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Sample records for anti-giardia lamblia serum

  1. Detection of anti-Giardia lamblia serum antibody among children of day care centers

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    Guimarães Semíramis

    2002-01-01

    Full Text Available OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF and enzyme-linked immunosorbent assay (ELISA for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3% showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3% and 100 (68% samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39% than IIF (70%. IIF also showed a higher concordance with microscopic examination than ELISA.

  2. Detection of anti-Giardia lamblia serum antibody among children of day care centers

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    Semíramis Guimarães

    2002-02-01

    Full Text Available OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF and enzyme-linked immunosorbent assay (ELISA for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3% showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3% and 100 (68% samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39% than IIF (70%. IIF also showed a higher concordance with microscopic examination than ELISA.

  3. Detection of anti-Giardia lamblia serum antibody among children of day care centers Detecção de anticorpos séricos anti-Giardia lamblia em crianças de creches

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    Semíramis Guimarães

    2002-02-01

    Full Text Available OBJETIVES: To detect anti-Giardia lamblia serum antibodies in healthy children attending public day care centers and to assess serological tests as tools for estimating the prevalence of G. lamblia in endemic areas. METHODS: Three separate stool specimens and filter paper blood samples were collected from 147 children ranging from 0 to 6 years old. Each stool sample was processed using spontaneous sedimentation and zinc sulfate flotation methods. Blood samples were tested by indirect immunofluorescence (IIF and enzyme-linked immunosorbent assay (ELISA for Giardia IgG. RESULTS AND CONCLUSIONS: Of 147 individuals tested, 93 (63.3% showed Giardia cysts in their feces. Using IIF and ELISA, serum antibodies were detected in 93 (63.3% and 100 (68% samples , respectively. Sensitivity of IIF and ELISA was 82% and 72%, respectively. However, ELISA revealed to be less specific (39% than IIF (70%. IIF also showed a higher concordance with microscopic examination than ELISA.OBJETIVOS: Detectar anticorpos séricos anti-Giardia lamblia entre crianças atendidas em creches e estimar a freqüência de infecção por Giardia lamblia em área endêmica. MÉTODOS: Foram coletadas três amostras de fezes de cada uma das 147 crianças de três creches da rede municipal de Botucatu, SP, com idade variando de 0 a 6 anos, e as amostras foram processadas pelos métodos de sedimentação espontânea e flutuação pelo sulfato de zinco. Amostras de sangue foram obtidas da polpa digital, coletadas em papel de filtro e testadas pelos métodos de imunofluorescência indireta (IFI e de reação imunoenzimática (Elisa para pesquisa de IgG anti-Giardia. RESULTADOS E CONCLUSÕES: De um total de 147 crianças, 93 (63,3% apresentaram cistos de Giardia nas fezes. Dos 147 eluatos testados, 93 (63,3% e 100 (68% foram positivos para Giardia em IFI e em Elisa, respectivamente. A sensibilidade de IFI foi de 82% e de Elisa, 72%. Contudo, Elisa foi menos específica (39% do que IFI (70

  4. In vitro anti-Giardia lamblia activity of 2-aryl-3-hydroxymethyl imidazo[1,2-a]pyridines and -pyrimidines, individually and in combination with albendazole.

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    Velázquez-Olvera, Stephanía; Salgado-Zamora, Héctor; Jiménez-Cardoso, Enedina; Campos-Aldrete, Maria-Elena; Pérez-González, Cuauhtémoc; Ben Hadda, Taibi

    2016-03-01

    Giardiasis is a major diarrheal disease found throughout the world, the causative agent being the flagellate protozoan Giardia intestinalis. Infection is more common in children than in adults. The appearance of drug resistance has complicated the treatment of several parasitic diseases, including giardiasis. Thus, the aim of this investigation was to make an in vitro evaluation of the antigiardia response of synthetic derivatives 2-aryl-3-hydroxymethylimidazo[1,2-a]pyridines 1 and -pyrimidines 2 against trophozoites of Giardia lamblia WB, in comparison with the reference drug, albendazole. Additionally, the synergistic action of albendazole in combination with each of the most active 2-aryl-3-hydroxymethyl imidazo[1,2-a]pyridines and pyrimidines was also assessed. Based on the IC50 values obtained, the best anti-Giardia activity was provided by the 3-hydroxymethyl-4-fluorophenylimidazo[1,2-a]pyrimidine derivative 2c and the corresponding imidazo[1,2-a]pyrimidine with the p-tolyl substituent 2d, followed by 2a and 2b. These four compounds showed effectiveness at a concentration similar to that of albendazole. Regarding synergism, the IC50 of the combination of albendazole with 2a, 2b or 2c gave the best anti-Giardia action, showing greater efficacy than albendazole alone. Hence, G. lamblia WB showed high susceptibility to some 2-aryl-3-hydroxymethyl imidazo[1,2-a] pyrimidines, which acted synergistically when used in combination with albendazole.

  5. Validity of silver, chitosan, and curcumin nanoparticles as anti-Giardia agents.

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    Said, D E; Elsamad, L M; Gohar, Y M

    2012-08-01

    This study was carried out to evaluate the anti parasitic potential of silver, chitosan, and curcumin nanoparticles as anti-Giardia agents. Non-treated infected control rats were inoculated with Giardia lamblia cysts in a dose of 2 × 10(5) cysts/rat. Experimental group was infected then treated with curcumin, curcumin nanoparticles, chitosan, chitosan nanoparticles, and silver nanoparticles as single or combined therapy. The number of Giardia cysts in stools and trophozoites in intestinal sections were detected. Toxicity of nanoparticles was evaluated by comparing hematological and histopathological parameters of the normal control group and treated non-infected control group. The amount of silver was also measured in the liver, kidney, small intestine, lung, and brain of rats treated with silver nanoparticles. The number of the parasites in stool and small intestinal sections decreased in treated infected rats compared with infected non-treated ones. The effect in the single therapy was better with nanoparticles, and the best effect was detected in nano-silver. The combined therapy gave better results than single. Combination between nanoparticles was better than the combination of nano-forms and native chitosan and curcumin. The best effect was detected in combinations of nano-silver and nano-chitosan but with no full eradication. In conclusion, the highest effect and complete cure was gained by combining the three nano-forms. The parasite was successfully eradicated from stool and intestine. None of the treatments exhibited any toxicity. Accumulated silver in different organs was within the safe limits.

  6. Immuno-PCR for detection of Giardia lamblia cysts in water.

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    Deng, Ming Jun; Ji, Xin Cheng; Xiao, Xi Zhi; Sun, Tao; Wu, Zhen Xing; Zheng, Xiao Long; Wang, Qun; Zhu, Lai Hua

    2014-01-01

    Giardia lamblia cysts at low concentrations were detected in water samples using a highly sensitive immunological-PCR (IPCR) method. Magnetic gold particles were precoated with monoclonal anti-Giardia antibodies, and Giardia lamblia cysts ranging from 1 to 100 cysts were diluted in 500 microL of water. A biotinylated detection antibody bound to the G. lamblia cysts was then linked by a streptavidin bridge to biotinylated Giardia-reporter DNA. After extensive washing, reporter DNA was released by denaturation, transferred to PCR tubes, amplified, electrophoresed, and visualized. An optimized immuno-PCR method detected as little as five G. lamblia cysts. To further evaluate the specificity and the clinical application of this IPCR assay for G. lamblia cysts, Entamoeba histolytica and Cryptosporidium parvum were also collected and detected by IPCR. The data demonstrated that this monoclonal antibody-based IPCR method is particularly useful for analysis of environmental water samples in which the densities of G. lamblia cysts is very low, and provides a platform capable of rapid screening of samples from drinking water, wells, rivers, lakes, and recreational swimming pools for trace levels of G. lamblia cysts.

  7. An integrated microfludic device for culturing and screening of Giardia lamblia.

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    Zheng, Guo-Xia; Zhang, Xue-Mei; Yang, Yu-Suo; Zeng, Shu-Rui; Wei, Jun-Feng; Wang, Yun-Hua; Li, Ya-Jie

    2014-02-01

    In vitro culturing of trophozoites was important for research of Giardia lamblia (G. lamblia), especially in discovery of anti-Giardia agents. The current culture methods mainly suffer from lab-intension or the obstacle in standardizing the gas condition. Thus, it could benefit from a more streamlined and integrated approach. Microfluidics offers a way to accomplish this goal. Here we presented an integrated microfluidic device for culturing and screening of G. lamblia. The device consisted of a polydimethylsiloxane (PDMS) microchip with an aerobic culture system. In the microchip, the functionality of integrated concentration gradient generator (CGG) with micro-scale cell culture enables dose-response experiment to be performed in a simple and reagent-saving way. The diffusion-based culture chambers allowed growing G. lamblia at the in vivo like environment. It notable that the highly air permeable material of parallel chambers maintain uniform anaerobic environment in different chambers easily. Using this device, G. lamblia were successfully cultured and stressed on-chip. In all cases, a dose-related inhibitory response was detected. The application of this device for these purposes represents the first step in developing a completely integrated microfluidic platform for high-throughput screening and might be expanded to other assays based on in vitro culture of G. lamblia with further tests.

  8. Plant species used in giardiasis treatment: ethnopharmacology and in vitro evaluation of anti-Giardia activity

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    Vanessa do A. Neiva

    2014-04-01

    Full Text Available The aim of this study was to compile the traditional knowledge about plants used for the treatment of giardiasis, and also to carry out experimental research to evaluate the anti-Giardia activity of five species.To reach this objective, 398 interviews were performed using a previously prepared questionnaire, followed by an in vitro evaluation of giardicidal potential of hydroalcoholic leaf extracts of Anacardium occidentale L., Chenopodium ambrosioides L., Passiflora edulis Sims, Psidium guajava L., and Stachytarpheta cayennensis(Rich. Vahl. Among the interviewed people, 55.53% reported the use of plants to treat diarrhea, the most severe symptom of giardiasis. The results indicated 36 species used by this population for these problems. The use of leaves (72.50% of a single plant (64.25% collected from backyards and gardens (44.34% and prepared by decoction were predominant. The majority of the interviewees (85.52% attributed their cure to the use of plants. In the experimental tests, all extracts inhibited the growth of Giardia lambliatrophozoites in different intensities: A. occidentale and P. guajava extracts elicited a moderate activity (250 ≤ IC50 ≤ 500 μg/ml, C. ambrosioides and S. cayennensis extracts evoked a high activity (100 ≤ IC50 ≤ 250 μg/ml, and P. edulis extract showed very high activity (IC50≤ 100 μg/ml. This study shows that an ethnopharmacological approach is useful in the selection of plant materials with potential giardicidal activity.

  9. Synthesis and Biological Evaluation of Metronidazole Derivatives as Anti-Giardia Agents

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    Bahador Shahriari

    2015-06-01

    Full Text Available Nitroimidazole derivatives such as Metronidazole (MTZ have been used as anti- protozoa and anti-anaerobic bacteria. In this study several derivatives of MTZ were synthesized and evaluated against Giardia lamblia cyst. MTZ were reacted with several alkyl halide to obtain o-alkyl MTZ derivatives, then products were purified and their chemical structures were confirmed by spectral analysis (1HNMR and Mass. In order to assess biological evaluation, all compounds were investigated against 25 Giardia samples isolated from Giardia-infected patients. Results showed that compound 2 had the most activity on cyst of Giardia in comparison with MTZ.

  10. Effects of the extract and glycoalkaloids of Solanum lycocarpum St. Hill on Giardia lamblia trophozoites

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    Gilmarcio Z Martins

    2015-01-01

    Full Text Available Background: Solanum lycocarpum has great importance for food and medicinal traditional use. Recently, it was also evidenced that extracts of S. lycocarpum St. Hill (Solanaceae and its glycoalkaloids, solamargine (Sg and solasonine (Sn, are active against flagellated protozoa. Objective: The aim was to assess the effects of the extract of S. lycocarpum and its glycoalkaloids, Sn, and Sg, on Giardia lamblia trophozoites. Materials and Methods: A crude extract (96% ethanol (EB of fruits of S. lycocarpum was prepared and fractionated by partition with 40% ethanol and n-hexane: Ethyl acetate. Glycoalkaloids, Sn, and Sg were recognized in the ethanol fraction (EF and further isolated by column chromatography. EB, EF, the isolated Sn and Sg and a mixture (1:1 of both glycoalkaloids were tested on cultures of G. lamblia trophozoites and macrophages. Results: EB, EF and glycoalkaloids of S. lycocarpum showed activity against Giardia (95.0 < Inhibitory concentration 50 [IC 50 ] ≤120.3 μg/mL. The mixture of glycoalkaloids (1:1 was more active (IC 50 = 13.23 μg/mL than each one individually, suggesting a synergic effect. Moreover, the mixture is nontoxic to macrophage cells. Conclusion: Results are optimistic concerning the anti-Giardia potential of the mixture Sn + Sg. Further studies, in vitro and in vivo, will be required to consolidate the usefulness of the mixture of Sn + Sg in view of a new therapeutic strategy for giardiasis.

  11. The Giardia lamblia genome.

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    Adam, R D

    2000-04-10

    Giardia lamblia is a protozoan parasite of humans and other mammals that is thought to be one of the most primitive extant eukaryotic organisms. Although distinctly eukaryotic, it is notable for its lack of mitochondria, nucleoli, and perixosomes. It has been suggested that Giardia spp. are pre-mitochondriate organisms, but the identification of genes in G. lamblia thought to be of mitochondrial origin has generated controversy regarding that designation. Giardi lamblia trophozoites have two nuclei that are identical in all ways that have been studied. They are polyploid with at least four, and perhaps eight or more, copies of each of five chromosomes per organism and have an estimated genome complexity of 1.2x10(7)bp of DNA, and GC content of 46%. There is evidence for recombination at the telomeres of some of the chromosomes, and multiple size variants of single chromosomes have been identified within cloned isolates. However, the internal regions of the chromosomes demonstrate no evidence of recombination. For example, there is no evidence for control of vsp gene expression by DNA recombination, and no evidence for rapid mutation in the vsp genes. Single pass sequences of approximately 9% of the G. lamblia genome have already been obtained. An ongoing genome project plans to obtain approximately 95% of the genome by a random approach, as well as a complete physical map using a bacterial artificial chromosome library. The results will facilitate a better understanding of the biology of Giardia spp. as well as their phylogenetic relationship to other primitive organisms.

  12. Sonoran propolis and some of its chemical constituents inhibit in vitro growth of Giardia lamblia trophozoites.

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    Alday-Provencio, Samuel; Diaz, Gabriela; Rascon, Lucila; Quintero, Jael; Alday, Efrain; Robles-Zepeda, Ramón; Garibay-Escobar, Adriana; Astiazaran, Humberto; Hernandez, Javier; Velazquez, Carlos

    2015-06-01

    Propolis is a cereus resin with a complex chemical composition that possesses a wide range of biological activities. The aim of this study was to evaluate the in vitro anti-Giardia lamblia activity of Sonoran propolis collected from three different areas of Sonoran Desert in northwestern Mexico (Caborca, Pueblo de Alamos, and Ures) and some of its chemical constituents. Additionally, we also analyzed the seasonal effect on the anti-G. lamblia activity of propolis. G. lamblia trophozoite cultures were treated with different concentrations of Sonoran propolis or chemical compounds during 48 h cell proliferation and cell viability were determined. Ures propolis showed the highest inhibitory activity against G. lamblia (IC50 63.8 ± 7.1 µg/mL) in a dose-dependent manner (Ures > Pueblo de Alamos > Caborca). Season had a significant effect on the in vitro anti-G. lamblia activity of Ures propolis. Summer propolis showed the highest inhibitory effect on the G. lamblia trophozoite growth (IC50 23.8 ± 2.3 µg/mL), followed by propolis collected during winter (IC50 59.2 ± 34.7 µg/mL), spring (IC50 102.5 ± 15.3 µg/mL), and autumn (IC50 125.0 ± 3.1 µg/mL). Caffeic acid phenethyl ester, an Ures propolis exclusive constituent, had the highest growth-inhibitory activity towards G. lamblia [IC50 63.1 ± 0.9 µg/mL (222.1 ± 3.2 µM)]. To our knowledge, this is the first study showing that caffeic acid phenethyl ester possesses antiparasitic activity against G. lamblia. Naringenin [IC50 125.7 ± 20.7 µg/mL (461.8 ± 76.3 µM)], hesperetin [IC50 149.6 ± 24.8 µg/mL (494.9 ± 82.2 µM)], and pinocembrin [IC50 174.4 ± 26.0 µg/mL (680.6 ± 101.7 µM)] showed weak anti-G. lamblia activity. On the other hand, chrysin and rutin did not show significant antiparasitic activity. In conclusion, our results suggest that Sonoran propolis and some of its chemical constituents had inhibitory effects on the

  13. Host defences against Giardia lamblia.

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    Lopez-Romero, G; Quintero, J; Astiazarán-García, H; Velazquez, C

    2015-08-01

    Giardia spp. is a protozoan parasite that inhabits the upper small intestine of mammals and other species and is the aetiological agent of giardiasis. It has been demonstrated that nitric oxide, mast cells and dendritic cells are the first line of defence against Giardia. IL-6 and IL-17 play an important role during infection. Several cytokines possess overlapping functions in regulating innate and adaptive immune responses. IgA and CD4(+) T cells are fundamental to the process of Giardia clearance. It has been suggested that CD4(+) T cells play a double role during the anti-Giardia immune response. First, they activate and stimulate the differentiation of B cells to generate Giardia-specific antibodies. Second, they act through a B-cell-independent mechanism that is probably mediated by Th17 cells. Several Giardia proteins that stimulate humoral and cellular immune responses have been described. Variant surface proteins, α-1 giardin, and cyst wall protein 2 can induce host protective responses to future Giardia challenges. The characterization and evaluation of the protective potential of the immunogenic proteins that are associated with Giardia will offer new insights into host-parasite interactions and may aid in the development of an effective vaccine against the parasite. © 2015 John Wiley & Sons Ltd.

  14. MODELING INACTIVATION OF GIARDIA LAMBLIA

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    Under the auspices of the Safe Drinking Water Act (SDWA)the U.S. EPA hasa promulgated the Surface Water Treatment Rule (SWTR) requiring public water systems using surface water to provide minimum disinfection to Control Giardia Lamblia, enteric virsues, and bacteria. The C-t con...

  15. MODELING INACTIVATION OF GIARDIA LAMBLIA

    Science.gov (United States)

    Under the auspices of the Safe Drinking Water Act (SDWA)the U.S. EPA hasa promulgated the Surface Water Treatment Rule (SWTR) requiring public water systems using surface water to provide minimum disinfection to Control Giardia Lamblia, enteric virsues, and bacteria. The C-t con...

  16. Identification of an immunogenic protein of Giardia lamblia using monoclonal antibodies generated from infected mice

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    Jael Quintero

    2013-08-01

    Full Text Available The humoral immune response plays an important role in the clearance of Giardia lamblia. However, our knowledge about the specific antigens of G. lamblia that induce a protective immune response is limited. The purpose of this study was to identify and characterise the immunogenic proteins of G. lamblia in a mouse model. We generated monoclonal antibodies (moAbs specific to G. lamblia (1B10, 2C9.D11, 3C10.E5, 3D10, 5G8.B5, 5F4, 4C7, 3C5 and 3C6 by fusing splenocytes derived from infected mice. Most of these moAbs recognised a band of ± 71 kDa (5G8 protein and this protein was also recognised by serum from the infected mice. We found that the moAbs recognised conformational epitopes of the 5G8 protein and that this antigen is expressed on the cell surface and inside trophozoites. Additionally, antibodies specific to the 5G8 protein induced strong agglutination (> 70-90% of trophozoites. We have thus identified a highly immunogenic antigen of G. lamblia that is recognised by the immune system of infected mice. In summary, this study describes the identification and partial characterisation of an immunogenic protein of G. lamblia. Additionally, we generated a panel of moAbs specific for this protein that will be useful for the biochemical and immunological characterisation of this immunologically interesting Giardia molecule.

  17. Binding of cholera toxin to Giardia lamblia.

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    McCardell, B. A.; Madden, J M; Stanfield, J T; Tall, B D; Stephens, M. J.

    1987-01-01

    Binding of cholera toxin to Giardia lamblia was demonstrated by two slightly different methods: an immunofluorescence technique using antibody to cholera toxin and anti-rabbit immunoglobulin G conjugated to fluorescein isothiocyanate, and a one-step fluorescence method in which G. lamblia was incubated with the B subunit of cholera toxin conjugated to fluorescein isothiocyanate.

  18. Serodiagnóstico de giardiosis: identificación de inmunoglobulina G anti-Giardia duodenalis en suero mediante ELISA

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    Sofía Duque

    2001-09-01

    Full Text Available El diagnóstico de la infección por Giardia duodenalis se basa en la identificación de quistes o trofozoítos en materia fecal, alcanzando una sensibilidad máxima de 85% en muestras seriadas, debido a la excreción intermitente de quistes. Aunque la determinación de anticuerpos no permite diferenciar entre una infección pasada y una reciente, si podría ayudar en el diagnóstico diferencial. El presente estudio tuvo como objetivos estandarizar y evaluar una prueba inmunoenzimática (ELISA para la detección de anticuerpos tipo IgG anti-Giardia duodenalis en sueros de pacientes con la infección. La preparación del antigeno se hizo a partir de trofozoitos cultivados que, inicialmente, fueron obtenidos del intestino de gerbils (Meriones unguiculatus infectados experimentalmente con quistes purificados a partir de muestras positivas de materia fecal humana. Para la estandarización y evaluación de la prueba, se emplearon 60 sueros de pacientes con giardiosis comprobada parasitológicamente (muestras positivas y 47 sueros de cordón umbilical recién cortado (muestras negativas. Como conjugado, se utilizó anti-lgG humana ligada a fosfatasa alcalina. La concentración óptima de antígeno de trofozoíto de G. duodenalis fue de 15 ug/ml y las diluciones óptimas de suero y conjugado fueron 1:25 y 1:400, respectivamente. El valor del punto de corte (absorbancia fue 0,300. Los parámetros de la prueba fueron: sensibilidad, 98,3% (intervalo de confianza de 95% (lC 95%: 89,9%-99,974; especificidad, 957% (IC 95%:84,3%-99,3%; valor predictivo positivo, 96,7% (IC 95%: 87,6%-99,4%, y valor predictivo negativo, 97,8% (IC 95%: 87,0%-99,9%. La prueba ELlSA descrita contribuirá a mejorar el diagnóstico de giardiosis y podrá ser empleada para estudios epidemiológicos de seroprevalencia.

  19. Diagnosis of Giardia lamblia infections by detection of parasite-specific antigens.

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    Janoff, E N; Craft, J C; Pickering, L K; Novotny, T; Blaser, M J; Knisley, C V; Reller, L B

    1989-03-01

    Antigen detection methods may facilitate diagnosis of Giardia lamblia in stool specimens. As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and immunoblotting, G. lamblia cysts and trophozoites share several antigens, especially in the 65-kilodalton and 30- to 34-kilodalton regions. By using blind methods, we compared results obtained by counterimmunoelectrophoresis using cyst-immune rabbit serum and by enzyme-linked immunosorbent assay (ELISA) using trophozoite-immune rabbit serum with results obtained by microscopic examination of a preserved, concentrated, and permanently stained stool specimen. Results were similar when these three methods were used to examine 118 stool specimens from clinical microbiology laboratories (53 specimens with G. lamblia) and specimens from 239 day-care-center toddlers (39 specimens with G. lamblia). Compared with microscopy, we found, for counterimmunoelectrophoresis and ELISA, respectively: sensitivity, 88 versus 94%; specificity, 97 versus 95%; positive predictive value, 86 versus 76%; negative predictive value, 98 versus 97%; and concordance, 89%. The false-positive rate by ELISA was 24% (10 of 42) in day-care-center toddlers but only 3% (1 of 32) in healthy adults (P less than 0.04) as corroborated by microscopy. This discrepancy suggests that the ELISA may be more sensitive than microscopy, which is considered the reference standard, and that results may be dependent, in part, on the epidemiology of the infection in the study subjects.

  20. Giardia lamblia infections in children in Ghana.

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    Anim-Baidoo, Isaac; Narh, Charles Akugbey; Oddei, Dora; Brown, Charles Addoquaye; Enweronu-Laryea, Christabel; Bandoh, Betty; Sampane-Donkor, Eric; Armah, George; Adjei, Andrew Anthony; Adjei, David Nana; Ayeh-Kumi, Patrick Ferdinand; Gyan, Ben Adu

    2016-01-01

    Though giardiasis is an important public health problem in Ghana, several aspects of its epidemiology, particularly the molecular epidemiology has not been investigated adequately. This could be a major hindrance to effective surveillance and control of giardiasis in the country. The study was carried out to determine the prevalence, risk factors and genotypes of Giardia lamblia infecting children at a paediatric hospital in Ghana. A total of 485 patients including 365 diarrhoea and 120 non-diarrhoea children were enrolled into the study. Stool samples were collected and analysed for parasite presence using microscopy, ELISA and PCR. Positive samples were subsequently characterized into assemblages by PCR-RFLP, and further confirmed with sequencing of the glutamate dehydrogenase (gdh) gene. Epidemiological data on demographic, clinical and behavioral features of the study subjects were also collected. Prevalence of G. lamblia infections in diarrhoea and non-diarrhoea children were 5.8% and 5% respectively (P>0.5). Sequence data confirmed Giardia lamblia assemblage B as the predominant genotype in both diarrhoea and non-diarrhoea cases. There was no significant association of G. lamblia infection with any of the epidemiological variables investigated. Our findings suggest that assemblage B could be the predominant genotype causing giardiasis in children. Increased public health education focusing on good sanitary practices, particularly among mothers and children, could decrease the risk of G. lamblia infection.

  1. A spliceosomal intron in Giardia lamblia

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    Nixon, Julie E. J.; Wang, Amy; Morrison, Hilary G.; McArthur, Andrew G.; Sogin, Mitchell L.; Loftus, Brendan J.; Samuelson, John

    2002-01-01

    Short introns occur in numerous protist lineages, but there are no reports of intervening sequences in the protists Giardia lamblia and Trichomonas vaginalis, which may represent the deepest known branches in the eukaryotic line of descent. We have discovered a 35-bp spliceosomal intron in a gene encoding a putative [2Fe-2S] ferredoxin of G. lamblia. The Giardia intron contains a canonical splice site at its 3′ end (AG), a noncanonical splice site at its 5′ end (CT), and a branch point sequence that fits the yeast consensus sequence of TACTAAC except for the first nucleotide (AACTAAC). We have also identified several G. lamblia genes with spliceosomal peptides, including homologues of eukaryote-specific spliceosomal peptides (Prp8 and Prp11), several DExH-box RNA-helicases that have homologues in eubacteria, but serve essential functions in the splicing of introns in eukaryotes, and 11 predicted archaebacteria-like Sm and like-Sm core peptides, which coat small nuclear RNAs. Phylogenetic analyses show the Giardia Sm core peptides are the products of multiple, ancestral gene duplications followed by divergence, but they retain strong similarity to Sm and like-Sm peptides of other eukaryotes. Although we have documented only a single intron in Giardia, it likely has other introns and fully functional, spliceosomal machinery. If introns were added during eukaryotic evolution (the introns-late hypothesis), then these results push back the date of this event before the branching of G. lamblia. PMID:11854456

  2. Morphological Studies of Nucleologenesis in Giardia lamblia.

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    Lara-Martínez, Reyna; De Lourdes Segura-Valdez, María; De La Mora-De La Mora, Ignacio; López-Velázquez, Gabriel; Jiménez-García, Luis Felipe

    2016-05-01

    The nucleolus is a nuclear organelle involved in ribosome biogenesis. In most eukaryotes this structure disperses during prophase through anaphase and reorganizes at telophase by a process known as nucleologenesis. This process involves new transcription of ribosomal DNA at the nucleolar organizer region and the formation of prenucleolar bodies fusing to it. In Giardia lamblia, for a long time considered the only anucleolated eukaryote, a very small nucleolus has been recently described. In order to evaluate whether nucleologenesis is also present in Giardia, we analyzed the distribution of nucleolar material during telophase using different light and electron microscopy techniques including silver staining for the nucleolar organizer. Results indicate that in G. lamblia, nucleolar elements persist mainly as an intranuclear peripheral organelle during all stages of division, including telophase, however, no prenucleolar bodies are detected in the nucleoplasm. Therefore, in the parasite, nucleolar material is present throughout cell division including telophase and formation of prenucleolar bodies may not be required for nucleologenesis.

  3. The proteome landscape of Giardia lamblia encystation.

    Science.gov (United States)

    Faso, Carmen; Bischof, Sylvain; Hehl, Adrian B

    2013-01-01

    Giardia lamblia is an intestinal protozoan parasite required to survive in the environment in order to be transmitted to a new host. To ensure parasite survival, flagellated trophozoites colonizing the small intestine differentiate into non-motile environmentally-resistant cysts which are then shed in the environment. This cell differentiation process called encystation is characterized by significant morphological remodeling which includes secretion of large amounts of cyst wall material. Although much is known about the transcriptional regulation of encystation and the synthesis and trafficking of cyst wall material, the investigation of global changes in protein content and abundance during G. lamblia encystation is still unaddressed. In this study, we report on the quantitative analysis of the G. lamblia proteome during encystation using tandem mass spectrometry. Quantification of more than 1000 proteins revealed major changes in protein abundance in early, mid and late encystation, notably in constitutive secretory protein trafficking. Early stages of encystation were marked by a striking decrease of endoplasmic reticulum-targeted variant-specific surface proteins and significant increases in cytoskeleton regulatory components, NEK protein kinases and proteins involved in protein folding and glycolysis. This was in stark contrast to cells in the later stages of encystation which presented a surprisingly similar proteome composition to non-encysting trophozoites. Altogether these data constitute the first quantitative atlas of the Giardia proteome covering the whole process of encystation and point towards an important role for post-transcriptional control of gene expression in Giardia differentiation. Furthermore, our data provide a valuable resource for the community-based annotation effort of the G. lamblia genome, where almost 70% of all predicted gene models remains "hypothetical".

  4. The proteome landscape of Giardia lamblia encystation.

    Directory of Open Access Journals (Sweden)

    Carmen Faso

    Full Text Available Giardia lamblia is an intestinal protozoan parasite required to survive in the environment in order to be transmitted to a new host. To ensure parasite survival, flagellated trophozoites colonizing the small intestine differentiate into non-motile environmentally-resistant cysts which are then shed in the environment. This cell differentiation process called encystation is characterized by significant morphological remodeling which includes secretion of large amounts of cyst wall material. Although much is known about the transcriptional regulation of encystation and the synthesis and trafficking of cyst wall material, the investigation of global changes in protein content and abundance during G. lamblia encystation is still unaddressed. In this study, we report on the quantitative analysis of the G. lamblia proteome during encystation using tandem mass spectrometry. Quantification of more than 1000 proteins revealed major changes in protein abundance in early, mid and late encystation, notably in constitutive secretory protein trafficking. Early stages of encystation were marked by a striking decrease of endoplasmic reticulum-targeted variant-specific surface proteins and significant increases in cytoskeleton regulatory components, NEK protein kinases and proteins involved in protein folding and glycolysis. This was in stark contrast to cells in the later stages of encystation which presented a surprisingly similar proteome composition to non-encysting trophozoites. Altogether these data constitute the first quantitative atlas of the Giardia proteome covering the whole process of encystation and point towards an important role for post-transcriptional control of gene expression in Giardia differentiation. Furthermore, our data provide a valuable resource for the community-based annotation effort of the G. lamblia genome, where almost 70% of all predicted gene models remains "hypothetical".

  5. Giardia lamblia: Molecular Studies of an Early Branching Eukaryote

    Science.gov (United States)

    The rapid advance in our understanding of the biology of Giardia lamblia over the last several years is due in part to the complete DNA sequencing of the 11.7 Mb genome of this diplomonad. Insight on the molecular nature of G. lamblia has been gained by searching the genome using query sequences fr...

  6. Altered lipid parameters in patients infected with Entamoeba histolytica, Entamoeba dispar and Giardia lamblia.

    Science.gov (United States)

    Bansal, D; Bhatti, H S; Sehgal, R

    2005-01-01

    Information on the effect of parasitic infections on lipid parameters is scarce. Certain parasites induce significant changes in lipid parameters, as demonstrated by the fact that substitution of lipid/cholesterol for serum in axenic culture medium (in vitro) and in experimental models (in vivo) supports vigorous growth of Entamoeba histolytica. Thus, significant changes in lipid parameters may be induced in an infected host. Blood samples are obtained from intestinal amoebiasis patients passing E. histolytica (n=8), E. dispar (n=15) or Giardia lamblia (n=9) cysts, or diagnosed with amoebic liver abscess (ALA; n=50) and from apparently normal healthy individuals (control group; n=30). Levels of total serum cholesterol, high-density lipoprotein and low-density lipoprotein are assessed using commercial kits. E. histolytica and E. dispar isolates are differentiated by hexokinase isoenzyme electrophoresis and by enzyme-linked immunosorbent assay (ELISA; Techlab) tests. Results show that E. histolytica, E. dispar and G. lamblia cyst passers had significantly lower levels of total serum cholesterol (73.42 +/- 2.24 mg/dL), compared to levels in ALA cases (101 +/- 2.85 mg/dL) and in controls (166.26 +/- 2.02 mg/dL). Further study of a greater number of cases is needed to explore the relevance of this finding.

  7. The efficacy of a Giardia lamblia vaccine in kittens.

    OpenAIRE

    Olson, M. E.; Morck, D. W.; Ceri, H.

    1996-01-01

    Twenty kittens were vaccinated with a Giardia lamblia vaccine prepared on a commercial scale on day 0 and boosted on day 21 (group 1); while 10 kittens received only saline (group 2). These kittens were challenged on day 35 with 10(6) Giardia lamblia trophozoites by a surgical intraduodenal injection. Three control kittens were not vaccinated and not challenged (group 3). Following challenge, Giardia vaccinated kittens had significantly fewer days in which abnormal stools were observed and re...

  8. A Lesson in Survival, by Giardia lamblia

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    Andrea A. S. Rópolo

    2010-01-01

    Full Text Available In the relationships between host and parasites, there is a cross-talk that involves diverse mechanisms developed by two different genetic systems during years of evolution. On the one hand, immunocompetent hosts have developed effective innate and acquired immune responses that are used to restrict or avoid parasitism. On the other hand, parasites evade the immune response, expressing different antigens on their surface or by using other specific mechanisms, such as nutrient depletion. In this review, we analyze the survival mechanisms used by the protozoan parasite Giardia lamblia during infection. In particular, we examine the multiple roles played by the enzyme arginine deiminase during colonization of the gut, also involving the parasite's mechanism of antigenic variation. Potential drug targets for the treatment of giardiasis are also discussed.

