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1.
Braz. j. biol ; 83: e247422, 2023. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1285631

ABSTRACT

Abstract Plasmodium falciparum resistance to Chloroquine (CQ) is a significant cause of mortality and morbidity worldwide. There is a paucity of documented data on the prevalence of CQ-resistant mutant haplotypes of Pfcrt and Pfmdr1 genes from malaria-endemic war effected Federally Administered Tribal Areas of Pakistan. The objective of this study was to investigate the prevalence of P. falciparum CQ-resistance in this area. Clinical isolates were collected between May 2017 and May 2018 from North Waziristan and South Waziristan agencies of Federally Administrated Trial Area. Subsequently, Giemsa-stained blood smears were examined to detect Plasmodium falciparum. Extraction of malarial DNA was done from microscopy positive P. falciparum samples, and P. falciparum infections were confirmed by nested PCR (targeting Plasmodium small subunit ribosomal ribonucleic acid (ssrRNA) genes). All PCR confirmed P. falciparum samples were sequenced by pyrosequencing to find out mutation in Pfcrt gene at codon K76T and in pfmdr1 at codons N86Y, Y184F, N1042D, and D1246Y. Out of 121 microscopies positive P. falciparum cases, 109 samples were positive for P. falciparum by nested PCR. Pfcrt K76T mutation was found in 96% of isolates, Pfmdr1 N86Y mutation was observed in 20%, and 11% harboured Y184F mutation. All samples were wild type for Pfmdr1 codon N1042D and D1246Y. In the FATA, Pakistan, the frequency of resistant allele 76T remained high despite the removal of CQ. However, current findings of the study suggest complete fixation of P. falciparum CQ-resistant genotype in the study area.


Resumo A resistência do Plasmodium falciparum à cloroquina (CQ) é uma causa significativa de mortalidade e morbidade em todo o mundo. Há uma escassez de dados documentados sobre a prevalência de haplótipos mutantes CQ-resistentes dos genes Pfcrt e Pfmdr1 da guerra endêmica da malária em áreas tribais administradas pelo governo federal do Paquistão. O objetivo deste estudo foi investigar a prevalência de resistência a CQ de P. falciparum nesta área. Isolados clínicos foram coletados entre maio de 2017 e maio de 2018 nas agências do Waziristão do Norte e do Waziristão do Sul da Área de Ensaio Administrada Federalmente. Posteriormente, esfregaços de sangue corados com Giemsa foram examinados para detectar Plasmodium falciparum. A extração do DNA da malária foi feita a partir de amostras de P. falciparum positivas para microscopia, e as infecções por P. falciparum foram confirmadas por nested PCR (visando genes de ácido ribonucleico ribossômico de subunidade pequena de Plasmodium (ssrRNA)). Todas as amostras de P. falciparum confirmadas por PCR foram sequenciadas por pirosequenciamento para descobrir a mutação no gene Pfcrt no códon K76T e em pfmdr1 nos códons N86Y, Y184F, N1042D e D1246Y. De 121 microscopias de casos positivos de P. falciparum, 109 amostras foram positivas para P. falciparum por nested PCR. A mutação Pfcrt K76T foi encontrada em 96% dos isolados, a mutação Pfmdr1 N86Y foi observada em 20% e 11% abrigou a mutação Y184F. Todas as amostras eram do tipo selvagem para o códon N1042D e D1246Y de Pfmdr1. No FATA, Paquistão, a frequência do alelo resistente 76T permaneceu alta apesar da remoção de CQ. No entanto, as descobertas atuais do estudo sugerem a fixação completa do genótipo resistente a CQ de P. falciparum na área de estudo.


Subject(s)
Plasmodium falciparum/genetics , Antimalarials/pharmacology , Pakistan , Membrane Transport Proteins/genetics , Drug Resistance/genetics , Protozoan Proteins/genetics , Chloroquine/pharmacology , Multidrug Resistance-Associated Proteins/genetics , Alleles
2.
Article in Chinese | WPRIM | ID: wpr-878979

ABSTRACT

With the emergence of drug resistance in Western medicine, the repeated administration of clinical first-line drugs becomes more severe. There are many factors leading to multidrug resistance(MDR), so it is very difficult to solve the problem. Since traditional Chinese medicine(TCM) has been used in the field of MDR in recent years, the research on the transporter-associated drug resistance and intervention of TCM has gradually become a hot spot. Therefore, in order to further explore the relationships among drug resistance, transporters, and TCM intervention, we review the relevant research progress in recent years and comb the achievements and limitations of this research at present. In the end, we put forward the research direction of changing body's ADME through the host's transporters and gastrointestinal flora, which provides new ideas for future research.


