parasites Plasmodium falciparum and Plasmodium vivax. Our earlier studies demonstrated that these enzymes are equipped with specific domains for defined functions and further suggested the mechanism of activation of cysteine proteases. The activities of these proteases are regulated by a new class of endogenous inhibitors of cysteine proteases (ICPs. Structural studies of the ICPs of Trypanosoma cruzi (chagasin and Plasmodium berghei (PbICP indicated that three loops (termed BC, DE, and FG are crucial for binding to target proteases. Falstatin, an ICP of P. falciparum, appears to play a crucial role in invasion of erythrocytes and hepatocytes. However, the mechanism of inhibition of cysteine proteases by falstatin has not been established. Our study suggests that falstatin is the first known ICP to function as a multimeric protein. Using site-directed mutagenesis, hemoglobin hydrolysis assays and peptide inhibition studies, we demonstrate that the BC loop, but not the DE or FG loops, inhibits cysteine proteases of P. falciparum and P. vivax via hydrogen bonds. These results suggest that the BC loop of falstatin acts as a hot-spot target for inhibiting malarial cysteine proteases. This finding suggests new strategies for the development of anti-malarial agents based on protease-inhibitor interactions.

Therapeutic NOTCH3 cysteine correction in CADASIL using exon skipping: in vitro proof of concept

NARCIS (Netherlands)

Rutten, J.W.; Dauwerse, H.G.; Peters, D.J.; Goldfarb, A.; Venselaar, H.; Haffner, C.; Ommen, G.J. van; Aartsma-Rus, A.M.; Oberstein, S.A.

2016-01-01

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, or CADASIL, is a hereditary cerebral small vessel disease caused by characteristic cysteine altering missense mutations in theNOTCH3gene.NOTCH3mutations in CADASIL result in an uneven number of cysteine

Global analysis of gene expression in response to L-Cysteine deprivation in the anaerobic protozoan parasite Entamoeba histolytica

Science.gov (United States)

2011-01-01

Background Entamoeba histolytica, an enteric protozoan parasite, causes amebic colitis and extra intestinal abscesses in millions of inhabitants of endemic areas. E. histolytica completely lacks glutathione metabolism but possesses L-cysteine as the principle low molecular weight thiol. L-Cysteine is essential for the structure, stability, and various protein functions, including catalysis, electron transfer, redox regulation, nitrogen fixation, and sensing for regulatory processes. Recently, we demonstrated that in E. histolytica, L-cysteine regulates various metabolic pathways including energy, amino acid, and phospholipid metabolism. Results In this study, employing custom-made Affymetrix microarrays, we performed time course (3, 6, 12, 24, and 48 h) gene expression analysis upon L-cysteine deprivation. We identified that out of 9,327 genes represented on the array, 290 genes encoding proteins with functions in metabolism, signalling, DNA/RNA regulation, electron transport, stress response, membrane transport, vesicular trafficking/secretion, and cytoskeleton were differentially expressed (≥3 fold) at one or more time points upon L-cysteine deprivation. Approximately 60% of these modulated genes encoded proteins of no known function and annotated as hypothetical proteins. We also attempted further functional analysis of some of the most highly modulated genes by L-cysteine depletion. Conclusions To our surprise, L-cysteine depletion caused only limited changes in the expression of genes involved in sulfur-containing amino acid metabolism and oxidative stress defense. In contrast, we observed significant changes in the expression of several genes encoding iron sulfur flavoproteins, a major facilitator super-family transporter, regulator of nonsense transcripts, NADPH-dependent oxido-reductase, short chain dehydrogenase, acetyltransferases, and various other genes involved in diverse cellular functions. This study represents the first genome-wide analysis of

Global analysis of gene expression in response to L-Cysteine deprivation in the anaerobic protozoan parasite Entamoeba histolytica

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Jeelani Ghulam

2011-05-01

Full Text Available Abstract Background Entamoeba histolytica, an enteric protozoan parasite, causes amebic colitis and extra intestinal abscesses in millions of inhabitants of endemic areas. E. histolytica completely lacks glutathione metabolism but possesses L-cysteine as the principle low molecular weight thiol. L-Cysteine is essential for the structure, stability, and various protein functions, including catalysis, electron transfer, redox regulation, nitrogen fixation, and sensing for regulatory processes. Recently, we demonstrated that in E. histolytica, L-cysteine regulates various metabolic pathways including energy, amino acid, and phospholipid metabolism. Results In this study, employing custom-made Affymetrix microarrays, we performed time course (3, 6, 12, 24, and 48 h gene expression analysis upon L-cysteine deprivation. We identified that out of 9,327 genes represented on the array, 290 genes encoding proteins with functions in metabolism, signalling, DNA/RNA regulation, electron transport, stress response, membrane transport, vesicular trafficking/secretion, and cytoskeleton were differentially expressed (≥3 fold at one or more time points upon L-cysteine deprivation. Approximately 60% of these modulated genes encoded proteins of no known function and annotated as hypothetical proteins. We also attempted further functional analysis of some of the most highly modulated genes by L-cysteine depletion. Conclusions To our surprise, L-cysteine depletion caused only limited changes in the expression of genes involved in sulfur-containing amino acid metabolism and oxidative stress defense. In contrast, we observed significant changes in the expression of several genes encoding iron sulfur flavoproteins, a major facilitator super-family transporter, regulator of nonsense transcripts, NADPH-dependent oxido-reductase, short chain dehydrogenase, acetyltransferases, and various other genes involved in diverse cellular functions. This study represents the first

Palmitoylation of the cysteine-rich endodomain of the SARS-coronavirus spike glycoprotein is important for spike-mediated cell fusion

International Nuclear Information System (INIS)

Petit, Chad M.; Chouljenko, Vladimir N.; Iyer, Arun; Colgrove, Robin; Farzan, Michael; Knipe, David M.; Kousoulas, K.G.

2007-01-01

The SARS-coronavirus (SARS-CoV) is the etiological agent of the severe acute respiratory syndrome (SARS). The SARS-CoV spike (S) glycoprotein mediates membrane fusion events during virus entry and virus-induced cell-to-cell fusion. The cytoplasmic portion of the S glycoprotein contains four cysteine-rich amino acid clusters. Individual cysteine clusters were altered via cysteine-to-alanine amino acid replacement and the modified S glycoproteins were tested for their transport to cell-surfaces and ability to cause cell fusion in transient transfection assays. Mutagenesis of the cysteine cluster I, located immediately proximal to the predicted transmembrane, domain did not appreciably reduce cell-surface expression, although S-mediated cell fusion was reduced by more than 50% in comparison to the wild-type S. Similarly, mutagenesis of the cysteine cluster II located adjacent to cluster I reduced S-mediated cell fusion by more than 60% compared to the wild-type S, while cell-surface expression was reduced by less than 20%. Mutagenesis of cysteine clusters III and IV did not appreciably affect S cell-surface expression or S-mediated cell fusion. The wild-type S was palmitoylated as evidenced by the efficient incorporation of 3 H-palmitic acid in wild-type S molecules. S glycoprotein palmitoylation was significantly reduced for mutant glycoproteins having cluster I and II cysteine changes, but was largely unaffected for cysteine cluster III and IV mutants. These results show that the S cytoplasmic domain is palmitoylated and that palmitoylation of the membrane proximal cysteine clusters I and II may be important for S-mediated cell fusion

Long-time treatment by low-dose N-acetyl-L-cysteine enhances proinflammatory cytokine expressions in LPS-stimulated macrophages.

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Tomokazu Ohnishi

Full Text Available N-acetyl-L-cysteine is known to act as a reactive oxygen species scavenger and used in clinical applications. Previous reports have shown that high-dose N-acetyl-L-cysteine treatment inhibits the expression of proinflammatory cytokines in activated macrophages. Here, we have found that long-time N-acetyl-L-cysteine treatment at low-concentration increases phosphorylation of extracellular signal-regulated kinase 1/2 and AKT, which are essential for the induction of proinflammatory cytokines including interleukin 1β and interleukin 6 in lipopolysaccharide-stimulated RAW264.7 cells. Furthermore, long-time N-acetyl-L-cysteine treatment decreases expressions of protein phosphatases, catalytic subunit of protein phosphatase-2A and dual specificity phosphatase 1. On the other hand, we have found that short-time N-acetyl-L-cysteine treatment at low dose increases p53 expression, which inhibits expressions of proinflammatory cytokines. These observations suggest that long-time low-dose N-acetyl-L-cysteine treatment increases expressions of proinflammatory cytokines through enhancement of kinase phosphorylation.

Functional cardiovascular action of L-cysteine microinjected into pressor sites of the rostral ventrolateral medulla of the rat.

Science.gov (United States)

Takemoto, Yumi

2014-04-01

The endogenous sulfur-containing amino acid L-cysteine injected into the cerebrospinal fluid space of the cisterna magna increases arterial blood pressure (ABP) and heart rate (HR) in the freely moving rat. The present study examined (1) cardiovascular responses to L-cysteine microinjected into the rostral ventrolateral medulla (RVLM), where a group of neurons regulate activities of cardiovascular sympathetic neurons and (2) involvement of ionotropic excitatory amino acid (iEAA) receptors in response. In the RVLM of urethane-anesthetized rats accessed ventrally and identified with pressor responses to L-glutamate (10 mM, 34 nl), microinjections of L-cysteine increased ABP and HR dose dependently (3-100 mM, 34 nl). The cardiovascular responses to L-cysteine (30 mM) were not attenuated by a prior injection of either antagonist alone, MK801 (20 mM, 68 nl) for the NMDA type of iEAA receptors, or CNQX (2 mM) for the non-NMDA type. However, inhibition of both NMDA and non-NMDA receptors with additional prior injection of either antagonist completely blocked those responses to L-cysteine. The results indicate that L-cysteine has functional cardiovascular action in the RVLM of the anesthetized rat, and the responses to L-cysteine involve both NMDA and non-NMDA receptors albeit in a mutually exclusive parallel fashion. The findings may suggest endogenous roles of L-cysteine indirectly via iEAA receptors in the neuronal network of the RVLM for cardiovascular regulation in physiological and pathological situations.

Eficácia anti-helmíntica comparativa da associação albendazole, levamisole e ivermectina à moxidectina em ovinos Comparative anthelmintic efficacy of the association albendazol, levamisol and ivermectin to moxidectin in sheep

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Carolina Buzzulini

2007-06-01

Full Text Available Avaliou-se a eficácia anti-helmíntica da associação de albendazole 2,0%, cloridrato de levamisole 2,55% e ivermectina 0,08% comparativamente à moxidectina 1% em ovinos naturalmente infectados. Foram selecionados 24 ovinos para a composição de três grupos experimentais com oito animais cada: T1, ovinos tratados com a associação albendazole, levamisole e ivermectina, na dosagem de 1 mL 4 kg-1 de peso corporal; T2, ovinos tratados com moxidectina, na dosagem de 1 mL 50 kg-1 de peso corporal e T3, ovinos sem tratamento anti-helmíntico. Foram realizadas contagens de ovos por grama de fezes (OPG no primeiro, terceiro, quinto e sétimo dia após os tratamentos. No sétimo dia todos os ovinos foram necropsiados e todos os helmintos encontrados no trato gastrintestinal foram quantificados e identificados quanto ao gênero e à espécie. A associação dos diferentes princípios ativos foi 100% eficaz no combate às espécies Cooperia punctata, C. pectinata, C. spatulata, Trichostrongylus axei, Oesophagostomum columbianum, Trichuris ovis, C. curticei e Strongyloides papillosus e, a moxidectina eliminou as seis primeiras espécies citadas. Contra Haemonchus contortus a associação apresentou eficácia superior (93% à moxidectina (51,4%. Ambas formulações foram eficazes contra Trichostrongylus colubriformis. A associação medicamentosa utilizada constitui alternativa no controle das nematodioses ovinas.The anthelmintic efficacy of 2.0% albendazol, 2.55% levamisol chloridrate and 0.08% ivermectin formulation to 1% moxidectin in sheep naturally infected with gastrointestinal nematodes was compared. Twenty-four animals were selected by faecal egg counts (FEC means, composing three experimental groups with eight sheep each: T1, sheep treated with albendazol, levamisol and ivermectin association; T2, sheep treated with 1% moxidectin and T3, untreated group. FEC was estimated at 1st, 3rd, 5th and 7th day post-treatment. All animals were

La epizootiología como herramienta para el control parasitario en ovinos Parasite epizootiology in the parasite control in sheep

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J Arece

2007-12-01

Full Text Available El parasitismo interno en los ovinos constituye una de las limitantes de mayor importancia en el desarrollo exitoso de la producción ovina. El primer paso para lograr un control parasitario está relacionado con el estudio de la epizootiología de los agentes etiológicos, y entre ellos el conocimiento de las especies que circulan en los rebaños. Para realizar este tipo de estudio se seleccionaron tres unidades de producción ovinas de la provincia de Matanzas, cuyos animales y sistema de manejo son representativos de los del resto de la provincia. Se encontró que los animales están infestados de forma simpátrica por dos especies del género Haemonchus (H. placei y H. contortus que superan el 90% del conteo fecal durante todo el año. También se encontró Trichostrongylus colubriformis y Oesophagostomum columbianum, en menor cuantía. Las mayores tasas de infestación se presentaron en el período lluvioso en las tres unidades, lo que muestra una estacionalidad evidente. Las crías se parasitaron en función del sistema de manejo implementado y las reproductoras paridas fueron las más susceptibles en un grupo de hembras incorporadas a la reproducción. Estos elementos son de vital importancia para la puesta en marcha de planes de control parasitario.Internal parasitism in sheep constitutes one of the most important limitations in the successful development of sheep production. The first step for achieving parasite control is related to the study of the epizootiology of the etiological agents, and among them the knowledge on the species that circulate in the herds. For performing this type of study, three sheep production units of Matanzas province were selected, which animals and management system are representative of the rest of the province. The animals were found to be infested in a sympatric way by two species of the Haemonchus genus (H. placei and H. contortus which exceed 90% of the fecal count during the whole year. Also

Low-quality birds do not display high-quality signals: The cysteine-pheomelanin mechanism of honesty

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Galván, Ismael; Wakamatsu, Kazumasa; Camarero, Pablo R; Mateo, Rafael; Alonso-Alvarez, Carlos

2015-01-01

The mechanisms that make that the costs of producing high-quality signals are unaffordable to low-quality signalers are a current issue in animal communication. The size of the melanin-based bib of male house sparrows Passer domesticus honestly signals quality. We induced the development of new bibs while treating males with buthionine-sulfoximine (BSO), a substance that depletes the levels of the antioxidant glutathione (GSH) and the amino acid cysteine, two elements that switch melanogenesis from eumelanin to pheomelanin. Final bib size is negatively related to pheomelanin levels in the bib feathers. BSO reduced cysteine and GSH levels in all birds, but improved phenotypes (bibs larger than controls) were only expressed by high-quality birds (BSO birds with largest bibs initially). Negative associations between final bib size and cysteine levels in erythrocytes, and between pheomelanin and cysteine levels, were observed in high-quality birds only. These findings suggest that a mechanism uncoupling pheomelanin and cysteine levels may have evolved in low-quality birds to avoid producing bibs of size not corresponding to their quality and greater relative costs. Indeed, greater oxidative stress in cells was not observed in low-quality birds. This may represent the first mechanism maintaining signal honesty without producing greater relative costs on low-quality signalers. PMID:25330349

Ga2O3 photocatalyzed on-line tagging of cysteine to facilitate peptide mass fingerprinting.

