scholarly journals Genetic features of P. falciparum parasites collected in 2012-2016 and anti-malaria resistance along China-Myanmar border

2020 ◽  
Author(s):  
Mei Li ◽  
Liu Hui ◽  
Tang Linhua ◽  
Yang Henglin ◽  
Maria Dorina Geluz Bustos ◽  
...  
Keyword(s):  
Yunnan Province ◽  
Tandem Repeats ◽  
Parasite Clearance ◽  
Nonparametric Tests ◽  
Chi Square ◽  
North West ◽  
Copy Numbers ◽  
Genetic Features ◽  
K13 Mutation

Abstract BackgroundThe therapeutic efficacy study (TES) of Dihydroartemisinin-Piperaquine (DHA-PIP) for uncomplicated P.falciparum patients had implemented during 2012-2016 along China (Yunnan province) -Myanmar border. The study focusing on the genetic features of falciparum parasites based on in vivo parasite clearance time (PCT) was investigated to explore if they had produced potential resistance to DHA and PIP at molecular level.MethodsThe genetic features were invested based on K13 propeller genotypes, Copy numbers of genes PfPM2 and Pfmdr1 and 9 microsatellite loci (Short Tandem Repeats, STR) flanking the K13 gene on chromosome 13. The (PCTs), copy numbers, expected heterozygosity ( He ), coefficient of gene differentiation (F st ) were calculated, compared and analyzed in Graph Prim5.0 and SPSS 23 with method of One-way Anova (Post Hoc test:LSD) or Nonparametric tests. Genetic diversity ( He and F st ) of parasites was estimated in Excel 2016 with method of GenAlEx 6.501.ResultsIn NW (North-West Yunnan province bordering with Myanmar) area, F446I prevalence was 58.96% (79/134) and no significant different PCT50 presented between any two K13 groups (Chi-Square=2.35, P=0.31, df=2). But the PCT50 of parasite isolates in this area (12.10, 10.72-13.47, n=136 hrs) was significantly shorter than that in SW (South-West Yunnan province bordering with Myanmar) area (P=0.036, t=-2.11, df=174) where isolates all showed wild K13 genotype. For the copy numbers of Pfmdr1 gene, 14.63% (18/123) and 62.5% (10/16) parasite isolates showed amplification (≥ 1.6) in NW and SW areas, but for those of PfM2 gene, none of them did. Based on STR data, He values of parasite isolates of 4 groups lined as ML group (Menglian County in SW), F446I group, Others (Non-F446I K13 mutation) group and W (wild K13 genotype) group from low to high. All of them showed significant with one another (Chi-Square=25.20, df=3, P=0.000). The mean F st value of 4 groups at 9 different loci (0.410±0.067) were significant higher than that of 3 NW groups (0.238±0.041) (Paired-T Test: t=-3.220,df=8,P=0.009). Highest mean F st (0.320±0.053) presented between ML group and W group when all mean F st values between W group and each of other 3 groups were compared.ConclusionsAccording to our study, we are inclined to conclude that no confirmed resistance of P.falciparum parasites to DHA and PIP occurred, although parasite isolates with prolonged PCTs were observed. Potentially resistant parasite isolates are widely spread along the Yunnan-Myanmar border based on the genetic index of K13, PfM2/3 and Pfmdr1, along with great genetic differentiation based on different areas and K13 mutations. F446I is the domain K13 mutation allele in NW area and seem to have emerged newly in those years.Trial registrationISRCTN, ISRCTN 11775446. Registered 13 April 2020 - Retrospectively registered, http://www.isrctn.com/ISRCTN11775446.

scholarly journals Genetic Features of P. falciparum Parasites Collected in 2012-2016 and Anti-Malaria Resistance Along China-Myanmar Border

2020 ◽  
Author(s):  
Mei Li ◽  
Hui Liu ◽  
Linhua Tang ◽  
Henglin Yang ◽  
Maria Dorina Geluz Bustos ◽  
...  
Keyword(s):  
Yunnan Province ◽  
Tandem Repeats ◽  
Artemisinin Resistance ◽  
Parasite Clearance ◽  
Pfmdr1 Gene ◽  
North West ◽  
Copy Numbers ◽  
Genetic Features ◽  
The Mean