Subject(s)
Drug Resistance, Multiple , Drugs, Chinese Herbal , Medicine, Chinese Traditional , Membrane Transport Proteins/genetics
3.
Braz. arch. biol. technol ; 63: e20180513, 2020. graf
Article in English | LILACS | ID: biblio-1132208

ABSTRACT

Abstract Silicon accumulation is known to improve tolerance of plants under both biotic and abiotic stress. Salinity stress is an inevitable crisis causing wide spread damage to rice leading to food insecurity. The influence of Si (1mM) on two rice cultivars cv. Ghanteswari (high accumulator) and cv.Badami (low accumulator) which differs in Si uptake potential under saline (10ds/m EC) and non- saline conditions were studied in nutrient culture. The Si transporter genes were isolated and characterized to determine their function in salinity tolerance. Under stress, there was an increase in Si accumulation, Na+/K+ ratio, electrolyte leakage, lipid peroxidation and antioxidant activities. On addition of silicon, the K+ uptake increased, membrane damage reduced and osmolytes balance improve under salinity. But, the level of resurgence was varied in both cultivars, due to their differential Si-accumulation. Molecular characterizations of Lsi1 protein revealed its involvement in the movement of ion and water and therefore prevent osmotic stress. The Lsi2 is responsible for removal of Na+, reducing salt toxicity. Silicon accumulation is responsible for maintenance of cell water status, osmotic balance and Na+ ion exclusion during high salinity. The variable relative expression of Lsi2 provides a possible explanation for differential genotypic uptake of silicon.


Subject(s)
Membrane Transport Proteins/genetics , Oryza/genetics , Silicon/metabolism , Gene Expression Regulation , Salinity , Salt Stress , Genotype
4.
Arch. argent. pediatr ; 115(3): 153-156, jun. 2017.
Article in English, Spanish | LILACS, BINACIS | ID: biblio-887326

ABSTRACT

Antecedentes. El síndrome de anemia megaloblástica sensible a la tiamina (TRMA, por sus siglas en inglés), también conocido como síndrome de Rogers, se caracteriza por presentar anemia megaloblástica, hipoacusia neurosensorial y diabetes mellitus. Las alteraciones en el transporte de la tiamina hacia las células se deben a mutaciones homocigotas o heterocigotas compuestas en el gen SLC19A2. Presentación de un caso. Presentamos el caso de una niña que manifestaba sordera neurosensorial tratada con una prótesis auditiva, diabetes con necesidad de insulina y anemia macrocítica, tratada con tiamina (100 mg/día). El nivel de hemoglobina mejoró hasta alcanzar 12,1 g/dl después de aumentar la dosis terapéutica de tiamina hasta 200 mg/día. Conclusión. Se debe evaluar a los pacientes con TRMA para detectar anemia megaloblástica, hipoacusia neurosensorial y diabetes mellitus. Se les debe dar seguimiento para determinar la respuesta de la enfermedad hematológica y de la diabetes después de la terapia con tiamina. La dosis terapéutica de tiamina puede aumentarse según la respuesta clínica. Debe proporcionarse asesoramiento genético.


Background. Thiamine-responsive megaloblastic anemia syndrome (TRMA), also known as Rogers syndrome, is characterized by megaloblastic anemia, sensorineural hearing loss, and diabetes mellitus. Disturbances of the thiamine transport into the cells results from homozygous or compound heterozygous mutations in the SLC19A2 gene. Case presentation. We report a girl which presented with sensorineural deafness treated with a hearing prosthesis, insulin requiring diabetes, macrocytic anemia, treated with thiamine (100 mg/day). Hemoglobin level improved to 12.1 g/dl after dose of thiamine therapy increased up to 200 mg/day. Conclusion. Patients with TRMA must be evaluated for megaloblastic anemia, sensorineural hearing loss, and diabetes mellitus. They must be followed for response of hematologic and diabetic after thiamine therapy. It should be kept in mind that dose of thiamine therapy may be increased according to the clinical response. Genetic counseling should be given.


Subject(s)
Humans , Female , Infant , Membrane Transport Proteins/genetics , Thiamine Deficiency/congenital , Thiamine Deficiency/genetics , Diabetes Mellitus/genetics , Hearing Loss, Sensorineural/genetics , Anemia, Megaloblastic/genetics , Mutation
5.
Braz. j. microbiol ; 48(2): 191-192, April.-June 2017.
Article in English | LILACS | ID: biblio-839376

ABSTRACT

Abstract Serratia marcescens is a Gram-negative rod intrinsically resistant to polymyxins and usually associated with wound, respiratory and urinary tract infections. The whole genome of the first GES-5-producing S. marcescens isolated from a Brazilian patient was sequenced using Ion Torrent PGM System. Besides blaGES-5, we were able to identify genes encoding for other β-lactamases, for aminoglycoside modifying enzymes and for an efflux pump to tetracyclines.