Science.gov (United States)

Qiao, Liang; Su, Fangzheng; Bi, Hongyan; Girault, Hubert H; Liu, Baohong

2011-09-01

β-Ga(2)O(3) is a wide-band-gap semiconductor having strong oxidation ability under light irradiation. Herein, the steel target plates modified with β-Ga(2)O(3) nanoparticles have been developed to carry out in-source photo-catalytic oxidative reactions for online peptide tagging during laser desorption/ionization mass spectrometry (LDI-MS) analysis. Under UV laser irradiation, β-Ga(2)O(3) can catalyze the photo-oxidation of 2-methoxyhydroquinone added to a sample mixture to 2-methoxy benzoquinone that can further react with the thiol groups of cysteine residues by Michael addition reaction. The tagging process leads to appearance of pairs of peaks with an m/z shift of 138.1Th. This online labelling strategy is demonstrated to be sensitive and efficient with a detection-limit at femtomole level. Using the strategy, the information on cysteine content in peptides can be obtained together with peptide mass, therefore constraining the database searching for an advanced identification of cysteine-containing proteins from protein mixtures. The current peptide online tagging method can be important for specific analysis of cysteine-containing proteins especially the low-abundant ones that cannot be completely isolated from other high-abundant non-cysteine-proteins. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Rapid and selective detection of cysteine based on its induced aggregates of cetyltrimethylammonium bromide capped gold nanoparticles

International Nuclear Information System (INIS)

Wang Jian; Li Yuanfang; Huang Chengzhi; Wu Tong

2008-01-01

A detection method of cysteine is reported in this contribution with water-soluble positively charged gold nanoparticles (Au-NPs) that were prepared by seed-mediated method and capped with cetyltrimethylammonium bromide (CTAB). In aqueous medium of pH 4.2, the CTAB-capped Au-NPs display greatly different features from those of generally prepared citrate-coated Au-NPs. It was found that in a medium of high salt concentration, the presence of cysteine could induce aggregation of CTAB-capped Au-NPs, while citrate-coated Au-NPs could get aggregation soon even if without the presence of cysteine. The cysteine-induced aggregates of CTAB-capped Au-NPs display strong plasmon resonance light scattering (PRLS) signals characterized at 566.0 nm when excited by a light beam, and the PRLS intensities of the aggregates are in proportion to the concentration of cysteine in the range of 0.01-0.40 μg mL -1 with the limit of detection (3σ) being 2.9 ng mL -1 . No amino acids in the samples interfere with the detection, and cysteine in artificial samples could be detected with the recovery between 95.3% and 105.9%, and R.S.D. is less than 3.6%

Cloning and characterization of a novel cysteine protease gene ...

Indian Academy of Sciences (India)

PRAKASH KUMAR

Cysteine proteases can be found in the animal and plant kingdoms as well as in some viruses and bacteria. They have been implemented in many ..... in developing resistance against pathogens and insects in other crops. Acknowledgments.

Kinetics and mechanism of reduction of iron(iii) kojic acid complex by hydroquinone and l-cysteine

International Nuclear Information System (INIS)

Hussain, Z.; Perviaz, M.; Kazmi, S.A.; Johnson, A.S.; Offiong, O.E.

2014-01-01

The effect of pH on the kinetics of reduction of iron(III) kojic acid complex by hydroquinone (H/sub 2/Q) and L-cysteine (L-Cys) was studied in the pH range of 2.34 - 4.03 for H/sub 2/Q and 3.04 - 5.5 for L-cysteine at ionic strength of 0.5 M and at 35 degree C. The pseudo-first order rate constants for the reduction of Fe(KA)3 by L-cysteine and hydroquinone increase linearly with increasing reductant concentration, indicating first-order kinetics in reductant concentration. However, whereas the rate of reduction by H2Q increases with increasing pH, an opposite trend was observed in the case of reduction by L-cysteine. Plausible rate laws and mechanisms have been proposed in line with these observations. Activation parameters (delta H no and delta S no) were evaluated for the reduction of iron (III) kojic acid complex by cysteine and the values obtained are 35.25 kJmol-1, -141.4 JK-1mol-1 and 28.14 kJmol-1 , 161.2 JK-1mol-1 for pH 4.5 and 3.52 respectively. (author)

A new simple phthalimide-based fluorescent probe for highly selective cysteine and bioimaging for living cells

Science.gov (United States)

Shen, Youming; Zhang, Xiangyang; Zhang, Youyu; Zhang, Chunxiang; Jin, Junling; Li, Haitao

2017-10-01

A new turn-on phthalimide fluorescent probe has designed and synthesized for sensing cysteine (Cys) based on excited state intramolecular proton transfer (ESIPT) process. It is consisted of a 3-hydroxyphthalimide derivative moiety as the fluorophore and an acrylic ester group as a recognition receptor. The acrylic ester acts as an ESIPT blocking agent. Upon addition of cystein, intermolecular nucleophilic attack of cysteine on acrylic ester releases the fluorescent 3-hydroxyphthalimide derivative, thereby enabling the ESIPT process and leading to enhancement of fluorescence. The probe displays high sensitivity, excellent selectivity and with large Stokes shift toward cysteine. The linear interval range of the fluorescence titration ranged from 0 to 1.0 × 10- 5 M and detection limit is low (6 × 10- 8 M). In addition, the probe could be used for bio-imaging in living cells.

Cyst(e)ine requirements in enterally fed very low birth weight preterm infants.

Science.gov (United States)

Riedijk, Maaike A; Voortman, Gardi; van Beek, Ron H T; Baartmans, Martin G A; Wafelman, Leontien S; van Goudoever, Johannes B

2008-03-01

Optimal nutrition is of utmost importance for the preterm infant's later health and developmental outcome. Amino acid requirements for preterm infants differ from those for term and older infants, because growth rates differ. Some nonessential amino acids, however, cannot be sufficiently synthesized endogenously. Cyst(e)ine is supposed to be such a conditionally essential amino acid in preterm infants. The objective of this study was to determine, at 32 and 35 weeks' postmenstrual age, cyst(e)ine requirements in fully enterally fed very low birth weight preterm infants with gestational ages of ine requirement was determined with the indicator amino acid oxidation technique ([1-(13)C]phenylalanine) after 24-hour adaptation. Fractional [1-(13)C]phenylalanine oxidation was established in 47 very low birth weight preterm infants (mean gestational age: 28 weeks +/- 1 week SD; birth weight: 1.07 kg +/- 0.21 kg SD). Increase in dietary cyst(e)ine intake did not result in a decrease in fractional [1-(13)C]phenylalanine oxidation. These data do not support the hypothesis that endogenous cyst(e)ine synthesis is limited in very low birth weight preterm infants with gestational ages of ine requirement is ine is probably not a conditionally essential amino acid in these infants.

Cysteine-grafted nonwoven geotextile: a new and efficient material for heavy metals sorption--Part B.

Science.gov (United States)

Vandenbossche, M; Vezin, H; Touati, N; Jimenez, M; Casetta, M; Traisnel, M

2014-10-01

The development of a new material designed to trap heavy metals from sediments or wastewater, based on a polypropylene non-woven covalently grafted with cysteine, has been reported in a previous paper (Part A). The non-woven was first functionalized with acrylic acid (AA) which is used as spacer, and then cysteine was immobilized on the substrate through covalent coupling in order to obtain the so-called PP-g-AA-cysteine. Some preliminary heavy metals adsorption tests gave interesting results: at 20 °C for 24 h and in a 1000 mg/L heavy metals solution, PP-g-AA-cysteine adsorbs 95 mg Cu/g PP (CuSO4 solution), 104 mg Cu/g PP (Cu(NO3)2 solution), 135 mg Pb/g PP (Pb(NO3)2 solution) and 21 mg Cr/g PP (Cr(NO3)3 solution). In this second part of the work, heavy metals sorption tests were carried out with Cu (II), Pb (II), and Cr (III) separately, in order to determine the sorption capacity of this new sorbent as a function of (i) the heavy metals concentration in the solution, (ii) the contact time with the solution, (iii) the pH and (iv) the ionic strength of the solution containing heavy metals. Moreover, the sorption capacity of PP-g-AA-Cysteine was studied using a polluted solution consisting of a mixture of these different heavy metals. An Electron Paramagnetic Resonance study was finally carried out in order to determine the coordination geometry in the environment of the copper trapped by the PP-g-AA-cysteine. Copyright © 2014 Elsevier Ltd. All rights reserved.

L-Cysteine Administration Attenuates Pancreatic Fibrosis Induced by TNBS in Rats by Inhibiting the Activation of Pancreatic Stellate Cell

Science.gov (United States)

Hu, GuoYong; Shen, Jie; Wang, Feng; Xu, Ling; Dai, WeiQi; Xiong, Jie; Ni, JianBo; Guo, ChuanYong; Wan, Rong; Wang, XingPeng

2012-01-01

Background and Aims Recent studies have shown that activated pancreatic stellate cells (PSCs) play a major role in pancreatic fibrogenesis. We aimed to study the effect of L-cysteine administration on fibrosis in chronic pancreatitis (CP) induced by trinitrobenzene sulfonic acid (TNBS) in rats and on the function of cultured PSCs. Methods CP was induced by TNBS infusion into rat pancreatic ducts. L-cysteine was administrated for the duration of the experiment. Histological analysis and the contents of hydroxyproline were used to evaluate pancreatic damage and fibrosis. Immunohistochemical analysis of α-SMA in the pancreas was performed to detect the activation of PSCs in vivo. The collagen deposition related proteins and cytokines were determined by western blot analysis. DNA synthesis of cultured PSCs was evaluated by BrdU incorporation. We also evaluated the effect of L-cysteine on the cell cycle and cell activation by flow cytometry and immunocytochemistry. The expression of PDGFRβ, TGFβRII, collagen 1α1 and α-SMA of PSCs treated with different concentrations of L-cysteine was determined by western blot. Parameters of oxidant stress were evaluated in vitro and in vivo. Nrf2, NQO1, HO-1, IL-1β expression were evaluated in pancreas tissues by qRT-PCR. Results The inhibition of pancreatic fibrosis by L-cysteine was confirmed by histological observation and hydroxyproline assay. α-SMA, TIMP1, IL-1β and TGF-β1 production decreased compared with the untreated group along with an increase in MMP2 production. L-cysteine suppressed the proliferation and extracellular matrix production of PSCs through down-regulating of PDGFRβ and TGFβRII. Concentrations of MDA+4-HNE were decreased by L-cysteine administration along with an increase in GSH levels both in tissues and cells. In addition, L-cysteine increased the mRNA expression of Nrf2, NQO1 and HO-1 and reduced the expression of IL-1β in L-cysteine treated group when compared with control group. Conclusion L-cysteine

Dynamics of postirradiation intracellular cysteine and aspartic proteinases profiles in proliferating and nonproliferating mammalian cells

International Nuclear Information System (INIS)

Korbelik, M.; Osmak, M.; Suhar, A.; Turk, V.; Skrk, J.

1990-01-01

Dynamics of postirradiation intracellular cysteine and aspartic proteinases profiles were examined in proliferating and nonproliferating Chinese hamster fibroblasts (V 79). The results show that there are significant alterations in cysteine and aspartic intracellular proteinases activity already in the early postirradiation period, which are different in proliferating and nonproliferating cells. Irradiation of the cells examined to low doses and up to 15 Gy induced an increase in cysteine proteinases activity in the early postexposure period, while at higher irradiation doses applied, the activity of these proteinases was decreased. These observations suggest that intracellular proteinases are actively participating in process involving recovery from radiation injury or cell killing. (orig.) [de

L-cysteine efflux in erythrocytes as a function of human age: correlation with reduced glutathione and total anti-oxidant potential.

Science.gov (United States)

Kumar, Prabhanshu; Maurya, Pawan Kumar

2013-06-01

Thiol compounds such as cysteine (Cys) and reduced glutathione (GSH) play an important role in human aging and age-related diseases. In erythrocytes, GSH is synthesized by glutamic acid, cysteine, and glycine, but the rate of GSH synthesis is determined only by the availability of L-cysteine. Cysteine supplementation has been shown to ameliorate several parameters that are known to degenerate during human aging. We have studied L-cysteine efflux in vitro in human erythrocytes as a function of age by suspending cells in solution containing 10 mM L-cysteine for uptake; later cells were re-suspended in phosphate-buffered saline (PBS)-glucose to allow efflux. Change in the free sulfhydryl (-SH) concentration was then measured to calculate the rate of efflux. The GSH/oxidized glutathione (GSSG) ratio was taken as a control to study the oxidation/reduction state of the erythrocyte. The total anti-oxidant potential of plasma was measured in terms of ferric reducing ability of plasma (FRAP) values. We have shown a significant (pL-cysteine in erythrocytes during human aging, and the GSH/GSSG ratio decreases as a function of human age. The decline in L-cysteine efflux during aging correlates with the decrease in GSH and the FRAP value. This finding may help to explain the shift in the redox status and low GSH concentration that might determine the rate of L-cysteine efflux observed in erythrocytes and an important factor in the development of oxidative stress in erythrocytes during aging.

Molecular cloning of a cysteine proteinase cDNA from the cotton boll weevil Anthonomus grandis (Coleoptera: Curculionidae).