  9. Giardia lamblia and Helicobater pylori Coinfection

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    R Shafie

    2009-03-01

    Full Text Available "nBackground: Giardia lamblia and Helicobacter pylori are two flagellate microorganisms that grow in duodenum and stom­ach. The aim of this study was to evaluate the prevalence of them in patients with dyspepsia and other GI disorders. "nMethods: In this cross-sectional study, co-infection of above-mentioned agents was investigated in a group of 130 patients [me­dian age of 40 yr (range=11-79 including 76 males (58.8%] with dyspepsia using three methods of duodenal aspiration sam­ple, duodenal biopsy samples and evaluation of stool samples."nResults: : From 105 patients (59 males, 46 females, median age 40 years, range 11-79 entering this study from 3 hospitals, 4 patients (3.8% had G. lamblia and 61 patients (58% had H. pylori. All 4 patients infected by Giardia had also H. pylori infec­tion. Tenesmus (3 out of 4 patients was the most common symptom in patients with H. pylori infection (48 out of 61 pa­tients was reflux. Other symptoms in patients infected with both organisms (4 patients included diarrhea (2 cases, weight loss (2 cases, and loss of appetite (1 case but no report of vomiting."nConclusion: In patients co-infected with Giardia, H.pylori differentiation by physical examination is not possible. So in those patients with positive Rapid Urease Test (RUT, stool examination for Giardia detection is recommended. In addition, met­ronidazole (broad spectrum, anti-protozoal drug can be useful in H. pylori infection.

  10. A systematic review of the effects of Iranian pharmaceutical plant extracts on Giardia lamblia

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    Hajar Ziaei Hezarjaribi

    2015-12-01

    Full Text Available This study aimed to provide a systematic review regarding anti-Giardia effect of different Iranian plant extracts used in vivo and in vitro on cysts and trophozoites. Many reports indicated that most of plant extracts used as anti-Giardia were obtained from Liliaceae, Apiaceae, Asteraceae, and Myrtaceae. These extracts included different fractions such as aqueous, alcoholic and chloroform extracts as well as Soxhlet extraction of juice or essence. The findings of this review showed that hydroalcoholic extract of asafoetida, Chenopodium botrys, and chloroformic extract of feverfew (Tanacetum parthenium have the maximum effect (100% efficacy on in vitro application against Giardia. However, the highest in vivo effect of 100% therapeutic significance was recorded for the extract of Allium sativum at 80 mg/mL concentration. Given the plant species richness of Iran in terms of herbal medicines with fewer side effects, it can be a good alternative to chemical drugs used to treat giardiasis.

  11. Giardiavirus-resistant Giardia lamblia lacks a virus receptor on the cell membrane surface.

    OpenAIRE

    1994-01-01

    Giardia lamblia virus (GLV) is a small nonenveloped double-stranded RNA virus that infects specifically the parasitic protozoan G. lamblia. Among the many collected strains of G. lamblia, a few turn out to be highly resistant to the virus infection. Two of these strains, Ac and JH, were subjected to electroporation with the RNA from GLV-infected G. lamblia WB strain. Subsequent studies indicated the presence of GLV double-stranded RNA and GLV protein in the electroporated and propagated cells...

  12. Gene duplication in the genome of parasitic Giardia lamblia

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    Flores Roberto

    2010-02-01

    Full Text Available Abstract Background Giardia are a group of widespread intestinal protozoan parasites in a number of vertebrates. Much evidence from G. lamblia indicated they might be the most primitive extant eukaryotes. When and how such a group of the earliest branching unicellular eukaryotes developed the ability to successfully parasitize the latest branching higher eukaryotes (vertebrates is an intriguing question. Gene duplication has long been thought to be the most common mechanism in the production of primary resources for the origin of evolutionary novelties. In order to parse the evolutionary trajectory of Giardia parasitic lifestyle, here we carried out a genome-wide analysis about gene duplication patterns in G. lamblia. Results Although genomic comparison showed that in G. lamblia the contents of many fundamental biologic pathways are simplified and the whole genome is very compact, in our study 40% of its genes were identified as duplicated genes. Evolutionary distance analyses of these duplicated genes indicated two rounds of large scale duplication events had occurred in G. lamblia genome. Functional annotation of them further showed that the majority of recent duplicated genes are VSPs (Variant-specific Surface Proteins, which are essential for the successful parasitic life of Giardia in hosts. Based on evolutionary comparison with their hosts, it was found that the rapid expansion of VSPs in G. lamblia is consistent with the evolutionary radiation of placental mammals. Conclusions Based on the genome-wide analysis of duplicated genes in G. lamblia, we found that gene duplication was essential for the origin and evolution of Giardia parasitic lifestyle. The recent expansion of VSPs uniquely occurring in G. lamblia is consistent with the increment of its hosts. Therefore we proposed a hypothesis that the increment of Giradia hosts might be the driving force for the rapid expansion of VSPs.

  13. Effects of Dietary Zinc Manipulation on Growth Performance, Zinc Status and Immune Response during Giardia lamblia Infection: A Study in CD-1 Mice

    Science.gov (United States)

    Iñigo-Figueroa, Gemma; Méndez-Estrada, Rosa O.; Quihui-Cota, Luis; Velásquez-Contreras, Carlos A.; Garibay-Escobar, Adriana; Canett-Romero, Rafael; Astiazarán-García, Humberto

    2013-01-01

    Associations between Giardia lamblia infection and low serum concentrations of zinc have been reported in young children. Interestingly, relatively few studies have examined the effects of different dietary zinc levels on the parasite-infected host. The aims of this study were to compare the growth performance and zinc status in response to varying levels of dietary zinc and to measure the antibody-mediated response of mice during G. lamblia infection. Male CD-1 mice were fed using 1 of 4 experimental diets: adequate-zinc (ZnA), low-zinc (ZnL), high-zinc (ZnH) and supplemented-zinc (ZnS) diet containing 30, 10, 223 and 1383 mg Zn/kg respectively. After a 10 days feeding period, mice were inoculated orally with 5 × 106 G. lamblia trophozoites and were maintained on the assigned diet during the course of infection (30 days). Giardia-free mice fed ZnL diets were able to attain normal growth and antibody-mediated response. Giardia-infected mice fed ZnL and ZnA diets presented a significant growth retardation compared to non-infected controls. Zinc supplementation avoided this weight loss during G. lamblia infection and up-regulated the host’s humoral immune response by improving the production of specific antibodies. Clinical outcomes of zinc supplementation during giardiasis included significant weight gain, higher anti-G. lamblia IgG antibodies and improved serum zinc levels despite the ongoing infection. A maximum growth rate and antibody-mediated response were attained in mice fed ZnH diet. No further increases in body weight, zinc status and humoral immune capacity were noted by feeding higher zinc levels (ZnS) than the ZnH diet. These findings probably reflect biological effect of zinc that could be of public health importance in endemic areas of infection. PMID:24002196

  14. Effects of Dietary Zinc Manipulation on Growth Performance, Zinc Status and Immune Response during Giardia lamblia Infection: A Study in CD-1 Mice

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    Humberto Astiazarán-García

    2013-09-01

    Full Text Available Associations between Giardia lamblia infection and low serum concentrations of zinc have been reported in young children. Interestingly, relatively few studies have examined the effects of different dietary zinc levels on the parasite-infected host. The aims of this study were to compare the growth performance and zinc status in response to varying levels of dietary zinc and to measure the antibody-mediated response of mice during G. lamblia infection. Male CD-1 mice were fed using 1 of 4 experimental diets: adequate-zinc (ZnA, low-zinc (ZnL, high-zinc (ZnH and supplemented-zinc (ZnS diet containing 30, 10, 223 and 1383 mg Zn/kg respectively. After a 10 days feeding period, mice were inoculated orally with 5 × 106 G. lamblia trophozoites and were maintained on the assigned diet during the course of infection (30 days. Giardia-free mice fed ZnL diets were able to attain normal growth and antibody-mediated response. Giardia-infected mice fed ZnL and ZnA diets presented a significant growth retardation compared to non-infected controls. Zinc supplementation avoided this weight loss during G. lamblia infection and up-regulated the host’s humoral immune response by improving the production of specific antibodies. Clinical outcomes of zinc supplementation during giardiasis included significant weight gain, higher anti-G. lamblia IgG antibodies and improved serum zinc levels despite the ongoing infection. A maximum growth rate and antibody-mediated response were attained in mice fed ZnH diet. No further increases in body weight, zinc status and humoral immune capacity were noted by feeding higher zinc levels (ZnS than the ZnH diet. These findings probably reflect biological effect of zinc that could be of public health importance in endemic areas of infection.

  15. Visualization of chromosomes in the binucleate intestinal parasite Giardia lamblia.

    Science.gov (United States)

    Shen, Hai E; Cao, Lei; Li, Ji; Tian, Xi Feng; Yang, Zhi Hong; Wang, Yue; Tian, Yu Na; Lu, Si Qi

    2011-11-01

    Mitosis of Giardia lamblia is a complex and rapid event that is poorly understood at the cellular and molecular levels. Therefore, we conducted this study to determine (1) whether the two nuclei have similar or different chromosomes, (2) the number of chromosomes of G. lamblia, and (3) the morphology and karyotype of the chromosomes. Trophozoites of the C2 and WB strains of G. lamblia were grown in modified TYI-S-33 medium at 37°C. The trophozoites were collected, and sample slides were prepared for conventional light and scanning electron microscopy. Light microscopy revealed five pairs of chromosomes. The chromosomes were approximately 0.64-0.94 μm long with a short rod-like shape and were usually arranged in pairs. Scanning electron microscopy yielded similar findings, and 10 chromosomes could be seen in each nucleus. Thus, the chromosome number of G. lamblia is 2n = 10. Chromosomes in pair 1 are submetacentric chromosomes, while pairs 2-5 are telocentric chromosomes. The present study shows that G. lamblia trophozoites have typical condensed chromosomes during mitosis and contains five pairs of chromosomes. The karyogram shows good fit to the formula 2n = 10 = 2sm + 8t revealed by scanning electron microscopy.

  16. Cytochrome b5 from Giardia lamblia.

    Science.gov (United States)

    Alam, Samiah; Yee, Janet; Couture, Manon; Takayama, Shin-ichi J; Tseng, Wan-Hsin; Mauk, A Grant; Rafferty, Steven

    2012-12-01

    The protozoan intestinal parasite Giardia lamblia lacks mitochondria and the ability to make haem yet encodes several putative haem-binding proteins, including three of the cytochrome b(5) family. We cloned one of these (gCYTb5-I) and expressed it within Escherichia coli as a soluble holoprotein. UV-visible and resonance Raman spectra of gCYTb5-I resemble those of microsomal cytochrome b(5), and homology modelling supports a structure in which a pair of invariant histidine residues act as axial ligands to the haem iron. The reduction potential of gCYTb5-I is -165 mV vs. SHE and is relatively low compared to most values (-110 to +80 mV) for this class of protein. The amino- and carboxy-terminal sequences that flank the central haem-binding core of the Giardia cytochromes are highly charged and differ from those of other family members. A core gCYTb5-I variant lacking these flanking sequences was also able to bind haem. The presence of one actual and two probable functional cytochromes b(5) in Giardia is evidence of uncharacterized cytochrome-mediated metabolic processes within this medically important protist.

  17. Importance of enolase in Giardia lamblia differentiation.

    Science.gov (United States)

    Castillo-Romero, Araceli; Davids, Barbara J; Lauwaet, Tineke; Gillin, Frances D

    2012-08-01

    The ability of Giardia to differentiate into cysts which survive in the environment and release the virulent trophozoites after ingestion in the small intestine is essential for transmission and disease. We examined the role of enolase, a glycolytic enzyme, in Giardia differentiation. The sequence of Giardia lamblia enolase (gEno) is most similar to enolases in Homo sapiens and Leishmania mexicana, and shows the conserved catalytic and metal-binding residues. We used an integration vector to stably express wild type and mutant gEno. In trophozoites, wild type gEno localized to the cell membrane, caudal flagella and cytosol. gEno is present on the wall of mature cysts, but not in encystation secretory vesicles (ESV). The expression of gEno with a deletion of residues G167-K169, or mutations H389Q/R390S significantly inhibited excystation while mutation of residue D257K had no effect. These results suggest a role for enolase in regulation of Giardia excystation.

  18. Tonic shock induces detachment of Giardia lamblia.

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    Wendy R Hansen

    Full Text Available BACKGROUND: The parasite Giardia lamblia must remain attached to the host small intestine in order to proliferate and subsequently cause disease. However, little is known about the factors that may cause detachment in vivo, such as changes in the aqueous environment. Osmolality within the proximal small intestine can vary by nearly an order of magnitude between host fed and fasted states, while pH can vary by several orders of magnitude. Giardia cells are known to regulate their volume when exposed to changes in osmolality, but the short-timescale effects of osmolality and pH on parasite attachment are not known. METHODOLOGY AND PRINCIPAL FINDINGS: We used a closed flow chamber assay to test the effects of rapid changes in media osmolality, tonicity, and pH on Giardia attachment to both glass and C2(Bbe-1 intestinal cell monolayer surfaces. We found that Giardia detach from both surfaces in a tonicity-dependent manner, where tonicity is the effective osmolality experienced by the cell. Detachment occurs with a characteristic time constant of 25 seconds (SD = 10 sec, n = 17 in both hypo- and hypertonic media but is otherwise insensitive to physiologically relevant changes in media composition and pH. Interestingly, cells that remain attached are able to adapt to moderate changes in tonicity. By exposing cells to a timed pattern of tonicity variations and adjustment periods, we found that it is possible to maximize the tonicity change experienced by the cells, overcoming the adaptive response and resulting in extensive detachment. CONCLUSIONS AND SIGNIFICANCE: These results, conducted with human-infecting Giardia on human intestinal epithelial monolayers, highlight the ability of Giardia to adapt to the changing intestinal environment and suggest new possibilities for treatment of giardiasis by manipulation of tonicity in the intestinal lumen.

  19. IDENTIFICATION OF A FLAVOBACTERIUM STRAIN VIRULENT AGAINT GIARDIA LAMBLIA CYSTS

    Science.gov (United States)

    We have isolated a bacterial strain capable of killing the cyst form of Giardia lamblia, from a Kentucky stream. This bacterium, designated Sun4, is a Gram negative, aerobic rod which produces a yellow pigment, but not of the flexirubin-type. Although true gliding motility has no...

  20. IDENTIFICATION OF A FLAVOBACTERIUM STRAIN VIRULENT AGAINT GIARDIA LAMBLIA CYSTS

    Science.gov (United States)

    We have isolated a bacterial strain capable of killing the cyst form of Giardia lamblia, from a Kentucky stream. This bacterium, designated Sun4, is a Gram negative, aerobic rod which produces a yellow pigment, but not of the flexirubin-type. Although true gliding motility has no...

  1. Giardia lamblia infection increases risk of chronic gastrointestinal disorders.

    Science.gov (United States)

    Dormond, Megan; Gutierrez, Ramiro L; Porter, Chad K

    2016-01-01

    Giardia lamblia is a common parasitic cause of infectious gastroenteritis in the United States and the world and may be linked to an increased risk of chronic gastrointestinal (GI) disorders. We sought to assess the risk of several chronic GI disorders following Giardia infection among active duty US military personnel. This study was designed as a retrospective cohort study in which active duty military personnel with documented G. lamblia infection were assessed for the subsequent risk of developing a chronic GI disorder including irritable bowel syndrome (IBS), dyspepsia and gastroesophageal reflux disease (GERD). Post-giardia chronic GI disorder risk was compared to risk in uninfected personnel matched on several demographic characteristics and medical encounter information. Data were obtained from the Defense Medical Surveillance System and exposures (1998-2009) with outcomes identified based on documented medical encounters with specific medical billing codes. Modified Poisson regression was used to evaluate the relationship between G. lamblia infection and chronic GI disorders. A total of 80 Giardia cases were identified for an estimated incidence of 0.55 cases per 100,000 person-years. Cases were matched to 294 unexposed subjects. After adjusting for important covariates, there was an increased risk of IBS (relative risk: 2.1, p = 0.03) associated with antecedent Giardia infection. These data add to a growing body of literature and demonstrate an increased risk of IBS after infection with G. lamblia.

  2. Analysis of Giardin expression during encystation of Giardia lamblia

    Science.gov (United States)

    The present study analyzed giardin transcription in trophozoites and cysts during encystation of Giardia lamblia. Encystment was induced using standard methods, and the number of trophozoites and cysts were counted at various time-points during encystation. At all time points, RNA from both stages...

  3. Evaluation of alpha-tubulin as an antigenic and molecular probe to detect Giardia lamblia.

    Science.gov (United States)

    Kim, Juri; Shin, Myeong Heon; Song, Kyoung-Ju; Park, Soon-Jung

    2009-09-01

    The alpha/beta-tubulin heterodimer is the basic subunit of microtubules in eukaryotes. Polyclonal antibodies specific to recombinant alpha-tubulin of Giardia lamblia were made, and found effective as a probe to specifically detect G. lamblia by immunofluorescence assays. Nucleotide sequences of alpha-tubulin genes were compared between G. lamblia WB and GS strains, prototypes of assemblage A and assemblage B, respectively. A set of primers was designed and used to amplify a portion of the alpha-tubulin gene from G. lamblia. PCR-RFLP analysis of this alpha-tubulin PCR product successfully differentiated G. lamblia into 2 distinct groups, assemblages A and B. The results indicate that alpha-tubulin can be used as a molecular probe to detect G. lamblia.

  4. PURE CULTURE METHOD: GIARDIA LAMBLIA FROM DIFFERENT STOOL SAMPLES

    Directory of Open Access Journals (Sweden)

    H.A YOUSEFI

    2000-03-01

    Full Text Available Introduction. Giardiasis is one of the health problems in the world including Iran. To determine the biochemical and biological problems and also identification of various strains, it is essential to obtain pure culture and then mass production of Giardia lamblia. The goal of this study was to isolate this protozoa purely.
    Methods. Giardia lamblia cysts were isolated from 50 stool samples by use of floating of a four - layer of sucrose method. The cysts were transfered to an inducing solution. Subsequently, they were cultured in a modified culture medium (TYIS-33. Following excystation of trophozoite and its multiplication, the parasite was caltured and purified.
    Findings. Excitation of trophozoite was observed in 40 samples (80 percent from which 22 samples (55 percent yielded pure culture. The doubling time was approximately 13hr and the peak of parasite was observed between third and fourth days.
    Conclusion. The proliferation and growth rate of Giardia lamblia have enabled us to use this method widely. Cystein and ascorbic acid which are present in the induction solution, have a key role in excystation of trophozoite. Purification and passage of samples has facilitated the culture of this parasite in vitro. Therefore this method has yielded better results in comparison with other studies. This is probably due to a decrease in the amount of bovine bile or using different strains of Giardia lamblia in the present study.

  5. Whole genome sequencing of clinical isolates of Giardia lamblia.

    Science.gov (United States)

    Hanevik, K; Bakken, R; Brattbakk, H R; Saghaug, C S; Langeland, N

    2015-02-01

    Clinical isolates from protozoan parasites such as Giardia lamblia are at present practically impossible to culture. By using simple cyst purification methods, we show that Giardia whole genome sequencing of clinical stool samples is possible. Immunomagnetic separation after sucrose gradient flotation gave superior results compared to sucrose gradient flotation alone. The method enables detailed analysis of a wide range of genes of interest for genotyping, virulence and drug resistance.

  6. Giardia lamblia infection after pancreas-kidney transplantation.

    Science.gov (United States)

    Kristensen, Ann Abkjaer; Horneland, Rune; Birn, Henrik; Svensson, My

    2016-01-18

    Infection is a common complication of solid organ transplantation. It is associated with an increased risk of acute cellular rejection and loss of graft function. The most common infections are due to bacteria and viruses, including transmission of cytomegalovirus from donor to recipient. In the past years, an increasing number of parasitic infections have been documented in transplant recipients. We describe the first reported case of intestinal Giardia lamblia transmission following simultaneous pancreas and kidney transplantation.

  7. Transient transfection and expression of firefly luciferase in Giardia lamblia.

    OpenAIRE

    1995-01-01

    We have developed a gene transfer system for the protozoan parasite Giardia lamblia. This organism is responsible for many cases of diarrhea worldwide and is considered to be one of the most primitive eukaryotes. Expression of a heterologous gene was detected in this parasite after electroporation with appropriate DNA constructs. We constructed a series of transfection plasmids using flanking sequences of the Giardia glutamate dehydrogenase (GDH) gene to drive expression of the firefly lucife...

  8. New insights regarding the biology of Giardia lamblia.

    Science.gov (United States)

    Carranza, Pedro G; Lujan, Hugo D

    2010-01-01

    Giardia lamblia is one of the most common causes of intestinal disease in humans. To adapt to environments both inside and outside of the host's small intestine, this protozoan parasite undergoes significant developmental changes during its life cycle. In this review, we analyze and discuss the most recent findings regarding the process of Giardia trophozoites differentiation into infective cysts as well as the mechanism of antigenic variation, which allows the parasite to cause chronic and recurrent infections in susceptible individuals.

  9. Transfection of the Giardia lamblia double-stranded RNA virus into giardia lamblia by electroporation of a single-stranded RNA copy of the viral genome.

    OpenAIRE

    1990-01-01

    The development of a genetic vector for protozoan parasites is a major hurdle yet to be crossed in the study of the molecular and cellular biology of these parasites. We have identified and isolated a double-stranded RNA virus (G. lamblia virus [GLV]) from certain strains of the intestinal parasitic protozoan Giardia lamblia (A. L. Wang and C. C. Wang, Mol. Biochem. Parasitol. 21:269-276, 1986), which is capable of infecting other virus-free strains of G. lamblia (R. L. Miller, A. L. Wang, an...

  10. STUDY OF LEPTIN CONCENTRATIONS IN CHILDREN INFECTED WITH ENTAMOEBA HISTOLYTICA\\DISPARE AND GIARDIA LAMBLIA PARASITES

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    SHAKER MOHAMMED JASIM

    2015-04-01

    Full Text Available Background: Leptin, a 16-kDa protein secreted from white adipocytes, has been implicated in the regulation of food intake, energy expenditure, and whole-body energy balance in humans, therefore may be a mediator of anorexia associated with acute and chronic inflammation. In all parasitic infections there is loss of appetite and anorexia. Objective: The aim of the present study is to analyze serum leptin concentration in children infected with intestinal parasites (Entamoeba histolytica\\dispare and Giardia lamblia and compare them with healthy controls. Patients and methods: A total of 70 males and females patients infected with intestinal parasites presented to the Central Teaching Hospital for Pediatric in Baghdad city - Iraq, during the period from April to September, 2013. The age of patients ranged from 5 to 15 years. Thirty healthy parasite-free individuals consider to be a control group. Stool samples were collected from patients for direct microscopic examination, then body weight, height and body mass index (BMI were measured for all patients and controls. Serum leptin concentrations were detected by immunoenzymometric assay using the DRG Company Leptin ELISA kit. Statistical analysis was made by Chi-Square test using SAS version 9.1. Results: In this study 70 Patients infected with intestinal parasites, 40 patients confirmed to be positive for E. histolytica\\dispare and 30 patients for G. lamblia. The results showed serum leptin levels were not statistically significant between patient: 2.234±0.13ng/ml and control: 2.171±0.23; p > 0.05, at the same time, highly differences noticed between two age groups of patients with concentration of leptin, related to that leptin levels are increased in aged paralleling changes in fat mass. In addition, we found significant differences between age, weight and BMI with concentration of leptin. but the height of patients were not statistically significant with concentration of leptin. Conclusion

  11. Flagellar generated flow mediates attachment of Giardia Lamblia

    Science.gov (United States)

    Picou, Theodore; Polackwich, Jamie; Burrola Gabilondo, Beatriz; McAllister, Ryan; Powers, Tom; Elmendorf, Heidi; Urbach, Jeff

    2011-11-01

    Giardia lamblia is a protozoan parasite responsible for widespread diarrheal disease in humans and animals worldwide. Attachment to the host intestinal mucosa and resistance to peristalsis is necessary for establishing infection, but the physical basis for this attachment is poorly understood. We report results from confocal fluorescence microscopy that demonstrate that the regular beating of the posterior flagella generate a flow through the ventral disk, a suction-cup shaped structure that is against the substrate during attachment. Finite element simulations show that the negative pressure generated by the flow is consistent with the measured force of attachement between the parasite and its substrate.

  12. [In vitro effect of osthole on ultrastructure of Giardia lamblia].

    Science.gov (United States)

    Li, Wen-Chao; Gu, You-Fang; Liu, Chang; Wu, Na; Luo, Wen-Wu; Gong, Peng-Tao; Li, He; Li, Jian-Hua; Zhang, Xi-Chen

    2014-06-01

    Giardia lamblia trophozoites were cultivated axenically in TYI-S-33 modified medium containing 1.345 mg/ml of osthole (24 h IC50). The parasites were observed by scanning and transmission electron microscopes after treated with osthole for 24 h. The surface of the trophozoites treated with osthole was rough. The surface of ventral sucker and median body had obvious lesions, the cell membrane was damaged and the content spilled out. There were a lot of vacuoles in the cytoplasm. And the nuclear was severely deformed with a serrated edge and marginated nuclear chromatin. The microtubules of sucker had partially disintegrated.

  13. [Molecular Detection of Giardia lamblia and Cryptosporidium Species in Pet Dogs].

    Science.gov (United States)

    GU, You-fang; WANG, Kai; LIU, De-yi; MEI, Nan; CHEN, Cheng; CHEN, Tao; HAN, Min-min; ZHOU, Li; CAO, Jia-tong; ZHANG, Heng; ZHANG, Xue-liang; FAN, Zi-lai; LI, Wen-chao

    2015-10-01

    To determine the prevalence of Giardia lamblia and Cryptosporidium species infection in pet dogs, and identify the G. lamblia assemblages and Cryptosporidium species. A total of 315 fresh fecal samples were collected from pet clinics in five counties of Anhui Province and in Hangzhou of Zhejiang Province. Hemi-nested-PCR targeting the GDH gene of G. lamblia and nested-PCR targeting the SSU rRNA gene of Cryptosporidium were performed in all the fecal samples. The PCR products were sequenced and analyzed using bioinformatics methods to identify the G. lamblia assemblages and Cryptosporidium species. The positive rates of G. lamblia and Cryptosporidium spp. infections in the 315 fecal samples were 3.2% (10/315) and 1.6% (5/315), respectively. Specifically, the two indicators were both significantly higher in dogs ≤12 months (17.8% and 11.1%, respectively) than in adult dogs (0.7% and 0.0%)(Plamblia assemblages were identified, assemblages B (n=6) and D (n=4). Sequence analysis of PCR products of the SSU rRNA gene showed that the five Cryptosporidium isolates were C. canis (n =5). The prevalences of G. lamblia and Cryptosporidium infection in pet dogs in Anhui and Zhejiang Provinces were 3.2 % and 1.6 %, respectively. The assemblages of G. lamblia in this study are of types B and D.

  14. Structural Basis for Inactivation of Giardia lamblia Carbamate Kinase by Disulfiram*

    Science.gov (United States)

    Galkin, Andrey; Kulakova, Liudmila; Lim, Kap; Chen, Catherine Z.; Zheng, Wei; Turko, Illarion V.; Herzberg, Osnat

    2014-01-01

    Carbamate kinase from Giardia lamblia is an essential enzyme for the survival of the organism. The enzyme catalyzes the final step in the arginine dihydrolase pathway converting ADP and carbamoyl phosphate to ATP and carbamate. We previously reported that disulfiram, a drug used to treat chronic alcoholism, inhibits G. lamblia CK and kills G. lamblia trophozoites in vitro at submicromolar IC50 values. Here, we examine the structural basis for G. lamblia CK inhibition of disulfiram and its analog, thiram, their activities against both metronidazole-susceptible and metronidazole-resistant G. lamblia isolates, and their efficacy in a mouse model of giardiasis. The crystal structure of G. lamblia CK soaked with disulfiram revealed that the compound thiocarbamoylated Cys-242, a residue located at the edge of the active site. The modified Cys-242 prevents a conformational transition of a loop adjacent to the ADP/ATP binding site, which is required for the stacking of Tyr-245 side chain against the adenine moiety, an interaction seen in the structure of G. lamblia CK in complex with AMP-PNP. Mass spectrometry coupled with trypsin digestion confirmed the selective covalent thiocarbamoylation of Cys-242 in solution. The Giardia viability studies in the metronidazole-resistant strain and the G. lamblia CK irreversible inactivation mechanism show that the thiuram compounds can circumvent the resistance mechanism that renders metronidazole ineffectiveness in drug resistance cases of giardiasis. Together, the studies suggest that G. lamblia CK is an attractive drug target for development of novel antigiardial therapies and that disulfiram, an FDA-approved drug, is a promising candidate for drug repurposing. PMID:24558036

  15. Structural basis for inactivation of Giardia lamblia carbamate kinase by disulfiram.

    Science.gov (United States)

    Galkin, Andrey; Kulakova, Liudmila; Lim, Kap; Chen, Catherine Z; Zheng, Wei; Turko, Illarion V; Herzberg, Osnat

    2014-04-11

    Carbamate kinase from Giardia lamblia is an essential enzyme for the survival of the organism. The enzyme catalyzes the final step in the arginine dihydrolase pathway converting ADP and carbamoyl phosphate to ATP and carbamate. We previously reported that disulfiram, a drug used to treat chronic alcoholism, inhibits G. lamblia CK and kills G. lamblia trophozoites in vitro at submicromolar IC50 values. Here, we examine the structural basis for G. lamblia CK inhibition of disulfiram and its analog, thiram, their activities against both metronidazole-susceptible and metronidazole-resistant G. lamblia isolates, and their efficacy in a mouse model of giardiasis. The crystal structure of G. lamblia CK soaked with disulfiram revealed that the compound thiocarbamoylated Cys-242, a residue located at the edge of the active site. The modified Cys-242 prevents a conformational transition of a loop adjacent to the ADP/ATP binding site, which is required for the stacking of Tyr-245 side chain against the adenine moiety, an interaction seen in the structure of G. lamblia CK in complex with AMP-PNP. Mass spectrometry coupled with trypsin digestion confirmed the selective covalent thiocarbamoylation of Cys-242 in solution. The Giardia viability studies in the metronidazole-resistant strain and the G. lamblia CK irreversible inactivation mechanism show that the thiuram compounds can circumvent the resistance mechanism that renders metronidazole ineffectiveness in drug resistance cases of giardiasis. Together, the studies suggest that G. lamblia CK is an attractive drug target for development of novel antigiardial therapies and that disulfiram, an FDA-approved drug, is a promising candidate for drug repurposing.

  16. Short Communication—The detection of nuclear matrix in most primitive presentexisting eukaryote,Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    DAIJIALING; JINGYANLI; 等

    1995-01-01

    The nuclear matrix of diplomonad Giardia lamblia was detected for the first time with DGD embedmentsectioning-embedment free electron microscopy after a series of specific extractions.The result showed that archaezoa Giardia Lamblia already possessed nuclear matrix within its two nuclei.The finest fibrils of the nuclear matrix of Giardia lamblia were measured to be about 11 to 13 nm in thickness.However,the nuclear lamina and nucleolus have never been observed.These results seem to suggest that nuclear matrix is an indispensable intranuclear structural component even in the primitive nucleus.

  17. Analysis of giardin expression during encystation of Giardia lamblia.

    Science.gov (United States)

    Jenkins, M C; O'Brien, C N; Macarisin, D; Miska, K; Fetterer, R; Fayer, R

    2012-12-01

    The present study analyzed giardin transcription in trophozoites and cysts during encystation of Giardia lamblia . Encystment was induced using standard methods, and the numbers of trophozoites and cysts were counted at various time points during the process. At all time points, RNA from both stages were assayed for levels of alpha2-, beta-, and delta-giardin mRNA as well as for cyst wall protein 3 (CWP3) mRNA using quantitative RT-PCR. In encystation medium, the number of G. lamblia trophozoites decreased, while the number of cysts increased between 0 and 72 hr. In trophozoites, alpha2- and beta-giardin transcription decreased over time, while delta-giardin transcription remained unchanged during the same time period. CWP3 transcription exhibited a slight increase in trophozoites at 8 hr, followed by a decrease at subsequent time points. Expression of alpha2-giardin increased at 48 hr in cysts followed by decreased expression at 72 hr, while beta- and delta-giardin expression was unchanged during encystation. CWP3 transcription gradually decreased from 24-72 hr in cysts. Consistent with previous studies, giardin proteins appeared to be disassembled into amorphous structures inside cysts during encystation. These findings represent the first analysis of giardin transcription in separate populations of trophozoites and cysts during encystation and indicate differential regulation of giardin mRNA expression by these developmental stages.

  18. Episomal and integrated maintenance of foreign DNA in Giardia lamblia.

    Science.gov (United States)

    Singer, S M; Yee, J; Nash, T E

    1998-04-01

    Giardia lamblia is an early diverging eukaryote which causes gastrointestinal disease throughout the world. Different subgroups of Giardia have been defined based on several biochemical and genetic criteria. We have developed a method for stably introducing DNA into the nuclei of the parasite using puromycin acetyltransferase (pac) as a dominant selectable marker. Transfected circular DNAs were maintained as episomes in the isolate WB, a representative of one Giardia subgroup. When input DNAs were linearized, integration was observed to occur by homologous recombination producing gene replacements in this isolate. In isolate GS, which represents a different subgroup, both linear and circular transfected DNAs were integrated into the genome by homologous recombination. In GS, linear DNA again produced gene replacements, while circular DNA produced duplicative integration events. The failure of GS to replicate episomes may reflect differences in the structure or recognition of DNA replication origins between these subgroups. A plasmid shuttle vector was also developed for expression of other genes in Giardia lamblia. Utilizing the green fluorescent protein as a reporter gene in the WB isolate, we show that gene expression from this vector correlated with plasmid copy number over a range of two orders of magnitude. Together these tools should greatly enhance our ability to study both the basic biology and the pathogenesis of this ubiquitous parasite.

  19. In Vitro Effects of Metronidazole and Albendazole on Giardia lamblia Isolated from Iranian Patients

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    A Dalimi

    2008-04-01

    Full Text Available Background: The aim of the present study was to evaluate the effects of metronidazole and albendazole against clinical isolates of Giardia lamblia in vitro. Methods: From all human samples of containing cysts, 10 isolates were successfully excysted in vitro. Trophozites viability was assessed by eosine 0.1% and cultured axenically in TYI-S-33 modified medium supplemented with heat inactivated bovine serum 10%. All cultures were incubated in 37°C for 24-48 h. After this time trophozoites were exposed to different concentration (0.05, 0.1, 2, 10, 50 µg/ml of drugs at 37º for 4 h.  The IC50 estimated between 0.1 and 10µg/ml for metronidazole and 0.062 and 0.1 µg/ml for albendazole. Results: Eight isolates were found susceptible to the metronidazole while all isolates were found susceptible to the albendazole. Statistical results indicated that there was significant difference (P<0.05 in the sensitivity to metronidazole and albendazole in all isolates.