Subject(s)
Humans , Serratia marcescens/enzymology , Serratia marcescens/genetics , beta-Lactamases/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/chemistry , Genome, Bacterial , Sequence Analysis, DNA , Membrane Transport Proteins/genetics , Serratia marcescens/isolation & purification , Transferases/metabolism , beta-Lactamases/genetics , Brazil
6.
Arch. endocrinol. metab. (Online) ; 60(4): 323-327, Aug. 2016. tab, graf
Article in English | LILACS | ID: lil-792946

ABSTRACT

ABSTRACT Objective Pendred syndrome (PS) is an autosomal recessive disorder characterised by sensorineural hearing loss and thyroid dyshormonogenesis. It is caused by biallelic mutations in the SLC26A4 gene encoding for pendrin. Hypothyroidism in PS can be present from birth and therefore diagnosed by neonatal screening. The aim of this study was to examine the SLC26A4 mutation spectrum and prevalence among congenital hypothyroidism (CH) patients in the Guangxi Zhuang Autonomous Region of China and to establish how frequently PS causes hearing impairment in our patients with CH. Subjects and methods Blood samples were collected from 192 CH patients in Guangxi Zhuang Autonomous Region, China, and genomic DNA was extracted from peripheral blood leukocytes. All exons of the SLC26A4 gene together with their exon-intron boundaries were screened by next-generation sequencing. Patients with SLC26A4 mutations underwent a complete audiological evaluation including otoscopic examination, audiometry and morphological evaluation of the inner ear. Results Next generation sequencing analysis of SLC26A4 in 192 CH patients revealed five different heterozygous variations in eight individuals (8/192, 4%). The prevalence of SLC26A4 mutations was 4% among studied Chinese CH. Three of the eight were diagnosed as enlargement of the vestibular aqueduct (EVA), no PS were found in our 192 CH patients. The mutations included one novel missense variant p.P469S, as well as four known missense variants, namely p.V233L, p.M147I, p.V609G and p.D661E. Of the eight patients identified with SLC26A4 variations in our study, seven patients showed normal size/location of thyroid gland, and one patients showed a decreased size one. Conclusions The prevalence of SLC26A4 pathogenic variants was 4% among studied Chinese patients with CH. Our study expanded the SLC26A4 mutation spectrum, provided the best estimation of SLC26A4 mutation rate for Chinese CH patients and indicated the rarity of PS as a cause of CH.


Subject(s)
Humans , Male , Female , Infant, Newborn , Membrane Transport Proteins/genetics , Genetic Testing/methods , Goiter, Nodular/genetics , Hearing Loss, Sensorineural/genetics , Mutation , Thyroxine/blood , Vestibular Aqueduct/abnormalities , Thyrotropin/blood , China/epidemiology , Prevalence , Cohort Studies , Neonatal Screening/methods , Sulfate Transporters , Goiter, Nodular/epidemiology , Hearing Loss, Sensorineural/epidemiology
7.
Braz. dent. j ; 27(3): 255-260, May-June 2016. tab, graf
Article in English | LILACS | ID: lil-782824

ABSTRACT

Abstract The periodontopathogen Aggregatibacter actinomycetemcomitans colonizes oral cavity by binding to and invading epithelial cells as well as by participating in biofilms formed on hard surfaces. Aae, an autotransporter protein, is implicated in bacterial adhesion to epithelial cells. Due to the multiple functions of bacterial autotransporter proteins, this study aimed to evaluate the role of aae in A. actinomycetemcomitans ability to adhere to both saliva-coated hydroxyapatite (SHA) and biofilm. An aae null mutant was constructed. Its hydrophobic properties as well as its ability to adhere to epithelial cells, SHA and to form biofilm were evaluated and compared with the parental strain, A. actinomycetemcomitans VT1169. The aae null mutant showed reduced hydrophobicity, as well as decreased binding to SHA and biofilm formation compared to the parental strain. These data suggest that aae mediates A. actinomycetemcomitans adhesion to epithelial cells and may be involved in biofilm formation and interaction with adsorbed salivary proteins.


Resumo O peridontopatógeno Aggregatibacter actinomycetemcomitans coloniza a cavidade oral aderindo e invadindo as células epiteliais e participando da formação de biofilme em superfícies duras. Aae, uma proteína autotransportadora está relacionada com a adesão bacteriana às células epiteliais. Devido às múltiplas funções desempenhadas por proteínas bacterianas autotransportadoras, este estudo teve como objetivo avaliar o papel de aae de A. actinomycetemcomitans tanto na capacidade de aderir à hidroxiapatita recoberta por saliva (SHA), quanto a de formar biofilme. Um mutante nulo aae foi construído. Suas propriedades hidrofóbicas, bem como a sia capacidade para aderir às células epiteliais, à SHA e para formar biofilme foram avaliadas e comparadas com a cepa -mãe, A. Actinomycetemcomitans VT1169. O mutante nulo aae apresentou redução de hidrofobicidade, assim como diminuição da adesão à SHA e na formação de biofilme, quando comparado à cepa parental. Estes dados sugerem que aae media a adesão de A. Actinomycetemcomitans às células epiteliais e pode também estar envolvida na formação de biofilme e na interação com proteínas salivares adsorvidas.