Science.gov (United States)

De Oliveira Neto, Osmundo Brilhante; Batista, João Aguiar Nogueira; Rigden, Daniel John; Franco, Octávio Luiz; Fragoso, Rodrigo Rocha; Monteiro, Ana Carolina Santos; Monnerat, Rose Gomes; Grossi-De-Sa, Maria Fátima

2004-06-01

The cotton boll weevil (Anthonomus grandis) causes severe cotton crop losses in North and South America. This report describes the presence of cysteine proteinase activity in the cotton boll weevil. Cysteine proteinase inhibitors from different sources were assayed against total A. grandis proteinases but, unexpectedly, no inhibitor tested was particularly effective. In order to screen for active inhibitors against the boll weevil, a cysteine proteinase cDNA (Agcys1) was isolated from A. grandis larvae using degenerate primers and rapid amplification of cDNA ends (RACE) techniques. Sequence analysis showed significant homologies with other insect cysteine proteinases. Northern blot analysis indicated that the mRNA encoding the proteinase was transcribed mainly in the gut of larvae. No mRNA was detected in neonatal larvae, pupae, or in the gut of the adult insect, suggesting that Agcys1 is an important cysteine proteinase for larvae digestion. The isolated gene will facilitate the search for highly active inhibitors towards boll weevil larvae that may provide a new opportunity to control this important insect pest.

Metabolic Design of Corynebacterium glutamicum for Production of l-Cysteine with Consideration of Sulfur-Supplemented Animal Feed.

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Joo, Young-Chul; Hyeon, Jeong Eun; Han, Sung Ok

2017-06-14

l-Cysteine is a valuable sulfur-containing amino acid widely used as a nutrition supplement in industrial food production, agriculture, and animal feed. However, this amino acid is mostly produced by acid hydrolysis and extraction from human or animal hairs. In this study, we constructed recombinant Corynebacterium glutamicum strains that overexpress combinatorial genes for l-cysteine production. The aims of this work were to investigate the effect of the combined overexpression of serine acetyltransferase (CysE), O-acetylserine sulfhydrylase (CysK), and the transcriptional regulator CysR on l-cysteine production. The CysR-overexpressing strain accumulated approximately 2.7-fold more intracellular sulfide than the control strain (empty pMT-tac vector). Moreover, in the resulting CysEKR recombinant strain, combinatorial overexpression of genes involved in l-cysteine production successfully enhanced its production by approximately 3.0-fold relative to that in the control strain. This study demonstrates a biotechnological model for the production of animal feed supplements such as l-cysteine using metabolically engineered C. glutamicum.

Cysteine reversal of the novel neuromuscular blocking drug CW002 in dogs: pharmacodynamics, acute cardiovascular effects, and preliminary toxicology.

Science.gov (United States)

Sunaga, Hiroshi; Malhotra, Jaideep K; Yoon, Edward; Savarese, John J; Heerdt, Paul M

2010-04-01

CW002 is a neuromuscular blocking drug that is inactivated by endogenous L-cysteine. This study determined the exogenous L-cysteine dose-response relationship for CW002 reversal along with acute cardiovascular effects and organ toxicity in dogs. Six dogs were each studied four times during isoflurane-nitrous oxide anesthesia and recording of muscle twitch, arterial pressure, and heart rate. CW002 (0.08 mg/kg or 9 x ED95) was injected, and the time to spontaneous muscle recovery was determined. CW002 was then administered again followed 1 min later by 10, 20, 50, or 100 mg/kg L-cysteine (1 dose/experiment). After twitch recovery, CW002 was given a third time to determine whether residual L-cysteine influenced duration. Preliminary toxicology was performed in an additional group of dogs that received CW002 followed by vehicle (n = 8) or 200 mg/kg L-cysteine (n = 8). Animals were awakened and observed for 2 or 14 days before sacrificing and anatomic, biochemical, and histopathologic analyses. L-cysteine at all doses accelerated recovery from CW002, with both 50 and 100 mg/kg decreasing median duration from more than 70 min to less than 5 min. After reversal, duration of a subsequent CW002 dose was also decreased in a dose-dependent manner. Over the studied dose range, L-cysteine had less than 10% effect on blood pressure and heart rate. Animals receiving a single 200-mg/kg dose of L-cysteine showed no clinical, anatomic, biochemical, or histologic evidence of organ toxicity. The optimal L-cysteine dose for rapidly reversing the neuromuscular blockade produced by a large dose of CW002 in dogs is approximately 50 mg/kg, which has no concomitant hemodynamic effect. A dose of 200 mg/kg had no evident organ toxicity.

Electrochemical immobilization of biomolecules on gold surface modified with monolayered L-cysteine

Energy Technology Data Exchange (ETDEWEB)

Honda, Mitsunori, E-mail: honda.mitsunori@jaea.go.jp; Baba, Yuji; Sekiguchi, Tetsuhiro; Shimoyama, Iwao; Hirao, Norie

2014-04-01

Immobilization of organic molecules on the top of a metal surface is not easy because of lattice mismatch between organic and metal crystals. Gold atoms bind to thiol groups through strong chemical bonds, and a self-assembled monolayer of sulfur-terminated organic molecules is formed on the gold surface. Herein, we suggested that a monolayer of L-cysteine deposited on a gold surface can act as a buffer layer to immobilize biomolecules on the metal surface. We selected lactic acid as the immobilized biomolecule because it is one of the simplest carboxyl-containing biomolecules. The immobilization of lactic acid on the metal surface was carried out by an electrochemical method in an aqueous environment under the potential range varying from − 0.6 to + 0.8 V. The surface chemical states before and after the electrochemical reaction were characterized using X-ray photoelectron spectroscopy (XPS). The N 1s and C 1s XPS spectra showed that the L-cysteine-modified gold surface can immobilize lactic acid via peptide bonds. This technique might enable the immobilization of large organic molecules and biomolecules. - Highlights: • Monolayer l-cysteine deposited on Au surface as a buffer layer to immobilize biomolecules. • Lactic acid as the immobilized biomolecule as it is simple carboxyl-containing biomolecule. • X-ray photoelectron spectroscopy (XPS) of surface chemical states, before and after. • L-cysteine-modified Au surface can immobilize lactic acid via peptide bonds.

Electrochemical immobilization of biomolecules on gold surface modified with monolayered L-cysteine

International Nuclear Information System (INIS)

Honda, Mitsunori; Baba, Yuji; Sekiguchi, Tetsuhiro; Shimoyama, Iwao; Hirao, Norie

2014-01-01

Immobilization of organic molecules on the top of a metal surface is not easy because of lattice mismatch between organic and metal crystals. Gold atoms bind to thiol groups through strong chemical bonds, and a self-assembled monolayer of sulfur-terminated organic molecules is formed on the gold surface. Herein, we suggested that a monolayer of L-cysteine deposited on a gold surface can act as a buffer layer to immobilize biomolecules on the metal surface. We selected lactic acid as the immobilized biomolecule because it is one of the simplest carboxyl-containing biomolecules. The immobilization of lactic acid on the metal surface was carried out by an electrochemical method in an aqueous environment under the potential range varying from − 0.6 to + 0.8 V. The surface chemical states before and after the electrochemical reaction were characterized using X-ray photoelectron spectroscopy (XPS). The N 1s and C 1s XPS spectra showed that the L-cysteine-modified gold surface can immobilize lactic acid via peptide bonds. This technique might enable the immobilization of large organic molecules and biomolecules. - Highlights: • Monolayer l-cysteine deposited on Au surface as a buffer layer to immobilize biomolecules. • Lactic acid as the immobilized biomolecule as it is simple carboxyl-containing biomolecule. • X-ray photoelectron spectroscopy (XPS) of surface chemical states, before and after. • L-cysteine-modified Au surface can immobilize lactic acid via peptide bonds

Selective chromogenic and fluorogenic peptide substrates for the assay of cysteine peptidases in complex mixtures.

Science.gov (United States)

Semashko, Tatiana A; Vorotnikova, Elena A; Sharikova, Valeriya F; Vinokurov, Konstantin S; Smirnova, Yulia A; Dunaevsky, Yakov E; Belozersky, Mikhail A; Oppert, Brenda; Elpidina, Elena N; Filippova, Irina Y

2014-03-15

This study describes the design, synthesis, and use of selective peptide substrates for cysteine peptidases of the C1 papain family, important in many biological processes. The structure of the newly synthesized substrates is Glp-Xaa-Ala-Y (where Glp=pyroglutamyl; Xaa=Phe or Val; and Y=pNA [p-nitroanilide], AMC [4-amino-7-methylcoumaride], or AFC [4-amino-7-trifluoromethyl-coumaride]). Substrates were synthesized enzymatically to guarantee selectivity of the reaction and optical purity of the target compounds, simplifying the scheme of synthesis and isolation of products. The hydrolysis of the synthesized substrates was evaluated by C1 cysteine peptidases from different organisms and with different functions, including plant enzymes papain, bromelain, ficin, and mammalian lysosomal cathepsins B and L. The new substrates were selective for C1 cysteine peptidases and were not hydrolyzed by serine, aspartic, or metallo peptidases. We demonstrated an application of the selectivity of the synthesized substrates during the chromatographic separation of a multicomponent set of digestive peptidases from a beetle, Tenebrio molitor. Used in combination with the cysteine peptidase inhibitor E-64, these substrates were able to differentiate cysteine peptidases from peptidases of other classes in midgut extracts from T. molitor larvae and larvae of the genus Tribolium; thus, they are useful in the analysis of complex mixtures containing peptidases from different classes. Published by Elsevier Inc.

A Serendipitous Formation of a Cysteine-bridged Disaccharide

African Journals Online (AJOL)

NICO

O-acetyl-b-D-glucopyranosylisothiouronium salt and the iodide or tosyl derivatives of L-serine,3 the desulfurization of disulfide- linked glycosyl cysteine derivatives,4 Lewis acid-catalyzed glycosylation,5,6 and solid phase glycosylation.7. Glycosylation of amino acids has previously relied on the use of amino acids protected ...

Protective effect of L-Cysteine upon leukopenic syndrome due to radiotherapy

Energy Technology Data Exchange (ETDEWEB)

Jingu, K; Matsuura, K [Kyushu Univ., Fukuoka (Japan). Faculty of Medicine; Bussaka, Y

1981-08-01

L-Cysteine, glutathione, inosine, and cepharanthin are the agents which have been widely used to prevent and treat the leukopenic syndrome following radiotherapy. Among these, the efficacy of inosine has been demonstrated in a double-blind comparative study. A double-blind controlled study of L-Cysteine compared to inosine and lactose-placebo was performed. Patients designated to receive radiotherapy for cancer of the lung, breast and uterine cervix were randomly allocated to 3 groups; namely, those to receive L-Cysteine 480 mg TID (CG group), inosine 1,800 mg TID (IN group) and placebo capsules (PL group) for 6 to 7 weeks. Among 159 cases, 152 were subjected to statistical analyses, and the latter consisted of 54 CG, 52 IN and 46 PL subjects. Any discrepancies among these 3 groups concerning sex, age, disease, WBC count, radiation procedure, or combined use of carcinostatics were negligible. According to the life-table analysis, the cumulative rates for the 3 groups were compared with respect to maintenance of WBC counts higher than 4,000/mm/sup 3/. Maintenance was best in the CG group, intermediate in the IN, and poorest in the PL group, the difference between CG and PL being statistically significant at the 5% level. Similar results were obtained in separate analyses of strata with and without concomitant carcinostatics. Furthermore, nearly the same results were obtained as those in the life-table analyses when data concerning efficacy and clinical usefulness as judged by physicians were analyzed. The present study indicates that the oral administration of L-Cysteine is safe and effective in preventing and treating leukopenic complications associated with radiotherapy.

Cysteine proteases and wheat (Triticum aestivum L) under drought: A still greatly unexplored association.

Science.gov (United States)

Botha, Anna-Maria; Kunert, Karl J; Cullis, Christopher A

2017-09-01

Bread wheat (Triticum aestivum L.) provides about 19% of global dietary energy. Environmental stress, such as drought, affects wheat growth causing premature plant senescence and ultimately plant death. A plant response to drought is an increase in protease-mediated proteolysis with rapid degradation of proteins required for metabolic processes. Among the plant proteases that are increased in their activity following stress, cysteine proteases are the best characterized. Very little is known about particular wheat cysteine protease sequences, their expression and also localization. The current knowledge on wheat cysteine proteases belonging to the five clans (CA, CD, CE, CF and CP) is outlined, in particular their expression and possible function under drought. The first successes in establishing an annotated wheat genome database are further highlighted which has allowed more detailed mining of cysteine proteases. We also share our thoughts on future research directions considering the growing availability of genomic resources of this very important food crop. Finally, we also outline future application of developed knowledge in transgenic wheat plants for environmental stress protection and also as senescence markers to monitor wheat growth under environmental stress conditions. © 2017 John Wiley & Sons Ltd.

Effect of cysteine and humic acids on bioavailability of Ag from Ag nanoparticles to a freshwater snail

Science.gov (United States)

Luoma, Samuel N.; Tasha Stoiber,; Croteau, Marie-Noele; Isabelle Romer,; Ruth Merrifeild,; Lead, Jamie

2016-01-01

Metal-based engineered nanoparticles (NPs) will undergo transformations that will affect their bioavailability, toxicity and ecological risk when released to the environment, including interactions with dissolved organic material. The purpose of this paper is to determine how interactions with two different types of organic material affect the bioavailability of silver nanoparticles (AgNPs). Silver uptake rates by the pond snail Lymnaea stagnalis were determined after exposure to 25 nmol l-1 of Ag as PVP AgNPs, PEG AgNPs or AgNO3, in the presence of either Suwannee River humic acid or cysteine, a high-affinity thiol-rich organic ligand. Total uptake rate of Ag from the two NPs was either increased or not strongly affected in the presence of 1 – 10 mg 1-1 humic acid. Humic substances contain relatively few strong ligands for Ag explaining their limited effects on Ag uptake rate. In contrast, Ag uptake rate was substantially reduced by cysteine. Three components of uptake from the AgNPs were quantified in the presence of cysteine using a biodynamic modeling approach: uptake of dissolved Ag released by the AgNPs, uptake of a polymer or large (>3kD) Ag-cysteine complex and uptake of the nanoparticle itself. Addition of 1:1 Ag:cysteine reduced concentrations of dissolved Ag, which contributed to, but did not fully explain the reductions in uptake. A bioavailable Ag-cysteine complex (> 3kD) appeared to be the dominant avenue of uptake from both PVP AgNPs and PEG AgNPs in the presence of cysteine. Quantifying the different avenues of uptake sets the stage for studies to assess toxicity unique to NPs.