  20. Infectious causation of chronic disease: Examining the relationship between Giardia lamblia infection and irritable bowel syndrome

    Institute of Scientific and Technical Information of China (English)

    Alice S Penrose; Eden V Wells; Allison E Aiello

    2007-01-01

    AIM: To evaluate whether a higher prevalence of Giardia lamblia infection is associated with an increase in irritable bowel syndrome (IBS) prescriptions at the county level in Michigan.METHODS: The Michigan Disease Surveillance System (MDSS) was used to ascertain both the numbers of Giardia lamblia infections as well as the total number of foodborne illnesses per population by county in Michigan during 2005. This was compared with Blue Cross Blue Shield (BCBS) of Michigan numbers of drug prescriptions for IBS per one thousand members per county in 2005.These data were also analyzed for associations with per capita income by county and the number of refugees entering each county in 2005.RESULTS: There were a total of 786 confirmed cases of Giardia lamblia reported to MDSS in 2005. During the same time period, the number of prescriptions for IBS varied from 0.5 per 1000 members up to 6.0 per 1000members per month. There was no trend towards higher numbers of IBS prescriptions in the counties with more Giardia lamblia infections. Per capita income was not associated with either IBS prescriptions or Giardiasis.There was a significant linear association between the number of refugees entering each county, and the number of Giardia lamblia cases per 100 000 population.CONCLUSION: In this ecological study, there was no association found between BCBS prescriptions for IBS and Giardia lamblia infections in Michigan counties. Our findings may have been influenced by the disparate number of refugees admitted per county.

  1. Reactivation of Giardia lamblia cysts after exposure to low-pressure UV irradiation.

    Science.gov (United States)

    Shin, Gwy-Am; Linden, Karl G

    2015-07-01

    In this study, we determined the repair capabilities of Giardia lamblia cysts when they were exposed to low-pressure (LP) UV and then 4 different repair conditions. A UV collimated beam apparatus was used to expose shallow suspensions of G. lamblia cysts in buffered reagent water (PBS, pH 7.2) to various doses of LP UV irradiation. After UV irradiation, samples were exposed to 4 repair conditions (light and dark repair conditions with 2 temperatures (25 °C and 37 °C) for each condition). The inactivation of G. lamblia cysts by LP UV was very extensive (∼ 5 log10) even with a low dose of LP UV (1 mJ/cm(2)). More importantly, there was significant restoration of infectivity in G. lamblia cysts when they were exposed to a low dose of LP UV and then to all the repair conditions tested. Overall, the results of this study indicate that G. lamblia cysts do have the ability to repair their UV-damaged DNA when they are exposed to low doses of LP UV irradiation. This is the first study to report the presence of repair in UV-irradiated G. lamblia cysts.

  2. DNA sequence analysis of the triose phosphate isomerase gene from isolates of Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    卢思奇; 文建凡; 李继红; 王凤云

    2002-01-01

    Objective To confirm the genetic relation between Giardia lamblia (G. lamblia) isolates from different geographic regions of China and other countries. Methods Genomic DNA were extracted from the trophozoites or cysts of Giardia lamblia. The triose phosphate isomerase (tim) gene was amplified using polymerase chain reaction (PCR) technique. PCR products were digested with endonuclease and sequenced. The data of sequencing were analyzed with the DNAstar software and compared with that of the isolates acquired from GenBank. Results Of nine isolates of Giardia lamblia from China (C1, C2, CH2 and CH3), Cambodia (CAM), Australia (A1 and A2) and America (BP and CDC), respectively, 3 (A1, A2 and CAM) fit into Group 1 (WB), 2 (CH2 and CH3)) into Group 2, and 4 (C1, C2, BP and CDC) into Group 3 (GS). The results confirmed the genetic relatedness of G. lamblia isolates from all over the world. Conclusion Genotyping isolates of G. Lamblia provides important information for establishing the phylogenetic relationship or for the epidemiological evaluation of the spreading of this organism.

  3. Identification of α-11 giardin as a flagellar and surface component of Giardia lamblia.

    Science.gov (United States)

    Kim, Juri; Lee, Hye Yeon; Lee, Mi-Ae; Yong, Tai-Soon; Lee, Kyu-Ho; Park, Soon-Jung

    2013-10-01

    Giardia lamblia is a protozoan pathogen with distinct cytoskeletal structures, including median bodies and eight flagella. In this study, we examined components comprising G. lamblia flagella. Crude flagellar extracts were prepared from G. lamblia trophozoites, and analyzed by two-dimensional (2-D) gel electrophoresis. The 19 protein spots were analyzed by MALDI-TOF mass spectrometry, identifying ten metabolic enzymes, six distinct giardins, Giardia trophozoite antigen 1, translational initiation factor eIF-4A, and an extracellular signal-regulated kinase 2. Among the identified proteins, we studied α-11 giardin which belongs to a group of cytoskeletal proteins specific to Giardia. Western blot analysis and real-time PCR indicated that expression of α-11 giardin is not significantly increased during encystation of G. lamblia. Immunofluorescence assays using anti-α-11 giardin antibodies revealed that α-11 giardin protein mainly localized to the plasma membranes and basal bodies of the anterior flagella of G. lamblia trophozoites, suggesting that α-11 giardin is a genuine component of the G. lamblia cytoskeleton.

  4. Identification of a Novel Microtubule-Binding Protein in Giardia lamblia.

    Science.gov (United States)

    Kim, Juri; Park, Soon-Jung

    2016-08-01

    Giardia lamblia is a protozoan that causes diarrheal diseases in humans. Cytoskeletal structures of Giardia trophozoites must be finely reorganized during cell division. To identify Giardia proteins which interact with microtubules (MTs), Giardia lysates were incubated with in vitro-polymerized MTs and then precipitated by ultracentifugation. A hypothetical protein (GL50803_8405) was identified in the precipitated fraction with polymerized MTs and was named GlMBP1 (G. lamblia microtubule-binding protein 1). Interaction of GlMBP1 with MTs was confirmed by MT binding assays using recombinant GlMBP1 (rGlMBP1). In vivo expression of GlMBP1 was shown by a real-time PCR and western blot analysis using anti-rGlMBP1 antibodies. Transgenic G. lamblia trophozoites were constructed by integrating a chimeric gene encoding hemagglutinin (HA)-tagged GlMBP1 into a Giardia chromosome. Immunofluorescence assays of this transgenic G. lamblia, using anti-HA antibodies, revealed that GlMBP1 mainly localized at the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. This result indicates that GlMBP1 is a component of the G. lamblia cytoskeleton.

  5. Giardia lamblia: intracellular localization of alpha8-giardin.

    Science.gov (United States)

    Wei, Chao Jun; Tian, Xi Feng; Adam, Rodney D; Lu, Si Qi

    2010-12-01

    Alpha8-giardin (α8-giardin) is a member of the multi-gene α-giardin family in the intestinal parasitic protozoan, Giardia lamblia. This gene family shares an ancestry with the annexin super family, whose common characteristic is calcium dependent binding to membranes that contain acidic phospholipids. In the present study, the antigenicity, hydrophilicity, flexibility, surface probability, and secondary structure of α8-giardin amino acids were predicted by bioinformatics applications. A specific anti-peptide antiserum, anti-P3, was used to determine the intracellular location of α8-giardin with confocal immunofluorescence microscopy and immunoelectron microscopy. The results indicated that α8-giardin was located on the plasma membrane and flagella, but not on the ventral disk. Reduction of α8-giardin transcript levels by ribozyme-mediated cleavage decreased trophozoite motility and growth rate, indicating the functional importance of α8-giardin to Giardia trophozoite biology.

  6. Therapeutic enhancement of newly derived bacteriocins against Giardia lamblia.

    Science.gov (United States)

    Amer, Eglal I; Mossallam, Shereen F; Mahrous, Hoda

    2014-11-01

    Trials for identifying efficient anti-giardial agents are still ongoing. Nowadays, bacteriocins have attracted the attention as potential antimicrobial compounds. For the first time, the current study evaluated the therapeutic efficacy of bacteriocins derived from newly isolated Egyptian strains of probiotics Lactobacilli; L. acidophilus (P106) and L. plantarum (P164) against Giardia lamblia. Bacteriocins' efficacy was evaluated both in vitro; by growth inhibition and adherence assays, and in vivo; through estimation of parasite density, intestinal histopathological examination and ultrastructural analysis of Giardia trophozoites. In vivo bacteriocins' clinical safety was assessed. In vitro results proved that 50 µg of L. acidophilus bacteriocin induced reduction of the mean Giardia lamblia trophozoites by 58.3 ± 4.04%, while at lower concentrations of 10 and 20 µg of both L. acidophilus and L. plantarum, non significant reduction of the mean parasite density was achieved. In vitro trophozoites adherence was susceptible to the tested bacteriocins at all studied concentrations with variable degrees, while the highest adherence reduction was demonstrated using 50 µg of L acidophilus bacteriocin. In vivo, oral inoculation of 50 µg/mouse L. acidophilus bacteriocin for 5 successive days resulted in a noteworthy decline of the intestinal parasite density, along with amelioration of intestinal pathology of infected mice. Ultrastructural examination proved thatfive doses of L. acidophilus bacteriocin showed marked changes in cellular architecture of the trophozoites with evident disorganization of the cell membrane, adhesive disc and cytoplasmic components. This is the first reported study of the safe anti-giardial efficacy of L. acidophilus (P106) derived bacteriocin, hence highlighting its great promise as a potential therapeutic safe alternative to existing commercial drugs.

  7. Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia.

    Science.gov (United States)

    Tan, Liping; Yu, Xingang; Abdullahi, Auwalu Yusuf; Wu, Sheng; Zheng, Guochao; Hu, Wei; Song, Meiran; Wang, Zhen; Jiang, Biao; Li, Guoqing

    2015-11-01

    Giardia lamblia is a zoonotic flagellate protozoan in the intestine of human and many mammals including dogs. To assess a threat of dog-derived G. lamblia to humans, the common dog-derived G. lamblia assemblages A, C, and D were genotyped by high-resolution melting (HRM) technology. According to β-giardin gene sequence, the qPCR-HRM primers BG5 and BG7 were designed. A series of experiments on the stability, sensitivity, and accuracy of the HRM method were also tested. Results showed that the primers BG5 and BG7 could distinguish among three assemblages A, C, and D, which Tm value differences were about 1 °C to each other. The melting curves of intra-assay reproducibility were almost coincided, and those of inter-assay reproducibility were much the same shape. The lowest detection concentration was about 5 × 10(-6)-ng/μL sample. The genotyping results from 21 G. lamblia samples by the HRM method were in complete accordance with sequencing results. It is concluded that the HRM genotyping method is rapid, stable, specific, highly sensitive, and suitable for clinical detection and molecular epidemiological survey of dog-derived G. lamblia.

  8. Sequence analysis and prokaryotic expression of Giardia lamblia α-18 giardin gene.

    Science.gov (United States)

    Wu, Sheng; Yu, Xingang; Abdullahi, Auwalu Yusuf; Hu, Wei; Pan, Weida; Shi, Xianli; Tan, Liping; Song, Meiran; Li, Guoqing

    2016-03-01

    To study the genetic variation and prokaryotic expression of α18 giardin gene of Giardia lamblia zoonotic assemblage A and host-specific assemblage F, the α18 genes were amplified from G. lamblia assemblages A and F by PCR and sequenced. The PCR product was cloned into the prokaryotic expression vector pET-28a(+) and the positive recombinant plasmid was transformed into Escherichia coli Rosetta (DE3) strain for the expression. The expressed α18 giardin fusion protein was validated by SDS-PAGE and Western blot analysis, and purified by Ni-Agarose resin. The putative sequence of α18 giardin amino acid was analyzed by bioinformatics software. Results showed that the α18 giardin gene was 861 bp in length, encoding 286 amino acids; it was 100% homologous between human-derived and dog-derived G. lamblia assemblage A, but it was 86.8% homologous with G. lamblia assemblage F (cat-derived). Giardin α18 was about 36 kDa in molecular weight, with good reactivity. Prediction based on in silico analyses: it had hydrophobicity, without signal peptide and transmembrane domain, and contained 11 alpha regions, 13 beta sheets, 1 beta turn and 7 random coils in secondary structure. The above information would lay the foundation for research about the subcellular localization and biological function of α18 giardin in G. lamblia.

  9. Isolation and partial characterization of an immunogenic antigen of Giardia lamblia.

    Science.gov (United States)

    Quintero, Jael; Valdez, Alejandra; Samaniego, Brenda; Lopez-Romero, Gloria; Astiazaran-Garcia, Humberto; Rascon, Lucila; Breci, Linda; Garibay-Escobar, Adriana; Robles-Zepeda, Ramón; Velazquez, Carlos

    2017-06-01

    Humoral and cellular immune responses play an important role during Giardia lamblia infection. Several Giardia proteins have been identified as immunogenic antigens based on their elicited humoral immune response. Poorly is known about Giardia antigens that stimulate a cellular immune response. The main purpose of this study was to isolate and partial characterize an immunogenic antigen (5G8) of G. lamblia. The 5G8 protein was isolated from G. lamblia trophozoite lysates by affinity chromatography using moAb 5G8-coupled CNBr-Sepharose. The isolated protein was analysed by electrospray tandem mass spectrometry (ESI-MS/MS), and by diverse bioinformatics tools (GiardiaDB, BLASTn, BLASTp and ExPASy). Additionally, several biochemical and immunological characteristics of the isolated protein were analysed. By ESI-MS/MS the amino acidic 5G8 sequence was deduced. The 5G8 antigen belongs to the VSP family proteins of G. lamblia. This protein is composed by one polypeptide chain (±71kDa). Using the algorithm SYFPHEITI, we identified candidate CD4(+) T-cell epitopes from the 5G8 antigen, which can elicit cell-mediated immune responses. In this study, we have identified a G. lamblia protein that induces a strong immune response in infected mice. The biochemical and immunological characterization of the immunogenic 5G8 antigen may contribute to the rational design of a Giardia vaccine. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  10. Chitin synthetase in encysting Giardia lamblia and Entamoeba invadens

    Energy Technology Data Exchange (ETDEWEB)

    Das, S.; Gillin, F.D.

    1987-05-01

    Giardia lamblia (Gl) and Entamoeba invadens (Ei) are protozoan parasites with two morphologic stages in their life cycles. Motile trophozoites colonize the intestine of humans and reptiles respectively. Water resistant cysts, which can survive outside the host, transmit infection. In vitro cyst formation of Ei from trophozoites has been reported, and the authors have recently induced in vitro encystation of Gl. Although the cyst walls of both parasites contain chitin, it synthesis by encysting trophozoites has not been reported. The authors now show that encystation conditions greatly increase chitin synthetase (CS) specific activity (incorporation of /sup 3/H GlcNAc from UDP-GlcNAc into TCA-or alcohol-precipitable material). Extracts of encysting Gl incorporated 3.6 nmol/mg protein in 5 hr compared to < 0.005 in controls. Extracts of encysting Fi incorporated 4.8 n mol/mg protein, compared to 1.7 in the control. CS activity of both parasites requires preformed chitin. The Gl enzyme requires a reducing agent, is inhibited by digitonin and the CS inhibitors, polyoxin D and Nikkomycin, but not by tunicamycin. The product is digested by chitinase. Ei enzyme does not require a reducing agent and is stimulated by 1 mg/ml digitonin, but inhibited by higher concentrations. These studies demonstrate CS enzymes which may play important roles in encystation of Gl and Ei.

  11. Genotyping of Giardia lamblia isolates from human in southern Iran.

    Science.gov (United States)

    Sarkari, B; Ashrafmansori, A; Hatam, G R; Motazedian, M H; Asgari, Q; Mohammadpour, I

    2012-09-01

    Giardia lamblia cysts isolated from human faeces in South of Iran were analyzed with PCR-restriction fragment length polymorphism (RFLP) assay, based on the detection of glutamate dehydrogenase (gdh) genes. Among 205 faecal samples from microscopically diagnosed giardiasis patients, the gdh gene was amplified from 172 cases with a semi nested PCR assay and typed by RFLP analysis. Of the 172 positive samples, 128 (74.41%) were typed as assemblage AII, 30 (17.44%) assemblage BIII, 6 (3.49%) assemblage BIV and in 8 (4.66%) isolates, mixed assemblages AII and BIV were detected. Clinical features were available for 52 successfully typed cases and the possible correlation of Giardia assemblages and clinical symptoms was evaluated. Both assemblages caused similar illness, but assemblage AII was significantly more frequently associated with abdominal pain, nausea and vomiting. Since these isolates, A and B, are of human origin, anthroponotic transmission of Giardia can be suggested for the route of infection in this region.

  12. Reconstruction of Sugar Metabolic Pathways of Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Jian Han

    2012-01-01

    Full Text Available Giardia lamblia is an “important” pathogen of humans, but as a diplomonad excavate it is evolutionarily distant from other eukaryotes and relatively little is known about its core metabolic pathways. KEGG, the widely referenced site for providing information of metabolism, does not yet include many enzymes from Giardia species. Here we identify Giardia’s core sugar metabolism using standard bioinformatic approaches. By comparing Giardia proteomes with known enzymes from other species, we have identified enzymes in the glycolysis pathway, as well as some enzymes involved in the TCA cycle and oxidative phosphorylation. However, the majority of enzymes from the latter two pathways were not identifiable, indicating the likely absence of these functionalities. We have also found enzymes from the Giardia glycolysis pathway that appear more similar to those from bacteria. Because these enzymes are different from those found in mammals, the host organisms for Giardia, we raise the possibility that these bacteria-like enzymes could be novel drug targets for treating Giardia infections.

  13. Prevalence Rate Of Giardia LambliaHelicobacter Pylori Co-Infections In Khartoum State Sudan

    Directory of Open Access Journals (Sweden)

    Nusaiba Fadul Mustafa Ahmed

    2015-08-01

    Full Text Available The study aimed to determine the prevalence rate of Giardia lamblia Helicobacter pylori co-infections in Khartoum State Sudan. A cross-sectional study was carried out during the period between May to December 2015. A total of 100 subjects were included in this study the age ranging between 1-80 years the mean age was 29 19 years old. Stool samples were taken from all subjects included in the study in addition to clinical and parasitological data were obtained and recorded. Out of 100 subjects 14 14 were positive for G. lamblia by using direct wet mount and 22 22 were positive by using formal ether concentration technique FECT p0.000. Out of 100 subjects 30 30 were positive for H. pylori when detected by using H. pylori antigen test. The study showed that the prevalence of G. lamblia was higher in females 11 11 than in males 3 3 p0.193. Also H. pylori prevalence was higher in females 17 17 than in males 13 13 p0.390. The prevalence rate of G. lamblia was higher 55 in the age group 16- 25 and 46-65 years old by using direct wet mount p0.053 while the prevalence rate of H. pylori was higher 9 9 in the age group 1-15 years old p0.424. The study revealed that the prevalence rate of G. lamblia and H. pylori co-infections were 5 9 by using direct wet mount and formal ether concentration technique respectively. This study indicated that the prevalence rate of G. lamblia and H. pylori in the study area were 14 30 respectively and co-infection was 5 with no significant value for co-infection p0.615.

  14. Virus-mediated expression of firefly luciferase in the parasitic protozoan Giardia lamblia.

    OpenAIRE

    1995-01-01

    Giardia lamblia, a prevalent human pathogen and one of the lineages that branched earliest from prokaryotes, can be infected with a double-stranded RNA virus, giardiavirus (GLV). The 6,277-bp viral genome has been previously cloned (A.L. Wang, H.-M. Yang, K.A. Shen, and C.C. Wang, Proc. Natl. Acad. Sci. USA 90:8595-8599, 1993; C.-H. Wu, C.C. Wang, H.M. Yang, and A.L. Wang, Gene, in press) and was converted to a transfection vector for G. lamblia in the present study. By flanking the firefly l...

  15. Proteomics of secretory and endocytic organelles in Giardia lamblia.

    Science.gov (United States)

    Wampfler, Petra B; Tosevski, Vinko; Nanni, Paolo; Spycher, Cornelia; Hehl, Adrian B

    2014-01-01

    Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs), mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs), for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER) which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694.

  16. Proteomics of secretory and endocytic organelles in Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Petra B Wampfler

    Full Text Available Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs, mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs, for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694.

  17. An Atypical Proprotein Convertase in Giardia lamblia Differentiation

    Science.gov (United States)

    Davids, B. J.; Gilbert, M. A.; Liu, Q.; Reiner, D. S.; Smith, A. J.; Lauwaet, T.; Lee, C.; McArthur, A. G.; Gillin, F. D.

    2010-01-01

    Proteolytic activity is important in the lifecycles of parasites and their interactions with hosts. Cysteine proteases have been best studied in Giardia, but other protease classes have been implicated in growth and/or differentiation. In this study, we employed bioinformatics to reveal the complete set of putative proteases in the Giardia genome. We identified 73 peptidase homologues distributed over 5 catalytic classes in the genome. Serial analysis of gene expression of the G. lamblia lifecycle found thirteen protease genes with significant transcriptional variation over the lifecycle, with only one serine protease transcript upregulated late in encystation. The translated gene sequence of this encystation-specific transcript was most similar to eukaryotic subtilisin-like proprotein convertases (SPC), although the typical catalytic triad was not identified. Epitope-tagged gSPC protein expressed in Giardia under its own promoter was upregulated during encystation with highest expression in cysts and it localized to encystation-specific secretory vesicles (ESV). Total gSPC from encysting cells produced proteolysis in gelatin gels that co-migrated with the epitope-tagged protease in immunoblots. Immuno-purified gSPC also had gelatinase activity. To test whether endogenous gSPC activity is involved in differentiation, trophozoites and cysts were exposed to the specific serine proteinase inhibitor 4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF). After 21 h encystation, a significant decrease in ESV was observed with 1 mM AEBSF and by 42 h the number of cysts was significantly reduced, but trophozoite growth was not inhibited. Concurrently, levels of cyst wall proteins 1 and 2, and AU1-tagged gSPC protein itself were decreased. Excystation of G. muris cysts was also significantly reduced in the presence of AEBSF. These results support the idea that serine protease activity is essential for Giardia encystation and excystation. PMID:21075147

  18. Proteomics of Secretory and Endocytic Organelles in Giardia lamblia

    Science.gov (United States)

    Wampfler, Petra B.; Tosevski, Vinko; Nanni, Paolo; Spycher, Cornelia; Hehl, Adrian B.

    2014-01-01

    Giardia lamblia is a flagellated protozoan enteroparasite transmitted as an environmentally resistant cyst. Trophozoites attach to the small intestine of vertebrate hosts and proliferate by binary fission. They access nutrients directly via uptake of bulk fluid phase material into specialized endocytic organelles termed peripheral vesicles (PVs), mainly on the exposed dorsal side. When trophozoites reach the G2/M restriction point in the cell cycle they can begin another round of cell division or encyst if they encounter specific environmental cues. They induce neogenesis of Golgi-like organelles, encystation-specific vesicles (ESVs), for regulated secretion of cyst wall material. PVs and ESVs are highly simplified and thus evolutionary diverged endocytic and exocytic organelle systems with key roles in proliferation and transmission to a new host, respectively. Both organelle systems physically and functionally intersect at the endoplasmic reticulum (ER) which has catabolic as well as anabolic functions. However, the unusually high degree of sequence divergence in Giardia rapidly exhausts phylogenomic strategies to identify and characterize the molecular underpinnings of these streamlined organelles. To define the first proteome of ESVs and PVs we used a novel strategy combining flow cytometry-based organelle sorting with in silico filtration of mass spectrometry data. From the limited size datasets we retrieved many hypothetical but also known organelle-specific factors. In contrast to PVs, ESVs appear to maintain a strong physical and functional link to the ER including recruitment of ribosomes to organelle membranes. Overall the data provide further evidence for the formation of a cyst extracellular matrix with minimal complexity. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000694. PMID:24732305

  19. MATRIX ASSISTED LASER DESORPTION/IONIZATION TIME OF FLIGHT MASS SPECTROMETRY BASED ANALYSIS OF GIARDIA LAMBLIA

    Science.gov (United States)

    Giardia lamblia is a zoonotic protozoan parasite that is a leading cause of drinking water related gastro-intestinal disease outbreaks worldwide. Due to the genotypic complexity and high prevalence of this parasite in the environment, numerous research studies are being done to ...

  20. Identification, characteristic and phylogenetic analysis of type Ⅱ DNA topoisomerase gene in Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    De HE; Jian Fan WEN; Wan Qun CHEN; Si Qi LU; De Dong XIN

    2005-01-01

    The genes encoding type Ⅱ DNA topoisomerases were investigated in Giardia lamblia genome, and a type ⅡA gene,GlTop 2 was identified. It is a single copy gene with a 4476 bp long ORF without intron. The deduced amino acid sequence shows strong homology to eukaryotic DNA Top 2. However, some distortions were found, such as six insertions in the ATPase domain and the central domain, a ~100 aa longer central domain; a ~200 aa shorter C-terminal domain containing rich charged residues. These features revealed by comparing with Top 2 of the host, human, might be helpful in exploiting drug selectivity for antigiardial therapy. Phylogenetic analysis of eukaryotic enzymes showed that kinetoplastids, plants, fungi, and animals were monophyletic groups, and the animal and fungi lineages shared a more recent common ancestor than either did with the plant lineage; microsporidia grouped with fungi. However, unlike many previous phylogenetic analyses, the "amitochondriate" G. lamblia was not the earliest branch but diverged after mitochondriate kinetoplastids in our trees. Both the finding of typical eukaryotic type ⅡA topoisomerase and the phylogenetic analysis suggest G. lamblia is not possibly as primitive as was regarded before and might diverge after the acquisition of mitochondria. This is consistent with the recent discovery of mitochondrial remnant organelles in G. lamblia.

  1. A comparison of the Giardia lamblia trophozoite and cyst transcriptome using microarrays

    Directory of Open Access Journals (Sweden)

    Widmer Giovanni

    2011-05-01

    Full Text Available Abstract Background Compared with many protists, Giardia lamblia has a simple life cycle alternating between cyst and trophozoite. Most research on the molecular biology of Giardia parasites has focused on trophozoites and the processes of excystation and encystation, whereas cysts have attracted less interest. The striking morphological differences between the dormant cyst and the rapidly dividing and motile trophozoite implies profound changes in the metabolism as the parasite encysts in the host's intestine and excysts upon ingestion by a new host. Results To investigate the magnitude of the transcriptional changes occurring during the G. lamblia life cycle we compared the transcriptome of G. lamblia trophozoites and cysts using single-color oligonucleotide microarrays. Cysts were found to possess a much smaller transcriptome, both in terms of mRNA diversity and abundance. Genes encoding proteins related to ribosomal functions are highly over-represented. The comparison of the transcriptome of cysts generated in culture or extracted from feces revealed little overlap, raising the possibility of significant biological differences between the two types of cysts. Conclusions The comparison of the G. lamblia cyst and trophozoite transcriptome showed that transcripts of most genes are present at a lower level in cysts. This global view of the cyst and trophozoite transcriptome complements studies focused on the expression of selected genes during trophozoite multiplication, encystation and excystation.

  2. Identification and characterization of a FYVE domain from the early diverging eukaryote Giardia lamblia.

    Science.gov (United States)

    Sinha, Abhishek; Mandal, Sananda; Banerjee, Sumana; Ghosh, Arjun; Ganguly, Sandipan; Sil, Alok Kumar; Sarkar, Srimonti

    2011-04-01

    The morphology of the endomembrane system of Giardia lamblia appears to be significantly different from higher eukaryotes. Therefore, the molecular mechanisms controlling vesicular trafficking are also likely to be altered. Since FYVE domain is a known regulator of endosomal trafficking, the authors used BLAST search to identify FYVE domain(s) in G. lamblia. A 990 amino acid long putative FYVE domain-containing ORF was identified, which contains all the conserved sequence elements in the ligand binding pocket. Phylogenetic analysis reveals that this domain is significantly diverged. The authors have shown that the corresponding gene is expressed in G. lamblia trophozoites and cysts. In spite of this phylogenetic divergence, in vitro biochemical assay indicates that this domain preferentially binds to phosphatidylinositol 3-phosphate {PtdIns(3)P}and in vivo expression of the GFP-tagged G. lamblia FYVE domain in S. cerevisiae, displays its selective localization to PtdIns(3)P-enriched endosomes. This is the first study to characterize a PtdIns(3)P effector protein in this early-diverged eukaryote.

  3. CLONING AND CHARACTERIZATION OF CDNA ENCODING GIARDIA LAMBLIA d-GIARDIN

    Science.gov (United States)

    A cDNA coding for d-giardin was cloned from Giardia lamblia trophozoites in order to localize the protein and study its function in mediating surface attachment. Recombinant d-giardin antigen was produced in Escherichia coli as a poly-histidine fusion protein and was purified by affinity chromatogr...

  4. A reprofiled drug, auranofin, is effective against metronidazole-resistant Giardia lamblia.

    Science.gov (United States)

    Tejman-Yarden, Noa; Miyamoto, Yukiko; Leitsch, David; Santini, Jennifer; Debnath, Anjan; Gut, Jiri; McKerrow, James H; Reed, Sharon L; Eckmann, Lars

    2013-05-01

    Giardiasis is one of the most common causes of diarrheal disease worldwide. Treatment is primarily with 5-nitro antimicrobials, particularly metronidazole. Resistance to metronidazole has been described, and treatment failures can occur in up to 20% of cases, making development of alternative antigiardials an important goal. To this end, we have screened a chemical library of 746 approved human drugs and 164 additional bioactive compounds for activity against Giardia lamblia. We identified 56 compounds that caused significant inhibition of G. lamblia growth and attachment. Of these, 15 were previously reported to have antigiardial activity, 20 were bioactive but not approved for human use, and 21 were drugs approved for human use for other indications. One notable compound of the last group was the antirheumatic drug auranofin. Further testing revealed that auranofin was active in the low (4 to 6)-micromolar range against a range of divergent G. lamblia isolates representing both human-pathogenic assemblages A and B. Most importantly, auranofin was active against multiple metronidazole-resistant strains. Mechanistically, auranofin blocked the activity of giardial thioredoxin oxidoreductase, a critical enzyme involved in maintaining normal protein function and combating oxidative damage, suggesting that this inhibition contributes to the antigiardial activity. Furthermore, auranofin was efficacious in vivo, as it eradicated infection with different G. lamblia isolates in different rodent models. These results indicate that the approved human drug auranofin could be developed as a novel agent in the armamentarium of antigiardial drugs, particularly against metronidazole-resistant strains.

  5. Cyst acquisition rate for Giardia lamblia in backcountry travelers to Desolation Wilderness, Lake Tahoe

    Science.gov (United States)

    Zell, S.C.; Sorenson, S.K.

    1993-01-01

    The objective of this study was to determine the incidence of Giardia lamblia acquisition in back-country travelers to a wilderness area, provide longitudinal follow-up on the incidence of symptomatic gastrointestinal illness and relate such information to concentrations of Giardia cysts in water samples from a high-use area. A prospective cohort non-interventional study of 41 healthy adult backcountry travelers from age 19 to 71 years in Desolation Wilderness, Lake Tahoe Basin was carried out. The incidence of Giardia cyst acquisition in backcountry travelers was only 5.7% (95% CI 0.17–20.2%). Mild, self-limiting gastrointestinal illness occurred in 16.7% of subjects (95% CI 4.9%–34.50%), none of whom demonstrated G. lamblia infection. Water sampling from three popular stream sites revealed cyst contamination to be generally at low levels with cyst concentrations in the single digit range for every 100 gallons filtered. G. lamblia contamination of water occurs, but at low levels. Acquisition of this parasite may be infrequent in backcountry recreationalists. Symptomatic gastrointestinal illness following wilderness travel can be due to other etiologies. Our findings may not be representative of all wilderness areas, but suggest that in the absence of documented G. lamblia infection, persons symptomatic following travel may suffer a self-limiting gastrointestinal illness. In such circumstances, empiric therapy for giardiasis is tempting but difficult to justify.

  6. Rapid clearance of Giardia lamblia DNA from the gut after successful treatment.

    Science.gov (United States)

    van den Bijllaardt, W; Overdevest, I T; Buiting, A G; Verweij, J J

    2014-11-01

    To assess the time it takes for a real-time PCR to become negative after treatment of a Giardia lamblia infection, we evaluated two consecutive follow-up samples from 75 infected patients. Approximately 1 week after treatment all samples tested negative, indicating rapid clearance of parasitic DNA after successful treatment.

  7. Differential dissolved protein expression throughout the life cycle of Giardia lamblia.

    Science.gov (United States)

    Lingdan, Li; Pengtao, Gong; Wenchao, Li; Jianhua, Li; Ju, Yang; Chengwu, Liu; He, Li; Guocai, Zhang; Wenzhi, Ren; Yujiang, Chen; Xichen, Zhang

    2012-12-01

    Giardia lamblia (G. lamblia) has a simple life cycle that alternates between a cyst and a trophozoite, and this parasite is an important human and animal pathogen. To increase our understanding of the molecular basis of the G. lamblia encystment, we have analyzed the soluble proteins expressed by trophozoites and cysts extracted from feces by quantitative proteomic analysis. A total of 63 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ) labeling, and were categorized as cytoskeletal proteins, a cell-cycle-specific kinase, metabolic enzymes and stress resistance proteins. Importantly, we demonstrated that the expression of seven proteins differed significantly between trophozoites and cysts. In cysts, the expression of three proteins (one variable surface protein (VSP), ornithine carbamoyltransferase (OTC), β-tubulin) increased, whereas the expression of four proteins (14-3-3 protein, α-tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), protein disulfide isomerase 2 (PDI-2)) decreased significantly when compared with the levels of these proteins in trophozoites. The mRNA expression patterns of four of these proteins (OTC, α-tubulin, GAPDH, VSP) were similar to the expression levels of the proteins. These seven proteins appear to play an important role in the completion of the life cycle of G. lamblia.