Subject(s)
Aggregatibacter actinomycetemcomitans/physiology , Bacterial Adhesion/physiology , Bacterial Proteins/physiology , Membrane Transport Proteins/physiology , Aggregatibacter actinomycetemcomitans/genetics , Bacterial Proteins/genetics , Biofilms , Gene Knockdown Techniques , Hydrophobic and Hydrophilic Interactions , Membrane Transport Proteins/genetics
8.
Braz. j. microbiol ; 47(1): 55-62, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-775121

ABSTRACT

Abstract Mercury, which is ubiquitous and recalcitrant to biodegradation processes, threatens human health by escaping to the environment via various natural and anthropogenic activities. Non-biodegradability of mercury pollutants has necessitated the development and implementation of economic alternatives with promising potential to remove metals from the environment. Enhancement of microbial based remediation strategies through genetic engineering approaches provides one such alternative with a promising future. In this study, bacterial isolates inhabiting polluted sites were screened for tolerance to varying concentrations of mercuric chloride. Following identification, several Pseudomonas and Klebsiella species were found to exhibit the highest tolerance to both organic and inorganic mercury. Screened bacterial isolates were examined for their genetic make-up in terms of the presence of genes (merP and merT) involved in the transport of mercury across the membrane either alone or in combination to deal with the toxic mercury. Gene sequence analysis revealed that the merP gene showed 86–99% homology, while the merT gene showed >98% homology with previously reported sequences. By exploring the genes involved in imparting metal resistance to bacteria, this study will serve to highlight the credentials that are particularly advantageous for their practical application to remediation of mercury from the environment.


Subject(s)
Humans , Klebsiella/metabolism , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Mercury/metabolism , Pseudomonas/metabolism , Water Pollutants, Chemical/metabolism , Drug Tolerance , Genes, Bacterial , India , Klebsiella/drug effects , Klebsiella/genetics , Molecular Sequence Data , Mercury/toxicity , Pseudomonas/drug effects , Pseudomonas/genetics , Sequence Analysis, DNA , Sequence Homology , Water Pollutants, Chemical/toxicity
9.
Mem. Inst. Oswaldo Cruz ; 110(3): 433-444, 05/2015. tab, graf
Article in English | LILACS | ID: lil-745976

ABSTRACT

Benznidazole (BZ) is one of the two drugs used for Chagas disease treatment. Nevertheless therapeutic failures of BZ have been reported, which were mostly attributed to variable drug susceptibility among Trypanosoma cruzi strains. ATP-binding cassette (ABC) transporters are involved in a variety of translocation processes and some members have been implicated in drug resistance. Here we report the characterisation of the first T. cruzi ABCG transporter gene, named TcABCG1, which is over-expressed in parasite strains naturally resistant to BZ. Comparison of TcABCG1 gene sequence of two TcI BZ-resistant strains with CL Brener BZ-susceptible strain showed several single nucleotide polymorphisms, which determined 11 amino acid changes. CL Brener transfected with TcI transporter genes showed 40-47% increased resistance to BZ, whereas no statistical significant increment in drug resistance was observed when CL Brener was transfected with the homologous gene. Only in the parasites transfected with TcI genes there was 2-2.6-fold increased abundance of TcABCG1 transporter protein. The analysis in wild type strains also suggests that the level of TcABCG1 transporter is related to BZ natural resistance. The characteristics of untranslated regions of TcABCG1 genes of BZ-susceptible and resistant strains were investigated by computational tools.


Subject(s)
Animals , Humans , ATP-Binding Cassette Transporters/genetics , Drug Resistance/genetics , Nitroimidazoles/pharmacology , Trypanocidal Agents/pharmacology , Trypanosoma cruzi/drug effects , Trypanosoma cruzi/genetics , DNA, Protozoan/genetics , Genotype , Membrane Transport Proteins/genetics , Parasitic Sensitivity Tests , Phylogeny
10.
Rev. latinoam. enferm ; 23(2): 259-266, Feb-Apr/2015. tab, graf
Article in English | LILACS, BDENF | ID: lil-747163

ABSTRACT

OBJECTIVE: to assess the clinical effect of topical treatment using Ulmo honey associated with oral ascorbic acid in patients with venous ulcers. METHOD: longitudinal and descriptive quantitative study. During one year, 18 patients were assessed who were clinically diagnosed with venous ulcer in different stages, male and female, adult, with a mean injury time of 13 months. Ulmo honey was topically applied daily. The dressing was applied in accordance with the technical standard for advanced dressings, combined with the daily oral consumptions of 500 mg of ascorbic acid. The monitoring instrument is the assessment table of venous ulcers. RESULTS: full healing was achieved in 100% of the venous ulcers. No signs of complications were observed, such as allergies or infection. CONCLUSION: the proposed treatment showed excellent clinical results for the healing of venous ulcers. The honey demonstrated debriding and non-adherent properties, was easy to apply and remove and was well accepted by the users. The described results generated a research line on chronic wound treatment. .