Avaliação do kit "TF-Test" para o diagnóstico das infecções por parasitas gastrintestinais em ovinos

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Giuliano Lumina

2006-08-01

Full Text Available Este estudo teve como objetivos padronizar o kit TF-Test para a quantificação de ovos de parasitas gastrintestinais de ovinos e compará-lo ao método de Gordon & Whitlock modificado (G&W. Vinte quatro cordeiros confinados foram infectados artificialmente com Haemonchus contortus, durante 12 semanas, até o abate, quando foram colhidas amostras fecais e realizada a identificação e contagem dos parasitas abomasais. Nestes animais, ovos de H. contortus foram detectados em 95,8% das amostras fecais por ambos os testes (P>;0,05. Os coeficientes de correlação (r entre a carga parasitária (CP e os valores de OPG obtidos pelos métodos de G&W e TF-Test foram, respectivamente, de r=0,52 e r=0,51 (dados não transformados e r=0,85 e r=0,87 (dados transformados em log. Outras 100 amostras fecais foram colhidas de ovinos naturalmente infectados. Nas amostras destes animais, os testes G&W e TF-Test propiciaram o diagnóstico de ovos de estrongilídeos em 85% e 86% das amostras, respectivamente (P>;0,05. Pelo TF-Test e pelo G&W, oocistos de Eimeria foram detectados em 33% e em 12% das amostras (P<0,001 e ovos de Strongyloides spp. em 15% e 5% das amostras, respectivamente (P<0,05. Ambos os testes foram precisos para o diagnóstico de estrongilídeos gastrintestinais, porém, o TF-Test foi superior para o diagnóstico de oocistos de Eimeria spp. e de ovos de Strongyloides spp., mas, por outro lado, subestimou o número de ovos de estrongilídeos presente nas amostras.

Heterologous Expression in Remodeled C. elegans: A Platform for Monoaminergic Agonist Identification and Anthelmintic Screening.

Science.gov (United States)

Law, Wenjing; Wuescher, Leah M; Ortega, Amanda; Hapiak, Vera M; Komuniecki, Patricia R; Komuniecki, Richard

2015-04-01

Monoamines, such as 5-HT and tyramine (TA), paralyze both free-living and parasitic nematodes when applied exogenously and serotonergic agonists have been used to clear Haemonchus contortus infections in vivo. Since nematode cell lines are not available and animal screening options are limited, we have developed a screening platform to identify monoamine receptor agonists. Key receptors were expressed heterologously in chimeric, genetically-engineered Caenorhabditis elegans, at sites likely to yield robust phenotypes upon agonist stimulation. This approach potentially preserves the unique pharmacologies of the receptors, while including nematode-specific accessory proteins and the nematode cuticle. Importantly, the sensitivity of monoamine-dependent paralysis could be increased dramatically by hypotonic incubation or the use of bus mutants with increased cuticular permeabilities. We have demonstrated that the monoamine-dependent inhibition of key interneurons, cholinergic motor neurons or body wall muscle inhibited locomotion and caused paralysis. Specifically, 5-HT paralyzed C. elegans 5-HT receptor null animals expressing either nematode, insect or human orthologues of a key Gαo-coupled 5-HT1-like receptor in the cholinergic motor neurons. Importantly, 8-OH-DPAT and PAPP, 5-HT receptor agonists, differentially paralyzed the transgenic animals, with 8-OH-DPAT paralyzing mutant animals expressing the human receptor at concentrations well below those affecting its C. elegans or insect orthologues. Similarly, 5-HT and TA paralyzed C. elegans 5-HT or TA receptor null animals, respectively, expressing either C. elegans or H. contortus 5-HT or TA-gated Cl- channels in either C. elegans cholinergic motor neurons or body wall muscles. Together, these data suggest that this heterologous, ectopic expression screening approach will be useful for the identification of agonists for key monoamine receptors from parasites and could have broad application for the identification

OCORRÊNCIA E DISTRIBUIÇÃO ESTACIONAL DE HELMINTOS GASTRINTESTINAIS DE CAPRINOS NO MUNICÍPIO DE TERESINA, PIAUÍ

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Eneide Santiago Girão

1992-08-01

Full Text Available Este trabalho objetiva verificar a ocorrência e distribuição estacional de helmintos gastrintestinais parasitas de caprinos durante as estações seca e chuvosa visando o estabelecimento de medidas adequadas para o controle da verminose. O experimento foi conduzido na base física da EMBRAPA/UEPAE de Teresina, no município de Teresina. Em um piquete de Brachiaria humidicola medindo dois hectares, foram colocados oito caprinos machos, entre seis a oito meses de idade. Cinco animais ficaram permanentes no piquete e não receberam tratamento anti-helmíntico durante todo o período (Testemunhas e três (incorporados ao trabalho a cada mês, após receberem três tratamentos anti-helmínticos, ficaram expostos à infecção helmíntica durante 30 dias no piquete (traçadores. Mensalmente, após 14 dias de estabulação, eram abatidos e necropsiados três caprinos traçadores e no final do experimento, os testemunhas para coleta, contagem e identificação dos helmintos. De janeiro/88 a fevereiro/90 foram necropsiados 70 caprinos traçadores e cinco testemunhas. Os helmintos identificados foram: Haemonchus contortus, Trichostrongylus colubriformis, Strongyloides papillosus, Moniezia expansa, Cooperia punctata, C. curticei, Oesophagostomum columbianum, Trichuris spp (T. ovis, Trichuris sp., T. skrjabini, Skrjabinema ovis, Cysticercus tenuicollis e Paramphistomum spp. As necropsias nos caprinos traçadores indicaram que ocorreu transmissão de helmintos durante o ano todo sendo em intensidades mais elevadas na época chuvosa e início da época seca e que o helminto de maior intensidade nos caprinos traçadores foi H. contortus, e nos testemunhas T. colubriformis.

Use of irradiation to produce vaccines and use of radioisotopes to study the pathophysiology and immunology of host-parasite relationships

International Nuclear Information System (INIS)

Holmes, P.H.; Abbott, E.M.; Parkins, J.J.

1991-01-01

A review is presented of the use of nuclear techniques to investigate various aspects of the control and study of parasitic diseases in domestic animals. The development of an irradiated vaccine against bovine lungworm disease has been very successful and many millions of calves have been vaccinated. The development of irradiated vaccines against other parasitic diseases has, for various reasons, been less successful. A major application of nuclear techniques in parasitology has been in the investigation of the pathophysiology of gastrointestinal helminth infections and particularly the aetiology of the anaemia and hypoalbuminaemia associated with such infections. These techniques have also been used to investigate the influence of nutrition on host-parasite relationships. Studies are described of the influence of protein nutrition on the establishment, pathophysiology and immunity of Haemonchus contortus infection of sheep. The results showed that the protein content of the diet per se did not influence the establishment of single primary infections. However, lambs given a low protein diet showed more severe clinical and pathophysiological changes, despite similar levels of gastric blood loss. Lambs given a high protein diet and subjected to repeated infections were more likely to develop resistance to reinfection than animals on a low protein diet. Dietary protein did not, however, appear to influence the ability of older, mature animals to respond to vaccination against H. contortus using gamma-irradiated larvae. These findings indicate the importance of nutrition on the pathogenesis of gastrointestinal nematode infections and could have considerable significance in field situations where suboptimal nutrition commonly occurs. There has also been extensive use of nuclear techniques to study the immunology of parasitic infections. Two examples are described, one using radiolabelled trypanosomes to measure immune clearance in vivo and the other using the in vivo

Anthelmintic Activity of a Herbal Formulation Against Gastrointestinal Nematodes of Sheep

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Muhammad Arfan Zaman*§, Zafar Iqbal, Muhammad Nisar Khan and Ghulam Muhammad1

2012-01-01

Full Text Available This study was carried out to evaluate the anthelmintic activity of a herbal formulation (HF based on aqueous extracts of leaves of Azadirachta indica and Nicotiana tabacum, flowers of Calotropis procera and seeds of Trachyspermum ammi. In vitro, eggs and adult Haemonchus contortus were exposed to different concentrations of HF following the standard procedures of egg hatch test (EHT; 50 to 0.024414 mg ml-1 and adult motility assay (AMA; 200-0.1953125mg ml-1, respectively. The reference drugs used in the study were oxfendazole (0.0056704 to 0.0000027 mg ml-1 and levamisole (1.50 mg ml-1 for EHT and AMA, respectively. In vivo, pre and post-treatment (4 mg, 2 mg and 500 µg kg-1 body weight fecal egg counts were determined following standard fecal egg count reduction test in sheep naturally parasitized with mixed species of gastrointestinal nematodes. In EHT, LC50 values of HF and oxfendazole (reference drug were 275.1 and 0.016 µg ml-1, respectively. In AMA, 100% mortality of H. contortus was observed 6 hr post-exposure to 3.125-200 mg ml-1 concentrations of HF and 2 hr post-exposure to levamisole. In vivo, maximum (96.2% fecal egg count (EPG reduction was recorded in sheep treated with HF @ 4 mg kg-1 body weight; whereas, 89.3% reduction in EPG was recorded in sheep treated with levamisole @ 7.5 mg kg-1 body weight. A graded dose response was noted in all the tests used in the present study to evaluate the anthelmintic activity of HF. Therefore, HF seems to be promising as an anthelmintic for animals. Large scale trials on efficacy and safety, however, are recommended before the HF is considered for commercialization in crude form.

Managing anthelmintic resistance in small ruminant livestock of resource-poor farmers in South Africa : review article

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A.F. Vatta

2006-06-01

Full Text Available Gastrointestinal parasitism is one of the most important disease complexes of sheep and goats impacting on the resource-poor livestock farmer. Of the responsible nematodes, Haemonchus contortus, a blood-sucking worm of the abomasum, poses possibly the greatest threat. Over the past several decades, the worm has been controlled through the use of anthelmintics, but the emergence of anthelmintic resistance has threatened this chemotherapeutic approach. In Africa, the overall prevalence of anthelmintic resistance has not been extensively investigated, particularly within the resource-poor farming sector, but resistance has been reported from at least 14 countries with most of the reports emanating from Kenya and South Africa and the majority concerning H. contortus. While levels of resistance under commercial sheep farming systems in South Africa is considered to be amongst the worst in the world, resistance has also been reported from the resource-poor farming sector. Increases in productivity and reproduction of livestock and the development of markets for sale of animals are seen by international funding bodies as a way out of poverty for communities that keep livestock. This must lead to the greater need for parasite control. At such times, the risk of levels of anthelmintic resistance escalating is much greater and there is therefore a need to look at alternatives to their use. Proposed strategies include the appropriate, but judicious use of anthelmintics by application of the FAMACHA(c system and the use of alternatives to anthelmintics such as strategic nutrient supplementation. It is also very clear that there is a strong demand for knowledge about animal diseases, including helminthosis, and their effective management in the resource-poor livestock farming communities. This is an important challenge to meet.

Use of irradiation to produce vaccines and use of radioisotopes to study the pathophysiology and immunology of host-parasite relationships

International Nuclear Information System (INIS)

Holmes, P.H.; Abbott, E.M.; Parkins, J.J.

1986-01-01

A review is presented of the use of nuclear techniques to investigate various aspects of the control and study of parasitic diseases in domestic animals. The development of an irradiated vaccine against bovine lungworm disease has been very successful and many millions of calves have been vaccinated. The development of irradiated vaccines against other parasitic diseases has, for various reasons, been less successful. A major application of nuclear techniques in parasitology has been in the investigation of the pathophysiology of gastrointestinal helminth infections and particularly the aetiology of the anaemia and hypoalbuminaemia associated with such infections. These techniques have also been used to investigate the influence of nutrition on host-parasite relationships. Studies are described of the influence of protein nutrition on the establishment, pathophysiology and immunity of Haemonchus contortus infection of sheep. The results showed that the protein content of the diet per se did not influence the establishment of single primary infections. However, lambs given a low protein diet showed more severe clinical and pathophysiological changes, despite similar levels of gastric blood loss. Lambs given a high protein diet and subjected to repeated infections were more likely to develop resistance to reinfection than animals on a low protein diet. Dietary protein did not, however, appear to influence the ability of older, mature animals to respond to vaccination against H. contortus using gamma-irradiated larvae. These findings indicate the importance of nutrition on the pathogenesis of gastrointestinal nematode infections and could have considerable significance in field situations where subtropical nutrition commonly occurs. There has also been extensive use of nuclear techniques to study the immunology of parasitic infections. Two examples are described, one using radiolabelled trypanosomes to measure immune clearance in vivo and the other using the in vivo

Heterologous Expression in Remodeled C. elegans: A Platform for Monoaminergic Agonist Identification and Anthelmintic Screening.

Directory of Open Access Journals (Sweden)

Wenjing Law

2015-04-01

Full Text Available Monoamines, such as 5-HT and tyramine (TA, paralyze both free-living and parasitic nematodes when applied exogenously and serotonergic agonists have been used to clear Haemonchus contortus infections in vivo. Since nematode cell lines are not available and animal screening options are limited, we have developed a screening platform to identify monoamine receptor agonists. Key receptors were expressed heterologously in chimeric, genetically-engineered Caenorhabditis elegans, at sites likely to yield robust phenotypes upon agonist stimulation. This approach potentially preserves the unique pharmacologies of the receptors, while including nematode-specific accessory proteins and the nematode cuticle. Importantly, the sensitivity of monoamine-dependent paralysis could be increased dramatically by hypotonic incubation or the use of bus mutants with increased cuticular permeabilities. We have demonstrated that the monoamine-dependent inhibition of key interneurons, cholinergic motor neurons or body wall muscle inhibited locomotion and caused paralysis. Specifically, 5-HT paralyzed C. elegans 5-HT receptor null animals expressing either nematode, insect or human orthologues of a key Gαo-coupled 5-HT1-like receptor in the cholinergic motor neurons. Importantly, 8-OH-DPAT and PAPP, 5-HT receptor agonists, differentially paralyzed the transgenic animals, with 8-OH-DPAT paralyzing mutant animals expressing the human receptor at concentrations well below those affecting its C. elegans or insect orthologues. Similarly, 5-HT and TA paralyzed C. elegans 5-HT or TA receptor null animals, respectively, expressing either C. elegans or H. contortus 5-HT or TA-gated Cl- channels in either C. elegans cholinergic motor neurons or body wall muscles. Together, these data suggest that this heterologous, ectopic expression screening approach will be useful for the identification of agonists for key monoamine receptors from parasites and could have broad application for

Low-energy electron-induced dissociation in condensed-phase L-cysteine I: Desorption of anions from chemisorbed films

International Nuclear Information System (INIS)

Alizadeh, E; Rowntree, P A; Massey, S; Sanche, L

2015-01-01

Among amino acids, cysteine has been widely studied, becoming a standard for molecular self-assembly experiments, because its mercapto group (-SH) allows the formation of self-assembled monolayers (SAMs) on metal surfaces. Dissociative electron attachment (DEA) on L-cysteine SAMs is investigated utilizing a time-of-flight mass spectrometer coupled with a low-energy electron gun. The results show that electrons with kinetic energies of 3 to 15 eV attach to L-cysteine producing anionic fragments of different masses (e.g., H - , O - , OH - , S - , SH - ) via dissociation of intermediate transient anions. The anion yield functions exhibited purely resonant behaviour with electron energies below 15 eV, indicating that the formation of transient anions is the predominant mechanism of production of anionic fragments from L-cysteine dissociation. (paper)

Metabolism, excretion, and pharmacokinetics of S-allyl-L-cysteine in rats and dogs.