  8. Significantly Diverged Did2/Vps46 Orthologues from the Protozoan Parasite Giardia lamblia.

    Science.gov (United States)

    Dutta, Somnath; Saha, Nabanita; Ray, Atrayee; Sarkar, Srimonti

    2015-09-01

    The endosomal compartment performs extensive sorting functions in most eukaryotes, some of which are accomplished with the help of the multivesicular body (MVB) sorting pathway. This pathway depends on the sequential action of complexes, termed the endosomal sorting complex required for transport (ESCRT). After successful sorting, the crucial step of recycling of the ESCRT complex components requires the activation of the AAA ATPase Vps4, and Did2/Vps46 plays an important role in this activation event. The endolysosomal system of the protozoan parasite Giardia lamblia appears to lack complexity, for instead of having distinct early endosomes, late endosomes and lysosomes, there are only peripheral vesicles (PVs) that are located close to the cell periphery. Additionally, comparative genomics studies predict the presence of only a subset of the ESCRT components in G. lamblia. Thus, it is possible that the MVB pathway is not functional in G. lamblia. To address this issue, the present study focused on the two putative orthologues of Did2/Vps46 of G. lamblia as their function is likely to be pivotal for a functional MVB sorting pathway. In spite of considerable sequence divergence, compared to other eukaryotic orthologues, the proteins encoded by both these genes have the ability to function as Did2/Vps46 in the context of the yeast ESCRT pathway. Furthermore, they also localized to the cellular periphery, where PVs are also located. Thus, this report is the first to provide experimental evidence indicating the presence of a functional ESCRT component in G. lamblia by characterizing the putative Did2/Vps46 orthologues.

  9. Giardia lamblia infection in patients with irritable bowel syndrome and dyspepsia: A prospective study

    Institute of Scientific and Technical Information of China (English)

    Barbara Grazioli; Francesco Luzza; Giovanni Matera; Costanza Laratta; Giuseppina Schipani; Giovanni Guarnieri; Ester Spiniello; Maria Imeneo; Andrea Amorosi; Alfredo Focà

    2006-01-01

    AIM: To evaluate the prevalence of Giardia lamblia (G.lamblia) infection in patients with irritable bowel syndrome (IBS) and dyspepsia and to establish which is the most accurate test to diagnose the infection in this setting.METHODS: One hundred and thirty-seven patients who consecutively attended the Outpatient Gastroenterology Clinic for the first time between January 2002 and December 2003 due to symptoms of IBS and/or dyspepsia were recruited. All patients underwent clinical evaluation,first-step haematology and chemistry tests, serologic assays for celiac disease, lactose-H2 breath test, abdominal ultrasonography, and esophagogastroduodenoscopy.Helicobacter pylori status was evaluated. In patients with symptoms of IBS older than 45 years, colonoscopy was also performed. In all patients, duodenal biopsies and stool samples were examined for trophozoites and cysts of G.lamblia by several methods.RESULTS: G. lamblia was identified in 9 patients. The following diagnoses were also made: IBS (100/137, 73%), functional dyspepsia (62/137, 45%), organic dyspepsia (33/137, 24%), and lactose intolerance (75/137, 55%). A significant association was found between giardiasis and Hpylori infection (x2=6.632,OR= 12.4, CI= 1.5-68.1). There were no symptoms that reliably allowed the recognition of giardiasis.Direct search of the parasite in duodenal biopsy and stool sample examinations gave concordant results in all cases while histological examination of duodenal biopsies displayed a low sensitivity (e.g., 22.2%).CONCLUSION: In this consecutive series, diagnosis of G. lamblia infection accounted for 6.5% of patients with IBS and dyspepsia. Duodenal biopsies for diagnosis of giardiasis may be unnecessary if stool sample examination is performed.

  10. Characterization of BIP protein of G. lamblia as a potential immunogen in a mouse infection model.

    Science.gov (United States)

    Lopez-Romero, Gloria; Garzon, Thania; Rascon, Raul; Valdez, Alejandra; Quintero, Jael; Arvizu-Flores, Aldo A; Garibay-Escobar, Adriana; Rascon, Lucila; Astiazarán-García, Humberto; Velazquez, Carlos

    2017-08-01

    Giardia lamblia is a protozoan parasite that causes one of the most common gastrointestinal diseases worldwide. To eliminate the parasite from the host intestine, it is necessary the activation of B-cell and T-cell dependent mechanisms. The knowledge about Giardia antigens that can stimulate the host immune response is limited. Recently, it has been described the Binding Immunoglobulin Protein (BIP) of G. lamblia (71kDa) as a potential immunogen. Additionally, our group has identified a highly immunogenic antigen (5G8 protein) of G. lamblia with a relative molecular mass of approximately 70kDa. There is some evidence suggesting that the 5G8 protein may activate both humoral and cellular immune responses. Based on these observations and preliminary mass spectrometry analyses, we hypothesized that the antigen 5G8 could be the BIP protein. In the present study, we characterize immunochemically the BIP protein of Giardia. Flow cytometric assays and western blotting were used to determine the expression profile of BIP and 5G8 antigens in Giardia trophozoites. The differences in expression profile indicated that BIP and 5G8 are not the same molecule. ELISA and Western blotting assays revealed that BIP protein was recognized by antibodies produced during G. lamblia infection in C3H/HeN mice. MTT assays did not reveal the activation of cellular immune response induced by BIP protein in vitro. In addition, we identified the potential B-cell and T-cell epitopes of G. lamblia BIP protein. This molecule is a conserved protein among Giardia strains and other pathogens. The complete immunological characterization of this antigen will contribute to a better understanding of the host-parasite interactions in Giardia infection. Copyright © 2017 Elsevier GmbH. All rights reserved.

  11. Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler's Diarrhea.

    Science.gov (United States)

    Shin, Ji-Hun; Lee, Sang-Eun; Kim, Tong Soo; Ma, Da-Won; Chai, Jong-Yil; Shin, Eun-Hee

    2016-10-01

    This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler's diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10(3) oocysts for C. parvum, >1×10(4) cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

  12. Identification of cis-acting signals in the giardiavirus (GLV) genome required for expression of firefly luciferase in Giardia lamblia.

    OpenAIRE

    1996-01-01

    Giardiavirus (GLV) is a 6,277-bp double-stranded RNA virus of Giardia lamblia, one of the earliest eukaryotic divergents from the prokaryotes. Our previous success in GLV-mediated transfection of G. lamblia has provided an effective way of monitoring the mechanisms underlining GLV gene replication and mRNA translation in this organism. Here we have investigated the cis-acting signals in the GLV genome that regulate replication, transcription, and translation of an inserted firefly luciferase ...

  13. Sensitive and rapid detection of Giardia lamblia infection in pet dogs using loop-mediated isothermal amplification.

    Science.gov (United States)

    Li, Jie; Wang, Peiyuan; Zhang, Aiguo; Zhang, Ping; Alsarakibi, Muhamd; Li, Guoqing

    2013-04-01

    Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10(-1) to 10(-5) ng/µl for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63℃ by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1α) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

  14. Lectin Activation in Giardia lamblia by Host Protease: A Novel Host-Parasite Interaction

    Science.gov (United States)

    Lev, Boaz; Ward, Honorine; Keusch, Gerald T.; Pereira, Miercio E. A.

    1986-04-01

    A lectin in Giardia lamblia was activated by secretions from the human duodenum, the environment where the parasite lives. Incubation of the secretions with trypsin inhibitors prevented the appearance of lectin activity, implicating proteases as the activating agent. Accordingly, lectin activation was also produced by crystalline trypsin and Pronase; other proteases tested were ineffective. When activated, the lectin agglutinated intestinal cells to which the parasite adheres in vivo. The lectin was most specific to mannose-6-phosphate and apparently was bound to the plasma membrane. Activation of a parasite lectin by a host protease represents a novel mechanism of hostparasite interaction and may contribute to the affinity of Giardia lamblia to the infection site.

  15. 蓝氏贾第鞭毛虫诊断%Diagnoses of Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    高正琴; 贺争鸣; 岳秉飞

    2015-01-01

    Objective Giardia lamblia is an important pathogen of zoonosis giardiasis , it poses a potential threat to the quality of SPF (specific pathogen-free) laboratory animals cannot be ignored.The aim of this study is to establish the method of rapid diagnosis of Giardia lamblia, and analyze the test results of 516 batches form 17 manufactures.Methods Direct microscopy, Giemsa-fast staining and multiplex polymerase chain reaction (multiplex PCR) were applied to detect Giardia lamblia.Results Numerous of Giardia lamblia trophozoites and cysts were detected in SPF laboratory animals by using direct microscopy and Giemsa-fast staining, and multiplex PCR were performed to identify 18S rDNA,β-giardin, TPI and GDH genes of DNA extracted from these trophozoites and cysts identified Giardia lamblia.Direct microscopy, Giemsa-fast staining, and multiplex PCR methods can be used to detect Giardia lamblia.Of the 2562 SPF laboratory animals studied, 22.9%(586/2562) were positive for Giardia lamblia.Conclusions Direct microscopy , Giemsa-fast staining , and multiplex PCR were effective techniques with high sensitivity and specificity for rapid diagnosis of Giardia lambliain.It is not satisfactory that the results of Giardia lamblia examination in 516 batches form 17 manufactures failed to meet the requirements 100%.%目的:蓝氏贾第鞭毛虫作为一种重要的人兽共患寄生虫病贾第虫病的病原,其对SPF实验动物质量造成的潜在威胁不容忽视。本研究的目的是建立蓝氏贾第鞭毛虫快速诊断方法,并对17个生产厂家516批2562只SPF实验动物检查结果进行分析。方法用直接镜检法、快速姬姆萨染色法和多重PCR方法,对蓝氏贾第鞭毛虫进行检测。结果在SPF实验动物中检出数量众多的蓝氏贾第鞭毛虫滋养体和包囊,鉴定出蓝氏贾第鞭毛虫18S rDNA、β-giardin、TPI和GDH基因。直接镜检法、快速姬姆萨染色法和多重PCR方法均能

  16. Prevalence of Giardia lamblia in Different Water Sources of District Nowshehra, Khyber Pakhtunkhwa Pakistan.

    Science.gov (United States)

    khan, Shaukat Ali; Ayaz, Sultan; Khan, Sanaullah; Anees, Muhammad; Khan, Amir Muhammad; Khan, Imran

    2012-12-01

    Giardia lamblia is a cosmopolitan parasite that occurs worldwide and generally effects gastrointestinal tract. Water played a media for transmission of Giardia to different hosts. A total of 300 water samples were examined from different water sources, i.e. tap, open well, bore well and drain waters and DNA was extracted by trizol method through prescribed protocol. DNA was amplified through PCR. The overall prevalence of G. lamblia was 27.66% (83/300). Among these 2.5% (1/40) in bore well water, 29% (29/100) open well, 18.83% (11/60) tap water and 42% (42/100) drain water. It is concluded from the study that Giardia is frequently found in all water sources and is the main cause of ill health.

  17. Giardia lamblia Nek1 and Nek2 kinases affect mitosis and excystation.

    Science.gov (United States)

    Smith, Alias J; Lauwaet, Tineke; Davids, Barbara J; Gillin, Frances D

    2012-04-01

    The NIMA-related serine/threonine kinases (Neks) function in the cell cycle and regulate ciliary and flagellar length. The Giardia lamblia genome encodes 198 Neks, of which 56 are predicted to be active. Here we believe that we report the first functional analysis of two G. lamblia Neks. The GlNek1 and GlNek2 kinase domains share 57% and 43% identity to the kinase domains of human Nek1 and Nek2, respectively. Both GlNeks are active in vitro, have dynamic relocalisation during the cell cycle, and are expressed throughout the life cycle, with GlNek1 being upregulated in cysts. Over-expression of inactive GlNek1 delays disassembly of the parental attachment disc and cytokinesis, whilst over-expression of either wild type GlNek1 or inactive mutant GlNek2 inhibits excystation.

  18. Nitroimidazole carboxamides as antiparasitic agents targeting Giardia lamblia, Entamoeba histolytica and Trichomonas vaginalis.

    Science.gov (United States)

    Jarrad, A M; Debnath, A; Miyamoto, Y; Hansford, K A; Pelingon, R; Butler, M S; Bains, T; Karoli, T; Blaskovich, M A T; Eckmann, L; Cooper, M A

    2016-09-14

    Diarrhoeal diseases caused by the intestinal parasites Giardia lamblia and Entamoeba histolytica constitute a major global health burden. Nitroimidazoles are first-line drugs for the treatment of giardiasis and amebiasis, with metronidazole 1 being the most commonly used drug worldwide. However, treatment failures in giardiasis occur in up to 20% of cases and development of resistance to metronidazole is of concern. We have re-examined 'old' nitroimidazoles as a foundation for the systematic development of next-generation derivatives. Using this approach, derivatisation of the nitroimidazole carboxamide scaffold provided improved antiparasitic agents. Thirty-three novel nitroimidazole carboxamides were synthesised and evaluated for activity against G. lamblia and E. histolytica. Several of the new compounds exhibited potent activity against G. lamblia strains, including metronidazole-resistant strains of G. lamblia (EC50 = 0.1-2.5 μM cf. metronidazole EC50 = 6.1-18 μM). Other compounds showed improved activity against E. histolytica (EC50 = 1.7-5.1 μM cf. metronidazole EC50 = 5.0 μM), potent activity against Trichomonas vaginalis (EC50 = 0.6-1.4 μM cf. metronidazole EC50 = 0.8 μM) and moderate activity against the intestinal bacterial pathogen Clostridium difficile (0.5-2 μg/mL, cf. metronidazole = 0.5 μg/mL). The new compounds had low toxicity against mammalian kidney and liver cells (CC50 > 100 μM), and selected antiparasitic hits were assessed for human plasma protein binding and metabolic stability in liver microsomes to demonstrate their therapeutic potential.

  19. Genotyping of Giardia lamblia and Entamoeba spp from river waters in Iran.

    Science.gov (United States)

    Mahmoudi, Mohammad Reza; Nazemalhosseini-Mojarad, Ehsan; Karanis, Panagiotis

    2015-12-01

    In this study, DNA from 55 surface and river water samples, which were collected from some water sources of Tehran and the Guilan Province, Iran, were extracted and examined for Entamoeba spp. and Giardia lamblia by PCR and genotyping. Twenty-seven samples, which were concentrated using the immunomagnetic separation technology (IMS) method, were examined for Giardia alone. Twenty-eight samples, which were concentrated using the sucrose flotation (SF) method, were examined for both Giardia and Entamoeba species. The results showed that 27/55 (17/27 and 10/28) (49 %), 4 /28 (14.28 %) and 3/28 (10.7 %) of the samples were positive for Giardia lamblia, Entamoeba spp and mixed infections (Entamoeba spp. and Giardia spp.), respectively. Sixteen out of 55 samples were negative. Entamoeba genus-specific PCR primers in single-round PCR were used to differentiate between the Entamoeba spp. (E. histolytica, E. dispar and E. moshkovskii). With respect to the 7 samples that were positive for Entamoeba, (14.28 %) 4 out of 28 were positive for E. moshkovskii, (7.14 %), 2 out of 28 were positive for E. histolytica and (3.57 %) 1 out of 28 was positive for E. dispar. Genus-specific PCR primers in a semi-nested PCR assay was performed to genotype Giardia species. Of the 27 samples that were positive for Giardia, 10 samples were sequences. All 10 successfully sequenced samples contained assemblage B of Giardia lamblia.This is first study to investigate the G. lamblia genotypes in the water supply of the Tehran and Guilan provinces, and it is the first study to investigate Entamoeba species in the water supplies of Iran. The investigated river water supplies, which are used for agriculture, camping and animal farming, were heavily contaminated by the human pathogenic Entamoeba and Giardia parasites. There is a potential risk of waterborne outbreaks in humans and animals.

  20. Rac Regulates Giardia lamblia Encystation by Coordinating Cyst Wall Protein Trafficking and Secretion

    OpenAIRE

    Jana Krtková; Elizabeth B Thomas; Germain C. M. Alas; Schraner, Elisabeth M.; Behjatnia, Habib R; Hehl, Adrian B.; Paredez, Alexander R.

    2016-01-01

    ABSTRACT Encystation of the common intestinal parasite Giardia lamblia involves the production, trafficking, and secretion of cyst wall material (CWM). However, the molecular mechanism responsible for the regulation of these sequential processes remains elusive. Here, we examined the role of GlRac, Giardia’s sole Rho family GTPase, in the regulation of endomembrane organization and cyst wall protein (CWP) trafficking. Localization studies indicated that GlRac is associated with the endoplasmi...

  1. Rac regulates giardia lamblia encystation by coordinating cyst wall protein trafficking and secretion

    OpenAIRE

    Krtková, Jana; Elizabeth B Thomas; Germain C. M. Alas; Schraner, Elisabeth M.; Behjatnia, Habib R; Hehl, Adrian B.; Paredez, Alexander R.

    2016-01-01

    UNLABELLED Encystation of the common intestinal parasite Giardia lamblia involves the production, trafficking, and secretion of cyst wall material (CWM). However, the molecular mechanism responsible for the regulation of these sequential processes remains elusive. Here, we examined the role of GlRac, Giardia's sole Rho family GTPase, in the regulation of endomembrane organization and cyst wall protein (CWP) trafficking. Localization studies indicated that GlRac is associated with the endoplas...

  2. Identification and Validation of Small-Gatekeeper Kinases as Drug Targets in Giardia lamblia

    OpenAIRE

    Hennessey, Kelly M.; Smith, Tess R.; Xu, Jennifer W.; Germain C. M. Alas; Ojo, Kayode K.; Merritt, Ethan A.; Paredez, Alexander R.

    2016-01-01

    Giardiasis is widely acknowledged to be a neglected disease in need of new therapeutics to address toxicity and resistance issues associated with the limited available treatment options. We examined seven protein kinases in the Giardia lamblia genome that are predicted to share an unusual structural feature in their active site. This feature, an expanded active site pocket resulting from an atypically small gatekeeper residue, confers sensitivity to “bumped” kinase inhibitors (BKIs), a class ...

  3. Molecular seasonality of Giardia lamblia in a cohort of Egyptian children: a circannual pattern.

    Science.gov (United States)

    Ismail, Mousa A M; El-Akkad, Dina M H; Rizk, Enas M A; El-Askary, Hala M; El-Badry, Ayman A

    2016-11-01

    Giardia lamblia (G. lamblia) is the most worldwide prevailing intestinal parasite, notorious for its broad range of seasonal and age-related prevalence. The potentially lethal nature of giardiasis makes it essential that the seasonality, the groups at risk, and other potential risk factors are identified. The present molecular epidemiological study was designed to determine the genetic diversity of G. lamblia infection, taking into account seasonal peaks, age distribution, and associated symptoms in a cohort of Egyptian diarrheic patients. Stool samples were collected from 1187 diarrheic patients attending outpatient clinics of Cairo University hospitals, of all age groups over a 12-month period. The patients were examined microscopically for fecal G. lamblia cysts, and/or trophozoites, and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays targeting beta giardin gene. PCR-positive samples were characterized molecularly by nPCR restriction fragment length polymorphism (RFLP) to determine Giardia assemblages. The findings revealed circannual prevalence of Giardia, with a seasonal pattern peaking in mid-summer and late winter, with the summer peak preceded by a peak in temperature. Infection was prevailing in 224 (18.9 %) cases, mainly assemblage B (81.2 %) followed by assemblage A (18.8 %). There were statistically significant associations between the detection of Giardia and flatulence, persistent diarrhea, vomiting, and abdominal pain, while gender and intermittent diarrhea showed no association. The pre-school age group was the most vulnerable. This is the first study of molecular characterization of Giardia to determine its circannual prevalence in Egypt, a finding which carries promising potential for the diagnosis, treatment, and elimination of the disease.

  4. Regulation of Cyst Wall Protein Promoters by Myb2 in Giardia lamblia*

    OpenAIRE

    2008-01-01

    Myb family transcription factors are important in regulating cell proliferation, differentiation, and cell cycle progression. Giardia lamblia differentiates into infectious cysts to survive outside of the host. During encystation, genes encoding cyst wall proteins (CWPs) are coordinately induced. We have identified an encystation-induced Myb2 protein, which binds to the promoter regions of the cwp genes and myb2 itself in vitro. To elucidate the role of Myb2 in G. ...

  5. The Giardia lamblia vsp gene repertoire: characteristics, genomic organization, and evolution

    OpenAIRE

    2010-01-01

    Abstract Background Giardia lamblia trophozoites colonize the intestines of susceptible mammals and cause diarrhea, which can be prolonged despite an intestinal immune response. The variable expression of the variant-specific surface protein (VSP) genes may contribute to this prolonged infection. Only one is expressed at a time, and switching expression from one gene to another occurs by an epigenetic mechanism. Results The WB Giardia isolate has been sequenced at 10× coverage and assembled i...

  6. Prevalence of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. in Libya: 2000–2015

    Science.gov (United States)

    Ghenghesh, Khalifa Sifaw; Ghanghish, Khaled; BenDarif, Elloulu T.; Shembesh, Khaled; Franka, Ezzadin

    2016-01-01

    Introduction The intestinal protozoa Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. are the causative agents of giardiasis, amebiasis, and cryptosporidiosis, respectively. Adequate knowledge of the geographical distribution of parasites and the demographic variables that influence their prevalence is important for effective control of infection in at-risk populations. Methods The data were obtained by an English language literature search of Medline and PubMed for papers using the search terms ‘intestinal parasites and Libya, G. lamblia and Libya, E. histolytica and Libya and Cryptosporidium and Libya’ for the period 2000–2015. Results The data obtained for the period 2000–2015 showed prevalence rates of 0.8–36.6% (mean 19.9%) for E. histolytica/dispar, 1.2–18.2% (mean 4.6%) for G. lamblia and 0.9–13% (mean 3.4%) for Cryptosporidium spp. among individuals in Libya with gastroenteritis (GE). On the other hand, prevalence rates of 0.8–16.3% (mean 8.3%), 1.8–28.8% (mean 4.8%), and 1.0–2.5% (mean=2.4), respectively, were observed for individuals without GE. The mean prevalence rate of E. histolytica/dispar was significantly higher among individuals with GE compared with those without GE (phistolytica, G. lamblia, and Cryptosporidium spp. may play a minor role in GE in Libya. The observed high prevalence rates of E. histolytica/dispar reported from Libya could be due mainly to the non-pathogenic E. dispar and E. moshkovskii. However, more studies are needed in the future using E. histolytica-specific enzyme immunoassays and/or molecular methods to confirm this observation. PMID:27363524

  7. Modified PEHPS Medium as an Alternative for the In Vitro Culture of Giardia lamblia

    Science.gov (United States)

    Vargas-Villarreal, Javier; Mata-Cárdenas, Benito D.; Hernández-García, Magda E.; Garza-González, Jesús N.; De La Garza-Salinas, Laura H.; González-Salazar, Francisco

    2014-01-01

    Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 103 trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 105Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia. PMID:24982905

  8. Prokaryotic Expression of α-13 Giardin Gene and Its Intracellular Localization in Giardia lamblia

    Science.gov (United States)

    Yu, Xingang; Pan, Weida; Shi, Xianli; Hu, Wei; Tan, Liping; Li, Kangxin; Wang, Zhen

    2017-01-01

    To study prokaryotic expression and subcellular localization of α-13 giardin in Giardia lamblia trophozoites, α-13 giardin gene was amplified and cloned into prokaryotic expression vector pET-28a(+). The positive recombinant plasmid was transformed into E. coli BL21(DE3) for expression by using IPTG and autoinduction expression system (ZYM-5052). The target protein was validated by SDS-PAGE and Western blotting and purified by Ni-NTA Resin. Rabbits were immunized with purified fusion proteins for preparation of polyclonal antibody; then the intracellular location of α-13 giardin was determined by fluorescence immunoassay. The results showed that the length of α-13 giardin gene was 1038 bp, encoding a polypeptide of 345 amino acids. The expressed product was a fusion protein with about 40 kDa largely present in soluble form. The target protein accounted for 21.0% of total proteins after being induced with IPTG, while it accounted for 28.8% with ZYM-5052. The anti-α13-giardin polyclonal antibody possessed good antigenic specificity as well as excellent binding activity with recombinant α-13 giardin. Immunofluorescence assays revealed that α-13 giardin was localized in the cytoplasm of G. lamblia trophozoite, suggesting that it is a cytoplasm-associated protein. The present study may lay a foundation for further functional research on α-13 giardin of G. lamblia. PMID:28286754

  9. Modified PEHPS medium as an alternative for the in vitro culture of Giardia lamblia.

    Science.gov (United States)

    Vargas-Villarreal, Javier; Mata-Cárdenas, Benito D; Hernández-García, Magda E; Garza-González, Jesús N; De La Garza-Salinas, Laura H; González-Salazar, Francisco

    2014-01-01

    Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 10(3) trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 10(5) Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia.

  10. Cloning, Expression and Characterization of Recombinant, NADH Oxidase from Giardia lamblia.

    Science.gov (United States)

    Castillo-Villanueva, Adriana; Méndez, Sara Teresa; Torres-Arroyo, Angélica; Reyes-Vivas, Horacio; Oria-Hernández, Jesús

    2016-02-01

    The NADH oxidase family of enzymes catalyzes the oxidation of NADH by reducing molecular O2 to H2O2, H2O or both. In the protozoan parasite Giardia lamblia, the NADH oxidase enzyme (GlNOX) produces H2O as end product without production of H2O2. GlNOX has been implicated in the parasite metabolism, the intracellular redox regulation and the resistance to drugs currently used against giardiasis; therefore, it is an interesting protein from diverse perspectives. In this work, the GlNOX gene was amplified from genomic G. lamblia DNA and expressed in Escherichia coli as a His-Tagged protein; then, the enzyme was purified by immobilized metal affinity chromatography, characterized, and its properties compared with those of the endogenous enzyme previously isolated from trophozoites (Brown et al. in Eur J Biochem 241(1):155-161, 1996). In comparison with the trophozoite-extracted enzyme, which was scarce and unstable, the recombinant heterologous expression system and one-step purification method produce a stable protein preparation with high yield and purity. The recombinant enzyme mostly resembles the endogenous protein; where differences were found, these were attributable to methodological discrepancies or artifacts. This homogenous, pure and functional protein preparation can be used for detailed structural or functional studies of GlNOX, which will provide a deeper understanding of the biology and pathogeny of G. lamblia.

  11. Prokaryotic Expression of α-13 Giardin Gene and Its Intracellular Localization in Giardia lamblia.

    Science.gov (United States)

    Yu, Xingang; Abdullahi, Auwalu Yusuf; Wu, Sheng; Pan, Weida; Shi, Xianli; Hu, Wei; Tan, Liping; Li, Kangxin; Wang, Zhen; Li, Guoqing

    2017-01-01

    To study prokaryotic expression and subcellular localization of α-13 giardin in Giardia lamblia trophozoites, α-13 giardin gene was amplified and cloned into prokaryotic expression vector pET-28a(+). The positive recombinant plasmid was transformed into E. coli BL21(DE3) for expression by using IPTG and autoinduction expression system (ZYM-5052). The target protein was validated by SDS-PAGE and Western blotting and purified by Ni-NTA Resin. Rabbits were immunized with purified fusion proteins for preparation of polyclonal antibody; then the intracellular location of α-13 giardin was determined by fluorescence immunoassay. The results showed that the length of α-13 giardin gene was 1038 bp, encoding a polypeptide of 345 amino acids. The expressed product was a fusion protein with about 40 kDa largely present in soluble form. The target protein accounted for 21.0% of total proteins after being induced with IPTG, while it accounted for 28.8% with ZYM-5052. The anti-α13-giardin polyclonal antibody possessed good antigenic specificity as well as excellent binding activity with recombinant α-13 giardin. Immunofluorescence assays revealed that α-13 giardin was localized in the cytoplasm of G. lamblia trophozoite, suggesting that it is a cytoplasm-associated protein. The present study may lay a foundation for further functional research on α-13 giardin of G. lamblia.

  12. Immunolocalization of α18- and α12-giardin in Giardia lamblia trophozoites.

    Science.gov (United States)

    Wu, Sheng; Pan, Weida; Shi, Xianli; Abdullahi, Auwalu Yusuf; Wang, Zhen; Yu, Xingang; Jiang, Biao; Li, Kangxin; Xu, Chang; Li, Guoqing

    2016-11-01

    To study subcellular localization of α18- and α12-giardin in Giardia lamblia trophozoites, the α18- and α12-giardin genes were amplified from G. lamblia assemblage A, respectively. The PCR products were cloned into the prokaryotic expression vector pET-28a(+), and the positive recombinant plasmids were transformed into E. coli Rosetta (DE3) strain for the expression, and expressed α18- and α12-giardin fusion protein were purified by Ni-Agarose resin, respectively. Mice were immunized with purified fusion proteins for preparation of polyclonal antibody, and then the subcellular localization of α18- and α12-giardin was determined by fluorescence immunoassay. Results showed that the concentrations of purified α18- and α12-giardin fusion proteins were 1.20 and 0.86 mg/ml, respectively. The titers of anti-α18- and anti-α12-giardin polyclonal antibody were both as high as 1:25600 dilutions. Immunofluorescent analysis showed that α18- and α12-giardin proteins were mainly localized at four pairs of flagella and the cytoplasm of G. lamblia trophozoites, suggesting that α18- and α12-giardin are the flagella and cytoplasm-associated proteins, respectively. The above information would lay the foundation for research about the crystal structure and biological function of α18- and α12-giardin.

  13. Can Giardia lamblia infection lower the risk of acute diarrhea among preschool children?

    Science.gov (United States)

    Muhsen, Khitam; Cohen, Dani; Levine, Myron M

    2014-04-01

    There are inconsistent findings concerning the role of Giardia lamblia in pediatric diarrhea. A prospective cohort study of the incidence of acute diarrhea among Israeli Arab preschool children offered the opportunity to examine the association between G. lamblia infection (at baseline) and subsequent diarrhea. Following baseline screening by light microscopy for the presence of Giardia in their stools, a cohort was assembled of 142 children who were followed between October 2003 and August 2004 for the incidence of diarrhea. Surveillance was performed through maternal interviews. At baseline, 21 children tested Giardia-positive. During the prospective surveillance, acute diarrhea occurred less often among Giardia-positive children (9.5%) than among children who were not infected with Giardia (26.5%). G. lamblia infection was associated with lower risk of acute diarrhea; adjusted odds ratio of 0.18 (95% confidence interval 0.04-0.93) (p = 0.041). This prospective study provides additional evidence that Giardia may lower the risk of subsequent acute diarrhea among preschool children.

  14. Modified PEHPS Medium as an Alternative for the In Vitro Culture of Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Javier Vargas-Villarreal

    2014-01-01

    Full Text Available Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 103 trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 105Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia.

  15. Phosphorylation of Serine 148 in Giardia lamblia End-binding 1 Protein is Important for Cell Division.

    Science.gov (United States)

    Kim, Juri; Lee, Hye-Yeon; Lee, Kyu-Ho; Park, Soon-Jung

    2017-07-01

    Giardia lamblia is a unicellular organism, showing a polarity with two nuclei and cytoskeletal structures. Accurate positioning of these organelles is essential for division of G. lamblia, which is poorly understood. Giardia lamblia end-binding 1 (GlEB1) protein and G. lamblia aurora kinase (GlAK) have been shown to modulate microtubule (MT) distribution during cytokinesis. A direct association between GlEB1 and GlAK was demonstrated. Like GlEB1, GlAK was also found at nuclear envelopes and median bodies of G. lamblia. In vitro kinase assays using Giardia lysates immunoprecipitated with anti-GlAK antibodies or recombinant GlAK suggested that GlEB1 is a substrate of GlAK. Site-directed mutagenesis indicated that threonine-205 in GlAK was auto-phosphorylated and that GlAK phosphorylated serine (Ser)-148 in GlEB1. Ectopic expression of a mutant GlEB1 (with conversion of Ser-148 into alanine of GlEB1) resulted in an increased number of Giardia cells with division defects. Treatment of G. lamblia with an AK inhibitor triggered cytokinesis defects, and ectopic expression of a phospho-mimetic mutant GlEB1 (with conversion of Ser-148 into aspartate) rescued the defects in Giardia cell division caused by the AK inhibitor. These results suggested that phosphorylation of GlEB1 played a role in cytokinesis in G. lamblia. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

  16. Simultaneous detection of Giardia lamblia and Cryptosporidium parvum (oo)cysts in soil using immunomagnetic separation and direct fluorescent antibody staining.

    Science.gov (United States)

    Orlofsky, Ezra; Gillor, Osnat; Melli, Ann; Miller, Woutrina; Wuertz, Stefan; Bernstein, Nirit; Shapiro, Karen

    2013-09-01

    An improved approach for simultaneous detection of Cryptosporidium parvum and Giardia lamblia (oo)cysts in soil is described. Recoveries>70% were obtained for concentrations>55 and 21 (oo)cysts g(-1) for C. parvum and G. lamblia, respectively. The limits of detection were determined to be<5 (oo)cysts g(-1) soil.

  17. Synthesis of 3-tetrazolylmethyl-4H-chromen-4-ones via Ugi-azide and biological evaluation against Entamoeba histolytica, Giardia lamblia and Trichomona vaginalis.

    Science.gov (United States)

    Cano, Pedro A; Islas-Jácome, Alejandro; González-Marrero, Joaquín; Yépez-Mulia, Lilian; Calzada, Fernando; Gámez-Montaño, Rocío

    2014-02-15

    The synthesis of novel 3-tetrazolylmethyl-4H-chromen-4-ones via an Ugi-azide multicomponent reaction and their biological evaluation against Entamoeba histolytica, Giardia lamblia and Trichomona vaginalis are described. Reported yields are moderate to good and biological results show that these compounds could be considered as candidates to anti-parasitic drugs, especially against G. lamblia.

  18. High-throughput multiplex quantitative polymerase chain reaction method for Giardia lamblia and Cryptosporidium species detection in stool samples.

    Science.gov (United States)

    Nurminen, Noora; Juuti, Rosa; Oikarinen, Sami; Fan, Yue-Mei; Lehto, Kirsi-Maarit; Mangani, Charles; Maleta, Kenneth; Ashorn, Per; Hyöty, Heikki

    2015-06-01

    Giardia lamblia and Cryptosporidium species belong to a complex group of pathogens that cause diseases hampering development and socioeconomic improvements in the developing countries. Both pathogens are recognized as significant causes of diarrhea and nutritional disorders. However, further studies are needed to clarify the role of parasitic infections, especially asymptomatic infections in malnutrition and stunting. We developed a high-throughput multiplex quantitative polymerase chain reaction (qPCR) method for G. lamblia and Cryptosporidium spp. detection in stool samples. The sensitivity and specificity of the method were ensured by analyzing confirmed positive samples acquired from diagnostics laboratories and participating in an external quality control round. Its capability to detect asymptomatic G. lamblia and Cryptosporidium spp. infections was confirmed by analyzing stool samples collected from 44 asymptomatic 6-month-old infants living in an endemic region in Malawi. Of these, five samples were found to be positive for G. lamblia and two for Cryptosporidium spp. In conclusion, the developed method is suitable for large-scale studies evaluating the occurrence of G. lamblia and Cryptosporidium spp. in endemic regions and for clinical diagnostics of these infections.