OBJETIVO: avaliar o efeito clínico de tratamento tópico com mel de Ulmo associado à administração oral de ácido ascórbico em pacientes portadores de úlceras venosas. MÉTODO: estudo quantitativo descritivo longitudinal. Um total de 18 pacientes adultos, ambos os sexos, clinicamente diagnosticados com úlcera venosa em diferentes estágios e com duração de 13 meses em média, foram avaliados pelo período de um ano. A aplicação tópica diária de mel de Ulmo foi realizada de acordo com a norma técnica de tratamento avançado combinada com o consumo diário de 500 mg de ácido ascórbico. O instrumento usado para monitoramento foi a tabela de avaliação de úlceras venosas. RESULTADOS: cicatrização completa foi observada em 100% das úlceras venosas. Não foram observados sinais de complicação tais como alergias ou infecção. CONCLUSÃO: o tratamento proposto apresentou resultados clínicos excelentes na cicatrização das úlceras venosas. Além de favorecer o debridamento, o mel não é aderente, é fácil de aplicar e remover, e é de fácil aceitação por parte dos usuários. Os resultados descritos geraram uma linha investigativa no tratamento de feridas crônicas. .


OBJETIVO: evaluar el efecto clínico del tratamiento con miel de Ulmo tópico asociado a ácido ascórbico oral en pacientes portadores de úlceras venosas. MÉTODO: estudio cuantitativo descriptivo longitudinal. Durante el período de un año se evaluaron 18 pacientes diagnosticados clínicamente de úlcera venosa en sus diferentes estadios, de ambos sexos, adultos, con 13 meses promedio de antigüedad de la lesión. Se realizó la aplicación tópica diaria de miel de Ulmo con curación según la norma técnica de curaciones avanzadas, combinada con el consumo oral diario de 500 mg de ácido ascórbico. El instrumento de seguimiento es la tabla de valoración de úlceras venosas. RESULTADOS: se logró la cicatrización total en el 100% de las úlceras venosas. No se observaron signos de complicación, tales como alergias o infección. CONCLUSIÓN: el tratamiento propuesto mostró excelentes resultados clínicos en la cicatrización de úlceras venosas, presentando la miel propiedades debridantes, no adherentes, fácil de aplicar, remover y aceptación del usuario. Los resultados descritos generaron una línea investigativa en el tratamiento de heridas crónicas. .


Subject(s)
Humans , Female , Adult , Middle Aged , Aged , Young Adult , Antineoplastic Agents/pharmacology , Breast Neoplasms/etiology , Genetic Predisposition to Disease , Germ-Line Mutation/genetics , Pharmacogenetics , Polymorphism, Single Nucleotide/genetics , Biomarkers, Tumor/genetics , Breast Neoplasms/drug therapy , Breast Neoplasms/epidemiology , Case-Control Studies , /genetics , DNA, Neoplasm/genetics , Follow-Up Studies , Genome-Wide Association Study , Genotype , Membrane Transport Proteins/genetics , Methyltransferases/genetics , Polymerase Chain Reaction , Prognosis , Registries , Risk Factors , United States/epidemiology
11.
Article in English | WPRIM | ID: wpr-130570

ABSTRACT

The aim of the study was to explore the possible molecular markers of chloroquine resistance in Plasmodium vivax isolates in Thailand. A total of 30 P. vivax isolates were collected from a malaria endemic area along the Thai-Myanmar border in Mae Sot district of Thailand. Dried blood spot samples were collected for analysis of Pvmdr1 and Pvcrt-o polymorphisms. Blood samples (100 mul) were collected by finger-prick for in vitro chloroquine susceptibility testing by schizont maturation inhibition assay. Based on the cut-off IC50 of 100 nM, 19 (63.3%) isolates were classified as chloroquine resistant P. vivax isolates. Seven non-synonymous mutations and 2 synonymous were identified in Pvmdr1 gene. Y976F and F1076L mutations were detected in 7 (23.3%) and 16 isolates (53.3%), respectively. Analysis of Pvcrt-o gene revealed that all isolates were wild-type. Our results suggest that chloroquine resistance gene is now spreading in this area. Monitoring of chloroquine resistant molecular markers provide a useful tool for future control of P. vivax malaria.