Science.gov (United States)

Amano, Hirotaka; Kazamori, Daichi; Itoh, Kenji; Kodera, Yukihiro

2015-05-01

The metabolism, excretion, and pharmacokinetics of S-allyl-l-cysteine (SAC), an active key component of garlic supplements, were examined in rats and dogs. A single dose of SAC was administered orally or i.v. to rats (5 mg/kg) and dogs (2 mg/kg). SAC was well absorbed (bioavailability >90%) and its four metabolites-N-acetyl-S-allyl-l-cysteine (NAc-SAC), N-acetyl-S-allyl-l-cysteine sulfoxide (NAc-SACS), S-allyl-l-cysteine sulfoxide (SACS), and l-γ-glutamyl-S-allyl-l-cysteine-were identified in the plasma and/or urine. Renal clearance values (l/h/kg) of SAC indicated its extensive renal reabsorption, which contributed to the long elimination half-life of SAC, especially in dogs (12 hours). The metabolism of SAC to NAc-SAC, principal metabolite of SAC, was studied in vitro and in vivo. Liver and kidney S9 fractions of rats and dogs catalyzed both N-acetylation of SAC and deacetylation of NAc-SAC. After i.v. administration of NAc-SAC, SAC appeared in the plasma and its concentration declined in parallel with that of NAc-SAC. These results suggest that the rate and extent of the formation of NAc-SAC are determined by the N-acetylation and deacetylation activities of liver and kidney. Also, NAc-SACS was detected in the plasma after i.v. administration of either NAc-SAC or SACS, suggesting that NAc-SACS could be formed via both N-acetylation of SACS and S-oxidation of NAc-SAC. In conclusion, this study demonstrated that the pharmacokinetics of SAC in rats and dogs is characterized by its high oral bioavailability, N-acetylation and S-oxidation metabolism, and extensive renal reabsorption, indicating the critical roles of liver and kidney in the elimination of SAC. Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics.

Cardiovascular actions of L-cysteine and L-cysteine sulfinic acid in the nucleus tractus solitarius of the rat.

Science.gov (United States)

Takemoto, Yumi

2014-07-01

The sulfur-containing excitatory amino acid (EAA) L-cysteine sulfinic acid (CSA), a neurotransmitter candidate, is endogenously synthesized from L-cysteine (Cys). Exogenous Cys administration into the brain produces cardiovascular effects; these effects likely occur via synaptic stimulation of central nervous system (CNS) neurons that regulate peripheral cardiovascular function. However, the cardiovascular responses produced by CNS Cys administration could result from CSA biosynthesized in synapse. The present study examined the role of CSA in Cys-induced cardiovascular responses within the nucleus tractus solitarius (NTS) of anesthetized rats. The NTS receives input from various visceral afferents that gate autonomic reflexes, including cardiovascular reflexes. Within the NTS, both Cys and CSA microinjections produced decrease responses in arterial blood pressure and heart rate that were similar to those produced by L-glutamate. Co-injection of the ionotropic EAA receptor antagonist kynurenic acid abolished Cys-, but not CSA-, induced cardiovascular responses. This finding suggests that only Cys-induced cardiovascular responses are mediated by kynurenate-sensitive receptors. This study provides the first demonstration that Cys- and CSA-induced cardiovascular responses occur via different mechanisms in the NTS of rats. Further, this study also indicates that Cys-induced cardiovascular responses do not occur via CSA. Thus, within the NTS, endogenous Cys and/or CSA might be involved in cardiovascular regulation.

Influence of cysteine and selenodicysteine on the uptake of zinc by Chlorella vulgaris Beijerinck

International Nuclear Information System (INIS)

Czauderna, M.; Samochocka, K.

1982-01-01

The uptake of zinc labelled with radioactive 65 Zn in the presence of cysteine and selenodicysteine by Chlorella vulgaris was examined. The concentration of zinc ions in the medium was 20 mg per 1. The uptake yield was found to be enhanced by selenodicysteine. At concentration of 10 - 7 -10 - 6 M the growth rate of Chlorella vulgaris was accelerated by the latter, provided that the specific activity of 65 Zn was 3.7 MBq/1. At this specific zinc activity cysteine increased the uptake yield during the initial 50 h of the incubation process. At specific 65 Zn-activity of 55.5 MBq/1 selenodicysteine and cysteine only slightly influenced the zinc uptake by Chlorella vulgaris. No increment in the biomass was observed at this specific zinc radioactivity. (author)

Reduction of mercury from mackerel fillet using combined solution of cysteine, EDTA, and sodium chloride.

Science.gov (United States)

Hajeb, P; Jinap, S

2012-06-13

An acidic solution containing mercury chelating agents to eliminate mercury in raw fish (mackerel) fillet was developed. The solution contained hydrochloric acid, sodium hydroxide, cysteine, EDTA, and NaCl. The optimum conditions for mercury reduction were achieved using response surface methodology (RSM) at cysteine concentration of 1.25%, EDTA of 275 mg/L, NaCl of 0.5%, pH of 3.75, and exposure time of 18 min. The optimized conditions produced a solution which can remove up to 91% mercury from raw fish fillet. Cysteine and EDTA were identified as potential chelating agents with the greatest potential for use. The solution can be employed in fish industries to reduce mercury in highly contaminated fish.

Polydopamine/Cysteine surface modified isoporous membranes with self-cleaning properties

KAUST Repository

Shevate, Rahul

2017-02-03

The major challenge in membrane filtration is fouling which reduces the membrane performance. Fouling is mainly due to the adhesion of foulants on the membrane surfaces. In this work, we studied the fouling behaviour of polystyrene-b-poly(4-vinylpyridine) (PS-b-P4VP) isoporous membrane and the mussel inspired polydopamine/L-cysteine isoporous zwitterionic membrane. Polystyrene-b-poly(4-vinylpyridine) (PS-b-P4VP) isoporous membranes were fabricated via self-assembly and non-solvent induced phase separation method. Subsequently, the isoporous membrane was modified by a mild mussel-inspired polydopamine (PDA) coating; the isoporous surface structure and the water flux was retained. Zwitterionic L-cysteine was further anchored on the PDA coated membranes via Michael addition reaction at pH 7 and 50 °C to alleviate their antifouling ability with foulants solution. The membranes were thoroughly characterized using X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and zeta potential measurements. Contact angle and dynamic scanning calorimetry (DSC) measurements were carried out to examine the hydrophilicity. The pH-responsive behaviour of the modified membrane remains unchanged and antifouling ability after PDA/L-cysteine functionalization was improved. The modified and unmodified isoporous membranes were tested using humic acid and natural organic matter model solutions at 0.5 bar feed pressure.

Cysteine- and glutathione-mediated uptake of lead and cadmium into Zea mays and Brassica napus roots

International Nuclear Information System (INIS)

Vadas, Timothy M.; Ahner, Beth A.

2009-01-01

This study examines a new mechanism for the uptake of Pb and Cd into Brassica napus and Zea mays roots. During hydroponic experiments, the uptake of Pb and Cd was enhanced in the presence of cysteine and glutathione, whereas no or very low uptake was observed in EDTA and penicillamine controls. Uptake rates were also enhanced after pre-exposure to cysteine or glutathione and inhibited in the presence of vanadate, suggesting a biological mechanism of uptake. Increasing concentrations of glutathione in solution resulted in decreasing Pb uptake rates, indicating competition for transport between free-glutathione and Pb-glutathione species. Pb uptake in the presence of increasing cysteine concentrations resulted in decreased uptake initially but linearly increasing uptake at higher concentrations. Experimentation showed concentration dependent Pb uptake rates. We speculate that there are specific transporters for these thiol ligands and describe what barriers remain for application of this novel transport mechanism in chelator-assisted phytoremediation. - Cysteine and glutathione mediate the transport of lead and cadmium into plant roots.

Cysteine- and glutathione-mediated uptake of lead and cadmium into Zea mays and Brassica napus roots

Energy Technology Data Exchange (ETDEWEB)

Vadas, Timothy M., E-mail: tvadas@umbc.ed [Department of Biological and Environmental Engineering, Cornell University, 320 Riley-Robb Hall, Ithaca, NY 14853 (United States); Ahner, Beth A., E-mail: baa7@cornell.ed [Department of Biological and Environmental Engineering, Cornell University, 320 Riley-Robb Hall, Ithaca, NY 14853 (United States)

2009-08-15

This study examines a new mechanism for the uptake of Pb and Cd into Brassica napus and Zea mays roots. During hydroponic experiments, the uptake of Pb and Cd was enhanced in the presence of cysteine and glutathione, whereas no or very low uptake was observed in EDTA and penicillamine controls. Uptake rates were also enhanced after pre-exposure to cysteine or glutathione and inhibited in the presence of vanadate, suggesting a biological mechanism of uptake. Increasing concentrations of glutathione in solution resulted in decreasing Pb uptake rates, indicating competition for transport between free-glutathione and Pb-glutathione species. Pb uptake in the presence of increasing cysteine concentrations resulted in decreased uptake initially but linearly increasing uptake at higher concentrations. Experimentation showed concentration dependent Pb uptake rates. We speculate that there are specific transporters for these thiol ligands and describe what barriers remain for application of this novel transport mechanism in chelator-assisted phytoremediation. - Cysteine and glutathione mediate the transport of lead and cadmium into plant roots.

Enhanced biocatalytic production of L-cysteine by Pseudomonas sp. B-3 with in situ product removal using ion-exchange resin.

Science.gov (United States)

Wang, Pu; He, Jun-Yao; Yin, Jiang-Feng

2015-03-01

Bioconversion of DL-2-amino-Δ(2)-thiazoline-4-carboxylic acid (DL-ATC) catalyzed by whole cells of Pseudomonas sp. was successfully applied for the production of L-cysteine. It was found, however, like most whole-cell biocatalytic processes, the accumulated L-cysteine produced obvious inhibition to the activity of biocatalyst and reduced the yield. To improve L-cysteine productivity, an anion exchange-based in situ product removal (ISPR) approach was developed. Several anion-exchange resins were tested to select a suitable adsorbent used in the bioconversion of DL-ATC for the in situ removal of L-cysteine. The strong basic anion-exchange resin 201 × 7 exhibited the highest adsorption capacity for L-cysteine and low adsorption for DL-ATC, which is a favorable option. With in situ addition of 60 g L(-1) resin 201 × 7, the product inhibition can be reduced significantly and 200 mmol L(-1) of DL-ATC was converted to L-cysteine with 90.4 % of yield and 28.6 mmol L(-1 )h(-1) of volumetric productivity. Compared to the bioconversion without the addition of resin, the volumetric productivity of L-cysteine was improved by 2.27-fold using ISPR method.

Drug susceptibility testing in microaerophilic parasites: Cysteine strongly affects the effectivities of metronidazole and auranofin, a novel and promising antimicrobial

Directory of Open Access Journals (Sweden)

David Leitsch

2017-12-01

Full Text Available The microaerophilic parasites Entamoeba histolytica, Trichomonas vaginalis, and Giardia lamblia annually cause hundreds of millions of human infections which are treated with antiparasitic drugs. Metronidazole is the most often prescribed drug but also other drugs are in use, and novel drugs with improved characteristics are constantly being developed. One of these novel drugs is auranofin, originally an antirheumatic which has been relabelled for the treatment of parasitic infections. Drug effectivity is arguably the most important criterion for its applicability and is commonly assessed in susceptibility assays using in vitro cultures of a given pathogen. However, drug susceptibility assays can be strongly affected by certain compounds in the growth media. In the case of microaerophilic parasites, cysteine which is added in large amounts as an antioxidant is an obvious candidate because it is highly reactive and known to modulate the toxicity of metronidazole in several microaerophilic parasites.In this study, it was attempted to reduce cysteine concentrations as far as possible without affecting parasite viability by performing drug susceptibility assays under strictly anaerobic conditions in an anaerobic cabinet. Indeed, T. vaginalis and E. histolytica could be grown without any cysteine added and the cysteine concentration necessary to maintain G. lamblia could be reduced to 20%. Susceptibilities to metronidazole were found to be clearly reduced in the presence of cysteine. With auranofin the protective effect of cysteine was extreme, providing protection to concentrations up to 100-fold higher as observed in the absence of cysteine. With three other drugs tested, albendazole, furazolidone and nitazoxanide, all in use against G. lamblia, the effect of cysteine was less pronounced. Oxygen was found to have a less marked impact on metronidazole and auranofin than cysteine but bovine bile which is standardly used in growth media for G

ASSESSMENT OF THE E 920 ADDITIVE (L - CYSTEINE IN RELATION TO SOME PROBLEMS OF MODERN FOOD INDUSTRY

Directory of Open Access Journals (Sweden)

Radiana Maria TAMBA BEREHOIU

2013-01-01

Full Text Available This paper aims to assess the current state of knowledge about the use of L - cysteine in food industry, regarding certain cultural, legal, technological, toxicological, and other aspects that influence the attitude of the consumerstowards food. Use of L - cysteine and its derivatives in bakery allows the optimizing of the technological characteristics of flours and their higher recovery, by using products with high added value. Use the E 920 additivein human food is subject to the cultural and religious controversy, due to the generalized process of obtaining this additive from animal products (keratin. Our study shows that these controversies will be overcome when industrialfermentative technologies of L - cysteine production will be generalized in the market. There exist no data on thepotential toxicity of L - cysteine in the usual doses which are used in the baking industry. The only threat to the status of E 920 as a safe additive is the excitotoxic potential, suggested in several recent studies. Also, there exists a potential for extending the use of L - cysteine in the food industry in order to reduce the contamination degree withcertain chemicals having carcinogen potential, such as acrylamide and mycotoxins.