Abstract Backgrounds The therapeutic efficacy studies (TES) of T(DHA-PIP) for uncomplicated P. falciparum patients were implemented during 2012-2016 along China (Yunnan province)-Myanmar border, which verified the high efficacy of DHA-PIP. The study focusing on the genetic features of falciparum parasites and basing on in vivo parasite clearance time (PCT) were carried out to explore if they had produced potential resistance to DHA and PIP at molecular level. Methods The genetic features were investigated based on K13 propeller genotypes, Copy numbers of genes pfPM2 and pfmdr1 and 9 microsatellite loci (Short Tandem Repeats, STR) flanking the K13 gene on chromosome 13. The PCT 50s basing on different K13 genotypes, sites, periods and copy numbers were compared.Results In NW (North-West Yunnan province bordering with Myanmar) area, F446I prevalence was 58.96% (79/134). No significant different PCT 50s presented among 3 K13 groups (Chi-Square=2.35, P=0.31, df=2) classified by K13 genotypes within NW area, but all of them were significantly shorter than that in SW (South-West Yunnan province bordering with Myanmar) area (P=0.036, t=-2.11, df=174) where isolates only showed wild K13 genotype. For the copy numbers of pfmdr1 gene, 14.63% (18/123) and 62.5% (10/16) parasite isolates showed double copies (≥ 1.6) in NW and SW areas separately, but for those of pfM2 gene, no parasite isolates did. Between isolates with single and double copies of pfmdr1 gene, no different PCT 50 presented. According to the mean He values, the four K13 groups were arranged as ML group (Menglian County in SW), F446I group, Others (Non-F446I K13 mutation) group and W (wild K13 genotype) group from low to high. The mean Fst values between ML and W groups were significantly higher than other 2 K13 group-W pairs (Paired-T Test: t=-2.659，df=8，P=0.029; t=-4.966, df=8，P=0.001). Conclusions According to our study, P. falciparum isolates in NW area and SW area are very different in genetic features. Artemisinin partial resistance, inferred from genetic marker, F446I, had independently appeared and spread in NW area during 2012-2016. With global application of ACTs, this pattern of artemisinin resistance producing was worth more attention. Fortunately, PIP-resistant feature basing on genetic analysis showed negative results in both areas. So, DHA-PIP was still recommended in antimalarial treatment along China-Myanmar border based on molecular data, which was agreed with the conclusion drawn from in vivo data obtained with same falciparum isolates. Trial registration: ISRCTN, ISRCTN 11775446. Registered 13 April 2020 - Retrospectively registered, http://www.isrctn.com/ISRCTN11775446.

scholarly journals Efficacy and safety of dihydroartemisinin-piperaquine for the treatment of uncomplicated Plasmodium falciparum and Plasmodium vivax malaria in Northern Papua and Jambi, Indonesia

2020 ◽  
Author(s):  
Din Syafruddin ◽  
Puji Budi Setia Asih ◽  
Ismail Ekoprayitno Rozi ◽  
Farahana Kresno Dewayanti ◽  
Suradi Wangsamuda ◽  
...  
Keyword(s):  
Vivax Malaria ◽  
Parasite Clearance ◽  
Single Copy ◽  
Efficacy And Safety ◽  
Once Daily ◽  
First Line Therapy ◽  
Copy Numbers ◽  
Kaplan Meier ◽  
Plasmodium Vivax Malaria

Dihydroartemisinin-piperaquine (DHA-PPQ) has been adopted as first-line therapy for uncomplicated falciparum malaria in Indonesia since 2010. The efficacy of DHA-PPQ was evaluated in 2 sentinel sites in Keerom District, Papua and Merangin District, Jambi Provinces from April 2017 to April 2018. Clinical and parasitological parameters were monitored over a 42-day period following the WHO standard in vivo protocol and subjects meeting the inclusion criteria were treated with DHA-PPQ once daily for 3 days, administered orally. In Keerom District, 6339 subjects were screened through active and passive cases detection. A total of 114 subjects infected by P. falciparum and 83 subjects infected by P. vivax agreed to take a part through written informed consent. Kaplan-Meier analysis of microscopy readings and PCR-corrected falciparum cases revealed a 93.1% (95%CI:86.4-97.2) and 97.9% (95%CI:92.7-99.7) DHA-PPQ efficacy, respectively and were classified as Adequate Clinical Parasitological Responses (ACPRs). For vivax malaria, the DHA-PPQ efficacy were 89% (95%CI: 80.2-94.9) and 100% (95%CI: 95.1-100) respectively. In Merangin District, 751 subjects were screened and 41 subjects infected by P. vivax were recruited. Microscopy reading and PCR-corrected analysis revealed a 97.4% (95%CI:86.2-99.9) and 100% (95%CI: 90.5-100) DHA-PPQ efficacy, respectively. No severe adverse events were found in both sites. In both sites, there was no delay in parasite clearance and no mutations in the PfK13 and PvK12 genes. Of the 6 recurrent P. falciparum found, 2 indicated recrudescent and 4 cases were re-infection. Analysis of the PfPM2 gene at day 0 and day of recurrence in recrudescent cases revealed the same single copy number, whereas 3 of the 4 re-infection cases carried 2-3 copy numbers. In conclusion, treatment of falciparum and vivax malaria cases with DHA-PPQ showed a high efficacy and safety. DHA-PPQ regimen is also efficacious against P. vivax cases in the absence of primaquine.