Subject(s)
Amino Acid Substitution , Antimalarials/pharmacology , Chloroquine/pharmacology , Drug Resistance , Humans , Inhibitory Concentration 50 , Malaria, Vivax/parasitology , Membrane Transport Proteins/genetics , Multidrug Resistance-Associated Proteins/genetics , Mutation, Missense , Myanmar , Parasitic Sensitivity Tests , Plasmodium vivax/drug effects , Protozoan Proteins/genetics , Thailand
12.
Article in English | WPRIM | ID: wpr-130563

ABSTRACT

The aim of the study was to explore the possible molecular markers of chloroquine resistance in Plasmodium vivax isolates in Thailand. A total of 30 P. vivax isolates were collected from a malaria endemic area along the Thai-Myanmar border in Mae Sot district of Thailand. Dried blood spot samples were collected for analysis of Pvmdr1 and Pvcrt-o polymorphisms. Blood samples (100 mul) were collected by finger-prick for in vitro chloroquine susceptibility testing by schizont maturation inhibition assay. Based on the cut-off IC50 of 100 nM, 19 (63.3%) isolates were classified as chloroquine resistant P. vivax isolates. Seven non-synonymous mutations and 2 synonymous were identified in Pvmdr1 gene. Y976F and F1076L mutations were detected in 7 (23.3%) and 16 isolates (53.3%), respectively. Analysis of Pvcrt-o gene revealed that all isolates were wild-type. Our results suggest that chloroquine resistance gene is now spreading in this area. Monitoring of chloroquine resistant molecular markers provide a useful tool for future control of P. vivax malaria.


Subject(s)
Amino Acid Substitution , Antimalarials/pharmacology , Chloroquine/pharmacology , Drug Resistance , Humans , Inhibitory Concentration 50 , Malaria, Vivax/parasitology , Membrane Transport Proteins/genetics , Multidrug Resistance-Associated Proteins/genetics , Mutation, Missense , Myanmar , Parasitic Sensitivity Tests , Plasmodium vivax/drug effects , Protozoan Proteins/genetics , Thailand
13.
Braz. j. microbiol ; 45(2): 509-514, Apr.-June 2014. tab
Article in English | LILACS | ID: lil-723107

ABSTRACT

The aim of this study was to investigate the phylogenetic background and to assess hlyD (involved in the secretion of haemolysin A) and intll (encoding a class 1 integrase) in Escherichia coli isolates derived from urinary and fecal specimens. A total of 200 E. coli isolates was collected from patients presenting with urinary tract infection (UTI) during September 2009 to September 2010 and screened for hlyD and intll genes by polymerase chain reaction (PCR). Phylogenetic analysis showed that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D) and that uropathogenic E. coli (UPEC) isolates mainly belong to groups B2 (54%) and D (34%) whereas group A (44%) and D (26%) are predominant among commensal E. coli isolates. In this study, hlyD was present in 26% of UPEC and 2% of commensal E. coli isolates. However, hemolytic activity was detected for 42% of UPEC and 6% of commensal E. coli isolates (p < 0.05). intll gene was more frequently expressed in UPEC (24%) in comparison with commensal E. coli isolates (12%). Resistance to aztreonam, co-trimoxazole and cefpodoxime were frequently found among UPEC isolates whereas commensal E. coli isolates were commonly resistant to co-trimoxazole, nalidixic acid and cefotaxime. Concluding, a considerable difference between UPEC and commensal E. coli isolates was observed regarding their phylogenetic groups, presence of class 1 integron and hlyD gene, hemolysin activity and resistance pattern. The detection of class 1 integrons and hlyD gene was higher among UPEC compared with commensal E. coli isolates. These findings may contribute for a better understanding of the factors involved in the pathogenesis of UPEC.


Subject(s)
Child , Child, Preschool , Female , Humans , Male , Escherichia coli Infections/microbiology , Escherichia coli/classification , Feces/microbiology , Genetic Variation , Phylogeny , Urinary Tract Infections/microbiology , Urine/microbiology , Anti-Bacterial Agents/pharmacology , Cluster Analysis , Drug Resistance, Bacterial , Escherichia coli Proteins/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/isolation & purification , Genotype , Hemolysin Proteins/genetics , Integrases/genetics , Membrane Transport Proteins/genetics , Polymerase Chain Reaction , Sequence Analysis, DNA
14.
Braz. j. microbiol ; 45(2): 661-665, Apr.-June 2014. ilus, tab
Article in English | LILACS | ID: lil-723133

ABSTRACT

The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90) of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi.


Subject(s)
Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Rhodococcus equi/genetics , Rhodococcus equi/metabolism , Biological Transport, Active , DNA, Bacterial/genetics , Ethidium/metabolism , Polymerase Chain Reaction
15.
Article in English | WPRIM | ID: wpr-17801