Revisiting the systemic lipopolysaccharide mediated neuroinflammation: Appraising the effect of l-cysteine mediated hydrogen sulphide on it

Directory of Open Access Journals (Sweden)

Abdulaziz S. Al-Saeedan

2018-05-01

Full Text Available The present research was ventured to examine the effect of l-cysteine on neuro-inflammation persuaded by peripheral lipopolysaccharides (LPS, 125 μg/kg, i.p. administration. No behavioral, biochemical, and inflammatory abnormality was perceived in the brain tissues of experimental animals after LPS administration. l-cysteine precipitated marginal symptoms of toxicity in the brain tissue. Similar pattern of wholesome effect of LPS were perceived when evaluated through the brain tissue fatty acid profile, histopathologically and NF-ĸBP65 protein expression. LPS was unsuccessful to alter the levels of hydrogen sulphide (H2S, cyclooxygenase (COX and lipoxygenase (LOX enzyme in brain tissue. LPS afforded significant peripheral toxicity, when figured out through inflammatory markers (COX, LOX, gaseous signaling molecules nitric oxide (NO, H2S, liver toxicity (SGOT, SGPT, and inflammatory transcription factor (NF-ĸBP65 and l-cysteine also provided a momentous protection against the same as well. The study inculcated two major finding, firstly LPS (i.p. cannot impart inflammatory changes to brain and secondly, l-cysteine can afford peripheral protection against deleterious effect of LPS (i.p. Keywords: Hydrogen sulphide, l-cysteine, Inflammation, Lipopolysaccharide, Neuroinflammation

Effects of DHA-enriched hen egg yolk and L-cysteine supplementation on quality of cryopreserved boar semen.

Science.gov (United States)

Chanapiwat, Panida; Kaeoket, Kampon; Tummaruk, Padet

2009-09-01

The objective of the present study was to determine the effects of docosahexaenoic acid (DHA)-enriched hen egg yolks and L-cysteine supplementation on the qualities of the cryopreserved boar semen. A total of 15 ejaculates from 5 Pietrain boars were divided into 4 groups according to the compositions of the freezing extenders used, that is, normal hen egg yolk (group I), DHA-enriched hen egg yolk (group II), normal hen egg yolk with 5 mmol L(-1) of cysteine supplementation (group III) and DHA-enriched hen egg yolk with 5 mmol L(-1) of cysteine supplementation (group IV). The semen was cryopreserved using controlled rate freezer and was thawed at 50 degrees C for 12 s. Progressive motility, sperm viability, acrosome integrity and functional integrity of sperm plasma membrane of the post-thawed semen were evaluated. The supplementation of L-cysteine in the freezing extender alone (group III) improved progressive motility (P semen qualities (P > 0.05). In conclusion, the supplementation of antioxidant L-cysteine alone or in combination with DHA-enriched hen egg yolk significantly improved the post-thawed semen qualities, especially progressive motility and acrosome integrity.

Neuroprotective effect of S-allyl-l-cysteine derivatives against endoplasmic reticulum stress-induced cytotoxicity is independent of calpain inhibition.

Science.gov (United States)

Imai, Toru; Kosuge, Yasuhiro; Saito, Hiroaki; Uchiyama, Taketo; Wada, Taira; Shimba, Shigeki; Ishige, Kumiko; Miyairi, Shinichi; Makishima, Makoto; Ito, Yoshihisa

2016-03-01

S-allyl-l-cysteine (SAC) is known to have neuroprotective properties. We synthesized various SAC derivatives and tested their effects on endoplasmic reticulum stress-induced neurotoxicity in cultured hippocampal neurons (HPNs). Among the compounds tested, S-propyl-l-cysteine (SPC) exhibited the strongest neuroprotective activity in HPNs, followed by S-ethyl-l-cysteine (SEC) and S-methyl-l-cysteine (SMC). Unlike SAC and SMC, SPC and SEC did not have inhibitory activity on μ-calpain, suggesting that the mechanism underlying the protective activity of SPC and SEC differs from that of SAC. Copyright © 2016 Japanese Pharmacological Society. Production and hosting by Elsevier B.V. All rights reserved.

Spectrophotometric determination of L-cysteine by using polyvinylpyrrolidone-stabilized silver nanoparticles in the presence of barium ions

Science.gov (United States)

Bamdad, Farzad; Khorram, Fateme; Samet, Maryam; Bamdad, Kourosh; Sangi, Mohammad Reza; Allahbakhshi, Fateme

2016-05-01

In this article a simple and selective colorimetric probe for cysteine determination using silver nano particles (AgNPS) is described. The determination process was based upon the surface plasmon resonance properties of polyvinylpyrrolidone-stabilized AgNPS. Interaction of AgNPS with cysteine molecules in the presence of barium ions induced a red shift in the surface plasmon resonance (SPR) maximum of AgNPs, as a result of nanoparticle aggregation. Consequently, yellow color of AgNP solution was changed to pink. The linear range for the determination of cysteine was 3.2-8.2 μM (R = 0.9965) with a limit of detection equal to 2.8 μM (3σ). The proposed method was successfully applied to the determination of cysteine in human plasma samples. Acceptable recovery results of the spiked samples confirmed the validity of the proposed method.

One-pot non-enzymatic formation of firefly luciferin in a neutral buffer from p-benzoquinone and cysteine.

Science.gov (United States)

Kanie, Shusei; Nishikawa, Toshio; Ojika, Makoto; Oba, Yuichi

2016-04-21

Firefly luciferin, the substrate for the bioluminescence reaction of luminous beetles, possesses a benzothiazole ring, which is rare in nature. Here, we demonstrate a novel one-pot reaction to give firefly luciferin in a neutral buffer from p-benzoquinone and cysteine without any synthetic reagents or enzymes. The formation of firefly luciferin was low in yield in various neutral buffers, whereas it was inhibited or completely prevented in acidic or basic buffers, in organic solvents, or under a nitrogen atmosphere. Labelling analysis of the firefly luciferin using stable isotopic cysteines showed that the benzothiazole ring was formed via the decarboxylation and carbon-sulfur bond rearrangement of cysteine. These findings imply that the biosynthesis of firefly luciferin can be developed/evolved from the non-enzymatic production of firefly luciferin using common primary biosynthetic units, p-benzoquinone and cysteine.

Attachment Site Cysteine Thiol pKa Is a Key Driver for Site-Dependent Stability of THIOMAB Antibody-Drug Conjugates.

Science.gov (United States)

Vollmar, Breanna S; Wei, Binqing; Ohri, Rachana; Zhou, Jianhui; He, Jintang; Yu, Shang-Fan; Leipold, Douglas; Cosino, Ely; Yee, Sharon; Fourie-O'Donohue, Aimee; Li, Guangmin; Phillips, Gail L; Kozak, Katherine R; Kamath, Amrita; Xu, Keyang; Lee, Genee; Lazar, Greg A; Erickson, Hans K

2017-10-18

The incorporation of cysteines into antibodies by mutagenesis allows for the direct conjugation of small molecules to specific sites on the antibody via disulfide bonds. The stability of the disulfide bond linkage between the small molecule and the antibody is highly dependent on the location of the engineered cysteine in either the heavy chain (HC) or the light chain (LC) of the antibody. Here, we explore the basis for this site-dependent stability. We evaluated the in vivo efficacy and pharmacokinetics of five different cysteine mutants of trastuzumab conjugated to a pyrrolobenzodiazepine (PBD) via disulfide bonds. A significant correlation was observed between disulfide stability and efficacy for the conjugates. We hypothesized that the observed site-dependent stability of the disulfide-linked conjugates could be due to differences in the attachment site cysteine thiol pK a . We measured the cysteine thiol pK a using isothermal titration calorimetry (ITC) and found that the variants with the highest thiol pK a (LC K149C and HC A140C) were found to yield the conjugates with the greatest in vivo stability. Guided by homology modeling, we identified several mutations adjacent to LC K149C that reduced the cysteine thiol pK a and, thus, decreased the in vivo stability of the disulfide-linked PBD conjugated to LC K149C. We also present results suggesting that the high thiol pK a of LC K149C is responsible for the sustained circulation stability of LC K149C TDCs utilizing a maleimide-based linker. Taken together, our results provide evidence that the site-dependent stability of cys-engineered antibody-drug conjugates may be explained by interactions between the engineered cysteine and the local protein environment that serves to modulate the side-chain thiol pK a . The influence of cysteine thiol pK a on stability and efficacy offers a new parameter for the optimization of ADCs that utilize cysteine engineering.

Chemoselective PEGylation of Cysteine Analogs of Human Basic ...

African Journals Online (AJOL)

Purpose: To improve the stability and bioactivity of human basic fibroblast growth factor (hbFGF) by site-specific pegylation. Methods: Four new mutants of hbFGF were designed with substituted Asp68, Lys77, Glu78 and Arg81 with cysteine with the aid of bioinformatics technique, and then cloned into pET21a plasmid, ...

Evaluation of cysteine ethyl ester as efficient inducer for glutathione overproduction in Saccharomyces spp.

Science.gov (United States)

Lorenz, Eric; Schmacht, Maximilian; Senz, Martin

2016-11-01

Economical yeast based glutathione (GSH) production is a process that is influenced by several factors like raw material and production costs, biomass production and efficient biotransformation of adequate precursors into the final product GSH. Nowadays the usage of cysteine for the microbial conversion into GSH is industrial state of practice. In the following study, the potential of different inducers to increase the GSH content was evaluated by means of design of experiments methodology. Investigations were executed in three natural Saccharomyces strains, S. cerevisiae, S. bayanus and S. boulardii, in a well suited 50ml shake tube system. Results of shake tube experiments were confirmed in traditional baffled shake flasks and finally via batch cultivation in lab-scale bioreactors under controlled conditions. Comprehensive studies showed that the usage of cysteine ethyl ester (CEE) for the batch-wise biotransformation into GSH led up to a more than 2.2 times higher yield compared to cysteine as inducer. Additionally, the intracellular GSH content could be significantly increased for all strains in terms of 2.29±0.29% for cysteine to 3.65±0.23% for CEE, respectively, in bioreactors. Thus, the usage of CEE provides a highly attractive inducing strategy for the GSH overproduction. Copyright © 2016 Elsevier Inc. All rights reserved.

Chemical detection of cysteine-rich circular petides in selected ...

African Journals Online (AJOL)

Cysteine-rich circular peptides (CRCs) comprise a large family of gene encoded and low molecular weight polypeptides that has recently engaged the attention of scientists. This class of peptides exhibit a continuous circular configuration and a cystine knot backbone, which defines their resilient nature-directed structural ...

Participation of intracellular cysteine proteinases, in particular cathepsin B, in degradation of collagen in periosteal tissue explants

NARCIS (Netherlands)

Creemers, L. B.; Hoeben, K. A.; Jansen, D. C.; Buttle, D. J.; Beertsen, W.; Everts, V.

1998-01-01

The involvement of cysteine proteinases in the degradation of soft connective tissue collagen was studied in cultured periosteal explants. Using cysteine proteinase inhibitors that were active intracellularly or extracellularly (Ep453 and Ep475, respectively), it was shown that over-all collagen

Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants - From the field to the test tube and back

DEFF Research Database (Denmark)

Jutta, Papenbrock; Anja, Riemenschneider; Kamp, Anja

2007-01-01

focussed mainly on the release of H2S as defence strategy. In field experiments using different Brassica napus genotypes it was shown that the genetic differ- ences among Brassica genotypes lead to differences in sulfur content and L-cysteine desulfhydrase activity. Another field ex- periment demonstrated...... that sulfur supply and infection with Pyrenopeziza brassica influenced L-cysteine desulfhydrase activity in Brassica napus. Cysteine-degrading enzymes such as cysteine desulfhydrases are hypothesized to be involved in H2S release. Several L- and D-cysteine-specific desulfhydrase candidates have been isolated...... in plants which might be involved in SIR, such as high levels of thiols, glucosinolates, cysteine-rich proteins, phytoalexins, elemental sulfur, or H2S. Probably more than one strategy is used by plants. Species- or even variety-dependent differences in the development of SIR are probably used. Our research...

Divergent unprotected peptide macrocyclisation by palladium-mediated cysteine arylation.

Science.gov (United States)

Rojas, Anthony J; Zhang, Chi; Vinogradova, Ekaterina V; Buchwald, Nathan H; Reilly, John; Pentelute, Bradley L; Buchwald, Stephen L

2017-06-01

Macrocyclic peptides are important therapeutic candidates due to their improved physicochemical properties in comparison to their linear counterparts. Here we detail a method for a divergent macrocyclisation of unprotected peptides by crosslinking two cysteine residues with bis-palladium organometallic reagents. These synthetic intermediates are prepared in a single step from commercially available aryl bis-halides. Two bioactive linear peptides with cysteine residues at i , i + 4 and i , i + 7 positions, respectively, were cyclised to introduce a diverse array of aryl and bi-aryl linkers. These two series of macrocyclic peptides displayed similar linker-dependent lipophilicity, phospholipid affinity, and unique volume of distributions. Additionally, one of the bioactive peptides showed target binding affinity that was predominantly affected by the length of the linker. Collectively, this divergent strategy allowed rapid and convenient access to various aryl linkers, enabling the systematic evaluation of the effect of appending unit on the medicinal properties of macrocyclic peptides.

Spectrophotometric determination of L-cysteine by using polyvinylpyrrolidone-stabilized silver nanoparticles in the presence of barium ions.