  19. In-vitro giardicidal effect of aqueous and alcoholic extracts of Asafoetida on Giardia lamblia cyst

    Directory of Open Access Journals (Sweden)

    Mohammadreza Rezaiemanesh

    2012-05-01

    Full Text Available Background and Aim: Giardia lamblia, Giardiasis causative protozoa, is one of the most common etiologic agents of diarrhea throughout the world especially in Iran. Asafoetida, an oleo-gum-resin (called Anjodan in Farsi obtained from an Iranian endemic herb, Ferula Assa-foetida has been used in treating of different diseases, particularly parasitic ones. The aim of this study was in-vitro evaluating of the effect of Asafoetida aqueous and alcoholic extracts on Giardia lamblia cysts. Materials and Methods: 500 µl of each of 1.25, 2.5, 5 and 20 mg/ml concentrtions of asafoetida aqueous and ethanol extracts was added to 500 µl of purified Giardia cysts,respectively. The mixtures were kept at 4, 24 and 37°C. The Giardicidal activity of the extracts was measured 1, 2, 3, 4 and 5 hour (s after exposure through 0.1% eosin dye staining and microscopic enumeration method. The gathered data was analyzed by means of one- way variance, Pearson's correlation coefficient and Independent T-test. Results: The highest Giardicidal effect of Asafoetida ethanol extract was 100% at 37°C, belonging to 20 mg/ml and in the 4th hour after experiment, while the maximum Giardicidal effect of Asafoetida aqueous extract was 57.23% at the same temperature and with the same concentration, in the 5th hour. There was a significant difference between Giardicidal effect of ethanol and water extracts with all concentrations and at different whiles and temperatures (P<0.005. Giardicidal effect of both extracts significantly increased due to rising the concentration, time and temperature (P<0.0001. Conclusion: Ethanol and aqueous extracts of asafetida have in-vitro Giardicidal effect on Giardia lamblia cysts. Ethanol extract has more Giardicidal effect.

  20. Viability and fate of Cryptosporidium parvum and Giardia lamblia in tubular anaerobic digesters.

    Science.gov (United States)

    Kinyua, Maureen N; Trimmer, John; Izurieta, Ricardo; Cunningham, Jeffrey; Ergas, Sarina J

    2016-06-01

    In many developing countries where pathogenic diseases of animal waste origin, such as giardiasis and cryptosporidiosis, are often prevalent, facilities are limited to treat livestock waste. However, household-scale anaerobic digesters are currently being promoted for bioenergy production from livestock manure. Since the effluent is often used as a fertilizer for food crops, it is critical to understand the effect of environmental conditions within household-scale digesters on the viability of Cryptosporidium parvum oocysts and Giardia lamblia cysts. In this study, key environmental parameters affecting (oo)cyst inactivation were measured in four tubular anaerobic digesters, which are a type of household-scale digester promoted for treatment of swine waste in rural Costa Rica. Interviews and participant observations were used to understand digester operation and maintenance procedures. Ambient temperatures (21-24°C), near-neutral pH, total ammonia nitrogen (TAN) concentrationsGiardia lamblia and Cryptosporidium parvum were 0.155 ± 0.041 and 0.054 ± 0.006 day(-1), respectively. Temperature and volatile fatty acids were the main factors contributing to Cryptosporidium parvum and Giardia lamblia inactivation. A mathematical model was developed that predicts the concentration of (oo)cysts in the liquid effluent of tubular digesters like those observed in Costa Rica. A mathematical model was developed that predicts the concentration of (oo)cysts in the liquid effluent of tubular digesters like those observed in Costa Rica. Two dimensionless groups can be used to predict the performance of the digesters for inactivating pathogens; both dimensionless groups depend upon the average HRT in the digester. This is the first study to combine mathematical modeling with qualitative analysis, field and laboratory studies to predict the concentrations of (oo)cysts in tubular digester effluents.

  1. Giardial triosephosphate isomerase as possible target of the cytotoxic effect of omeprazole in Giardia lamblia.

    Science.gov (United States)

    Reyes-Vivas, Horacio; de la Mora-de la Mora, Ignacio; Castillo-Villanueva, Adriana; Yépez-Mulia, Lilian; Hernández-Alcántara, Gloria; Figueroa-Salazar, Rosalia; García-Torres, Itzhel; Gómez-Manzo, Saúl; Méndez, Sara T; Vanoye-Carlo, América; Marcial-Quino, Jaime; Torres-Arroyo, Angélica; Oria-Hernández, Jesús; Gutiérrez-Castrellón, Pedro; Enríquez-Flores, Sergio; López-Velázquez, Gabriel

    2014-12-01

    Giardiasis is highly prevalent in the developing world, and treatment failures with the standard drugs are common. This work deals with the proposal of omeprazole as a novel antigiardial drug, focusing on a giardial glycolytic enzyme used to follow the cytotoxic effect at the molecular level. We used recombinant technology and enzyme inactivation to demonstrate the capacity of omeprazole to inactivate giardial triosephosphate isomerase, with no adverse effects on its human counterpart. To establish the specific target in the enzyme, we used single mutants of every cysteine residue in triosephosphate isomerase. The effect on cellular triosephosphate isomerase was evaluated by following the remnant enzyme activity on trophozoites treated with omeprazole. The interaction of omeprazole with giardial proteins was analyzed by fluorescence spectroscopy. The susceptibility to omeprazole of drug-susceptible and drug-resistant strains of Giardia lamblia was evaluated to demonstrate its potential as a novel antigiardial drug. Our results demonstrate that omeprazole inhibits giardial triosephosphate isomerase in a species-specific manner through interaction with cysteine at position 222. Omeprazole enters the cytoplasmic compartment of the trophozoites and inhibits cellular triosephosphate isomerase activity in a dose-dependent manner. Such inhibition takes place concomitantly with the cytotoxic effect caused by omeprazole on trophozoites. G. lamblia triosephosphate isomerase (GlTIM) is a cytoplasmic protein which can help analyses of how omeprazole works against the proteins of this parasite and in the effort to understand its mechanism of cytotoxicity. Our results demonstrate the mechanism of giardial triosephosphate isomerase inhibition by omeprazole and show that this drug is effective in vitro against drug-resistant and drug-susceptible strains of G. lamblia.

  2. Characterization of microtubule-binding and dimerization activity of Giardia lamblia end-binding 1 protein.

    Science.gov (United States)

    Kim, Juri; Nagami, Sara; Lee, Kyu-Ho; Park, Soon-Jung

    2014-01-01

    End-binding 1 (EB1) proteins are evolutionarily conserved components of microtubule (MT) plus-end tracking protein that regulate MT dynamics. Giardia lamblia, with two nuclei and cytoskeletal structures, requires accurate MT distribution for division. In this study, we show that a single EB1 homolog gene of G. lamblia regulates MT dynamics in mitosis. The haemagglutinin-tagged G. lamblia EB1 (GlEB1) localizes to the nuclear envelopes and median bodies, and is transiently present in mitotic spindles of dividing cells. Knockdown of GlEB1 expression using the morpholinos-based anti-EB1 oligonucleotides, resulted in a significant defect in mitosis of Giardia trophozoites. The MT-binding assays using recombinant GlEB1 (rGlEB1) proteins demonstrated that rGlEB1102-238, but not rGlEB11-184, maintains an MT-binding ability comparable with that of the full length protein, rGlEB11-238. Size exclusion chromatography showed that rGlEB1 is present as a dimer formed by its C-terminal domain and a disulfide bond. In vitro-mutagenesis of GlEB1 indicated that an intermolecular disulfide bond is made between cysteine #13 of the two monomers. Complementation assay using the BIM1 knockout mutant yeast, the yeast homolog of mammalian EB1, indicated that expression of the C13S mutant GlEB1 protein cannot rescue the mitotic defect of the BIM1 mutant yeast. These results suggest that dimerization of GlEB1 via the 13th cysteine residues plays a role during mitosis in Giardia.

  3. Differential gene expression in Giardia lamblia under oxidative stress: significance in eukaryotic evolution.

    Science.gov (United States)

    Raj, Dibyendu; Ghosh, Esha; Mukherjee, Avik K; Nozaki, Tomoyoshi; Ganguly, Sandipan

    2014-02-10

    Giardia lamblia is a unicellular, early branching eukaryote causing giardiasis, one of the most common human enteric diseases. Giardia, a microaerophilic protozoan parasite has to build up mechanisms to protect themselves against oxidative stress within the human gut (oxygen concentration 60 μM) to establish its pathogenesis. G. lamblia is devoid of the conventional mechanisms of the oxidative stress management system, including superoxide dismutase, catalase, peroxidase, and glutathione cycling, which are present in most eukaryotes. NADH oxidase is a major component of the electron transport chain of G. lamblia, which in concurrence with disulfide reductase, protects oxygen-labile proteins such as pyruvate: ferredoxin oxidoreductase against oxidative stress by sustaining a reduced intracellular environment. It also contains the arginine dihydrolase pathway, which occurs in a number of anaerobic prokaryotes, includes substrate level phosphorylation and adequately active to make a major contribution to ATP production. To study differential gene expression under three types of oxidative stress, a Giardia genomic DNA array was constructed and hybridized with labeled cDNA of cells with or without stress. The transcriptomic data has been analyzed and further validated using real time PCR. We identified that out of 9216 genes represented on the array, more than 200 genes encoded proteins with functions in metabolism, oxidative stress management, signaling, reproduction and cell division, programmed cell death and cytoskeleton. We recognized genes modulated by at least ≥ 2 fold at a significant time point in response to oxidative stress. The study has highlighted the genes that are differentially expressed during the three experimental conditions which regulate the stress management pathway differently to achieve redox homeostasis. Identification of some unique genes in oxidative stress regulation may help in new drug designing for this common enteric parasite prone to

  4. Identification and Validation of Small-Gatekeeper Kinases as Drug Targets in Giardia lamblia.

    Science.gov (United States)

    Hennessey, Kelly M; Smith, Tess R; Xu, Jennifer W; Alas, Germain C M; Ojo, Kayode K; Merritt, Ethan A; Paredez, Alexander R

    2016-11-01

    Giardiasis is widely acknowledged to be a neglected disease in need of new therapeutics to address toxicity and resistance issues associated with the limited available treatment options. We examined seven protein kinases in the Giardia lamblia genome that are predicted to share an unusual structural feature in their active site. This feature, an expanded active site pocket resulting from an atypically small gatekeeper residue, confers sensitivity to "bumped" kinase inhibitors (BKIs), a class of compounds that has previously shown good pharmacological properties and minimal toxicity. An initial phenotypic screen for biological activity using a subset of an in-house BKI library found that 5 of the 36 compounds tested reduced trophozoite growth by at least 50% at a concentration of 5 μM. The cellular localization and the relative expression levels of the seven protein kinases of interest were determined after endogenously tagging the kinases. Essentiality of these kinases for parasite growth and infectivity were evaluated genetically using morpholino knockdown of protein expression to establish those that could be attractive targets for drug design. Two of the kinases were critical for trophozoite growth and attachment. Therefore, recombinant enzymes were expressed, purified and screened against a BKI library of >400 compounds in thermal stability assays in order to identify high affinity compounds. Compounds with substantial thermal stabilization effects on recombinant protein were shown to have good inhibition of cell growth in wild-type G. lamblia and metronidazole-resistant strains of G. lamblia. Our data suggest that BKIs are a promising starting point for the development of new anti-giardiasis therapeutics that do not overlap in mechanism with current drugs.

  5. Development of T m -shift genotyping method for detection of cat-derived Giardia lamblia.

    Science.gov (United States)

    Pan, Weida; Fu, Yeqi; Abdullahi, Auwalu Yusuf; Wang, Mingwei; Shi, Xianli; Yang, Fang; Yu, Xingang; Yan, Xinxin; Zhang, Pan; Hang, Jianxiong; Li, Guoqing

    2017-04-01

    To develop T m -shift genotyping method for detection of cat-derived Giardia lamblia, two sets of primers with two GC-rich tails of unequal length attached to their 5'-end were designed according to two SNPs (BG434 and BG170) of β-giardin (bg) gene, and specific PCR products were identified by inspection of a melting curve on real-time PCR thermocycler. A series of experiments on the stability, sensitivity, and accuracy of T m -shift method was tested, and clinical samples were also detected. The results showed that two sets of primers based on SNP could distinguish accurately between assemblages A and F. Coefficient of variation of T m values of assemblage A and F was 0.14 and 0.07% in BG434 and 0.10 and 0.11% in BG170, respectively. The lowest detection concentration was 4.52 × 10(-5) and 4.88 × 10(-5) ng/μL samples of assemblage A and F standard plasmids. The T m -shift genotyping results of ten DNA samples from the cat-derived G. lamblia were consistent with their known genotypes. The detection rate of clinical samples by T m -shift was higher than that by microscopy, and their genotyping results were in complete accordance with sequencing results. It is concluded that the T m -shift genotyping method is rapid, specific, and sensitive and may provide a new technological mean for molecular detection and epidemiological investigation of the cat-derived G. lamblia.

  6. Detection and molecular characterisation of Giardia lamblia genotypes in Sharjah, United Arab Emirates.

    Science.gov (United States)

    ElBakri, Ali; Samie, Amidou; Bessong, Pascal; Potgieter, Natasha; Odeh, Ra'ed Abu

    2014-08-01

    No data are available on Giardia lamblia genotypes in the United Arab Emirates (UAE). This study aimed to identify G. lamblia from DNA extracted from human stool samples to gain information on its prevalence and to perform molecular analysis on isolates collected from expatriates from different localities residing in Sharjah, UAE. In total, 111 healthy expatriates residing in Sharjah were screened for G. lamblia using nested PCR amplification of the small subunit ribosomal RNA (ssu-rRNA) gene. Positive samples were genotyped using a nested PCR amplifying the triosephosphate isomerase (tpi) gene to differentiate between the two human assemblages (A and B). A subset of the PCR products (n=23) were sequenced and their phylogenetic relationships were determined. Of the 111 participants, 67 (60.4%) were identified as positive for the ssu-rRNA gene. When genotyped for the tpi gene, 18.9% (21/111) were of assemblage A, 17.1% (19/111) belonged to assemblage B and 5.4% (6/111) showed patterns compatible with mixed infections. A strong correlation between the presence of diarrhoea and assemblage B was observed (χ(2)=10.553; p=0.001). Moreover, an association was also observed between mixed infections (A+B) and diarrhoea (χ(2)=8.899; p=0.003). No correlation between age, gender and geographic origin of the infected individual was noted. Phylogenetic analysis showed three clusters for the tpi gene. No relationship between the clusters and the origin of samples was noted. This study is the first to determine the infection rate and genotypic composition of Giardia in Sharjah, UAE. © The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. An In Vitro Evaluation of Ozonized Organic Extra-Virgin Olive Oil on Giardia Lamblia Cysts.

    Science.gov (United States)

    Boland-Nazar, Najmeh Sadat; Eslamirad, Zahra; Sarmadian, Hossein; Ghasemikhah, Reza

    2016-11-01

    Giardia lamblia is a common intestinal parasite that has been reported all over the world. This study was conducted to evaluate the effect of ozonized organic extra-virgin olive oil on the cyst of G. lamblia. The olive oil was ozonized based on international standards and confirmed by the world health organization (WHO) at various times in a generator. The ozone concentration of olive oil was adjusted at 32, 64, 96, 128, 160 mg/g based on ozone absorption. Giardia lamblia cysts were isolated from heavily infected stool samples and the sucrose gradient flotation technique. Five groups of triple tubes containing Giardia cysts were exposed to olive oil with 32, 64, 96, 128, 160 ozone concentrations, and the sixth and seventh groups were exposed to non-ozonized olive oil and normal saline, respectively. The tubes were placed at room temperature, and every four hours, the mortality of the Giardia cysts was assessed. The results showed that the first five groups' mortality rate of Giardia cysts reached 100% in 100 hours. An increasing concentration of ozone in olive oil leads to an increase in the mortality rate of Giardia cysts. The results showed a significant difference in the mean time of the mortality in all the groups (P ≤ 0.05). Furthermore, the higher fatality effect of ozonized organic extra-virgin olive oil (Ozonized Olive Oil = OZO) was proved in comparison with metronidazole in vitro. We concluded that ozonized organic extra-virgin olive oil was a growth inhibitor of Giardia cysts, and concerning its compatibility with a biological system, it is recommended for further clinical trials.

  8. Prevalence of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. in Libya: 2000-2015.

    Science.gov (United States)

    Ghenghesh, Khalifa Sifaw; Ghanghish, Khaled; BenDarif, Elloulu T; Shembesh, Khaled; Franka, Ezzadin

    2016-01-01

    The intestinal protozoa Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. are the causative agents of giardiasis, amebiasis, and cryptosporidiosis, respectively. Adequate knowledge of the geographical distribution of parasites and the demographic variables that influence their prevalence is important for effective control of infection in at-risk populations. The data were obtained by an English language literature search of Medline and PubMed for papers using the search terms 'intestinal parasites and Libya, G. lamblia and Libya, E. histolytica and Libya and Cryptosporidium and Libya' for the period 2000-2015. The data obtained for the period 2000-2015 showed prevalence rates of 0.8-36.6% (mean 19.9%) for E. histolytica/dispar, 1.2-18.2% (mean 4.6%) for G. lamblia and 0.9-13% (mean 3.4%) for Cryptosporidium spp. among individuals in Libya with gastroenteritis (GE). On the other hand, prevalence rates of 0.8-16.3% (mean 8.3%), 1.8-28.8% (mean 4.8%), and 1.0-2.5% (mean=2.4), respectively, were observed for individuals without GE. The mean prevalence rate of E. histolytica/dispar was significantly higher among individuals with GE compared with those without GE (plamblia, and Cryptosporidium spp. may play a minor role in GE in Libya. The observed high prevalence rates of E. histolytica/dispar reported from Libya could be due mainly to the non-pathogenic E. dispar and E. moshkovskii. However, more studies are needed in the future using E. histolytica-specific enzyme immunoassays and/or molecular methods to confirm this observation.

  9. Identification and Validation of Small-Gatekeeper Kinases as Drug Targets in Giardia lamblia

    Science.gov (United States)

    Hennessey, Kelly M.; Smith, Tess R.; Xu, Jennifer W.; Alas, Germain C. M.; Ojo, Kayode K.; Merritt, Ethan A.

    2016-01-01

    Giardiasis is widely acknowledged to be a neglected disease in need of new therapeutics to address toxicity and resistance issues associated with the limited available treatment options. We examined seven protein kinases in the Giardia lamblia genome that are predicted to share an unusual structural feature in their active site. This feature, an expanded active site pocket resulting from an atypically small gatekeeper residue, confers sensitivity to “bumped” kinase inhibitors (BKIs), a class of compounds that has previously shown good pharmacological properties and minimal toxicity. An initial phenotypic screen for biological activity using a subset of an in-house BKI library found that 5 of the 36 compounds tested reduced trophozoite growth by at least 50% at a concentration of 5 μM. The cellular localization and the relative expression levels of the seven protein kinases of interest were determined after endogenously tagging the kinases. Essentiality of these kinases for parasite growth and infectivity were evaluated genetically using morpholino knockdown of protein expression to establish those that could be attractive targets for drug design. Two of the kinases were critical for trophozoite growth and attachment. Therefore, recombinant enzymes were expressed, purified and screened against a BKI library of >400 compounds in thermal stability assays in order to identify high affinity compounds. Compounds with substantial thermal stabilization effects on recombinant protein were shown to have good inhibition of cell growth in wild-type G. lamblia and metronidazole-resistant strains of G. lamblia. Our data suggest that BKIs are a promising starting point for the development of new anti-giardiasis therapeutics that do not overlap in mechanism with current drugs. PMID:27806042

  10. Variation in the ribosome interacting loop of the Sec61α from Giardia lamblia.

    Science.gov (United States)

    Sinha, Abhishek; Ray, Atrayee; Ganguly, Sandipan; Ghosh Dastidar, Shubhra; Sarkar, Srimonti

    2015-09-30

    The interaction between the ribosome and the endoplasmic reticulum-located Sec61 protein translocon is mediated through an arginine residue of Sec61α, which is conserved in all prokaryotic and eukaryotic orthologues characterized to date. Using in silico approaches we report that instead of arginine, this ribosome-interaction function is most likely discharged by a lysine residue in the protist Giardia lamblia. This functional substitution of the R with a K in GlSec61α may have taken place to accommodate a G-rich rRNA.

  11. Optical map of the genotype A1 WB C6 Giardia lamblia genome isolate.

    Science.gov (United States)

    Perry, D Alexander; Morrison, Hilary G; Adam, Rodney D

    2011-12-01

    The Giardia lamblia genome consists of 12 Mb divided among 5 chromosomes ranging in size from approximately 1 to 4 Mb. The assembled contigs of the genotype A1 isolate, WB, were previously mapped along the 5 chromosomes on the basis of hybridization of plasmid clones representing the contigs to chromosomes separated by PFGE. In the current report, we have generated an MluI optical map of the WB genome to improve the accuracy of the physical map. This has allowed us to correct several assembly errors and to better define the extent of the subtelomeric regions that are not included in the genome assembly.

  12. Prevalence of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. in Libya: 2000–2015

    Directory of Open Access Journals (Sweden)

    Khalifa Sifaw Ghenghesh

    2016-06-01

    Full Text Available Introduction: The intestinal protozoa Entamoeba histolytica, Giardia lamblia, and Cryptosporidium spp. are the causative agents of giardiasis, amebiasis, and cryptosporidiosis, respectively. Adequate knowledge of the geographical distribution of parasites and the demographic variables that influence their prevalence is important for effective control of infection in at-risk populations. Methods: The data were obtained by an English language literature search of Medline and PubMed for papers using the search terms ‘intestinal parasites and Libya, G. lamblia and Libya, E. histolytica and Libya and Cryptosporidium and Libya’ for the period 2000–2015. Results: The data obtained for the period 2000–2015 showed prevalence rates of 0.8–36.6% (mean 19.9% for E. histolytica/dispar, 1.2–18.2% (mean 4.6% for G. lamblia and 0.9–13% (mean 3.4% for Cryptosporidium spp. among individuals in Libya with gastroenteritis (GE. On the other hand, prevalence rates of 0.8–16.3% (mean 8.3%, 1.8–28.8% (mean 4.8%, and 1.0–2.5% (mean=2.4, respectively, were observed for individuals without GE. The mean prevalence rate of E. histolytica/dispar was significantly higher among individuals with GE compared with those without GE (p<0.0000001, OR=2.74. No significant difference in prevalence rate of the three organisms was found according to gender, but most of infections were observed in children aged 10 years or younger. Conclusion: The reviewed data suggest that E. histolytica, G. lamblia, and Cryptosporidium spp. may play a minor role in GE in Libya. The observed high prevalence rates of E. histolytica/dispar reported from Libya could be due mainly to the non-pathogenic E. dispar and E. moshkovskii. However, more studies are needed in the future using E. histolytica-specific enzyme immunoassays and/or molecular methods to confirm this observation.

  13. Rac Regulates Giardia lamblia Encystation by Coordinating Cyst Wall Protein Trafficking and Secretion.

    Science.gov (United States)

    Krtková, Jana; Thomas, Elizabeth B; Alas, Germain C M; Schraner, Elisabeth M; Behjatnia, Habib R; Hehl, Adrian B; Paredez, Alexander R

    2016-08-23

    Encystation of the common intestinal parasite Giardia lamblia involves the production, trafficking, and secretion of cyst wall material (CWM). However, the molecular mechanism responsible for the regulation of these sequential processes remains elusive. Here, we examined the role of GlRac, Giardia's sole Rho family GTPase, in the regulation of endomembrane organization and cyst wall protein (CWP) trafficking. Localization studies indicated that GlRac is associated with the endoplasmic reticulum (ER) and the Golgi apparatus-like encystation-specific vesicles (ESVs). Constitutive GlRac signaling increased levels of the ER marker PDI2, induced ER swelling, reduced overall CWP1 production, and promoted the early maturation of ESVs. Quantitative analysis of cells expressing constitutively active hemagglutinin (HA)-tagged GlRac (HA-Rac(CA)) revealed fewer but larger ESVs than control cells. Consistent with the phenotype of premature maturation of ESVs in HA-Rac(CA)-expressing cells, constitutive GlRac signaling resulted in increased CWP1 secretion and, conversely, morpholino depletion of GlRac blocked CWP1 secretion. Wild-type cells unexpectedly secreted large quantities of CWP1 into the medium, and free CWP1 was used cooperatively during cyst formation. These results, in part, could account for the previously reported observation that G. lamblia encysts more efficiently at high cell densities. These studies of GlRac show that it regulates encystation at several levels, and our findings support its coordinating role as a regulator of CWP trafficking and secretion. The central role of GlRac in regulating membrane trafficking and the cytoskeleton, both of which are essential to Giardia parasitism, further suggests its potential as a novel target for drug development to treat giardiasis. The encystation process is crucial for the transmission of giardiasis and the life cycle of many protists. Encystation for Giardia lamblia involves the assembly of a protective cyst wall

  14. [DETERMINATION OF GIARDIA DUODENALIS GENOTYPES IN LAMBLIA-INFESTED PEOPLE IN THE CITY OF MOSCOW].

    Science.gov (United States)

    Kurnosova, O P; Odoevskaya, I M; Krasavchenko, K S

    2015-01-01

    Our first experience in genotyping Giardia from Moscow residents, has shown that 4 and 2 of seven samples belong to G. duodenalis genotype A and genotype B, respectively; one sample was negative during amplification with two types of primers. Genotyping was Carried out using the specific primers TPIA and TPIB to the gene encoding for the enzyme triosephosphate isomerase from the parasite. Thus, further such investigations using a larger number of samples will be able to complement the epidemiology of Lamblia infection in Moscow residents.

  15. Structure of the prolyl-tRNA synthetase from the eukaryotic pathogen Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Larson, Eric T.; Kim, Jessica E.; Napuli, Alberto J.; Verlinde, Christophe L. M. J.; Fan, Erkang; Zucker, Frank H.; Van Voorhis, Wesley C.; Buckner, Frederick S.; Hol, Wim G. J.; Merritt, Ethan A., E-mail: merritt@u.washington.edu [Medical Structural Genomics of Pathogenic Protozoa, (United States); University of Washington, Seattle, WA 98195 (United States)

    2012-09-01

    The structure of Giardia prolyl-tRNA synthetase cocrystallized with proline and ATP shows evidence for half-of-the-sites activity, leading to a corresponding mixture of reaction substrates and product (prolyl-AMP) in the two active sites of the dimer. The genome of the human intestinal parasite Giardia lamblia contains only a single aminoacyl-tRNA synthetase gene for each amino acid. The Giardia prolyl-tRNA synthetase gene product was originally misidentified as a dual-specificity Pro/Cys enzyme, in part owing to its unexpectedly high off-target activation of cysteine, but is now believed to be a normal representative of the class of archaeal/eukaryotic prolyl-tRNA synthetases. The 2.2 Å resolution crystal structure of the G. lamblia enzyme presented here is thus the first structure determination of a prolyl-tRNA synthetase from a eukaryote. The relative occupancies of substrate (proline) and product (prolyl-AMP) in the active site are consistent with half-of-the-sites reactivity, as is the observed biphasic thermal denaturation curve for the protein in the presence of proline and MgATP. However, no corresponding induced asymmetry is evident in the structure of the protein. No thermal stabilization is observed in the presence of cysteine and ATP. The implied low affinity for the off-target activation product cysteinyl-AMP suggests that translational fidelity in Giardia is aided by the rapid release of misactivated cysteine.

  16. Detecting Cryptosporidium parvum and Giardia lamblia by coagulation concentration and real-time PCR quantification

    Institute of Scientific and Technical Information of China (English)

    Huining ZHANG; Xiaohu Zhang; Shuting ZHANG; Bo WEI; Qipei JIANG; Xin YU

    2013-01-01

    Rapid and sensitive pathogen detection methods are essential for the effective functioning of the water treatment industry, yet for many pathogens, effective detection and removal methods are still lacking. Cryptosporidium parvum oocysts and Giardia lamblia cysts are two of the most common waterborne pathogens currently infecting the water supply. In this study, a new method was developed for the detection of Cryptosporidium parvum oocysts and Giardia lamblia cysts. The method includes multi-steps as coagulation concentration of (oo)cysts in water, the dissolution of the resulting flocs into a small volume using acid, filtration of the (oo)cysts solution, and DNA extraction, purification, and examination using real-time PCR. The method was tested using spiked tap water and reservoir water as references, and the mean recovery ranged from 19.6% to 97.6% for oocysts, and from 51.4% to 98.7% for cysts. The method is economical and convenient, and is especially suitable for relatively high turbidity surface water.

  17. Histone methyltransferase 1 regulates the encystation process in the parasite Giardia lamblia.

    Science.gov (United States)

    Salusso, Agostina; Zlocowski, Natacha; Mayol, Gonzalo F; Zamponi, Nahuel; Rópolo, Andrea S

    2017-08-01

    In eukaryotes, histone lysine methylation is associated with either active or repressed chromatin states, depending on the status of methylation. Even when the amino-terminus of Giardia lamblia histones diverges from other organisms, these regions contain lysine residues that are potential targets for methylation. When we examined the role of the histone methyltransferase 1 (HMT1) in the regulation of the encystation process by giardial histone methyltransferase 1 (GlHMT1) overexpression or downregulation, we observed an increase or a decrease in cyst production, respectively, compared to wild-type trophozoites. A time-lapse analysis of encystation showed that overexpression of GlHMT1 induced an earlier and faster process than in wild-type cells together with an upregulation of mRNA expression of cyst wall proteins. Subcellular localization studies indicated that GlHMT1-hemaglutinin was mainly associated with the nuclear and perinuclear region in both growing and encysting parasites, in agreement with bioinformatics analyses showing that GlHMT-1 possesses nuclear localization signals in addition to the classical SU(var)3-9, Enhancer-of-Zeste, Trithorax (SET), and post-SET domains. Altogether, these findings suggest that the function of HMT1 is critical for the success and timing of the encystation process, and reinforce the idea that epigenetic marks are critical for cyst formation in G. lamblia. © 2017 Federation of European Biochemical Societies.

  18. Adhesion of Cryptosporidium parvum and Giardia lamblia to solid surfaces: the role of surface charge and hydrophobicity

    NARCIS (Netherlands)

    Dai, X.; Boll, J.; Hayes, M.E.; Aston, D.E.

    2004-01-01

    Adhesion of Cryptosporidium parvum and Giardia lamblia to four materials of different surface charge and hydrophobicity was investigated. Glass beads were used with and without three polymer coatings: aminosilines (A0750), fluorosilines (T2494), an amino cationic polymer. Surface charge density and

  19. A Multiplex PCR for Simultaneous Detection of Three Zoonotic Parasites Ancylostoma ceylanicum, A. caninum, and Giardia lamblia Assemblage A.

    Science.gov (United States)

    Hu, Wei; Wu, Sheng; Yu, Xingang; Abullahi, Auwalu Yusuf; Song, Meiran; Tan, Liping; Wang, Zhen; Jiang, Biao; Li, Guoqing

    2015-01-01

    Ancylostoma ceylanicum, A. caninum, and Giardia lamblia assemblage A are common intestinal parasites of dogs and cats; they can also infect humans, causing parasitic zoonoses. In this study, a multiplex PCR method was developed for simultaneous identification and detection of those three zoonotic parasites. Three pairs of specific primers were designed based on ITS sequence of A. ceylanicum and A. caninum and TPI gene of G. lamblia available in the GenBank. The multiplex PCR reaction system was established by optimizing the reaction condition, and a series of tests on the sensitivity, specificity, and clinical application were also conducted. Results showed that three target fragments were amplified specifically; the detection limit was 10 eggs for both A. ceylanicum and A. caninum, 72 pg DNA for G. lamblia. Of 112 clinical fecal samples, 34.8% and 17.8% samples were positive for A. caninum and A. ceylanicum, respectively, while only 2.7% samples were positive for G. lamblia assemblage A. It is concluded that the established multiplex PCR assay is a convenient, rapid, cost-effective, and high-efficiency method for molecular detection and epidemiological investigation of three zoonotic parasites.

  20. An ancient spliceosomal intron in the ribosomal protein L7a gene (Rpl7a of Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Gray Michael W

    2005-08-01

    Full Text Available Abstract Background Only one spliceosomal-type intron has previously been identified in the unicellular eukaryotic parasite, Giardia lamblia (a diplomonad. This intron is only 35 nucleotides in length and is unusual in possessing a non-canonical 5' intron boundary sequence, CT, instead of GT. Results We have identified a second spliceosomal-type intron in G. lamblia, in the ribosomal protein L7a gene (Rpl7a, that possesses a canonical GT 5' intron boundary sequence. A comparison of the two known Giardia intron sequences revealed extensive nucleotide identity at both the 5' and 3' intron boundaries, similar to the conserved sequence motifs recently identified at the boundaries of spliceosomal-type introns in Trichomonas vaginalis (a parabasalid. Based on these observations, we searched the partial G. lamblia genome sequence for these conserved features and identified a third spliceosomal intron, in an unassigned open reading frame. Our comprehensive analysis of the Rpl7a intron in other eukaryotic taxa demonstrates that it is evolutionarily conserved and is an ancient eukaryotic intron. Conclusion An analysis of the phylogenetic distribution and properties of the Rpl7a intron suggests its utility as a phylogenetic marker to evaluate particular eukaryotic groupings. Additionally, analysis of the G. lamblia introns has provided further insight into some of the conserved and unique features possessed by the recently identified spliceosomal introns in related organisms such as T. vaginalis and Carpediemonas membranifera.

  1. Adhesion of Cryptosporidium parvum and Giardia lamblia to solid surfaces: the role of surface charge and hydrophobicity

    NARCIS (Netherlands)

    Dai, X.; Boll, J.; Hayes, M.E.; Aston, D.E.

    2004-01-01

    Adhesion of Cryptosporidium parvum and Giardia lamblia to four materials of different surface charge and hydrophobicity was investigated. Glass beads were used with and without three polymer coatings: aminosilines (A0750), fluorosilines (T2494), an amino cationic polymer. Surface charge density and

  2. A Systematic Review and Meta-analysis of the Association Between Giardia lamblia and Endemic Pediatric Diarrhea in Developing Countries

    Science.gov (United States)

    Muhsen, Khitam; Levine, Myron M.