ABSTRACT

LR11, also known as SorLA or SORL1, is a type-I membrane protein from which a large extracellular part, soluble LR11 (sLR11), is released by proteolytic shedding on cleavage with a disintegrin and metalloproteinase 17 (ADAM17). A shedding mechanism is presumed to have a key role in the functions of LR11, but the evidence for this has not yet been demonstrated. Tetraspanin CD9 has been recently shown to regulate the ADAM17-mediated shedding of tumor necrosis factor-alpha and intercellular adhesion molecule-1 on the cell surface. Here, we investigated the role of CD9 on the shedding of LR11 in leukocytes. LR11 was not expressed in THP-1 monocytes, but it was expressed and released in phorbol 12-myristate 13-acetate (PMA)-induced THP-1 macrophages (PMA/THP-1). Confocal microscopy showed colocalization of LR11 and CD9 proteins on the cell surface of PMA/THP-1. Ectopic neo-expression of CD9 in CCRF-SB cells, which are LR11-positive and CD9-negative, reduced the amount of sLR11 released from the cells. In contrast, incubation of LR11-transfected THP-1 cells with neutralizing anti-CD9 monoclonal antibodies increased the amount of sLR11 released from the cells. Likewise, the PMA-stimulated release of sLR11 increased in THP-1 cells transfected with CD9-targeted shRNAs, which was negated by treatment with the metalloproteinase inhibitor GM6001. These results suggest that the tetraspanin CD9 modulates the ADAM17-mediated shedding of LR11 in various leukemia cell lines and that the association between LR11 and CD9 on the cell surface has an important role in the ADAM17-mediated shedding mechanism.


Subject(s)
ADAM Proteins/metabolism , Tetraspanin 29/genetics , Cell Line, Tumor , Humans , LDL-Receptor Related Proteins/genetics , Leukocytes/metabolism , Macrophages/metabolism , Membrane Transport Proteins/genetics , Proteolysis
16.
Indian J Exp Biol ; 2013 Dec; 51(12): 1130-1136
Article in English | IMSEAR | ID: sea-150302

ABSTRACT

The genomic and cDNA sequences of BnSUT1C were isolated from B. napus. Combination of cDNA and genomic DNA sequences revealed that the BnSUT1C gene contained three exons and two introns. The cDNA encodes a protein of 513 amino acids with a calculated molecular mass of 54.7 kDa and an isoelectric point of 9.12. It exhibits typical features of sucrose transporter with 12 trans-membranes spanning domains. BnSUT1C showed highly homologous with AtSUC1 and AtSUC5. A histidine residue, which is conserved across all functional sucrose transporter proteins in higher plants, is located at position 66 of the BnSUT1C. Two putative pollen-specific cis-elements, AGAAA and GTGA motifs, are located in 5′-upstream of BnSUT1C. The spatial and temporal expression patterns carried out by semi-quantitative RT-PCR and Real-Time PCR, which indicated that BnSUT1C predominantly expressed in later developmental stages of anther, as tapetal cells began to shrink and collapse. BnSUT1C could mediate the uptake of sucrose in the pollen and retrieval of tapetal degenerated products during pollen maturation.


Subject(s)
Amino Acid Sequence , Arabidopsis/genetics , Base Sequence , Brassica napus/genetics , Cloning, Molecular , DNA/genetics , DNA, Complementary/genetics , Membrane Transport Proteins/genetics , Membrane Transport Proteins/isolation & purification , Plant Proteins/genetics , Plant Proteins/isolation & purification , RNA, Messenger/genetics , Sequence Homology, Amino Acid , Transcriptome
17.
Indian J Biochem Biophys ; 2013 Dec; 50(6): 500-504
Article in English | IMSEAR | ID: sea-150263

ABSTRACT

Some compounds in the garlic inhibit cholesterol synthesis, resulting in lowering of serum cholesterol and triglycerides and increase in HDL level. However, the mechanism of this specific effect is not fully understood. In the small intestine, ATP-binding cassette transporters G5, G8 and A1 (ABCG5, ABCG8 and ABCA1), as well as Niemann-Pick C1 like 1 (NPC1L1) protein have important roles in cholesterol metabolism. In this study, we evaluated the beneficial effect of aqueous extract of garlic on lipid profile and also expression of npc1l1, abca1, abcg5 and abcg8 genes in the intestine of N-Marry mice fed a high cholesterol diet as a possible mechanism of garlic effect. Twenty-four mice were randomly divided into three groups: Group 1: hypercholesterolmic (received chow + 2% cholesterol + 0.5% cholic acid); Group 2: garlic (received chow + 4% (w/w) garlic extract + 2% cholesterol + 0.5% cholic acid); and Group 3: received chow only. After one month, mice were anesthetized and blood was collected from their heart. The jejunum was removed, washed with PBS and entrocytes were scraped and used for the experiments. Serum lipids were measured enzymatically and expression of mRNA levels for the above-mentioned proteins was determined by semi-quantitative RT-PCR. Garlic extract significantly reduced serum lipids (p<0.05), compared with the hypercholesterolemic group. Expression of the intestinal npc1l1 was significantly decreased (p<0.01) in the garlic group, compared with the chow group, while abcg5 (p<0.01), abcg8 (p<0.01) and abca1 (p<0.05) expressions were significantly increased. In conclusion, this study reveals a possible mechanism for the beneficial effects of the garlic in lowering serum lipids by decreasing the intestinal lipid absorption and increasing excretion of cholesterol back into the intestinal lumen.