Science.gov (United States)

Bamdad, Farzad; Khorram, Fateme; Samet, Maryam; Bamdad, Kourosh; Sangi, Mohammad Reza; Allahbakhshi, Fateme

2016-05-15

In this article a simple and selective colorimetric probe for cysteine determination using silver nano particles (AgNPS) is described. The determination process was based upon the surface plasmon resonance properties of polyvinylpyrrolidone-stabilized AgNPS. Interaction of AgNPS with cysteine molecules in the presence of barium ions induced a red shift in the surface plasmon resonance (SPR) maximum of AgNPs, as a result of nanoparticle aggregation. Consequently, yellow color of AgNP solution was changed to pink. The linear range for the determination of cysteine was 3.2-8.2 μM (R=0.9965) with a limit of detection equal to 2.8 μM (3σ). The proposed method was successfully applied to the determination of cysteine in human plasma samples. Acceptable recovery results of the spiked samples confirmed the validity of the proposed method. Copyright © 2016 Elsevier B.V. All rights reserved.

A study of low-energy ion induced radiolysis of thiol-containing amino acid cysteine in the solid and aqueous solution states

Energy Technology Data Exchange (ETDEWEB)

Ke Zhigang [Key Laboratory of Ion Beam Bio-engineering, Institute of Plasma Physics of Chinese Academy of Sciences, 350 Shushanhu Road, P.O. Box 1126, Hefei 230031 (China); Huang Qing, E-mail: huangq@ipp.ac.c [Key Laboratory of Ion Beam Bio-engineering, Institute of Plasma Physics of Chinese Academy of Sciences, 350 Shushanhu Road, P.O. Box 1126, Hefei 230031 (China); Dang Bingrong [Institute of Modern Physics, Chinese Academy of Sciences, 509 Nanchang Road, Lanzhou 730000 (China); Lu Yilin [Key Laboratory of Ion Beam Bio-engineering, Institute of Plasma Physics of Chinese Academy of Sciences, 350 Shushanhu Road, P.O. Box 1126, Hefei 230031 (China); Department of Physics, Anhui University, Hefei 230031 (China); Yuan Hang; Zhang Shuqing; Yu Zengliang [Key Laboratory of Ion Beam Bio-engineering, Institute of Plasma Physics of Chinese Academy of Sciences, 350 Shushanhu Road, P.O. Box 1126, Hefei 230031 (China)

2010-09-15

The radiolysis of cysteine under plasma discharge and irradiation of low-energy ion beam was investigated. The damage of cysteine in aqueous solution under discharge was assessed via the acid ninhydrin reagent and the yield of cystine produced from the reaction was analyzed by FTIR. In addition, the generation of hydrogen sulfide was also identified. The destruction of solid cysteine under low-energy ion beam irradiation was estimated via monitoring IR bands of different functional groups (-SH, -NH{sub 3}, -COO{sup -}) of cysteine, and the production of cystine from ion-irradiated solid cysteine after dissolution in water was also verified. These results may help us to understand the inactivation of sulphydryl enzymes under direct and indirect interaction with the low-energy ion irradiation.

l-Cysteine improves antioxidant enzyme activity, post-thaw quality and fertility of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

Science.gov (United States)

Iqbal, S; Riaz, A; Andrabi, S M H; Shahzad, Q; Durrani, A Z; Ahmad, N

2016-11-01

The effects of l-cysteine in extender on antioxidant enzymes profile during cryopreservation, post-thaw quality parameters and in vivo fertility of Nili-Ravi buffalo bull spermatozoa were studied. Semen samples from 4 buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of l-cysteine (0.0, 0.5, 1.0, 2.0 and 3.0 mm) and frozen in 0.5-ml French straws. The antioxidative enzymes [catalase, super oxide dismutase and total glutathione (peroxidase and reductase)] were significantly higher (P l-cysteine as compared to other groups. Post-thaw total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm s -1 ), straight line velocity (μm s -1 ), curvilinear velocity (μm s -1 ), beat cross frequency (Hz), viable spermatozoa with intact plasmalemma (%), acrosome and DNA integrity (%) were higher with the addition of 2.0 mm l-cysteine as compared to other groups (P l-cysteine than in the control. In conclusion, the addition of 2.0 mm l-cysteine in extender improved the antioxidant enzymes profile, post-thaw quality and in vivo fertility of Nili-Ravi buffalo bull spermatozoa. © 2016 Blackwell Verlag GmbH.

Parasites gastro-intestinaux d'antilopes et de buffles (Syncerus caffer brachyceros du ranch de gibier de Nazinga au Burkina Faso

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Belem AMG.

2009-01-01

Full Text Available Gastro-intestinal parasites of antelopes and buffalos (Syncerus caffer brachyceros from the Nazinga game ranch in Burkina Faso. Parasitological survey done on the digestive tracts of antelopes [roans (Hippotragus equinus koba, hartebeests (Alcelaphus buselaphus major, defassa waterbucks (Kobus ellipsiprymnus defassa, oribis (Ourebia ourebi quadriscopa] and buffalos (Syncerus caffer brachyceros from the Nazinga game ranch in Burkina Faso allowed the identification of gastrointestinal parasites and the estimation of their prevalence and load. Nine different nematode species and three cestode species were found. Females and larvae of Cooperia were not identified up to the species level and were recognized as Cooperia spp. The nematode species found were: Haemonchus contortus, Trichostrongylus axei, Cooperia curticei, Cooperia spatulata, Skrjabinema sp., Trichostrongylus colubriformis, Trichuris ovis, Bunostomum phlebotomum, and Oesophagostomum sp. Cestodes’ species were: Moniezia expansa, Avitellina centripunctata, and Stilesia globipunctata. It was also noted without a thorough investigation the presence of Setaria labiato-papillosa in the abdominal cavity and paramphistomes in the rumen. In spite of the presence of parasites on the antelopes and buffalos, their killing out percentages (47.5 to 54.7% indicated a good physical status.

Genetic control of acquired resistance to gastrointestinal nematode parasites in sheep

International Nuclear Information System (INIS)

Windon, R.G.; Wagland, B.M.; Dineen, J.K.

1988-01-01

A radiation attenuated larval vaccine has been used to evaluate the role of genetic components of the immune response in the control of Trichostrongylus colubriformis in sheep. Through selection based on age dependent responsiveness, lines of sheep have been established in which lambs are either high or low responders to vaccination and challenge infection. The estimated and realized heritabilities for the selected trait are 0.35-0.41. Significant interline differences were demonstrated and, within the lines, ewe lambs were consistently more responsive than rams or wethers. The effect of selection was not antigenically specific, since high responsiveness to T. colubriformis was associated with increased responsiveness to other related (T. rugatus) and unrelated parasites (Ostertagia circumcincta and Haemonchus contortus). In addition, high responders had a more vigorous reaction against naturally acquired infections than low responders. The general immunological competence was also increased in high responders; this was shown by the levels of serum complement fixing antibody to larval antigens after vaccination and challenge, in vitro blastogenic responses stimulated by larval antigens and the phagocytic function of peripheral leucocytes. No production penalty (weight gain and wool growth) was associated with heightened responsiveness. (author). 10 refs, 1 fig

Effect of Musa spp. extract on eggs and larvae of gastrointestinal nematodes from infected sheep

Directory of Open Access Journals (Sweden)

Natalie Neuwirt

2015-12-01

Full Text Available Helminthes are listed as one of the main problems facing the development of goat and sheep production. Haemonchus contortus is the specie that causes greatest negative impact in ranching. Resistance to anti-parasitic drugs and demand for residue-free animal-derived food products has elevated the importance of herbal treatments. The aim of this study was to develop an extract of Musa spp. and assess by in vitro testing, the anthelmintic effect on eggs and larvae in the gastrointestinal nematodes in sheep. Stool samples from sheep naturally infected were used to obtain eggs and larvae and was then followed by a test of hatchability and a larval migration inhibition test. In vitro tests on the inhibition of larval hatchability at concentrations of 160 and 180 mg mL-1 of larval extracts and inhibition of migration at concentrations of 800 and 1000 mg mL-1 were observed. The results indicate that the use of banana leaf has an anthelmintic effect and that in vivo studies on the applicability of this technology to the field should be made to further understanding and bring more information to what has already been revealed in this study.

Linear distribution of nematodes in the gastrointestinal tract of tracer lambs.

Science.gov (United States)

Makovcová, Katerina; Langrová, Iva; Vadlejch, Jaroslav; Jankovská, Ivana; Lytvynets, Andriy; Borkovcová, Marie

2008-12-01

Forty-eight tracer lambs were killed in 2004-2007. The abomasum, duodenum, small intestine (jejunum and ileum), colon and caecum were collected and processed for parasites enumeration and identification-mucosal scrapings of both abomasums and intestines were digested. Out of 48 gastrointestinal tracts examined, all were found to be positive for nematode infection. Seventeen species of gastrointestinal nematodes were recovered: Bunostomum trigonocephalum, Cooperia curticei, Haemonchus contortus, Chabertia ovina, Nematodirus battus, Nematodirus filicollis, Oesophagostomum venulosum, Teladorsagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Strongyloides papillosus, Trichuris ovis, Trichuris globulosa, Trichuris skrjabini and Skrjabinema ovis. All species were searched for in the entire gastrointestinal tract. Six species of nematodes were recovered from abnormal sites, naturally in small numbers of lambs as well as in small amounts: Nematodirus battus in the abomasums (6.67% of lambs), N. filicollis in the caecum and in the colon (%4 and 8%, respectively), T. axei in the colon (9.52%), T. colubriformis in the colon (13.89%), T. vitrinus in the caecum (16.67%), in the colon (20.00%) and in the abomasum (3.33%). T. ovis was found in one case in the small intestine.

Transport of S-cysteine conjugates in LL-PK1 cells and its role in toxicity

International Nuclear Information System (INIS)

Schaeffer, V.; Stevens, J.

1986-01-01

In order to study its role in S-cysteine conjugate toxicity, the transport of the nephrotoxic cysteine conjugate, S-1,2-dichlorovinyl-L-cysteine (L-DCVC), was investigated in the kidney cell line, LLC-PK1. When incubated with [ 35 S]-DCVC, accumulation of radioactivity within the cells was linear for at least 5 min with 92% of the 35 S present as unmetabolized L-DCVC. Kinetic analysis indicated two saturable uptake systems; Km = 6.8 μM and 1.6 mM; V/sub max/ = .048 and 1.4 nmol/min/mg protein. Both systems were Na + -independent and were inhibited by amino acid transport system L substrates but not substrates of systems A, ASC or organic anion transport. L-DCVC uptake at 5 μM and 500 μM was significantly greater in subconfluent cells than in confluent cultures by 5 and 2.8 fold, respectively. The presence of the non-toxic conjugates S-methyl-(SMC), S-ethyl-(SEC), S-benzyl-L- cysteine (SBC) and the D isomer of DCVC blocked the toxicity and inhibited the transport of L-DCVC in LLC-PK1 cells in the rank order of SBC > SEC congruent to D-DCVC > SMC. The metabolism of L-DCVC, previously shown to be required for cytotoxicity, was only slightly inhibited by these conjugates and did not correlate with their relative protection against toxicity. This study suggests that L-DCVC is transported into these cells by the system L transporter and that protection against toxicity by non-toxic S-cysteine conjugates is due to the inhibition of transport

Study of the histochemical detection of cysteine desulfhydrase in the vitellin sac of birds (1961)

International Nuclear Information System (INIS)

Chapeville, F.; Khau Van Kien, L.

1961-01-01

We have developed a method for the histochemical detection of cysteine desulfhydrase in the vitellin sac of the chicken embryo. The enzyme is localized in the presence of a lead salt by lead sulphide formed in situ from hydrogen sulphide liberated from the cysteine. The micrographs obtained are histological and show the presence of the enzyme in the different types of endoderm cell. (authors) [fr

Impact of cysteine variants on the structure, activity, and stability of recombinant human α-galactosidase A

Science.gov (United States)

Qiu, Huawei; Honey, Denise M; Kingsbury, Jonathan S; Park, Anna; Boudanova, Ekaterina; Wei, Ronnie R; Pan, Clark Q; Edmunds, Tim

2015-01-01

Recombinant human α-galactosidase A (rhαGal) is a homodimeric glycoprotein deficient in Fabry disease, a lysosomal storage disorder. In this study, each cysteine residue in rhαGal was replaced with serine to understand the role each cysteine plays in the enzyme structure, function, and stability. Conditioned media from transfected HEK293 cells were assayed for rhαGal expression and enzymatic activity. Activity was only detected in the wild type control and in mutants substituting the free cysteine residues (C90S, C174S, and the C90S/C174S). Cysteine-to-serine substitutions at the other sites lead to the loss of expression and/or activity, consistent with their involvement in the disulfide bonds found in the crystal structure. Purification and further characterization confirmed that the C90S, C174S, and the C90S/C174S mutants are enzymatically active, structurally intact and thermodynamically stable as measured by circular dichroism and thermal denaturation. The purified inactive C142S mutant appeared to have lost part of its alpha-helix secondary structure and had a lower apparent melting temperature. Saturation mutagenesis study on Cys90 and Cys174 resulted in partial loss of activity for Cys174 mutants but multiple mutants at Cys90 with up to 87% higher enzymatic activity (C90T) compared to wild type, suggesting that the two free cysteines play differential roles and that the activity of the enzyme can be modulated by side chain interactions of the free Cys residues. These results enhanced our understanding of rhαGal structure and function, particularly the critical roles that cysteines play in structure, stability, and enzymatic activity. PMID:26044846

The toxic effects of l-Cysteine-capped cadmium sulfide nanoparticles on the aquatic plant Spirodela polyrrhiza

International Nuclear Information System (INIS)

Khataee, Alireza; Movafeghi, Ali; Nazari, Fatemeh; Vafaei, Fatemeh; Dadpour, Mohammad Reza; Hanifehpour, Younes; Joo, Sang Woo

2014-01-01

Plants play an important role in the fate of nanoparticles in the environment through their uptake, bioaccumulation, and transfer to trophic chains. However, the impacts of nanoparticles on plants as essential components of all ecosystems are not well documented. In the present study, the toxic effects of l-Cysteine-capped CdS nanoparticles on Spirodela polyrrhiza as an aquatic higher plant species were studied. l-Cysteine-capped CdS nanoparticles were synthesized using hydrothermal method and their characteristics were determined by XRD, SEM, HR-TEM, and FT-IR techniques. The diameter of majority of synthesized nanoparticles was about 15–20 nm. Subsequently, the uptake of l-Cysteine-capped CdS nanoparticles by the plant species was confirmed using epifluorescence microscopy. The activity of peroxidase and superoxide dismutase as antioxidant enzymes was assayed and the relative frond number was calculated in the presence of different concentrations of l-Cysteine-capped CdS nanoparticles. The obtained results revealed the toxic effects of the synthesized nanoparticles on S. polyrrhiza, leading to growth reduction and significant changes in antioxidant enzymes’ activity.Graphical Abstract