    2012-01-01

    We performed a systematic literature review and meta-analysis examining the association between diarrhea in young children in nonindustrialized settings and Giardia lamblia infection. Eligible were case/control and longitudinal studies that defined the outcome as acute or persistent (>14 days) diarrhea, adjusted for confounders and lasting for at least 1 year. Data on G. lamblia detection (mainly in stools) from diarrhea patients and controls without diarrhea were abstracted. Random effects model meta-analysis obtained pooled odds ratios (ORs) and 95% confidence intervals (CIs). Twelve nonindustrialized-setting acute pediatric diarrhea studies met the meta-analysis inclusion criteria. Random-effects model meta-analysis of combined results (9774 acute diarrhea cases and 8766 controls) yielded a pooled OR of 0.60 (95% CI, .38–.94; P = .03), indicating that G. lamblia was not associated with acute diarrhea. However, limited data suggest that initial Giardia infections in early infancy may be positively associated with diarrhea. Meta-analysis of 5 persistent diarrhea studies showed a pooled OR of 3.18 (95% CI, 1.50–6.76; P Giardia with that syndrome. The well-powered Global Enteric Multicenter Study (GEMS) is prospectively addressing the association between G. lamblia infection and diarrhea in children in developing countries. PMID:23169940

  3. Detection of Giardia lamblia Antigens in Human Fecal Specimens by a Solid-Phase Qualitative Immunochromatographic Assay▿

    OpenAIRE

    Garcia, Lynne S.; Garcia, John Paul

    2006-01-01

    The SIMPLE-READ Giardia rapid assay (Medical Chemical Corporation) is a solid-phase qualitative immunochromatographic assay that detects Giardia lamblia in aqueous extracts of human fecal specimens. Testing 106 Giardia-positive and 104 Giardia-negative stool specimens yielded a sensitivity of 97.2% and a specificity of 100% for the SIMPLE-READ Giardia rapid assay.

  4. Effect of olive leaf, Satureja khuzestanica, and Allium sativum extracts on Giardia lamblia cysts compared with metronidazole in vitro.

    Science.gov (United States)

    Fallahi, Sh; Rostami, A; Delfan, B; Pournia, Y; Rashidipour, M

    2016-12-01

    Giardia lamblia is one of the common causes of worldwide diarrhea in children. Appropriate medicinal treatment for giardiasis is available but there are some evidences of drug resistance, insufficient efficacy, and unpleasant side effects. In order to reach a more natural drug with suitable efficacy and the lowest side effects, the effects of the hydroalcoholic extracts of olive leaf, Satureja khuzestanica, and Allium sativum on G. lamblia cysts were evaluated in vitro, as well as antigiardial effect of the extracts was compared with metronidazole as the drug of choice. 2 and 5 mg of the plants extracts and powder of metronidazole 250 mg pills were added to 1 ml of G. lamblia cysts suspension (containing 5,000 cyst/ml normal saline), and the percentages of bioavailability of G. lamblia cysts were examined at the 2nd and 4th h after exposure and in 4 and 37 °C temperatures using eosin 0.1 % and a haemocytometer. The data were analyzed by multiway ANOVA test, Tukey's test, and the SPSS software, version 18. The examinations demonstrated that olive leaf extract had the most fatality rate on G. lamblia cysts in vitro (37.90 ± 7.01 %), followed by the extract of S. khuzestanica (32.52 ± 9.07 %). Metronidazole 250 mg pills had relatively effective fatality rate on G. lamblia cysts in vitro (28.75 ± 10.30 %), whereas A. sativum (garlic) had the lowest fatality effect on G. lamblia cysts in vitro (22.65 ± 10.47 %). With respect to higher fatality effect of olive leaf and S. khuzestanica extracts compared with metronidazole in vitro, these plants can be used as suitable candidates to make new antigiardial drugs with low side effects and without drug resistance in the treatment of giardiasis in children.

  5. Evolutionarily divergent spliceosomal snRNAs and a conserved non-coding RNA processing motif in Giardia lamblia

    Science.gov (United States)

    Hudson, Andrew J.; Moore, Ashley N.; Elniski, David; Joseph, Joella; Yee, Janet; Russell, Anthony G.

    2012-01-01

    Non-coding RNAs (ncRNAs) have diverse essential biological functions in all organisms, and in eukaryotes, two such classes of ncRNAs are the small nucleolar (sno) and small nuclear (sn) RNAs. In this study, we have identified and characterized a collection of sno and snRNAs in Giardia lamblia, by exploiting our discovery of a conserved 12 nt RNA processing sequence motif found in the 3′ end regions of a large number of G. lamblia ncRNA genes. RNA end mapping and other experiments indicate the motif serves to mediate ncRNA 3′ end formation from mono- and di-cistronic RNA precursor transcripts. Remarkably, we find the motif is also utilized in the processing pathway of all four previously identified trans-spliced G. lamblia introns, revealing a common RNA processing pathway for ncRNAs and trans-spliced introns in this organism. Motif sequence conservation then allowed for the bioinformatic and experimental identification of additional G. lamblia ncRNAs, including new U1 and U6 spliceosomal snRNA candidates. The U6 snRNA candidate was then used as a tool to identity novel U2 and U4 snRNAs, based on predicted phylogenetically conserved snRNA–snRNA base-pairing interactions, from a set of previously identified G. lamblia ncRNAs without assigned function. The Giardia snRNAs retain the core features of spliceosomal snRNAs but are sufficiently evolutionarily divergent to explain the difficulties in their identification. Most intriguingly, all of these snRNAs show structural features diagnostic of U2-dependent/major and U12-dependent/minor spliceosomal snRNAs. PMID:23019220

  6. Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler’s Diarrhea

    Science.gov (United States)

    Shin, Ji-Hun; Lee, Sang-Eun; Kim, Tong Soo; Ma, Da-Won; Chai, Jong-Yil; Shin, Eun-Hee

    2016-01-01

    This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler’s diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×103 oocysts for C. parvum, >1×104 cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples. PMID:27853120

  7. vsp Gene Expression by Giardia lamblia Clone GS/M-83-H7 during Antigenic Variation In Vivo and In Vitro

    OpenAIRE

    2001-01-01

    Giardia lamblia infections are associated with antigenic variation of the parasite, which is generated by a continuous change of the variant-specific surface proteins (VSPs). Many investigations on the process of antigenic variation were based on the use of G. lamblia clone GS/M-83-H7, which expresses VSP H7 as its major surface antigen. In the present study, mice were infected with the aforementioned clonal line to investigate vsp gene expression during the complex process of antigenic varia...

  8. In-vitro susceptibility of Giardia lamblia to albendazole, mebendazole and other chemotherapeutic agents.

    Science.gov (United States)

    Cedillo-Rivera, R; Muñoz, O

    1992-09-01

    The susceptibility of a strain of Giardia lamblia to benzimidazole carbamates, 5-nitroimidazoles, nitrofurans and other drugs was studied in vitro. Albendazole was the most active compound, with a 50% inhibitory concentration (IC50) of 0.01 mg/L and a minimal lethal concentration (MLC) of less than 0.04 mg/L; the IC50 of mebendazole was 0.06 mg/L and the MLC less than 0.5 mg/L. Among the 5-nitroimidazoles tested, ornidazole was the most effective (IC50 0.12 mg/L); tinidazole, metronidazole, secnidazole and hemezole were less active. Nifuroxazide, etofamide and nalidixic acid exhibited modest anti-giardial activity; quinfamide did not inhibit the growth of the parasite at a concentration of 200 mg/L. Albendazole and mebendazole are promising candidates for clinical use and should be further evaluated.

  9. Identification of Giardia lamblia Assemblage E in Humans Points to a New Anthropozoonotic Cycle.

    Science.gov (United States)

    Fantinatti, Maria; Bello, Alexandre R; Fernandes, Octavio; Da-Cruz, Alda M

    2016-10-15

    Giardia lamblia is a pathogen transmitted by water and food that causes infection worldwide. Giardia genotypes are classified into 8 assemblages (A-H). Assemblages A and B are detected in humans, but they are potentially zoonotic because they infect other mammalian hosts. Giardia in samples from 44 children was genotyped. Conserved fragments of the genes encoding β-giardin and glutamate dehydrogenase were sequenced and their alignment were carried out with sequences deposited in GenBank. As expected for Rio de Janeiro, the majority of samples were related to assemblage A. Surprisingly, assemblage E was detected in 15 samples. Detection of assemblage E in humans suggests a new zoonotic route of Giardia transmission. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  10. Human Memory CD4+ T Cell Immune Responses against Giardia lamblia.

    Science.gov (United States)

    Saghaug, Christina Skår; Sørnes, Steinar; Peirasmaki, Dimitra; Svärd, Staffan; Langeland, Nina; Hanevik, Kurt

    2015-09-16

    The intestinal protozoan parasite Giardia lamblia may cause severe prolonged diarrheal disease or pass unnoticed as an asymptomatic infection. T cells seem to play an important role in the immune response to Giardia infection, and memory responses may last years. Recently, TH17 responses have been found in three animal studies of Giardia infection. The aim of this study was to characterize the human CD4(+) T cell responses to Giardia. Peripheral blood mononuclear cells (PBMCs) were obtained from 21 returning travelers with recent or ongoing giardiasis and 12 low-risk healthy controls and stimulated in vitro with Giardia lamblia proteins. Production of tumor necrosis factor alpha (TNF-α), gamma interferon, interleukin-17A (IL-17A), IL-10, and IL-4 was measured in CD4(+) effector memory (EM) T cells after 24 h by flow cytometry. After 6 days of culture, activation and proliferation were measured by flow cytometry, while an array of inflammatory cytokine levels in supernatants were measured with multiplex assays. We found the number of IL-17A-producing CD4(+) EM T cells, as well as that of cells simultaneously producing both IL-17A and TNF-α, to be significantly elevated in the Giardia-exposed individuals after 24 h of antigen stimulation. In supernatants of PBMCs stimulated with Giardia antigens for 6 days, we found inflammation-associated cytokines, including 1L-17A, as well as CD4(+) T cell activation and proliferation, to be significantly elevated in the Giardia-exposed individuals. We conclude that symptomatic Giardia infection in humans induces a CD4(+) EM T cell response of which IL-17A production seems to be an important component.

  11. Field portable mobile phone based fluorescence microscopy for detection of Giardia lamblia cysts in water samples

    Science.gov (United States)

    Ceylan Koydemir, Hatice; Gorocs, Zoltan; McLeod, Euan; Tseng, Derek; Ozcan, Aydogan

    2015-03-01

    Giardia lamblia is a waterborne parasite that causes an intestinal infection, known as giardiasis, and it is found not only in countries with inadequate sanitation and unsafe water but also streams and lakes of developed countries. Simple, sensitive, and rapid detection of this pathogen is important for monitoring of drinking water. Here we present a cost-effective and field portable mobile-phone based fluorescence microscopy platform designed for automated detection of Giardia lamblia cysts in large volume water samples (i.e., 10 ml) to be used in low-resource field settings. This fluorescence microscope is integrated with a disposable water-sampling cassette, which is based on a flow-through porous polycarbonate membrane and provides a wide surface area for fluorescence imaging and enumeration of the captured Giardia cysts on the membrane. Water sample of interest, containing fluorescently labeled Giardia cysts, is introduced into the absorbent pads that are in contact with the membrane in the cassette by capillary action, which eliminates the need for electrically driven flow for sample processing. Our fluorescence microscope weighs ~170 grams in total and has all the components of a regular microscope, capable of detecting individual fluorescently labeled cysts under light-emitting-diode (LED) based excitation. Including all the sample preparation, labeling and imaging steps, the entire measurement takes less than one hour for a sample volume of 10 ml. This mobile phone based compact and cost-effective fluorescent imaging platform together with its machine learning based cyst counting interface is easy to use and can even work in resource limited and field settings for spatio-temporal monitoring of water quality.

  12. Impaired parasite attachment as fitness cost of metronidazole resistance in Giardia lamblia.

    Science.gov (United States)

    Tejman-Yarden, Noa; Millman, Maya; Lauwaet, Tineke; Davids, Barbara J; Gillin, Frances D; Dunn, Linda; Upcroft, Jacqueline A; Miyamoto, Yukiko; Eckmann, Lars

    2011-10-01

    Infections with the diarrheagenic protozoan pathogen Giardia lamblia are most commonly treated with metronidazole (Mz). Treatment failures with Mz occur in 10 to 20% of cases and Mz resistance develops in the laboratory, yet clinically, Mz-resistant (Mz(r)) G. lamblia has rarely been isolated from patients. To understand why clinical Mz(r) isolates are rare, we questioned whether Mz resistance entails fitness costs to the parasite. Our studies employed several newly generated and established isogenic Mz(r) cell lines with stable, high-level resistance to Mz and significant cross-resistance to tinidazole, nitazoxanide, and furazolidone. Oral infection of suckling mice revealed that three of five Mz(r) cell lines could not establish infection, while two Mz(r) cell lines infected pups, albeit with reduced efficiencies. Failure to colonize resulted from a diminished capacity of the parasite to attach to the intestinal mucosa in vivo and to epithelial cells and plastic surfaces in vitro. The attachment defect was related to impaired glucose metabolism, since the noninfectious Mz(r) lines consumed less glucose, and glucose promoted ATP-independent parasite attachment in the parental lines. Thus, resistance of Giardia to Mz is accompanied by a glucose metabolism-related attachment defect that can interfere with colonization of the host. Because glucose-metabolizing pathways are important for activation of the prodrug Mz, it follows that a fitness trade-off exists between diminished Mz activation and reduced infectivity, which may explain the observed paucity of clinical Mz(r) isolates of Giardia. However, the data also caution that some forms of Mz resistance do not markedly interfere with in vivo infectivity.

  13. Rac Regulates Giardia lamblia Encystation by Coordinating Cyst Wall Protein Trafficking and Secretion

    Directory of Open Access Journals (Sweden)

    Jana Krtková

    2016-08-01

    Full Text Available Encystation of the common intestinal parasite Giardia lamblia involves the production, trafficking, and secretion of cyst wall material (CWM. However, the molecular mechanism responsible for the regulation of these sequential processes remains elusive. Here, we examined the role of GlRac, Giardia’s sole Rho family GTPase, in the regulation of endomembrane organization and cyst wall protein (CWP trafficking. Localization studies indicated that GlRac is associated with the endoplasmic reticulum (ER and the Golgi apparatus-like encystation-specific vesicles (ESVs. Constitutive GlRac signaling increased levels of the ER marker PDI2, induced ER swelling, reduced overall CWP1 production, and promoted the early maturation of ESVs. Quantitative analysis of cells expressing constitutively active hemagglutinin (HA-tagged GlRac (HA-RacCA revealed fewer but larger ESVs than control cells. Consistent with the phenotype of premature maturation of ESVs in HA-RacCA-expressing cells, constitutive GlRac signaling resulted in increased CWP1 secretion and, conversely, morpholino depletion of GlRac blocked CWP1 secretion. Wild-type cells unexpectedly secreted large quantities of CWP1 into the medium, and free CWP1 was used cooperatively during cyst formation. These results, in part, could account for the previously reported observation that G. lamblia encysts more efficiently at high cell densities. These studies of GlRac show that it regulates encystation at several levels, and our findings support its coordinating role as a regulator of CWP trafficking and secretion. The central role of GlRac in regulating membrane trafficking and the cytoskeleton, both of which are essential to Giardia parasitism, further suggests its potential as a novel target for drug development to treat giardiasis.

  14. Giardia lamblia.

    OpenAIRE

    Rodríguez Padilla, Eva María

    2015-01-01

    Las infecciones parasitarias son un verdadero problema para la salud pública especialmente en países con niveles socioeconómicos bajos. La infección por Giardia es la responsable de producir un 2-3% de las diarreas del viajero, muy relacionada también con la diarrea infecciosa no viral en niños. El reservorio de Giardia es el hombre, ya sea enfermo o en estado de portador asintomático, la infección puede ser frecuente entre animales domésticos, mamíferos salvajes y aves....

  15. UPF1, a conserved nonsense-mediated mRNA decay factor, regulates cyst wall protein transcripts in Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Yi-Hsiu Chen

    Full Text Available The Giardia lamblia cyst wall is required for survival outside the host and infection. Three cyst wall protein (cwp genes identified to date are highly up-regulated during encystation. However, little is known of the molecular mechanisms governing their gene regulation. Messenger RNAs containing premature stop codons are rapidly degraded by a nonsense-mediated mRNA decay (NMD system to avoid production of non-functional proteins. In addition to RNA surveillance, NMD also regulates thousands of naturally occurring transcripts through a variety of mechanisms. It is interesting to know the NMD pathway in the primitive eukaryotes. Previously, we have found that the giardial homologue of a conserved NMD factor, UPF1, may be functionally conserved and involved in NMD and in preventing nonsense suppression. In this study, we tested the hypothesis that NMD factors can regulate some naturally occurring transcripts in G. lamblia. We found that overexpression of UPF1 resulted in a significant decrease of the levels of CWP1 and cyst formation and of the endogenous cwp1-3, and myb2 mRNA levels and stability. This indicates that NMD could contribute to the regulation of the cwp1-3 and myb2 transcripts, which are key to G. lamblia differentiation into cyst. Interestingly, we also found that UPF1 may be involved in regulation of eight other endogenous genes, including up-regulation of the translation elongation factor gene, whose product increases translation which is required for NMD. Our results indicate that NMD factor could contribute to the regulation of not only nonsense containing mRNAs, but also mRNAs of the key encystation-induced genes and other endogenous genes in the early-diverging eukaryote, G. lamblia.

  16. Coexistence of sense and anti-sense mRNAs of variant surface protein in Giardia lamblia trophozoites.

    Science.gov (United States)

    Guo, Junli; Zheng, Wenyu; Wang, Yuehua; Li, Yao; Lu, Siqi; Feng, Xianmin

    2014-02-14

    A strategy of the parasitic protozoan Giardia lamblia to evade attack from the host immune system is periodic changes of its surface antigen, a member of the variant surface protein (VSP) family. A post-transcriptional gene silencing mechanism has been proposed to explain the presence of only one among many possible VSPs at any time. To investigate this phenomenon further, we extracted total RNA from cultured trophozoites of the G. lamblia C2 isolate, and cDNA was reverse-transcribed from the RNA. Sense and anti-sense VSPs were amplified from the total cDNA using nested PCR with primers designed from the 3'-conserved region and the known 5' or 3' end of the cDNA library. Sequence analyses of the amplified products revealed more than 34 full-length antisense VSPs and a smear of sense VSPs. Sequence alignments and comparisons revealed that these VSPs contained variable N-termini and conserved C-termini, and could be classified into 5 clades based on the sizes and variations of the N-terminal sequence. All antisense VSPs existed in the sense forms, but no corresponding antisense VSP existed for sense RNA (snsRNA) 16. The coexistence of sense and antisense VSP mRNAs in cultured G. lamblia supports the post-transcriptional regulation of VSP expression. We propose that VSPs transcribed simultaneously in the sense and antisense forms form double-stranded RNAs (dsRNAs) which are degraded by the Dicer endonuclease, while a VSP without an antisense transcription (e.g., snsRNA16) will be expressed on the surface of Giardia. In addition, in the course of this investigation VSPs were identified that were previously not known. PCR-based amplification of specific sense and antisense VSP cDNAs can be used to identify the specific VSP on G. lamblia trophozoites, which is easier than using specific monoclonal antibody approaches.

  17. Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    Jun-Li Guo; Jie Jiang; Wen-Yu Zheng; Ming-Luan Li; Xi-Feng Tian; Xian-Min Feng; Yue-Hua Wang; Xiao-Hong Ju; Yue-Qiong Kong

    2012-01-01

    Objective: To construct rapidly a full-length cDNA library from nanogram amounts total RNA of Giardia lamblia (G. lamblia) trophozoites stocked in RNA stabilization reagent. Methods: Total RNA of Giardia was extracted using Trizol reagent. A full-length cDNA library of G. lamblia trophozoites was constructed by a long-distance PCR (LD-PCR) method. The recombinant rate and the coverage rate of full-length clones of the library were evaluated. The inserted fragments were identified and sequenced by PCR amplification. Results: The titer of cDNA library was 3.85 ×107 pfu/mL. The length of inserted fragments ranged from 0.4 to 2.5 kb, and the recombination efficiency accounted for 100% (20/20). The coverage rate of full-length clones is high (17/20). Conclusions: The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature. The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA.

  18. A novel pax-like protein involved in transcriptional activation of cyst wall protein genes in Giardia lamblia.

    Science.gov (United States)

    Wang, Yi-Ting; Pan, Yu-Jiao; Cho, Chao-Cheng; Lin, Bo-Chi; Su, Li-Hsin; Huang, Yu-Chang; Sun, Chin-Hung

    2010-10-15

    Giardia lamblia differentiates into infectious cysts to survive outside of the host. It is of interest to identify factors involved in up-regulation of cyst wall proteins (CWPs) during this differentiation. Pax proteins are important regulators of development and cell differentiation in Drosophila and vertebrates. No member of this gene family has been reported to date in yeast, plants, or protozoan parasites. We have identified a pax-like gene (pax1) encoding a putative paired domain in the G. lamblia genome. Epitope-tagged Pax1 localized to nuclei during both vegetative growth and encystation. Recombinant Pax1 specifically bound to the AT-rich initiator elements of the encystation-induced cwp1 to -3 and myb2 genes. Interestingly, overexpression of Pax1 increased cwp1 to -3 and myb2 gene expression and cyst formation. Deletion of the C-terminal paired domain or mutation of the basic amino acids of the paired domain resulted in a decrease of the transactivation function of Pax1. Our results indicate that the Pax family has been conserved during evolution, and Pax1 could up-regulate the key encystation-induced genes to regulate differentiation of the protozoan eukaryote, G. lamblia.

  19. Automated 3D detection and classification of Giardia lamblia cysts using digital holographic microscopy with partially coherent source

    Science.gov (United States)

    El Mallahi, A.; Detavernier, A.; Yourassowsky, C.; Dubois, F.

    2012-06-01

    Over the past century, monitoring of Giardia lamblia became a matter of concern for all drinking water suppliers worldwide. Indeed, this parasitic flagellated protozoan is responsible for giardiasis, a widespread diarrhoeal disease (200 million symptomatic individuals) that can lead immunocompromised individuals to death. The major difficulty raised by Giardia lamblia's cyst, its vegetative transmission form, is its ability to survive for long periods in harsh environments, including the chlorine concentrations and treatment duration used traditionally in water disinfection. Currently, there is a need for a reliable, inexpensive, and easy-to-use sensor for the identification and quantification of cysts in the incoming water. For this purpose, we investigated the use of a digital holographic microscope working with partially coherent spatial illumination that reduces the coherent noise. Digital holography allows one to numerically investigate a volume by refocusing the different plane of depth of a hologram. In this paper, we perform an automated 3D analysis that computes the complex amplitude of each hologram, detects all the particles present in the whole volume given by one hologram and refocuses them if there are out of focus using a refocusing criterion based on the integrated complex amplitude modulus and we obtain the (x,y,z) coordinates of each particle. Then the segmentation of the particles is processed and a set of morphological and textures features characteristic to Giardia lamblia cysts is computed in order to classify each particles in the right classes.

  20. A Comparison of Microscopy and Enzyme Linked Immunosorbent Assay for Diagnosis of Giardia lamblia in Human Faecal Specimens.

    Science.gov (United States)

    Jahan, Noor; Khatoon, Razia; Ahmad, Siraj

    2014-11-01

    Giardia lamblia, a flagellate protozoa, is a common causative agent of parasitic diarrhoeal diseases of humans. Laboratory diagnosis mainly consists of direct microscopic examination of stool specimen for trophozoite and cysts of Giardia. However, due to intermittent faecal excretion of parasite, the case may be miss diagnosed and the patient may continue excreting the parasite and infecting others. Therefore, other mode of diagnosis should be looked for, which overcome the above drawbacks of microscopy used alone for diagnosis. The present study was done to evaluate the efficacy of RIDASCREEN Giardia (ELISA) test in comparison to direct microscopy in the diagnosis of Giardia lamblia in stool specimens from patients with diarrhea and other gastrointestinal symptoms. A total of 1680 patients were included in the study and three faecal specimens were taken from each patient which was divided into two parts. One part was used for direct wet mount examination and second part was used to put ELISA by using RIDASCREEN Giardia test. Out of 1680 stool samples, 380 specimens (22.6%) were found to be positive for Giardia lamblia. Maximum cases were detected by RIDASCREEN Giardia (ELISA) test with sensitivity of 100% and specificity of 91.5%. Maximum cases of giardiasis were detected in children less than 10 y of age (12.8%). RIDASCREEN Giardia test is a rapid and effective method with high sensitivity and specificity and detects Giardia antigens in stool specimens even when the count of parasite is low, thus reducing the chances of missing even the asymptomatic cases.

  1. Three-dimensional structure of a protozoal double-stranded RNA virus that infects the enteric pathogen Giardia lamblia.

    Science.gov (United States)

    Janssen, Mandy E W; Takagi, Yuko; Parent, Kristin N; Cardone, Giovanni; Nibert, Max L; Baker, Timothy S

    2015-01-15

    Giardia lamblia virus (GLV) is a small, nonenveloped, nonsegmented double-stranded RNA (dsRNA) virus infecting Giardia lamblia, the most common protozoan pathogen of the human intestine and a major agent of waterborne diarrheal disease worldwide. GLV (genus Giardiavirus) is a member of family Totiviridae, along with several other groups of protozoal or fungal viruses, including Leishmania RNA viruses and Trichomonas vaginalis viruses. Interestingly, GLV is more closely related than other Totiviridae members to a group of recently discovered metazoan viruses that includes penaeid shrimp infectious myonecrosis virus (IMNV). Moreover, GLV is the only known protozoal dsRNA virus that can transmit efficiently by extracellular means, also like IMNV. In this study, we used transmission electron cryomicroscopy and icosahedral image reconstruction to examine the GLV virion at an estimated resolution of 6.0 Å. Its outermost diameter is 485 Å, making it the largest totivirus capsid analyzed to date. Structural comparisons of GLV and other totiviruses highlighted a related "T=2" capsid organization and a conserved helix-rich fold in the capsid subunits. In agreement with its unique capacity as a protozoal dsRNA virus to survive and transmit through extracellular environments, GLV was found to be more thermoresistant than Trichomonas vaginalis virus 1, but no specific protein machinery to mediate cell entry, such as the fiber complexes in IMNV, could be localized. These and other structural and biochemical findings provide a basis for future work to dissect the cell entry mechanism of GLV into a "primitive" (early-branching) eukaryotic host and an important enteric pathogen of humans. Numerous pathogenic bacteria, including Corynebacterium diphtheriae, Salmonella enterica, and Vibrio cholerae, are infected with lysogenic bacteriophages that contribute significantly to bacterial virulence. In line with this phenomenon, several pathogenic protozoa, including Giardia lamblia

  2. Retention of Escherichia coli, Giardia lamblia cysts and Ascaris lumbricoides eggs in agricultural soils irrigated by untreated wastewater.

    Science.gov (United States)

    Landa-Cansigno, O; Durán-Álvarez, J C; Jiménez-Cisneros, B

    2013-10-15

    In central Mexico, agricultural irrigation reusing Mexico City's municipal wastewater has been occurring for the last century, resulting in the recharge of the local aquifer. However, groundwater of this zone is of good quality, indicating that the microorganisms contained in wastewater are retained by soil after infiltration. This study aims to assess the capacity of three agricultural soils to retain three microorganisms frequently found in wastewater, namely Escherichia coli (E. coli), Giardia lamblia (G. lamblia) cysts and Ascaris lumbricoides (A. lumbricoides) eggs, through batch sorption-desorption assays. The tested soils were: an organic-clayey soil (C-OM), a clayey soil (C-om) and a sandy soil (c-om). For the three soils, sorption equilibrium of E. coli was reached before 1 h, while for G. lamblia cysts and A. lumbricoides eggs, sorption equilibrium took 2.5 h. Sorption of E. coli was better described by the Freundlich model than by the Langmuir one. Higher retention of bacteria was observed in the C-om soil (KF = 4340) than in the C-OM and c-om ones (KF = 1821 and 0.01, respectively). Regarding G. lamblia cysts and A. lumbricoides eggs, data could not be fitted to the tested sorption models. For both organisms, retention was lower in the C-OM soil than in the C-om and c-om ones. In the desorption tests, a sudden liberation of E. coli from soils was observed, probably due to bacterial re-growth. Desorption of G. lamblia was higher in the sandy soil than in the clayey ones; desorption was not increased when a surfactant was applied to the soil, suggesting that hydrophobic interactions are not necessarily responsible for retention of the cysts onto the tested soils. For A. lumbricoides eggs, desorption using NaOCl solution suggested that retention was caused by interactions between the mineral fraction of the soil and the external walls of eggs. This study showed that the three target microorganisms are retained by the tested soils and that

  3. Experimental verification of the identity of variant-specific surface proteins in Giardia lamblia trophozoites.

    Science.gov (United States)

    Li, Wei; Saraiya, Ashesh A; Wang, Ching C

    2013-05-21

    The cell membrane of a Giardia lamblia trophozoite is covered with a single species of variant-specific surface protein (VSP) that is replaced by another VSP every 6 to 13 generations of cell growth, possibly for an evasion of host immunity. Experimentally, only six VSP species have been verified to localize to the cell membrane thus far. By assuming that VSP contains multiple CXXC motifs, 219 vsp genes were annotated in GiardiaDB of the WB isolate. By further assuming that VSP possesses both CXXC motifs and a CRGKA tail at the C terminus, Adam et al. (BMC Genomics 11:424, 2010) identified a total of 303 potential vsp genes in Giardia WB. The discrepancies between these two assumed VSP identities have caused some confusion. Here, we used experimental approaches to further verify what is required of the structures of a VSP to localize to the surface of cell membrane. The data led to the following conclusions. (i) The C-terminal CRGKA sequence is not essential for localizing VSPs to the cell membrane. (ii) A "motif 1" of 45 residues, consisting of two CXXCs separated by 12 to 15 amino acid residues, located close to the C terminus and a hydrophobic "motif 2" of 38 residues at the C terminus are both essential and sufficient for localizing the protein to the cell membrane. (ii) An N-terminal sequence upstream from motif 1 is not required for targeting VSPs to the cell membrane. By these criteria, we are able to identify 73 open reading frames as the putative vsp genes in Giardia. IMPORTANCE The intestinal pathogen Giardia lamblia expresses only one variant-specific surface protein (VSP) on the cell membrane surface at a given time, but it changes spontaneously every 6 to 13 generations of growth, presumably for evading the host immunity. Only 6 VSPs have been empirically shown to localize to the cell membrane surface thus far. Here, we used mutations of VSPs and methods of identifying their locations in Giardia cells and found that a "motif 1" of 45 residues

  4. Excretory-secretory products of Giardia lamblia induce interleukin-8 production in human colonic cells via activation of p38, ERK1/2, NF-κB and AP-1.

    Science.gov (United States)

    Lee, H-Y; Hyung, S; Lee, N Y; Yong, T-S; Han, S-H; Park, S-J

    2012-04-01

    Giardia lamblia, a pathogen causing diarrhoeal outbreaks, is interesting how it triggers immune response in the human epithelial cells. This study defined the crucial roles of signalling components involved in G. lamblia-induced cytokine production in human epithelial cells. Incubation of the gastrointestinal cell line HT-29 with G. lamblia GS trophozoites triggered production of interleukin (IL)-1β, IL-8 and tumour necrosis factor (TNF)-α. IL-8 production was not significantly decreased by physically separating the HT-29 cells and G. lamblia GS trophozoites. Indeed, treatment of HT-29 with G. lamblia excretory-secretory products (ESP) induced IL-8 production. Electrophoretic mobility gel shift and transfection assays using mutagenized IL-8 promoter reporter plasmids indicated that IL-8 production by G. lamblia ESP occurs through activation of two transcriptional factors, nuclear factor kappaB (NF-κB) and activator protein 1 (AP-1) in HT-29 cells. In addition, activation of two mitogen-activated protein kinases (MAPKs), p38 and ERK1/2, was also detected in the HT-29 cells stimulated with G. lamblia ESP. Selective inhibition of these MAPKs resulted in decreased production of ESP-induced IL-8. These results indicate that activation of p38, ERK1/2 MAPK, NF-κB and AP-1 comprises the signalling pathway responsible for IL-8 production by G. lamblia ESP.

  5. Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife.

    Science.gov (United States)

    Bednarska, Malgorzata; Bajer, Anna; Sinski, Edward; Girouard, Autumn S; Tamang, Leena; Graczyk, Thaddeus K

    2007-02-01

    Fecal samples of five terrestrial mammalian wildlife species stored at 4 degrees C or at -20 degrees C for up to 36 months have been tested for human zoonotic enteric parasites (i.e., Cryptosporidium parvum and Giardia lamblia) using combined fluorescent in situ hybridization (FISH) and direct fluorescent antibody techniques. The prevalence of C. parvum and G. lamblia varied from 20 to 63% (mean, 45.8%) and from 13 to 100% (mean, 53.2%), respectively. The prevalence of C. parvum and G. lamblia infections was higher in small rodents (mean, 68.5%) than in other wildlife (mean, 21%). Overall, 31.1% of animals were coinfected, and coinfections were more prevalent in small rodents (mean, 52%) than in other wildlife species (mean, 13.2%). The present study has shown that the FISH assay can be retrospectively applied to fecal samples for the identification of C. parvum oocysts, but is less suitable for the identification of G. lamblia cysts in such samples. Terrestrial mammalian wildlife, particularly small rodents, can contribute to watershed contamination with C. parvum oocysts and G. lamblia cysts. To control contamination, the management of pristine watersheds used for drinking water purposes should incorporate control measures for terrestrial wildlife, especially field rodents residing within such watersheds.

  6. High-speed microscopic imaging of flagella motility and swimming in Giardia lamblia trophozoites.

    Science.gov (United States)

    Lenaghan, Scott C; Davis, Corinne A; Henson, William R; Zhang, Zhili; Zhang, Mingjun

    2011-08-23

    We report, in this paper, several findings about the swimming and attachment mechanisms of Giardia lamblia trophozoites. These data were collected using a combination of a high-contrast CytoViva imaging system and a particle image velocimetry camera, which can capture images at speeds greater than 800 frames/s. Using this system, we discovered that, during rapid swimming of Giardia trophozoites, undulations of the caudal region contributed to forward propulsion combined with the beating of the flagella pairs. It was also discovered, in contrast to previous studies with 10 times slower image sampling technique, that the anterior and posterolateral flagella beat with a clearly defined power stroke and not symmetrical undulations. During the transition from free swimming to attachment, trophozoites modified their swimming behavior from a rapid rotating motion to a more stable planar swimming. While using this planar swimming motion, the trophozoites used the flagella for propulsion and directional control. In addition to examination of the posterolateral and anterior flagella, a model to describe the motion of the ventral flagella was derived, indicating that the ventral flagella beat in an expanding sine wave. In addition, the structure of the ventrocaudal groove creates boundary conditions that determine the form of beating of the ventral flagella. The results from this study indicate that Giardia is able to simultaneously generate both ciliary beating and typical eukaryotic flagellar beating using different pairs of flagella.