Novel concatameric heparin-binding peptides reverse heparin and low-molecular-weight heparin anticoagulant activities in patient plasma in vitro and in rats in vivo

Blood ◽  
2004 ◽  
Vol 103 (4) ◽  
pp. 1356-1363 ◽  
Author(s):  
Barbara P. Schick ◽  
David Maslow ◽  
Adrianna Moshinski ◽  
James D. San Antonio
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight Heparin ◽  
Tandem Repeats ◽  
Factor Xa ◽  
Low Molecular Weight ◽  
Heparin Binding ◽  
High Affinity ◽  
Binding Peptides

Abstract Patients given unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) for prophylaxis or treatment of thrombosis sometimes suffer serious bleeding. We showed previously that peptides containing 3 or more tandem repeats of heparin-binding consensus sequences have high affinity for LMWH and neutralize LMWH (enoxaparin) in vivo in rats and in vitro in citrate. We have now modified the (ARKKAAKA)n tandem repeat peptides by cyclization or by inclusion of hydrophobic tails or cysteines to promote multimerization. These peptides exhibit high-affinity binding to LMWH (dissociation constant [Kd], ≈ 50 nM), similar potencies in neutralizing anti–Factor Xa activity of UFH and enoxaparin added to normal plasma in vitro, and efficacy equivalent to or greater than protamine. Peptide (ARKKAAKA)3VLVLVLVL was most effective in all plasmas from enoxaparin-treated patients, and was 4- to 20-fold more effective than protamine. Several other peptide structures were effective in some patients' plasmas. All high-affinity peptides reversed inhibition of thrombin-induced clot formation by UFH. These peptides (1 mg/300 g rat) neutralized 1 U/mL anti–Factor Xa activity of enoxaparin in rats within 1 to 2 minutes. Direct blood pressure and heart rate measurements showed little or no hemodynamic effect. These heparin-binding peptides, singly or in combination, are potential candidates for clinical reversal of UFH and LMWH in humans.

Detection of Protein Changes in Serum of Workers following Inhalation Exposure to Toluene Diisocyanate Vapors

1992 ◽  
Vol 8 (6) ◽  
pp. 407-413 ◽  
Author(s):  
Adam B. Czuppon ◽  
Boleslaw Marczynski ◽  
Xaver Baur
Keyword(s):  
Serum Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Inhalation Exposure ◽  
Staining Intensity ◽  
Peak Height ◽  
Toluene Diisocyanate ◽  
Serum Samples ◽  
Chi Square ◽  
Challenge Tests

Serum samples of 10 workers undergoing occupational type inhalative challenge tests by toluene diisocyanate (TDI) were investigated by anion-exchange fast-protein-liquid-chromatography (FPLC) and polyacrylamide-gel electrophoresis (PAGE-SDS). Their serum chromatography profiles were compared to those of 20 unexposed individuals. The peak height of the first prealbumin peak in sera of workers after inhalative challenge tests was significantly different (p > 0, 01 Chi-square test) compared to that obtained before exposure and to that of unexposed subjects. In addition, qualitative changes of these peaks were also noted in sera of workers exposed to TDI. In the cases of exposed individuals, that peak was more diffuse with some shoulders and less symmetric in appearance. Similarly, PAGE-SDS of the serum proteins, followed by silver nitrate staining, revealed a different banding pattern after in vivo TDI exposure. One of the serum components at approximately 15 kD showed an increase of staining intensity after exposure (n = 10), compared to unexposed subjects or to patients before exposure. This serum fraction has not yet been identified. The results here demonstrate that it is possible to detect changes of serum proteins in TDI-exposed individuals within a relatively short analysis time. This could be useful for biological monitoring of exposure, since no method for such is yet available.