Subject(s)
ATP Binding Cassette Transporter 1/genetics , ATP-Binding Cassette Transporters/genetics , Animals , Garlic/chemistry , Gene Expression Regulation/drug effects , Hypercholesterolemia/genetics , Intestines/drug effects , Intestines/metabolism , Lipids/blood , Lipoproteins/genetics , Membrane Transport Proteins/genetics , Mice , Plant Extracts/pharmacology , Water/chemistry
18.
Mem. Inst. Oswaldo Cruz ; 108(8): 947-961, 6/dez. 2013. tab, graf
Article in English | LILACS | ID: lil-697142

ABSTRACT

The development and rapid spread of chloroquine resistance (CQR) in Plasmodium falciparum have triggered the identification of several genetic target(s) in the P. falciparum genome. In particular, mutations in the Pfcrt gene, specifically, K76T and mutations in three other amino acids in the region adjoining K76 (residues 72, 74, 75 and 76), are considered to be highly related to CQR. These various mutations form several different haplotypes and Pfcrt gene polymorphisms and the global distribution of the different CQR- Pfcrt haplotypes in endemic and non-endemic regions of P. falciparum malaria have been the subject of extensive study. Despite the fact that the Pfcrt gene is considered to be the primary CQR gene in P. falciparum , several studies have suggested that this may not be the case. Furthermore, there is a poor correlation between the evolutionary implications of the Pfcrt haplotypes and the inferred migration of CQR P. falciparum based on CQR epidemiological surveillance data. The present paper aims to clarify the existing knowledge on the genetic basis of the different CQR- Pfcrt haplotypes that are prevalent in worldwide populations based on the published literature and to analyse the data to generate hypotheses on the genetics and evolution of CQR malaria.


Subject(s)
Humans , Antimalarials/pharmacology , Chloroquine/pharmacology , Drug Resistance/genetics , Haplotypes/genetics , Malaria, Falciparum/parasitology , Membrane Transport Proteins/genetics , Plasmodium falciparum/genetics , Protozoan Proteins/genetics , DNA, Protozoan/genetics , Malaria, Falciparum/drug therapy , Polymorphism, Genetic , Plasmodium falciparum/drug effects
19.
Braz. j. infect. dis ; 17(2): 125-130, Mar.-Apr. 2013. ilus, tab
Article in English | LILACS | ID: lil-673188

ABSTRACT

Several studies have been conducted in recent years to elucidate the structure, function and significance of AcrB, MarA, SoxS and RamA in Salmonella enterica. In this study, the relative quantification of acrB, soxS, marA and ramA genes expression was evaluated in 14 strains of S. enterica, with or without accompanying mutations in the quinolone resistance-determining regions of the gyrA gene, that were exposed to ciprofloxacin during the exponential growth phase. The presence of ciprofloxacin during the log phase of bacterial growth activated the genes marA, soxS, ramA and acrB in all S. enterica strains analyzed in this study. The highest expression levels for acrB were observed in strains with gyrA mutation, and marA showed the highest expression in the strains without mutation. Considering only the strains with ciprofloxacin minimum inhibitory concentration values < 0.125 [1]g/mL (sensitive to ciprofloxacin), the most expressed gene in the strains both with and without mutations was acrB. In the strains with ciprofloxacin minimum inhibitory concentration values > 0.125 [1]g/mL (low susceptibility), with and without mutations in gyrA, the most expressed gene was marA. In this study, we observed that strains resistant to nalidixic acid may express genes associated with the efflux pump and the expression of the AcrAB-TolC pump genes seems to occur independently of mutations in gyrA.


Subject(s)
Humans , Bacterial Proteins/genetics , Drug Resistance, Bacterial/genetics , Mutation/genetics , Salmonella enterica/drug effects , Salmonella enterica/genetics , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Ciprofloxacin/pharmacology , Gene Expression Regulation, Bacterial , Genes, Bacterial/genetics , Microbial Sensitivity Tests , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , RNA, Bacterial/genetics , Trans-Activators/genetics , Trans-Activators/metabolism
20.
Clinics ; 68(2): 277-280, 2013.
Article in English | LILACS | ID: lil-668817

ABSTRACT

In this study, we investigated the associations between single-nucleotide polymorphisms in GAB2 (rs2373115), GSK3B (rs6438552) and SORL1 (rs641120) and Alzheimer's disease (AD), both alone and in combination with the APOE*4 allele.


Subject(s)
Aged , Female , Humans , Male , Adaptor Proteins, Signal Transducing/genetics , Alzheimer Disease/genetics , /genetics , Cytoskeletal Proteins/genetics , LDL-Receptor Related Proteins/genetics , Membrane Transport Proteins/genetics , Nuclear Proteins/genetics , Polymorphism, Single Nucleotide/genetics , Age of Onset , Risk Factors
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