  7. What Entamoeba histolytica and Giardia lamblia tell us about the evolution of eukaryotic diversity

    Indian Academy of Sciences (India)

    J Samuelson

    2002-11-01

    Entamoeba histolytica and Giardia lamblia are microaerophilic protists, which have long been considered models of ancient pre-mitochondriate eukaryotes. As transitional eukaryotes, amoebae and giardia appeared to lack organelles of higher eukaryotes and to depend upon energy metabolism appropriate for anaerobic conditions, early in the history of the planet. However, our studies have shown that amoebae and giardia contain splicoeosomal introns, ras-family signal-transduction proteins, ATP-binding casettes (ABC)-family drug transporters, Golgi, and a mitochondrion-derived organelle (amoebae only). These results suggest that most of the organelles of higher eukaryotes were present in the common ancestor of all eukaryotes, and so dispute the notion of transitional eukaryotic forms. In addition, phylogenetic studies suggest many of the genes encoding the fermentation enzymes of amoebae and giardia derive from prokaryotes by lateral gene transfer (LGT). While LGT has recently been shown to be an important determinant of prokaryotic evolution, this is the first time that LGT has been shown to be an important determinant of eukaryotic evolution. Further, amoebae contain cyst wall-associated lectins, which resemble, but are distinct from lectins in the walls of insects (convergent evolution). Giardia have a novel microtubule-associated structure which tethers together pairs of nuclei during cell division. It appears then that amoebae and giardia tell us less about the origins of eukaryotes and more about the origins of eukaryotic diversity.

  8. Bax function in the absence of mitochondria in the primitive protozoan Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Adrian B Hehl

    Full Text Available Bax-induced permeabilization of the mitochondrial outer membrane and release of cytochrome c are key events in apoptosis. Although Bax can compromise mitochondria in primitive unicellular organisms that lack a classical apoptotic machinery, it is still unclear if Bax alone is sufficient for this, or whether additional mitochondrial components are required. The protozoan parasite Giardia lamblia is one of the earliest branching eukaryotes and harbors highly degenerated mitochondrial remnant organelles (mitosomes that lack a genome. Here we tested whether human Bax expressed in Giardia can be used to ablate mitosomes. We demonstrate that these organelles are neither targeted, nor compromised, by Bax. However, specialized compartments of the regulated secretory pathway are completely ablated by Bax. As a consequence, maturing cyst wall proteins that are sorted into these organelles are released into the cytoplasm, causing a developmental arrest and cell death. Interestingly, this ectopic cargo release is dependent on the carboxy-terminal 22 amino acids of Bax, and can be prevented by the Bax-inhibiting peptide Ku70. A C-terminally truncated Bax variant still localizes to secretory organelles, but is unable to permeabilize these membranes, uncoupling membrane targeting and cargo release. Even though mitosomes are too diverged to be recognized by Bax, off-target membrane permeabilization appears to be conserved and leads to cell death completely independently of mitochondria.

  9. Effects of metronidazole analogues on Giardia lamblia: experimental infection and cell organization.

    Science.gov (United States)

    Busatti, Haendel G N O; Alves, Ricardo J; Santana-Anjos, Karla G; Gil, Frederico F; Cury, Marcia C; Vannier-Santos, Marcos A; Gomes, Maria A

    2013-02-01

    The chemotherapeutic agents used for the treatment of giardiasis are often associated with adverse side effects and are refractory cases, due to the development of resistant parasites. Therefore the search for new drugs is required. We have previously reported the giardicidal effects of metronidazole (MTZ) and its analogues (MTZ-Ms, MTZ-Br, MTZ-N(3), and MTZ-I) on the trophozoites of Giardia lamblia. Now we evaluated the activity of some giardicidal MTZ analogues in experimental infections in gerbils and its effects on the morphology and ultrastructural organization of Giardia. The giardicidal activity in experimental infections showed ED(50) values significantly lower for MTZ-I and MTZ-Br when compared to MTZ. Transmission electron microscopy was employed to approach the mechanism(s) of action of MTZ analogues upon the protozoan. MTZ analogues were more active than MTZ in changing significantly the morphology and ultrastructure of the parasite. The analogues affected parasite cell vesicle trafficking, autophagy, and triggered differentiation into cysts. These results coupled with the excellent giardicidal activity and lower toxicity demonstrate that these nitroimidazole derivates may be important therapeutic alternatives for combating giardiasis. In addition, our results suggest a therapeutic advantage in obtaining synthetic metronidazole analogues for screening of activities against other infectious agents.

  10. Modeling and analysis of propulsion in the multiflagellated micoorganism Giardia lamblia

    Science.gov (United States)

    Lenaghan, Scott C.; Chen, Jun; Zhang, Mingjun

    2013-07-01

    The goal of this work was to analyze the propulsion of multiflagellated microorganisms, and to draw insight to the underlying physics and biology of the movement. Giardia lamblia was chosen as the model organism due to its unique ability to mechanically attach to various surfaces, its rapid movement, and its fine control over steering and navigation. In this work, a mechanics model was utilized to study the mechanics and propulsive contribution of the ventral and anterior flagella in Giardia. It was discovered that energy is supplied mainly at the proximal portion of these flagella, supporting the hypothesis that a decreasing adenosine triphosphate (ATP) gradient along the length of the flagella would not affect the motion observed. Similarly, the elasticity of the flagella allows the energy input at the proximal portion to be transferred to the distal portion, where the majority of thrust is generated. Specifically, we found that the ventral flagella are the driving force for planar propulsion and turning, while the anterior flagella are used for steering and control.

  11. Whipple's disease concomitant with candida esophagitis and subsequent Giardia lamblia coinfection.

    Science.gov (United States)

    Sulis, Giorgia; Villanacci, Vincenzo; Missale, Guido; Salemme, Marianna; Castelli, Francesco; Caligaris, Silvio

    2014-10-01

    Whipple's disease is a rare systemic condition resulting from a chronic infection by Tropheryma whipplei. Clinical presentation can be widely heterogeneous, often leading to delayed diagnosis and treatment. Furthermore, little is known about the underlying pathogenic mechanisms, although several abnormalities in immune cell function have been observed. We describe the case of a 51-year-old woman presenting with dysphagia, odynophagia, long-lasting low-grade fever, and malabsorption syndrome who underwent an upper gastrointestinal endoscopy showing esophageal candidiasis. On the same occasion, duodenal biopsies were also performed, with evidence of mucosal infiltration of periodic acid-Schiff-positive and CD68+ foamy macrophages at microscopic examination. Such findings were suggestive of Whipple's disease, as also confirmed by molecular analysis by PCR for T. whipplei. No specific risk factors were identified in our patient that could explain the occurrence of an opportunistic infection such as candida esophagitis, thus leading to the hypothesis of a direct correlation with Whipple's disease. Interestingly, a Giardia lamblia coinfection was subsequently identified, which is consistent with an underlying immune deficit although still undefined.

  12. Detection of Giardia lamblia and Cryptosporidium parvum by direct immunofluorescence assay in stool specimen.

    Science.gov (United States)

    Rahman, M M; Hossain, M A; Paul, S K; Ahmed, S; Islam, A; Ehsan, M A; Alam, M M; Kabir, M R; Sarkar, S R

    2014-07-01

    Giardia and Cryptosporidium are the pathogens which transmitted through contaminated soil and contaminated water are significant causes of diarrhea and nutritional disorders in institutional and community peoples. Children and immune compromise persons are more vulnerable for these infections. Both Giardiasis and Cryptosporidiosis were included in 2004 as WHO Neglected Disease. So this is a major public health problem in developing countries. The present study was carried out to detect the Giardia and Cryptosporidium from diarrheic or patient having loose stool by Direct Immunofluorescence assay. The study was conducted during July 20012 to February 2013 and the work was done in Mymensingh Medical College in the department of Microbiology and in Bangladesh Agricultural University in the department of Veterinary Medicine. A total of 100 loose stools were collected from school children of different area and hospital under sadar upazilla, Mymensingh. The detection of Giardia lamblia and Cryptosporidium parvum showed the individual prevalence 8% and 4% respectively. The highest cyst/oocyst count was 85,000 and 1,000/gm of stool and the lowest being 100 and 50/gm of stool for Giardiasis and Cryptosporidiosis respectively. The detection rate of Giardia and Cryptosporidium by Immunofluorescence assay was relatively higher than the previous study done in Bangladesh and this was the first report from Bangladesh over human stool specimen using Immunofluorescence assay. So, Immunofluorescence assay could be adapted for rapid and accurate detection of Giardia and Cryptosporidium.

  13. Protein SUMOylation is Involved in Cell-cycle Progression and Cell Morphology in Giardia lamblia.

    Science.gov (United States)

    Di Genova, Bruno M; da Silva, Richard C; da Cunha, Júlia P C; Gargantini, Pablo R; Mortara, Renato A; Tonelli, Renata R

    2017-07-01

    The unicellular protozoa Giardia lamblia is a food- and waterborne parasite that causes giardiasis. This illness is manifested as acute and self-limited diarrhea and can evolve to long-term complications. Successful establishment of infection by Giardia trophozoites requires adhesion to host cells and colonization of the small intestine, where parasites multiply by mitotic division. The tight binding of trophozoites to host cells occurs by means of the ventral adhesive disc, a spiral array of microtubules and associated proteins such as giardins. In this work we show that knock down of the Small Ubiquitin-like MOdifier (SUMO) results in less adhesive trophzoites, decreased cell proliferation and deep morphological alterations, including at the ventral disc. Consistent with the reduced proliferation, SUMO knocked-down trophozoites were arrested in G1 and in S phases of the cell cycle. Mass spectrometry analysis of anti-SUMO immunoprecipitates was performed to identify SUMO substrates possibly involved in these events. Among the identified SUMOylation targets, α-tubulin was further validated by Western blot and confirmed to be a SUMO target in Giardia trophozoites. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

  14. The minimal kinome of Giardia lamblia illuminates early kinase evolution and unique parasite biology

    Science.gov (United States)

    2011-01-01

    Background The major human intestinal pathogen Giardia lamblia is a very early branching eukaryote with a minimal genome of broad evolutionary and biological interest. Results To explore early kinase evolution and regulation of Giardia biology, we cataloged the kinomes of three sequenced strains. Comparison with published kinomes and those of the excavates Trichomonas vaginalis and Leishmania major shows that Giardia's 80 core kinases constitute the smallest known core kinome of any eukaryote that can be grown in pure culture, reflecting both its early origin and secondary gene loss. Kinase losses in DNA repair, mitochondrial function, transcription, splicing, and stress response reflect this reduced genome, while the presence of other kinases helps define the kinome of the last common eukaryotic ancestor. Immunofluorescence analysis shows abundant phospho-staining in trophozoites, with phosphotyrosine abundant in the nuclei and phosphothreonine and phosphoserine in distinct cytoskeletal organelles. The Nek kinase family has been massively expanded, accounting for 198 of the 278 protein kinases in Giardia. Most Neks are catalytically inactive, have very divergent sequences and undergo extensive duplication and loss between strains. Many Neks are highly induced during development. We localized four catalytically active Neks to distinct parts of the cytoskeleton and one inactive Nek to the cytoplasm. Conclusions The reduced kinome of Giardia sheds new light on early kinase evolution, and its highly divergent sequences add to the definition of individual kinase families as well as offering specific drug targets. Giardia's massive Nek expansion may reflect its distinctive lifestyle, biphasic life cycle and complex cytoskeleton. PMID:21787419

  15. Eight unique basal bodies in the multi-flagellated diplomonad Giardia lamblia.

    Science.gov (United States)

    McInally, Shane G; Dawson, Scott C

    2016-01-01

    Giardia lamblia is an intestinal parasitic protist that causes significant acute and chronic diarrheal disease worldwide. Giardia belongs to the diplomonads, a group of protists in the supergroup Excavata. Diplomonads are characterized by eight motile flagella organized into four bilaterally symmetric pairs. Each of the eight Giardia axonemes has a long cytoplasmic region that extends from the centrally located basal body before exiting the cell body as a membrane-bound flagellum. Each basal body is thus unique in its cytological position and its association with different cytoskeletal features, including the ventral disc, axonemes, and extra-axonemal structures. Inheritance of these unique and complex cytoskeletal elements is maintained through basal body migration, duplication, maturation, and their subsequent association with specific spindle poles during cell division. Due to the complex composition and inheritance of specific basal bodies and their associated structures, Giardia may require novel basal body-associated proteins. Thus, protists such as Giardia may represent an undiscovered source of novel basal body-associated proteins. The development of new tools that make Giardia genetically tractable will enable the composition, structure, and function of the eight basal bodies to be more thoroughly explored.

  16. Amplification, expression, and packaging of a foreign gene by giardiavirus in Giardia lamblia.

    Science.gov (United States)

    Yu, D C; Wang, A L; Wang, C C

    1996-12-01

    Giardia lamblia is an intestinal protozoan parasite and one of the earliest eukaryotic divergents. The trophozoite multiplies via asexual binary fission and lacks all natural means of lateral gene transfer. A system is developed here for long-term expression of a foreign gene in this organism by exploiting recombinant virions derived from the giardiavirus (GLV), a double-stranded RNA virus that infects many Giardia isolates. An in vitro transcript of the cloned GLV cDNA, comprising the firefly luciferase-encoding region flanked by 5' and 3' fragments of GLV positive-strand RNA, was electroporated into GLV-infected trophozoites. Luciferase activity in electroporated cells peaked on day 2 at levels 6 orders of magnitude above background. Expression of this foreign gene remained at 80% of its peak level after 30 days in the absence of selective pressure. The chimeric RNA was replicated as double-stranded RNA and packaged into virus-like particles. The recombinant virions were partially purified from the wild-type helper virus by CsCl equilibrium density-gradient centrifugation and used to superinfect Giardia trophozoites. At multiplicities of infection of 100 or higher, these chimeric virions were able to initiate new rounds of expression of luciferase activity in the superinfected cells. Thus, the engineered virion can be successfully used to introduce and efficiently express a heterologous gene in this eukaryotic microorganism.

  17. Assembly and export of a Toxoplasma microneme complex in Giardia lamblia.

    Science.gov (United States)

    Gaechter, Verena; Hehl, Adrian B

    2005-11-01

    The microneme proteins of Toxoplasma gondii belong to a large family of adhesins of apicomplexan parasites involved in motility and host cell invasion. During secretory transport, soluble micronemes associate with membrane-bound carriers/escorters and become exposed on the parasite surface as complexes with an array of adhesive domains. Previously, we have exploited the intestinal protozoan Giardia lamblia as an expression system to produce correctly folded and unglycosylated monomeric surface proteins of T. gondii. Here, we report assembly and export of a trimeric microneme (MIC1/4/6) adhesin complex from Toxoplasma. Co-expressed, recombinant microneme proteins were used to investigate structural requirements for microneme complex formation. In addition, export of a microneme subunit induced development of novel Golgi-like compartments demonstrating the existence of post endoplasmic reticulum structures involved in constitutive secretion in this 'Golgi-less' cell. Recreation of the trimeric microneme escorter-cargo system in Giardia is a versatile tool to analyse universal requirements for complex assembly, receptor-ligand interactions and Golgi neogenesis in the basal Giardia secretory system.

  18. The Giardia lamblia vsp gene repertoire: characteristics, genomic organization, and evolution

    Directory of Open Access Journals (Sweden)

    Nash Theodore E

    2010-07-01

    Full Text Available Abstract Background Giardia lamblia trophozoites colonize the intestines of susceptible mammals and cause diarrhea, which can be prolonged despite an intestinal immune response. The variable expression of the variant-specific surface protein (VSP genes may contribute to this prolonged infection. Only one is expressed at a time, and switching expression from one gene to another occurs by an epigenetic mechanism. Results The WB Giardia isolate has been sequenced at 10× coverage and assembled into 306 contigs as large as 870 kb in size. We have used this assembly to evaluate the genomic organization and evolution of the vsp repertoire. We have identified 228 complete and 75 partial vsp gene sequences for an estimated repertoire of 270 to 303, making up about 4% of the genome. The vsp gene diversity includes 30 genes containing tandem repeats, and 14 vsp pairs of identical genes present in either head to head or tail to tail configurations (designated as inverted pairs, where the two genes are separated by 2 to 4 kb of non-coding DNA. Interestingly, over half the total vsp repertoire is present in the form of linear gene arrays that can contain up to 10 vsp gene members. Lastly, evidence for recombination within and across minor clades of vsp genes is provided. Conclusions The data we present here is the first comprehensive analysis of the vsp gene family from the Genotype A1 WB isolate with an emphasis on vsp characterization, function, evolution and contributions to pathogenesis of this important pathogen.

  19. Modeling and analysis of propulsion in the multiflagellated micoorganism Giardia lamblia.

    Science.gov (United States)

    Lenaghan, Scott C; Chen, Jun; Zhang, Mingjun

    2013-07-01

    The goal of this work was to analyze the propulsion of multiflagellated microorganisms, and to draw insight to the underlying physics and biology of the movement. Giardia lamblia was chosen as the model organism due to its unique ability to mechanically attach to various surfaces, its rapid movement, and its fine control over steering and navigation. In this work, a mechanics model was utilized to study the mechanics and propulsive contribution of the ventral and anterior flagella in Giardia. It was discovered that energy is supplied mainly at the proximal portion of these flagella, supporting the hypothesis that a decreasing adenosine triphosphate (ATP) gradient along the length of the flagella would not affect the motion observed. Similarly, the elasticity of the flagella allows the energy input at the proximal portion to be transferred to the distal portion, where the majority of thrust is generated. Specifically, we found that the ventral flagella are the driving force for planar propulsion and turning, while the anterior flagella are used for steering and control.

  20. A contiguous compartment functions as endoplasmic reticulum and endosome/lysosome in Giardia lamblia.

    Science.gov (United States)

    Abodeely, Marla; DuBois, Kelly N; Hehl, Adrian; Stefanic, Sasa; Sajid, Mohammed; DeSouza, Wanderley; Attias, Marcia; Engel, Juan C; Hsieh, Ivy; Fetter, Richard D; McKerrow, James H

    2009-11-01

    The dynamic evolution of organelle compartmentalization in eukaryotes and how strictly compartmentalization is maintained are matters of ongoing debate. While the endoplasmic reticulum (ER) is classically envisioned as the site of protein cotranslational translocation, it has recently been proposed to have pluripotent functions. Using transfected reporter constructs, organelle-specific markers, and functional enzyme assays, we now show that in an early-diverging protozoan, Giardia lamblia, endocytosis and subsequent degradation of exogenous proteins occur in the ER or in an adjacent and communicating compartment. The Giardia endomembrane system is simple compared to those of typical eukaryotes. It lacks peroxisomes, a classical Golgi apparatus, and canonical lysosomes. Giardia orthologues of mammalian lysosomal proteases function within an ER-like tubulovesicular compartment, which itself can dynamically communicate with clathrin-containing vacuoles at the periphery of the cell to receive endocytosed proteins. These primitive characteristics support Giardia's proposed early branching and could serve as a model to study the compartmentalization of endocytic and lysosomal functions into organelles distinct from the ER. This system also may have functional similarity to the retrograde transport of toxins and major histocompatibility complex class I function in the ER of mammals.

  1. Comparison between immunomagnetic separation, coupled with immunofluorescence, and the techniques of Faust et al. and of Lutz for the diagnosis of Giardia lamblia cysts in human feces

    Directory of Open Access Journals (Sweden)

    Souza Doris Sobral Marques

    2003-01-01

    Full Text Available In the present study, the performance of Immunomagnetic Separation technique, coupled with Immunofluorescence (IMS-IFA, was compared with the FAUST et al. and Lutz parasitological techniques for the detection of Giardia lamblia cysts in human feces. One hundred and twenty-seven samples were evaluated by the three techniques at the same time showing a rate of cyst detection of 27.5% by IMS-IFA and 15.7% by both Faust et al. and Lutz techniques. Data analysis showed a higher sensitivity of IMS-IFA for the detection of G. lamblia cysts in comparison with the techniques of FAUST et al. and Lutz. The use of this methodology as a routine procedure enables the processing of many samples simultaneously, in order to increase recovery rate of G. lamblia cysts and reduce the time of sample storage.

  2. Epidemiología molecular de Giardia lamblia en comunidades urbanas y rurales de Buenos Aires y Mendoza, Argentina

    OpenAIRE

    Molina, Nora Beatriz

    2009-01-01

    Giardia lamblia (G. duodenalis; G. intestinalis), es un parásito del intestino delgado que afecta principalmente a niños y produce cuadros clínicos de variada intensidad desde la infección asintomática hasta cuadros graves de diarrea crónica con síndrome de malabsorción. Si bien, en los últimos años se ha incrementado el conocimiento de la fisiopatogenia de la infección por Giardia, hasta el momento, los factores que determinan la variabilidad de las manifestaciones clínicas de giardiosis no ...

  3. Macrophages expressing arginase 1 and nitric oxide synthase 2 accumulate in the small intestine during Giardia lamblia infection.

    Science.gov (United States)

    Maloney, Jenny; Keselman, Aleksander; Li, Erqiu; Singer, Steven M

    2015-06-01

    Nitric oxide (NO) has been shown to inhibit Giardia lamblia in vitro and in vivo. This study sought to determine if Giardia infection induces arginase 1 (ARG1) expression in host macrophages to reduce NO production. Stimulations of RAW 264.7 macrophage-like cells with Giardia extract induced arginase activity. Real-time PCR and immunohistochemistry showed increased ARG1 and nitric oxide synthase 2 (NOS2) expression in mouse intestine following infection. Flow cytometry demonstrated increased numbers of macrophages positive for both ARG1 and NOS2 in lamina propria following infection, but there was no evidence of increased expression of ARG1 in these cells.

  4. Molecular Characterization of Giardia lamblia: First Report of Assemblage B in Human Isolates from Rio de Janeiro (Brazil).

    Science.gov (United States)

    Faria, Clarissa Perez; Zanini, Graziela Maria; Dias, Gisele Silva; da Silva, Sidnei; Sousa, Maria do Céu

    2016-01-01

    Despite the high prevalence of giardiasis, the genetic characterization of Giardia lamblia has been poorly documented in Brazil and molecular epidemiology research has only been conducted in the last few years. The aim of this study was to determine the prevalence of different G. lamblia assemblages and detect mixed infections among patients with giardiasis. The cross-section survey was conducted among patients attending the FIOCRUZ in Rio de Janeiro. In order to discriminate the genetic assemblages/sub-assemblages, G. lamblia isolates were characterized by PCR-RFLP and qPCR using four loci genes (bg, gdh, tpi and orfC4). Of the 65 positive samples, 41 (63.1%) were successfully amplified by nested-PCR of bg and gdh genes. Among them, 16 were typed as sub-assemblage AII, 7 as BIII, 4 as BIV and 8 as a mixture of BIII and BIV. After the analysis by qPCR assay, a total of 55 (84.6%) samples were amplified using at least one locus: bg gene was amplified in 38 (58.5%) samples, gdh in 41 (63.1%), tpi in 39 (60%), and orfC4 in 39 (60%). Multilocus genotyping results showed that 29 (52.7%) samples belonged to Assemblage A and 26 (47.3%) samples belonged to Assemblage B. In 2011 and 2012, 20 (74.1%) samples belonged to Assemblage A and 7 (25.9%) belonged to Assemblage B. In subsequent years (2013-2015) there was a predominance of Assemblage B, 19 (67.9%) versus 9 (32.1%) Assemblage A. This is the first time that Assemblage B of G. lamblia was reported in human clinical samples from Rio de Janeiro (Brazil) and is the first report about genetic characterization using four genes. The qPCR assemblage-specific showed no mixed infections by Assemblages A and B. A switch in genetic profile over the years was observed, firstly predominance of Assemblage A and lastly of Assemblage B.

  5. Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system.

    Science.gov (United States)

    Kumar, Thulasi; Abd Majid, Mohamad Azlan; Onichandran, Subashini; Jaturas, Narong; Andiappan, Hemah; Salibay, Cristina C; Tabo, Hazel A L; Tabo, Norbel; Dungca, Julieta Z; Tangpong, Jitbanjong; Phiriyasamith, Sucheep; Yuttayong, Boonyaorn; Polseela, Raxsina; Do, Binh Nhu; Sawangjaroen, Nongyao; Tan, Tian-Chye; Lim, Yvonne A L; Nissapatorn, Veeranoot

    2016-01-13

    Access to clean and safe drinking water that is free from pathogenic protozoan parasites, especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans, is still an issue in Southeast Asia (SEA). This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA, using real-time polymerase chain reaction (qPCR) assays. A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia (53), Thailand (120), the Philippines (33), and Vietnam (15). A physicochemical analysis was conducted. The water samples were processed in accordance with the US Environmental Protection Agency's methods 1622/1623.1, microscopically observed and subsequently screened using qPCR assays. Cryptosporidium oocysts were detected in treated water samples from the Philippines (1/10), with a concentration of 0.06 ± 0.19 oocyst/L, and untreated water samples from Thailand (25/93), Malaysia (17/44), and the Philippines (11/23), with concentrations ranging from 0.13 ± 0.18 to 0.57 ± 1.41 oocyst/L. Giardia cysts were found in treated water samples from the Philippines (1/10), with a concentration of 0.02 ± 0.06 cyst/L, and in untreated water samples from Thailand (20/93), Vietnam (5/10), Malaysia (22/44), and the Philippines (16/23), with concentrations ranging from 0.12 ± 0.3 to 8.90 ± 19.65 cyst/L. The pathogens C. parvum and G. lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene, respectively. C. parvum was detected in untreated water samples from the Philippines (1/23) and Malaysia (2/44), whilst, G. lamblia detected was detected in treated water samples from the Philippines (1/10) and in untreated water samples from Thailand (21/93), Malaysia (12/44), and the Philippines (17/23). Nitrate concentration was found to have a high positive correlation with (oo)cyst (0.993). The presence of

  6. Selective condensation drives partitioning and sequential secretion of cyst wall proteins in differentiating Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Christian Konrad

    2010-04-01

    Full Text Available Controlled secretion of a protective extracellular matrix is required for transmission of the infective stage of a large number of protozoan and metazoan parasites. Differentiating trophozoites of the highly minimized protozoan parasite Giardia lamblia secrete the proteinaceous portion of the cyst wall material (CWM consisting of three paralogous cyst wall proteins (CWP1-3 via organelles termed encystation-specific vesicles (ESVs. Phylogenetic and molecular data indicate that Diplomonads have lost a classical Golgi during reductive evolution. However, neogenesis of ESVs in encysting Giardia trophozoites transiently provides basic Golgi functions by accumulating presorted CWM exported from the ER for maturation. Based on this "minimal Golgi" hypothesis we predicted maturation of ESVs to a trans Golgi-like stage, which would manifest as a sorting event before regulated secretion of the CWM. Here we show that proteolytic processing of pro-CWP2 in maturing ESVs coincides with partitioning of CWM into two fractions, which are sorted and secreted sequentially with different kinetics. This novel sorting function leads to rapid assembly of a structurally defined outer cyst wall, followed by slow secretion of the remaining components. Using live cell microscopy we find direct evidence for condensed core formation in maturing ESVs. Core formation suggests that a mechanism controlled by phase transitions of the CWM from fluid to condensed and back likely drives CWM partitioning and makes sorting and sequential secretion possible. Blocking of CWP2 processing by a protease inhibitor leads to mis-sorting of a CWP2 reporter. Nevertheless, partitioning and sequential secretion of two portions of the CWM are unaffected in these cells. Although these cysts have a normal appearance they are not water resistant and therefore not infective. Our findings suggest that sequential assembly is a basic architectural principle of protective wall formation and requires

  7. Regulation of a Myb transcription factor by cyclin-dependent kinase 2 in Giardia lamblia.

    Science.gov (United States)

    Cho, Chao-Cheng; Su, Li-Hsin; Huang, Yu-Chang; Pan, Yu-Jiao; Sun, Chin-Hung

    2012-02-03

    The protozoan Giardia lamblia parasitizes the human small intestine to cause diseases. It undergoes differentiation into infectious cysts by responding to intestinal stimulation. How the activated signal transduction pathways relate to encystation stimulation remain largely unknown. During encystation, genes encoding cyst wall proteins (CWPs) are coordinately up-regulated by a Myb2 transcription factor. Because cell differentiation is linked to cell cycle regulation, we tried to understand the role of cell cycle regulators, cyclin-dependent kinases (Cdks), in encystation. We found that the recombinant Myb2 was phosphorylated by Cdk-associated complexes and the levels of phosphorylation increased significantly during encystation. We have identified a putative cdk gene (cdk2) by searching the Giardia genome database. Cdk2 was found to localize in the cytoplasm with higher expression during encystation. Interestingly, overexpression of Cdk2 resulted in a significant increase of the levels of cwp gene expression and cyst formation. In addition, the Cdk2-associated complexes can phosphorylate Myb2 and the levels of phosphorylation increased significantly during encystation. Mutations of important catalytic residues of Cdk2 resulted in a significant decrease of kinase activity and ability of inducing cyst formation. Addition of a Cdk inhibitor, purvalanol A, significantly decreased the Cdk2 kinase activity and the levels of cwp gene expression and cyst formation. Our results suggest that the Cdk2 pathway may be involved in phosphorylation of Myb2, leading to activation of the Myb2 function and up-regulation of cwp genes during encystation. The results provide insights into the use of Cdk inhibitory drugs in disruption of Giardia differentiation into cysts.

  8. A detailed look at the cytoskeletal architecture of the Giardia lamblia ventral disc.

    Science.gov (United States)

    Brown, Joanna R; Schwartz, Cindi L; Heumann, John M; Dawson, Scott C; Hoenger, Andreas

    2016-04-01

    Giardia lamblia is a protistan parasite that infects and colonizes the small intestine of mammals. It is widespread and particularly endemic in the developing world. Here we present a detailed structural study by 3-D negative staining and cryo-electron tomography of a unique Giardia organelle, the ventral disc. The disc is composed of a regular array of microtubules and associated sheets, called microribbons that form a large spiral, held together by a myriad of mostly unknown associated proteins. In a previous study we analyzed by cryo-electron tomography the central microtubule portion (here called disc body) of the ventral disc and found a large portion of microtubule associated inner (MIPs) and outer proteins (MAPs) that render these microtubules hyper-stable. With this follow-up study we expanded our 3-D analysis to different parts of the disc such as the ventral and dorsal areas of the overlap zone, as well as the outer disc margin. There are intrinsic location-specific characteristics in the composition of microtubule-associated proteins between these regions, as well as large differences between the overall architecture of microtubules and microribbons. The lateral packing of microtubule-microribbon complexes varies substantially, and closer packing often comes with contracted lateral tethers that seem to hold the disc together. It appears that the marginal microtubule-microribbon complexes function as outer, laterally contractible lids that may help the cell to clamp onto the intestinal microvilli. Furthermore, we analyzed length, quantity, curvature and distribution between different zones of the disc, which we found to differ from previous publications.

  9. Crystal structure of a macrophage migration inhibitory factor from Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Buchko, Garry W.; Abendroth, Jan; Robinson, Howard; Zhang, Yanfeng; Hewitt, Stephen N.; Edwards, Tom E.; Van Voorhis, Wesley C.; Myler, Peter J.

    2013-06-15

    Macrophage migration inhibitory factor (MIF) is a eukaryotic cytokine that affects a broad spectrum of immune responses and its activation/inactivation is associated with numerous diseases. During protozoan infections MIF is not only expressed by the host, but, has also been observed to be expressed by some parasites and released into the host. To better understand the biological role of parasitic MIF proteins, the crystal structure of the MIF protein from Giardia lamblia (Gl-MIF), the etiological agent responsible for giardiasis, has been determined at 2.30 Å resolution. The 114-residue protein adopts an α/β fold consisting of a four-stranded β-sheet with two anti-parallel α-helices packed against a face of the β-sheet. An additional short β-strand aligns anti-parallel to β4 of the β-sheet in the adjacent protein unit to help stabilize a trimer, the biologically relevant unit observed in all solved MIF crystal structures to date, and form a discontinuous β-barrel. The structure of Gl-MIF is compared to the MIF structures from humans (Hs-MIF) and three Plasmodium species (falciparum, berghei, and yoelii). The structure of all five MIF proteins are generally similar with the exception of a channel that runs through the center of each trimer complex. Relative to Hs-MIF, there are differences in solvent accessibility and electrostatic potential distribution in the channel of Gl-MIF and the Plasmodium-MIFs due primarily to two “gate-keeper” residues in the parasitic MIFs. For the Plasmodium MIFs the gate-keeper residues are at positions 44 (Y==>R) and 100 (V==>D) and for Gl-MIF it is at position 100 (V==>R). If these gate-keeper residues have a biological function and contribute to the progression of parasitemia they may also form the basis for structure-based drug design targeting parasitic MIF proteins.

  10. Structural and evolutionary characteristics of pyruvate phosphate dikinase in Giardia lamblia and other amitochondriate protozoa

    Institute of Scientific and Technical Information of China (English)

    Feng Xianmin; Yang Chunlin; Zheng Wenyu; Wen Jianfan

    2014-01-01

    Background Pyruvate phosphate dikinase (PPDK) reversibly catalyzes the interconversion of phosphoenolpyruvate (PEP) and pyruvic acid,leading to catabolism and adenosine triphosphate (ATP) synthesis or gluconeogenesis and ATP consumption.Molecular modeling of PPDKs from divergent organisms demonstrates that the orientation of the phosphorylatable histidine residue within the central domain of PPDK determines whether this enzyme promotes catabolism or gluconeogenesis.The goal of this study was to determine whether PDDK from Giardia underwent adaptive evolution in order to produce more energy under anaerobic conditions.Methods A total of 123 PPDK sequences from protozoans,proteobacteria,plants,and algae were selected,based upon sequence similarities to Giardia lamblia PPDK and Zea mays PPDK.Three-dimensional (3-D) models were generated for PPDKs from divergent organisms and were used to compare the orientation of the phosphorylatable histidine residue within the central domain of PPDKs.These PPDKs were compared using a maximum-likelihood tree.Results For PPDK from Giardia,as well as from other anaerobic protozoans,the central domain tilted toward the N-terminal nucleotide-binding domain,indicating that this enzyme catalyzed ATP synthesis.Furthermore,the orientation of this central domain was determined by interactions between the N-and C-terminal domains.Phylogenetic analysis of the N-and C-terminal sequences of PPDKs from different species suggested that PPDK has likely undergone adaptive evolution in response to differences in environmental and metabolic conditions.Conclusion These results suggested that PPDK in anaerobic organisms is functionally adapted to generate energy more efficiently in an anaerobic environment.