The toxic effects of l-Cysteine-capped cadmium sulfide nanoparticles on the aquatic plant Spirodela polyrrhiza

Energy Technology Data Exchange (ETDEWEB)

Khataee, Alireza, E-mail: ar_khataee@yahoo.com [University of Tabriz, Research Laboratory of Advanced Water and Wastewater Treatment Processes, Department of Applied Chemistry, Faculty of Chemistry (Iran, Islamic Republic of); Movafeghi, Ali [University of Tabriz, Department of Plant Biology, Faculty of Natural Sciences (Iran, Islamic Republic of); Nazari, Fatemeh [University of Tabriz, Research Laboratory of Advanced Water and Wastewater Treatment Processes, Department of Applied Chemistry, Faculty of Chemistry (Iran, Islamic Republic of); Vafaei, Fatemeh [University of Tabriz, Department of Plant Biology, Faculty of Natural Sciences (Iran, Islamic Republic of); Dadpour, Mohammad Reza [University of Tabriz, Department of Horticultural Science, Faculty of Agriculture (Iran, Islamic Republic of); Hanifehpour, Younes; Joo, Sang Woo, E-mail: swjoo@yu.ac.kr [Yeungnam University, School of Mechanical Engineering (Korea, Republic of)

2014-12-15

Plants play an important role in the fate of nanoparticles in the environment through their uptake, bioaccumulation, and transfer to trophic chains. However, the impacts of nanoparticles on plants as essential components of all ecosystems are not well documented. In the present study, the toxic effects of l-Cysteine-capped CdS nanoparticles on Spirodela polyrrhiza as an aquatic higher plant species were studied. l-Cysteine-capped CdS nanoparticles were synthesized using hydrothermal method and their characteristics were determined by XRD, SEM, HR-TEM, and FT-IR techniques. The diameter of majority of synthesized nanoparticles was about 15–20 nm. Subsequently, the uptake of l-Cysteine-capped CdS nanoparticles by the plant species was confirmed using epifluorescence microscopy. The activity of peroxidase and superoxide dismutase as antioxidant enzymes was assayed and the relative frond number was calculated in the presence of different concentrations of l-Cysteine-capped CdS nanoparticles. The obtained results revealed the toxic effects of the synthesized nanoparticles on S. polyrrhiza, leading to growth reduction and significant changes in antioxidant enzymes’ activity.Graphical Abstract.

Preparation and characterization of a cysteine based DTPA derivative and its immunoconjugate for radioimmunotherapy

International Nuclear Information System (INIS)

Lee, S. Y.; Hong, Y. D.; Choi, S. J.

2007-01-01

Recently, radioimmunotherapy (RIT), which uses a monoclonal antibody in addition to a radionuclide to deliver radiation to the sites of a disease, has been extensively studied in this population. To label an antibody with radionuclides it is necessary to introduce a bifunctional chelating agent (BFCA) such a DTPA since it can not be directly labeled to a radionuclide. Therefore, developing a better BFCA for chelating biomolecule and radionuclide has been of major interest in developing radioimmunotherapeutic agents. Thereby, we describe the entantiospecific synthesis of a DTPA analogue which is derived from L-cysteine via bis N-alkylation. And the prepared DTPA derivative was conjugated with human Immunoglobulin G, and a characterization of the immunoconjugate was carried out. N, N-Bis[(tert-butoxycarbonyl)methyl]-2-ethanolamine, N, N-Bis[(tert-butoxycarbony)methyl]-2-bromoethyl-amine, 2-(4-N-Boc-aminophenyl) ethanol, 1-(4-N-Boc-aminophenyl)-2-bromoethane, S-((4-N-Boc-arninophenyl)-1-ethyl)-cysteine methylester, S-(N-Boc-aminophenyl)-Cys(tBu4-DTPA) methylester, -aminophenylethyl-Cys-DTPA, isothiocyanate-cysteine-DTPA, Immunoconjugation with IgG. The optimal molar DTPA derivative to IgG conjugation ratio was 1: 1. At higher amounts of DTPA derivative, amounts of unbounded DTPA derivative increased, and the immunoactivities of immunoconjugates reduced. Gel electrophoresis analysis of the immunoconjugates showed no degradation products or other impurities. This demonstrates the stability of the IgG in DTPA derivative. We established the preparation of an amino acid based DTPA by producing 4-Ethylaniline-DTPA-L-Cysteine. At the same molar this DTPA derivative to IgG, the immunoconjugate has stable molecular structure. In conclusion, 4-Ethylaniline-DTPA-L-Cysteine as a BFCA will show good properties for preparing a specific regional delivery system such as in radiopharmaceuticals, as a radiotracer, and NMR contrasting agents

Effect of mixing low palatable grasses of heteropogon contortus with ipil ipil leaves on digestibility in goats

International Nuclear Information System (INIS)

Bashir, M.; Qamar, I.A.; Babar, R.

2015-01-01

The study was conducted at the National Agricultural Research Centre, Islamabad, Pakistan during 2012 to find out the effect of mixing low palatable grasses of Heteropogon contortus (HC), with tree leaves of Leucaena leucocephala (Ipil ipil, II) in the ratio of 75:25, 50:50, 25:75, along with sole species on their digestibility in small ruminants. Goats fed II/sub 100%/, HC/sub 25%/ II/sub 75%/, HC/sub 50%/ II/sub 50%/, HC/sub 75%/ II/sub 25%/ and HC/sub 100%/ had similar dry matter (DM), crude protein (CP) and crude fibre (CF) consumption among all the treatments. The digestibility percentage of dry matter intake (DMI) varied among the treatments ranging from 68.25% to 41.66%. Mixtures of low palatable grass and Ipil ipil were in general more digestible with more than 65% dry matter digestibility. The lowest digestibility of dry matter (41.66%) was observed in HC/sub 100%/. A similar trend was noted for CP digestibility. However, reverse trend was observed in digestibility of CF where highest digestibility was recorded in HC100% and lowest in II100%. It can be concluded that grass and ipil ipil leaf mixture are better regarding forage quality and nutrient digestibility and can be recommended as animal feed. (author)

Ultrasensitive Detection of Cu2+ Using a Microcantilever Sensor Modified with L-Cysteine Self-Assembled Monolayer.

Science.gov (United States)

Xu, Xiaohe; Zhang, Na; Brown, Gilbert M; Thundat, Thomas G; Ji, Hai-Feng

2017-10-01

A microcantilever was modified with a self-assembled monolayer (SAM) of L-cysteine for the sensitively and selectively response to Cu(II) ions in aqueous solution. The microcantilever undergoes bending due to sorption of Cu(II) ions. The interaction of Cu(II) ions with the L-cysteine on the cantilever is diffusion controlled and does not follow a simple Langmuir adsorption model. A concentration of 10 -10  M Cu(II) was detected in a fluid cell using this technology. Other cations, such as Ni 2+ , Zn 2+ , Pb 2+ , Cd 2+ , Ca 2+ , K + , and Na + , did not respond with a significant deflection, indicating that this L-cysteine-modified cantilever responded selectively and sensitively to Cu(II).

Isolation of recombinant cysteine dioxygenase protein from Trichophyton mentagrophytes

Czech Academy of Sciences Publication Activity Database

Kašperová, A.; Kunert, J.; Horynová, M.; Weigl, E.; Sebela, M.; Lenobel, René; Raška, M.

2011-01-01

Roč. 54, č. 5 (2011), E456-E462 ISSN 0933-7407 R&D Projects: GA ČR GA301/08/1649 Institutional research plan: CEZ:AV0Z50380511 Keywords : Cysteine dioxygenase * dermatophytes * recombinant protein * keratinolytic fungi * cDNA Subject RIV: CE - Biochemistry Impact factor: 2.247, year: 2011

Effect of L-cysteine on remote organ injury in rats with severe acute pancreatitis induced by bile-pancreatic duct obstruction.

Science.gov (United States)

Yang, Li-Juan; Wan, Rong; Shen, Jia-Qing; Shen, Jie; Wang, Xing-Peng

2013-08-01

Remote organ failure occurs in cases of acute pancreatitis (AP); however, the reports on AP induced by pancreatic duct obstruction are rare. In this study we determined the effect of L-cysteine on pancreaticobiliary inflammation and remote organ damage in rats after pancreaticobiliary duct ligation (PBDL). AP was induced by PBDL in rats with 5/0 silk. Sixty rats were randomly divided into 4 groups. Groups A and B were sham-operated groups that received injections of saline or L-cysteine (10 mg/kg) intraperitoneally (15 rats in each group). Groups C and D were PBDL groups that received injections of saline or L-cysteine (10 mg/kg) intraperitoneally (15 rats in each group). The tissue samples of the pancreas and remote organs such as the lung, liver, intestine and kidney were subsequently examined for pathological changes under a light microscope. The samples were also stored for the determination of malondialdehyde and glutathione levels. Blood urea nitrogen (BUN), plasma amylase, ALT and AST levels were determined spectrophotometrically using an automated analyzer. Also, we evaluated the effect of L-cysteine on remote organ injury in rats with AP induced by retrograde infusion of 3.5% sodium taurocholate (NaTc) into the bile-pancreatic duct. Varying degrees of injury in the pancreas, lung, liver, intestine and kidney were observed in the rats 24 hours after PBDL. The severity of injury to the lung, liver and intestine was attenuated, while injury status was not changed significantly in the pancreas and kidney after L-cysteine treatment. Oxidative stress was also affected by L-cysteine in PBDL-treated rats. The concentration of tissue malondialdehyde decreased in the pancreas and remote organs of PBDL and L-cysteine administrated rats, and the concentration of glutathione increased more significantly than that of the model control group. However, L-cysteine administration reduced the severity of injury in remote organs but not in the pancreas in rats with Na

Unusual specific heat of almost dry L-cysteine and L-cystine amino acids.

Science.gov (United States)

Ishikawa, M S; Lima, T A; Ferreira, F F; Martinho, H S

2015-03-01

A detailed quantitative analysis of the specific heat in the 0.5- to 200-K temperature range for almost dry L-cysteine and its dimer, L-cystine, amino acids is presented. We report the occurrence of a sharp first-order transition at ∼76 K for L-cysteine associated with the thiol group ordering which was successfully modeled with the two-dimensional Ising model. We demonstrated that quantum rotors, two-level systems (TLS), Einstein oscillators, and acoustic phonons (the Debye model) are essential ingredients to correctly describe the overall experimental data. Our analysis pointed out the absence of the TLS contribution to the low temperature specific heat of L-cysteine. This result was similar to that found in other noncrystalline amorphous materials, e.g., amorphous silicon, low density amorphous water, and ultrastable glasses. L-cystine presented an unusual nonlinear acoustic dispersion relation ω(q)=vq0.95 and a Maxwell-Boltzmann-type distribution of tunneling barriers. The presence of Einstein oscillators with ΘE∼70 K was common in both systems and adequately modeled the boson peak contributions.

Studies on sister chromatid exchange (SCE) induction by γ-irradiation and protective effect of L-cysteine in HeLa cells

International Nuclear Information System (INIS)

Shah, V.C.; Mittal, S.C.

1987-01-01

Effect of different doses of γ-irradiation on SCE induction in unifiliarly 5-bromo 2-deoxyuridine substituted DNA was studied in various phases of cell cycle. Changes in γ-irradiation induced SCE frequency was measured by post-irradiation treatment with antimutagen L-cysteine. Perturbation in cellular proliferation kinetics due to γ-irradiation and γ-irradiation plus L-cysteine was also studied. It was observed that γ-irradiation is an efficient inducer of SCE and is most effective in S phase. L-cysteine also causes SCE induction which is slightly higher than the spontaneous level of SCEs found in HeLa cells. However, post-irradiation addition of L-cysteine reduces SCE frequency in γ-irradiated cultures and this reduction is maximum in G 1 phase irradiated cells. γ-irradiation delayed the mitosis considerably and this delay continued to increase with increasing doses. L-cysteine reduced the delay in cell cycle caused by γ-irradiation. (orig.) [de

Detection of Acetaldehyde in the Esophageal Tissue among Healthy Male Subjects after Ethanol Drinking and Subsequent L-Cysteine Intake.

Science.gov (United States)

Okata, Hideki; Hatta, Waku; Iijima, Katsunori; Asanuma, Kiyotaka; Tsuruya, Atsuki; Asano, Naoki; Koike, Tomoyuki; Hamada, Shin; Nakayama, Toru; Masamune, Atsushi; Shimosegawa, Tooru

2018-04-01

Ethanol is oxidized by alcohol dehydrogenase to acetaldehyde, a recognized carcinogen for the esophagus. However, no previous study has measured the acetaldehyde levels in the esophageal tissue. L-cysteine has been shown to reduce the acetaldehyde levels in the saliva; however, it is unknown whether L-cysteine intake affects the acetaldehyde concentration in the esophageal tissue. The aim of this study was to measure the acetaldehyde concentration in the esophageal tissue after ethanol drinking and evaluate the effect of L-cysteine intake on the acetaldehyde levels in the esophagus. We enrolled 10 male subjects with active acetaldehyde dehydrogenase-2*1/*1 (ALDH2*1/*1) genotype and 10 male subjects with the inactive acetaldehyde dehydrogenase-2*1/*2 (ALDH2*1/*2) genotype, the mean ages of whom were 25.6 and 27.9 years, respectively. In this prospective, single-blind, placebo-controlled study using L-cysteine and placebo lozenges (first and second examination), saliva and blood were collected before and after ethanol drinking. Esophageal tissue was obtained by endoscopic biopsy at 60 minutes after drinking, and the acetaldehyde and ethanol concentrations were measured. The acetaldehyde concentration of the saliva was significantly lower in those taking L-cysteine than in those taking the placebo. Acetaldehyde in the esophageal tissue was detected only in those taking L-cysteine lozenges. There were no correlations between the acetaldehyde concentrations in the esophageal tissue and saliva or blood. In conclusion, we detected acetaldehyde in the human esophageal tissue after ethanol drinking. Unexpectedly, intake of L-cysteine lozenges appears to contribute to detection of acetaldehyde in the esophageal tissue.

Optimization of Maillard Reaction in Model System of Glucosamine and Cysteine Using Response Surface Methodology.