scholarly journals Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo

2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4257
Author(s):  
Alane Pains Oliveira do Monte ◽  
João Bosco Loiola Filho ◽  
Thais Thatiane Dos Santos Souza ◽  
Mayara De Souza Miranda ◽  
Lívia Correia Magalhães ◽  
...  
Keyword(s):  
Embryo Quality ◽  
Control Group ◽  
Chi Square ◽  
Mass Occurrence ◽  
Chi Square Test ◽  
Vitrification Solution ◽  
Cryoprotectant Solution ◽  
Pellucid Zone

<p>This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced <em>in vivo</em>. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P &lt; 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.</p>

scholarly journals Identification of two intermediates during processing of profilaggrin to filaggrin in neonatal mouse epidermis.

1984 ◽  
Vol 99 (4) ◽  
pp. 1372-1378 ◽  
Author(s):  
K A Resing ◽  
K A Walsh ◽  
B A Dale
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Tandem Repeats ◽  
Peptide Mapping ◽  
Two Dimensional ◽  
Major Event ◽  
Mouse Epidermis ◽  
Cell Stage ◽  
Processing Steps

A major event in the keratinization of epidermis is the production of the histidine-rich protein filaggrin (26,000 mol wt) from its high molecular weight (greater than 350,000) phosphorylated precursor (profilaggrin). We have identified two nonphosphorylated intermediates (60,000 and 90,000 mol wt) in NaSCN extracts of epidermis from C57/Bl6 mice by in vivo pulse-chase studies. Results of peptide mapping using a two-dimensional technique suggest that these intermediates consist of either two or three copies of filaggrin domains. Each of the intermediates has been purified. The ratios of amino acids in the purified components are unusual and essentially identical. The data are discussed in terms of a precursor containing tandem repeats of similar domains. In vivo pulse-chase experiments demonstrate that the processing of the high molecular weight phosphorylated precursor involves dephosphorylation and proteolytic steps through three-domain and two-domain intermediates to filaggrin. These processing steps appear to occur as the cell goes through the transition cell stage to form a cornified cell.

Repeated CT elements bound by zinc finger proteins control the absolute and relative activities of the two principal human c-myc promoters

1993 ◽  
Vol 13 (9) ◽  
pp. 5710-5724
Author(s):  
E DesJardins ◽  
N Hay
Keyword(s):  
Zinc Finger ◽  
Tandem Repeats ◽  
Zinc Finger Protein ◽  
Consensus Sequence ◽  
Regulatory Region ◽  
Maximal Activity ◽  
Single Copy ◽  
Promoter Usage

Transcription of the human proto-oncogene c-myc is governed by two tandem principal promoters, termed P1 and P2. In general, the downstream promoter, P2, is predominant, which is in contrast to the promoter occlusion phenomenon usually observed in genes containing tandem promoters. A shift in human c-myc promoter usage has been observed in some tumor cells and in certain physiological conditions. However, the mechanisms that regulate promoter usage are not well understood. The present studies identify regulators which are required to promote transcription from both human c-myc promoters, P1 and P2, and have a role in determining their relative activities in vivo. A novel regulatory region located 101 bp upstream of P1 was characterized and contains five tandem repeats of the consensus sequence CCCTCCCC (CT element). The integrity of the region containing all five elements is required to promote transcription from P1 and for maximal activity from P2 in vivo. A single copy of this same element, designated CT-I2, also appears in an inverted orientation 53 bp upstream of the P2 transcription start site. This element has an inhibitory effect on P1 transcription and is required for P2 transcription. The transcription factor Sp1 was identified as the factor that binds specifically to the tandem CT elements upstream of P1 and to the CT-I2 element upstream of P2. In addition, the recently cloned zinc finger protein ZF87, or MAZ, was also able to bind these same elements in vitro. The five tandem CT elements can be functionally replaced by a heterologous enhancer that only in the absence of CT-I2 reverses the promoter usage, similar to what is observed in the translocated c-myc allele of Burkitt's lymphoma cells.

Association of the brain anion exchanger, AE3, with the repeat domain of ankyrin

1993 ◽  
Vol 105 (4) ◽  
pp. 1137-1142 ◽  
Author(s):  
C.W. Morgans ◽  
R.R. Kopito
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Anion Exchanger ◽  
Deletion Mutant ◽  
Tandem Repeats ◽  
Structural Homology ◽  
Terminal Domain ◽  
Repeat Domain ◽  
The Brain

The 89 kDa NH2-terminal domain of erythrocyte ankyrin is composed almost entirely of 22 tandem repeats of a 33 amino acid sequence and constitutes the binding site for the cytoplasmic NH2-terminal domain of the erythrocyte anion exchanger, AE1. We have developed an assay to evaluate the in vivo interaction between a fragment of ankyrin corresponding to this domain (ANK90) and two non-erythroid anion exchangers, AE2 and AE3, that share considerable structural homology with AE1. Association was assessed by co-immunoprecipitation of ANK90-anion exchanger complexes from detergent extracts of cells cotransfected with plasmids encoding the ankyrin fragment and the anion exchanger or mutants thereof. ANK90 was co-immunoprecipitated with AE1 but not with an AE1 deletion mutant lacking the cytoplasmic NH2-terminal domain. Using this assay, we show that the brain anion exchanger AE3, but not the closely related homologue, AE2, is capable of binding to ankyrin.

Transcription of spacer sequences flanking the rat 45S ribosomal DNA gene

1987 ◽  
Vol 7 (1) ◽  
pp. 314-325
Author(s):  
C A Harrington ◽  
D M Chikaraishi
Keyword(s):  
Ribosomal Dna ◽  
Transcriptional Activity ◽  
Tandem Repeats ◽  
Initiation Site ◽  
Rrna Synthesis ◽  
Rna Transcripts ◽  
Pair Sequence ◽  
Polymerase I

The transcriptional activity of spacer sequences flanking the rat 45S ribosomal DNA (rDNA) gene were studied. Nascent RNA labeled in in vitro nuclear run-on reactions hybridized with both 5' and 3' spacer regions. The highest level of hybridization was seen with an rDNA fragment containing tandem repeats of a 130-base-pair sequence upstream of the 45S rRNA initiation site. Synthesis of RNA transcripts homologous to this internally repetitious spacer region was insensitive to high levels of alpha-amanitin, suggesting that it is mediated by RNA polymerase I. Analysis of steady-state RNA showed that these transcripts were present at extremely low levels in vivo relative to precursor rRNA transcripts. In contrast, precursor and spacer run-on RNAs were synthesized at similar levels. This suggests that spacer transcripts are highly unstable in vivo; therefore, it may be the process of transcription rather than the presence of spacer transcripts that is functionally important. Transcription in this upstream rDNA region may be involved in regulation of 45S rRNA synthesis in rodents, as has been suggested previously for frog rRNA. In addition, the presence of transcriptional activity in other regions of the spacer suggests that some polymerase I molecules may transcribe through the spacer from one 45S gene to the next on rodent rDNA.

Use of insecticides and other capturing techniques of bush cricket (Tettigonia viridissima): dietary implications among population of Tubah, Cameroon

2019 ◽  
Vol 5 (3) ◽  
pp. 163-169
Author(s):  
J. Sonchieu ◽  
H.C. Roméo ◽  
Y. Wenceslas ◽  
N.K. Mireille
Keyword(s):  
Local Population ◽  
Protein Supplementation ◽  
Dietary Factors ◽  
Cooking Methods ◽  
Chemical Toxicity ◽  
Good Source ◽  
Chi Square ◽  
North West ◽  
West Region ◽  
Structured Questionnaire

Worldwide consumption of insects is today regarded as a good source of protein supplementation. This study aims at exploring the dietary factors, cooking, preservation techniques, and harvesting methods of longhorn grasshoppers (Tettigonia viridissima). The survey was conducted from September 2016 to January 2017 in Tubah council (North-West Region, Cameroon) using a semi-structured questionnaire, which was administered to two hundred T. viridissima harvesters. Chi-square was used to compare variables in a particular group for related measured factors. The main objective of the questionnaire was to find out how longhorn grasshoppers were harvested and consumed in the community. The harvesting of grasshoppers included three major techniques: hand picking (53%), trapping (29%) and use of insecticides (18%). Insecticides used (cypermethrin and deltamethrin) are from class II chemical toxicity. Dietary factors focused on: cooking methods (frying, roasting and boiling) and methods of preservation (sun drying and refrigerating). For the local population, grasshoppers constitute a good source of proteins from the month of September to January, but its safety is a concern as far as insecticides are used in harvesting them. Therefore, the population should be educated on the health risks linked to this malpractice method and the local and national authorities informed.

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