  11. Serum sickness

    Science.gov (United States)

    ... the problem should be stopped. Avoid using that medicine or antiserum in the future. ... Call your provider if you received medicine or antiserum in the last 4 weeks and have symptoms of serum sickness.

  12. Expression, purification, crystallization and preliminary X-ray diffraction analysis of α-11 giardin from Giardia lamblia

    Energy Technology Data Exchange (ETDEWEB)

    Pathuri, Puja; Nguyen, Emily Tam [Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697 (United States); Luecke, Hartmut, E-mail: hudel@uci.edu [Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697 (United States); Department of Physiology and Biophysics, University of California, Irvine, CA 92697 (United States); Department of Information and Computer Sciences, University of California, Irvine, CA 92697 (United States)

    2006-11-01

    α-11 giardin from the intestinal protozoan parasite, G. lamblia has been cloned, expressed, purified and crystallized under two different conditions and in two different space groups. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group and crystals obtained in the second condition diffracted to 2.93 Å and belong to a primitive monoclinic space group. α-11 Giardin, a protein from the annexin superfamily, is a 35.0 kDa protein from the intestinal protozoan parasite Giardia lamblia which triggers a form of diarrhea called giardiasis. Here, the cloning, expression, purification and the crystallization of α-11 giardin under two different conditions and in two different space groups is reported. Crystals from the first condition diffracted to 1.1 Å and belong to a primitive orthorhombic space group, while crystals from the second condition, which included calcium in the crystallization solution, diffracted to 2.93 Å and belong to a primitive monoclinic space group. Determination of the detailed atomic structure of α-11 giardin will provide a better insight into its biological function and might establish whether this class of proteins is a potential drug target against giardiasis.

  13. Identification and evolutionary implication of four novel box H/ACA snoRNAs from Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    LUO Jun; ZHOU Hui; CHEN Chongjian; LI Yan; CHEN Yueqin; QU Lianghu

    2006-01-01

    From a specialized cDNA library, four novel box H/ACA snoRNAs, named GLsR22,GLsR23, GLsR24 and GLsR25, were identified from the primitive eukaryote, Giardia lamblia. Bioinformatics analyses indicated that all of them can be potentially folded into double hairpins, the typical secondary structures of box H/ACA snoRNAs. GLsR24 and GLsR25 are predicted to guide the site-specific pseudouridylation at U1753 and U2396 on 23S rRNA,respectively, while GLsR22 and GLsR23 belong to the family of "orphan" snoRNAs. All of the four novel snoRNAs are encoded by single copy genes and located in small intergenic regions. Interestingly,compared with the counterparts previously reported in Archaea and other unicellular protozoan, the box H/ACA snoRNAs identified from G. lamblia have unique structural features, implying that snoRNAs evolved from prokaryotes to eukaryotes in different ways.

  14. DNA topoisomerase II is involved in regulation of cyst wall protein genes and differentiation in Giardia lamblia.

    Science.gov (United States)

    Lin, Bo-Chi; Su, Li-Hsin; Weng, Shih-Che; Pan, Yu-Jiao; Chan, Nei-Li; Li, Tsai-Kun; Wang, Hsin-Chih; Sun, Chin-Hung

    2013-01-01

    The protozoan Giardia lamblia differentiates into infectious cysts within the human intestinal tract for disease transmission. Expression of the cyst wall protein (cwp) genes increases with similar kinetics during encystation. However, little is known how their gene regulation shares common mechanisms. DNA topoisomerases maintain normal topology of genomic DNA. They are necessary for cell proliferation and tissue development as they are involved in transcription, DNA replication, and chromosome condensation. A putative topoisomerase II (topo II) gene has been identified in the G. lamblia genome. We asked whether Topo II could regulate Giardia encystation. We found that Topo II was present in cell nuclei and its gene was up-regulated during encystation. Topo II has typical ATPase and DNA cleavage activity of type II topoisomerases. Mutation analysis revealed that the catalytic important Tyr residue and cleavage domain are important for Topo II function. We used etoposide-mediated topoisomerase immunoprecipitation assays to confirm the binding of Topo II to the cwp promoters in vivo. Interestingly, Topo II overexpression increased the levels of cwp gene expression and cyst formation. Microarray analysis identified up-regulation of cwp and specific vsp genes by Topo II. We also found that the type II topoisomerase inhibitor etoposide has growth inhibition effect on Giardia. Addition of etoposide significantly decreased the levels of cwp gene expression and cyst formation. Our results suggest that Topo II has been functionally conserved during evolution and that Topo II plays important roles in induction of the cwp genes, which is key to Giardia differentiation into cysts.

  15. Giardia lamblia low-density lipoprotein receptor-related protein is involved in selective lipoprotein endocytosis and parasite replication.

    Science.gov (United States)

    Rivero, Maria R; Miras, Silvana L; Quiroga, Rodrigo; Rópolo, Andrea S; Touz, Maria C

    2011-03-01

    As Giardia lamblia is unable to synthesize cholesterol de novo, this steroid might be obtained from the host's intestinal milieu by endocytosis of lipoproteins. In this work, we identified a putative Giardia lamblia low-density lipoprotein receptor-related proteins (GlLRP), a type I membrane protein, which shares the substrate N-terminal binding domain and a FXNPXY-type endocytic motif with human LRPs. Expression of tagged GlLRP showed that it was localized predominantly in the endoplasmic reticulum, lysosomal-like peripheral vacuoles and plasma membrane. However, the FXNPXY-deleted GlLRP was retained at the plasma membrane suggesting that it is abnormally transported and processed. The low-density lipoprotein and chylomicrons interacted with GlLRP, with this interaction being necessary for lipoprotein internalization and cell proliferation. Finally, we show that GlLRP binds directly to the medium subunit of Giardia adaptor protein 2, indicating that receptor-mediated internalization occurs through an adaptin mechanism.

  16. Characterization of Giardia lamblia genotypes in dogs from Tucson, Arizona using SSU-rRNA and β-giardin sequences.

    Science.gov (United States)

    Johansen, K M; Castro, N S; Lancaster, K E; Madrid, E; Havas, A; Simms, J; Sterling, C R

    2014-01-01

    The objective of this study was to determine if human genotypes of Giardia lamblia could be found in canine companion animals from urban and peri-urban environments in Tucson, Arizona. Canine fecal samples collected from the Humane Society of Southern Arizona between July 2006 and April 2009 were screened for G. lamblia infection using immunofluorescent microscopy and confirmed by polymerase chain reaction (PCR). Of the 672 samples screened, 196 were found positive by IFA and 185 of those positive were successfully amplified through PCR. Sequencing analysis showed samples were primarily of the C or D genotypes (n =154), or showing a mix of the C and D genotypes (n =10). One sample showed a mixed infection of a human genotype (A) and a dog-specific genotype (C). These data are consistent with previous studies showing dog specific genotypes to be dominant in environments where dog-to-dog transmission is likely to occur, and provides further evidence that multiple genes should be targeted for more accurate genotype characterization.

  17. Prevalence and Risk Factors of Giardia lamblia and Blastocystis hominis Infections in Children Under Ten Years Old, Hamadan, Iran

    Directory of Open Access Journals (Sweden)

    Sedighi

    2015-05-01

    Full Text Available Background Nowadays, parasitic infections are a major health problem throughout the world, particularly in the developing countries. Objectives Considering the high susceptibility of children against parasitic infections, the current study aimed to determine the prevalence and associated risk factors of intestinal parasitic infections among children less than 10 years old in urban and rural areas of Hamadan district. Patients and Methods The current study was conducted on 395 children (214 males and 181 females, referred to urban and rural health centers in Hamadan district in 2013. Stool samples were examined by formalin-ether concentration technique, and trichrome and modified Ziehl-Neelsen staining methods. The results were analyzed by chi-square test. Results Of the 395 studied children, 112 (28.4% were infected with intestinal parasites. Blastocystis hominis was the most frequently detected parasite with the prevalence of 18.5%, followed by Giardia lamblia (10.9%, Entamoeba coli (2.8%, Dientamoeba fragilis (0.8%, Iodamoeba buetschlii (0.8%, Chilomastix mesnili (0.5%, Cryptosporidium spp. (0.5%, Endolimax nana (0.3% and Entamoeba hartmanni (0.3%. No cases of infection with helminth parasites were found. Conclusions The results of the study showed a high prevalence of Giardia lamblia and Blastocystis hominis in rural areas compared to urban regions. Therefore it is necessary to promote the public health awareness in the rural population, in order to reduce the frequency of parasitic infections.

  18. Hydrologic and vegetative removal of Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii Surrogate microspheres in coastal wetlands.

    Science.gov (United States)

    Hogan, Jennifer N; Daniels, Miles E; Watson, Fred G; Oates, Stori C; Miller, Melissa A; Conrad, Patricia A; Shapiro, Karen; Hardin, Dane; Dominik, Clare; Melli, Ann; Jessup, David A; Miller, Woutrina A

    2013-03-01

    Constructed wetland systems are used to reduce pollutants and pathogens in wastewater effluent, but comparatively little is known about pathogen transport through natural wetland habitats. Fecal protozoans, including Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii, are waterborne pathogens of humans and animals, which are carried by surface waters from land-based sources into coastal waters. This study evaluated key factors of coastal wetlands for the reduction of protozoal parasites in surface waters using settling column and recirculating mesocosm tank experiments. Settling column experiments evaluated the effects of salinity, temperature, and water type ("pure" versus "environmental") on the vertical settling velocities of C. parvum, G. lamblia, and T. gondii surrogates, with salinity and water type found to significantly affect settling of the parasites. The mesocosm tank experiments evaluated the effects of salinity, flow rate, and vegetation parameters on parasite and surrogate counts, with increased salinity and the presence of vegetation found to be significant factors for removal of parasites in a unidirectional transport wetland system. Overall, this study highlights the importance of water type, salinity, and vegetation parameters for pathogen transport within wetland systems, with implications for wetland management, restoration efforts, and coastal water quality.

  19. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens

    NARCIS (Netherlands)

    Schuurman, T.; Lankamp, P.; van Belkum, A.; Kooistra-Smid, M.; van Zwet, A.

    2007-01-01

    Giardia lamblia is one of the most common intestinal parasites worldwide, with microscopy being the diagnostic reference standard for use with human stools. However, microscopy is time-consuming, labour-intensive and lacks sensitivity when single stools are examined. In the present study, microscopy

  20. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley

    National Research Council Canada - National Science Library

    Díaz-Cinco Martha Elvia; Ballesteros-Vázquez Martha Nydia; Pérez-Morales Rosalba; Mata-Haro Verónica

    2002-01-01

    Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos...

  1. Giardia lamblia binding immunoglobulin protein triggers maturation of dendritic cells via activation of TLR4-MyD88-p38 and ERK1/2 MAPKs.

    Science.gov (United States)

    Lee, H-Y; Kim, J; Noh, H J; Kim, H-P; Park, S-J

    2014-12-01

    Much remains unknown about the mammalian immune response to Giardia lamblia, a protozoan pathogen that causes diarrhoeal outbreaks. We fractionated protein extracts of G. lamblia trophozoites by Viva-spin centrifugation, DEAE ion exchange and gel filtration chromatography. Resultant fractions were screened for antigenic molecules by western blots analysis using anti-G. lamblia antibodies (Abs), resulting in identification of G. lamblia binding immunoglobulin protein (GlBiP). Maturation of mouse dendritic cells (DCs) in response to recombinant GlBiP (rGlBiP) was detected by increased expression of surface molecules such as CD80, CD86 and MHC class II; these mature DCs, produced pro-inflammatory cytokines (TNF-α, IL-12 and IL-6). Especially, the truncated rGlBiP containing the heat-shock protein 70 domain-induced cytokine production from mouse DCs. rGlBiP-induced DC activation was initiated by TLR4 in a MyD88-dependent way and occurred through activation of p38 and ERK1/2 MAPKs as well as increased activity of NF-κB and AP-1. Moreover, CD4(+) T cells stimulated with rGlBiP-treated DCs produced high levels of IL-2 and IFN-γ. Together, our results suggest that GlBiP contributes to maturation of DCs via activation of TLR4-MyD88-p38, ERK1/2 MAPK, NF-κB and AP-1.

  2. Development of a combined in vitro cell culture--quantitative PCR assay for evaluating the disinfection performance of pulsed light for treating the waterborne enteroparasite Giardia lamblia.

    Science.gov (United States)

    Garvey, Mary; Stocca, Alessia; Rowan, Neil

    2014-09-01

    Giardia lamblia is a flagellated protozoan parasite that is recognised as a frequent cause of water-borne disease in humans and animals. We report for the first time on the use of a combined in vitro HCT-8 cell culture-quantitative PCR assay for evaluating the efficacy of using pulsed UV light for treating G. lamblia parasites. Findings showed that current methods that are limited to using vital stains before and after cyst excystation are not appropriate for monitoring or evaluating cyst destruction post PUV-treatments. Use of the human ileocecal HCT-8 cell line was superior to that of the human colon Caco-2 cell line for in vitro culture and determining PUV sensitivity of treated cysts. G. lamblia cysts were also shown to be more resistant to PUV irradiation compared to treating similar numbers of Cryptosporidium parvum oocysts. These observations also show that the use of this HCT-8 cell culture assay may replace use of animal models for determining disinfection performances of PUV for treating both C. parvum and G. lamblia.

  3. Comparison of microscopy, real-time PCR and a rapid immunoassay for the detection of Giardia lamblia in human stool specimens

    NARCIS (Netherlands)

    Schuurman, T.; Lankamp, P.; van Belkum, A.; Kooistra-Smid, M.; van Zwet, A.

    2007-01-01

    Giardia lamblia is one of the most common intestinal parasites worldwide, with microscopy being the diagnostic reference standard for use with human stools. However, microscopy is time-consuming, labour-intensive and lacks sensitivity when single stools are examined. In the present study,

  4. Clinical and microbiological features of dientamoebiasis in patients suspected of suffering from a parasitic gastrointestinal illness: A comparison of Dientamoeba fragilis and Giardia lamblia infections

    NARCIS (Netherlands)

    O. Vandenberg; R. Peek; H. Souayah; A. Dediste; M. Buset; R. Scheen; P. Retore; G. Zissis; T. van Gool

    2006-01-01

    Objectives: To describe the clinical and microbiological features of Dientomoeba fragilis and Giordia lamblia infected patients, and to analyze the genetic variation of D. fragilis strains. Methods: For a period of two years, all stool samples collected from patients suspected of having a parasitic

  5. Comparação de quatro métodos laboratoriais para diagnóstico da Giardia lamblia em fezes de crianças residentes em Belém, Pará Evaluation of four techniques for diagnosis of Giardia lamblia in children's stool from Belém city, Para State, Brazil

    Directory of Open Access Journals (Sweden)

    Ricardo Luiz Dantas Machado

    2001-02-01

    Full Text Available Relatamos a comparação de quatro metodologias para o diagnóstico da Giardia lamblia em material fecal de crianças, Belém/PA. A Hematoxilina Férrica e o método direto apresentaram menor positividade, enquanto que o Método de Faust continua uma boa escolha para o diagnóstico e o Ensaio imunoenzimático melhora a qualidade da detecção deste parasito.We report the evaluation of four techniques for Giardia lamblia diagnosis in children's stool. The Iron haematoxilin staining and direct examination with lugol showed lower positivity, while the method of Faust et al. Continues to be a good option for G. lamblia diagnosis and Immunoenzymatic assay increases the detection of this parasite.

  6. Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia

    Science.gov (United States)

    Marcial-Quino, Jaime; Gómez-Manzo, Saúl; Fierro, Francisco; Vanoye-Carlo, America; Rufino-González, Yadira; Sierra-Palacios, Edgar; Castillo-Villanueva, Adriana; Castillo-Rodríguez, Rosa Angélica; Rodríguez-Bustamante, Eduardo; Arreguin-Espinosa, Roberto; Reyes-Vivas, Horacio

    2016-01-01

    Stem-loop quantitative reverse transcription PCR (RT-qPCR) is a molecular technique used for identification and quantification of individual small RNAs in cells. In this work, we used a Universal ProbeLibrary (UPL)-based design to detect—in a rapid, sensitive, specific, and reproducible way—the small nucleolar RNA (snoRNA) GlsR17 and its derived miRNA (miR2) of Giardia lamblia using a stem-loop RT-qPCR approach. Both small RNAs could be isolated from both total RNA and small RNA samples. Identification of the two small RNAs was carried out by sequencing the PCR-amplified small RNA products upon ligation into the pJET1.2/blunt vector. GlsR17 is constitutively expressed during the 72 h cultures of trophozoites, while the mature miR2 is present in 2-fold higher abundance during the first 48 h than at 72 h. Because it has been suggested that miRNAs in G. lamblia have an important role in the regulation of gene expression, the use of the stem-loop RT-qPCR method could be valuable for the study of miRNAs of G. lamblia. This methodology will be a powerful tool for studying gene regulation in G. lamblia, and will help to better understand the features and functions of these regulatory molecules and how they work within the RNA interference (RNAi) pathway in G. lamblia. PMID:27999395

  7. Stem-Loop RT-qPCR as an Efficient Tool for the Detection and Quantification of Small RNAs in Giardia lamblia

    Directory of Open Access Journals (Sweden)

    Jaime Marcial-Quino

    2016-12-01

    Full Text Available Stem-loop quantitative reverse transcription PCR (RT-qPCR is a molecular technique used for identification and quantification of individual small RNAs in cells. In this work, we used a Universal ProbeLibrary (UPL-based design to detect—in a rapid, sensitive, specific, and reproducible way—the small nucleolar RNA (snoRNA GlsR17 and its derived miRNA (miR2 of Giardia lamblia using a stem-loop RT-qPCR approach. Both small RNAs could be isolated from both total RNA and small RNA samples. Identification of the two small RNAs was carried out by sequencing the PCR-amplified small RNA products upon ligation into the pJET1.2/blunt vector. GlsR17 is constitutively expressed during the 72 h cultures of trophozoites, while the mature miR2 is present in 2-fold higher abundance during the first 48 h than at 72 h. Because it has been suggested that miRNAs in G. lamblia have an important role in the regulation of gene expression, the use of the stem-loop RT-qPCR method could be valuable for the study of miRNAs of G. lamblia. This methodology will be a powerful tool for studying gene regulation in G. lamblia, and will help to better understand the features and functions of these regulatory molecules and how they work within the RNA interference (RNAi pathway in G. lamblia.

  8. Stop codon recognition in the early-diverged protozoans Giardia lamblia and Trichomonas vaginalis.

    Science.gov (United States)

    Chai, Baofeng; Li, Cui; Yu, Jingfei; Hao, Yanrong; Guo, Ping; Shen, Quan

    2015-07-01

    Two classes of polypeptide release factors (RFs) are responsible for maintaining accuracy in translation termination; however, their detailed mechanism of action and evolutionary history of these factors remain elusive. The structure and function of RFs vary in bacteria and eukaryotes, a fact that is suggestive of evolutionary changes in the translation termination system. Giardia lamblia (Diplomonada) and Trichomonas vaginalis (Parabasalia) are considered as early-diverged eukaryotes. The class II release factor, eRF3, of Giardia (Gl-eRF3) appears to have only one domain that corresponds to EF-1α and lacks the N-terminal domain, similar to that of eRF3 of other organisms. In the present study, we show that the chimeric molecules Gl/Sc eRF1 and Tv/Sc eRF1, which are composed of the N-terminal domain of Gl-eRF1 or Tv-eRF1, fused to the core domain (M and C domain) of Saccharomyces cerevisiae eRF1 (Sc-eRF1), resulting in loss of the RF properties of the N-terminal domain. This suggests that the conformation of eRF1 for stop codon recognition in Giardia and Trichomonas varies from the eRF1s of other eukaryotes, including ciliates and yeast. Further studies using intra-N-terminal chimeras of eRF1 indicated that the combination of the GTS loop and NIKS motif from Gl-eRF1 and the Y-C-F motif from Sc-eRF1within the N terminal domain of hybrid eRF1 could restore UGA, but not UAG and UGA recognition. In contrast, the combination of the GTS loop and the NIKS motif of Sc-eRF1 and the Y-C-F motif of Gl-eRF1 could restore UAG and UAA recognition, but not UGA recognition. Thus, these results confirm the findings of previous studies that three motifs in eRF1 are necessary for discrimination of the three bases of stop codons. The NIKS motif is responsible for recognition of the first two bases of UAA and UAG, and the Y-C-F motif identifies the second base of UGA by Gl-eRF1. Amino acid residue substitutions in Gl/Sc-eRF1 by corresponding residues of Sc-eRF1 could change and even

  9. Comparative characterisation of two nitroreductases from Giardia lamblia as potential activators of nitro compounds.

    Science.gov (United States)

    Müller, Joachim; Rout, Samuel; Leitsch, David; Vaithilingam, Jathana; Hehl, Adrian; Müller, Norbert

    2015-08-01

    Giardia lamblia is a protozoan parasite that causes giardiasis, a diarrhoeal disease affecting humans and various animal species. Nitro drugs such as the nitroimidazole metronidazole and the nitrothiazolide nitazoxanide are used for treatment of giardiasis. Nitroreductases such as GlNR1 and GlNR2 may play a role in activation or inactivation of these drugs. The aim of this work is to characterise these two enyzmes using functional assays. For respective analyses recombinant analogues from GlNR1 and GlNR2 were produced in Escherichia coli. E. coli expressing GlNR1 and GlNR2 alone or together were grown in the presence of nitro compounds. Furthermore, pull-down assays were performed using HA-tagged GlNR1 and GlNR2 as baits. As expected, E. coli expressing GlNR1 were more susceptible to metronidazole under aerobic and semi-aerobic and to nitazoxanide under semi-aerobic growth conditions whereas E. coli expressing GlNR2 were susceptible to neither drug. Interestingly, expression of both nitroreductases gave the same results as expression of GlNR2 alone. In functional assays, both nitroreductases had their strongest activities on the quinone menadione (vitamin K3) and FAD, but reduction of nitro compounds including the nitro drugs metronidazole and nitazoxanide was clearly detected. Full reduction of 7-nitrocoumarin to 7-aminocoumarin was preferentially achieved with GlNR2. Pull-down assays revealed that GlNR1 and GlNR2 interacted in vivo forming a multienzyme complex. These findings suggest that both nitroreductases are multifunctional. Their main biological role may reside in the reduction of vitamin K analogues and FAD. Activation by GlNR1 or inactivation by GlNR2 of nitro drugs may be the consequence of a secondary enzymatic activity either yielding (GlNR1) or eliminating (GlNR2) toxic intermediates after reduction of these compounds.

  10. Characteristics and risk factors for symptomatic Giardia lamblia infections in Germany

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    Stark Klaus

    2010-01-01

    Full Text Available Abstract Background In developed countries, giardiasis is considered a travel related disease. However, routine surveillance data from Germany indicate that >50% of infections were acquired indigenously. We studied the epidemiological characteristics of symptomatic Giardia infections acquired in Germany and abroad, and verified the proportion of cases acquired in Germany in order to investigate risk factors for sporadic autochthonous Giardia infections. Methods We identified Giardia cases notified by 41 local health authorities between February 2007 and January 2008 and interviewed them on their clinical symptoms, underlying morbidities, travel abroad and potential risk factors for the disease. We conducted a case-control-study including laboratory-confirmed (microscopy or antigen-test autochthonous Giardia cases with clinical manifestations (diarrhoea, cramps, bloating and randomly selected controls from the local population registry matched by county of residence and age-group (0-5, 6-19, ≥20 years. Secondary cases, controls with diarrhoea and persons who had travelled outside Germany in the three weeks prior to disease onset (exposure period were excluded. We calculated adjusted odds ratios (aOR with 95% confidence intervals (CI using conditional logistic regression. Results Of 273 interviewed cases, 131 (48% had not travelled abroad during the defined exposure period. Of these 131, 85 (65% were male, 68 (54% were living in communities with >100,000 inhabitants and 107 (83% were aged 20 years or older. We included 120 cases and 240 controls in the case-control study. Cases were more likely to be male (aOR 2.5 CI 1.4-4.4, immunocompromised (aOR 15.3 CI 1.8-127 and daily consumers of green salad (aOR 2.9 CI 1.2-7.2. Contact with animals (pets/farm animals and exposure to surface water (swimming/water sports were not associated with symptomatic disease. Conclusions A substantial proportion of Giardia lamblia cases in Germany are indigenously

  11. Functional redundancy of two Pax-like proteins in transcriptional activation of cyst wall protein genes in Giardia lamblia.

    Science.gov (United States)

    Chuang, Shen-Fung; Su, Li-Hsin; Cho, Chao-Cheng; Pan, Yu-Jiao; Sun, Chin-Hung

    2012-01-01

    The protozoan Giardia lamblia differentiates from a pathogenic trophozoite into an infectious cyst to survive outside of the host. During encystation, genes encoding cyst wall proteins (CWPs) are coordinately induced. Pax family transcription factors are involved in a variety of developmental processes in animals. Nine Pax proteins have been found to play an important role in tissue and organ development in humans. To understand the progression from primitive to more complex eukaryotic cells, we tried to identify putative pax genes in the G. lamblia genome and found two genes, pax1 and pax2, with limited similarity. We found that Pax1 may transactivate the encystation-induced cwp genes and interact with AT-rich initiatior elements that are essential for promoter activity and transcription start site selection. In this study, we further characterized Pax2 and found that, like Pax1, Pax2 was present in Giardia nuclei and it may specifically bind to the AT-rich initiator elements of the encystation-induced cwp1-3 and myb2 genes. Interestingly, overexpression of Pax2 increased the cwp1-3 and myb2 gene expression and cyst formation. Deletion of the C-terminal paired domain or mutation of the basic amino acids of the paired domain resulted in a decrease of nuclear localization, DNA-binding activity, and transactivation activity of Pax2. These results are similar to those found in the previous Pax1 study. In addition, the profiles of gene expression in the Pax2 and Pax1 overexpressing cells significantly overlap in the same direction and ERK1 associated complexes may phosphorylate Pax2 and Pax1, suggesting that Pax2 and Pax1 may be downstream components of a MAPK/ERK1 signaling pathway. Our results reveal functional redundancy between Pax2 and Pax1 in up-regulation of the key encystation-induced genes. These results illustrate functional redundancy of a gene family can occur in order to increase maintenance of important gene function in the protozoan organism G. lamblia.

  12. Functional redundancy of two Pax-like proteins in transcriptional activation of cyst wall protein genes in Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Shen-Fung Chuang

    Full Text Available The protozoan Giardia lamblia differentiates from a pathogenic trophozoite into an infectious cyst to survive outside of the host. During encystation, genes encoding cyst wall proteins (CWPs are coordinately induced. Pax family transcription factors are involved in a variety of developmental processes in animals. Nine Pax proteins have been found to play an important role in tissue and organ development in humans. To understand the progression from primitive to more complex eukaryotic cells, we tried to identify putative pax genes in the G. lamblia genome and found two genes, pax1 and pax2, with limited similarity. We found that Pax1 may transactivate the encystation-induced cwp genes and interact with AT-rich initiatior elements that are essential for promoter activity and transcription start site selection. In this study, we further characterized Pax2 and found that, like Pax1, Pax2 was present in Giardia nuclei and it may specifically bind to the AT-rich initiator elements of the encystation-induced cwp1-3 and myb2 genes. Interestingly, overexpression of Pax2 increased the cwp1-3 and myb2 gene expression and cyst formation. Deletion of the C-terminal paired domain or mutation of the basic amino acids of the paired domain resulted in a decrease of nuclear localization, DNA-binding activity, and transactivation activity of Pax2. These results are similar to those found in the previous Pax1 study. In addition, the profiles of gene expression in the Pax2 and Pax1 overexpressing cells significantly overlap in the same direction and ERK1 associated complexes may phosphorylate Pax2 and Pax1, suggesting that Pax2 and Pax1 may be downstream components of a MAPK/ERK1 signaling pathway. Our results reveal functional redundancy between Pax2 and Pax1 in up-regulation of the key encystation-induced genes. These results illustrate functional redundancy of a gene family can occur in order to increase maintenance of important gene function in the protozoan

  13. A group of Giardia lamblia variant-specific surface protein (VSP) genes with nearly identical 5' regions.

    Science.gov (United States)

    Yang, Y; Adam, R D

    1995-12-01

    The surfaces of Giardia lamblia trophozoites contain one of a set of variant-specific surface proteins. The genes encoding these proteins are highly conserved at the 3' terminus, but frequently demonstrate little similarity in the remainder of the coding region. This report describes a family of vsp genes highly similar to a repeat-containing vsp gene (vspC5) at the 5' coding and flanking regions, but which diverge abruptly from vspC5 in the first repeat and do not themselves contain full copies of the repeat. This observation suggests the possibility that recombination among different vsp genes may have played a role in development of the vsp gene repertoire.

  14. Immunodominant proteins α-1 giardin and β-giardin are expressed in both assemblages A and B of Giardia lamblia.

    Science.gov (United States)

    Feliziani, Constanza; Merino, María C; Rivero, María R; Hellman, Ulf; Pistoresi-Palencia, María C; Rópolo, Andrea S

    2011-10-19

    To date, eight assemblages of Giardia lamblia have been described, but only assemblages A and B are known to infect humans. Despite the fact that the genomic, biological, and clinical differences found between these two assemblages has raised the possibility that they may be considered different species, there is relatively limited information on their phenotypic differences. In the present study, we developed monoclonal antibodies against alpha-1 and beta giardin, two immunodominant proteins produced during G. lamblia infection, and studied their expression and localization in WB (assemblage A) and GS trophozoites (assemblage B). The polyclonal antibodies generated against WB trophozoites, particularly those recognizing intracellular proteins as well as the proteins present at the plasma membrane (variable-specific surface proteins), showed cross-reactivity with intracellular proteins in GS trophozoites. The use of monoclonal antibodies against beta giardin indicated ventral disc localization, particularly at the periphery in WB trophozoites. Interestingly, although beta giardin was also restricted to the ventral disc in GS trophozoites, the pattern of localization clearly differed in this assemblage. On the other hand, monoclonal antibodies against alpha-1 giardin showed plasma membrane localization in both assemblages with the bare area of GS trophozoites also being distinguished. Moreover, the same localization at the plasma membrane was observed in Portland-1 (Assemblage A) and in P15 (Assemblage E) trophozoites. We found differences in localization of the beta giardin protein between assemblages A and B, but the same pattern of localization of alpha-1 giardin in strains from Assemblages A, B and E. These findings reinforce the need for more studies based on phenotypic characteristics in order to disclose how far one assemblage is from the other.

  15. The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target.

    Science.gov (United States)

    Kunz, Stefan; Balmer, Vreni; Sterk, Geert Jan; Pollastri, Michael P; Leurs, Rob; Müller, Norbert; Hemphill, Andrew; Spycher, Cornelia

    2017-09-01

    Giardiasis is an intestinal infection correlated with poverty and poor drinking water quality, and treatment options are limited. According to the Center for Disease Control and Prevention, Giardia infections afflict nearly 33% of people in developing countries, and 2% of the adult population in the developed world. This study describes the single cyclic nucleotide-specific phosphodiesterase (PDE) of G. lamblia and assesses PDE inhibitors as a new generation of anti-giardial drugs. An extensive search of the Giardia genome database identified a single gene coding for a class I PDE, GlPDE. The predicted protein sequence was analyzed in-silico to characterize its domain structure and catalytic domain. Enzymatic activity of GlPDE was established by complementation of a PDE-deficient Saccharomyces cerevisiae strain, and enzyme kinetics were characterized in soluble yeast lysates. The potency of known PDE inhibitors was tested against the activity of recombinant GlPDE expressed in yeast and against proliferating Giardia trophozoites. Finally, the localization of epitope-tagged and ectopically expressed GlPDE in Giardia cells was investigated. Giardia encodes a class I PDE. Catalytically important residues are fully conserved between GlPDE and human PDEs, but sequence differences between their catalytic domains suggest that designing Giardia-specific inhibitors is feasible. Recombinant GlPDE hydrolyzes cAMP with a Km of 408 μM, and cGMP is not accepted as a substrate. A number of drugs exhibit a high degree of correlation between their potency against the recombinant enzyme and their inhibition of trophozoite proliferation in culture. Epitope-tagged GlPDE localizes as dots in a pattern reminiscent of mitosomes and to the perinuclear region in Giardia. Our data strongly suggest that inhibition of G. lamblia PDE activity leads to a profound inhibition of parasite proliferation and that GlPDE is a promising target for developing novel anti-giardial drugs.

  16. Identification and analysis of the RNA degrading complexes and machinery of Giardia lamblia using an in silico approach

    Science.gov (United States)

    2011-01-01

    Background RNA degradation is critical to the survival of all cells. With increasing evidence for pervasive transcription in cells, RNA degradation has gained recognition as a means of regulating gene expression. Yet, RNA degradation machinery has been studied extensively in only a few eukaryotic organisms, including Saccharomyces cerevisiae and humans. Giardia lamblia is a parasitic protist with unusual genomic traits: it is binucleated and tetraploid, has a very compact genome, displays a theme of genomic minimalism with cellular machinery commonly comprised of a reduced number of protein components, and has a remarkably large population of long, stable, noncoding, antisense RNAs. Results Here we use in silico approaches to investigate the major RNA degradation machinery in Giardia lamblia and compare it to a broad array of other parasitic protists. We have found key constituents of the deadenylation and decapping machinery and of the 5'-3' RNA degradation pathway. We have similarly found that all of the major 3'-5' RNA degradation pathways are present in Giardia, including both exosome-dependent and exosome-independent machinery. However, we observe significant loss of RNA degradation machinery genes that will result in important differences in the protein composition, and potentially functionality, of the various RNA degradation pathways. This is most apparent in the exosome, the central mediator of 3'-5' degradation, which apparently contains an altered core configuration in both Giardia and Plasmodium, with only four, instead of the canonical six, distinct subunits. Additionally the exosome in Giardia is missing both the Rrp6, Nab3, and Nrd1 proteins, known to be key regulators of noncoding transcript stability in other cells. Conclusions These findings suggest that although the full complement of the major RNA degradation mechanisms were present - and likely functional - early in eukaryotic evolution, the composition and function of the complexes is more

  17. GIS-based analysis of the fate of waste-related pathogens Cryptosporidium parvum, Giardia lamblia and Escherichia coli in a tropical canal network.

    Science.gov (United States)

    Diallo, Mamadou B C; Anceno, Alfredo J; Tawatsupa, Benjawan; Tripathi, Nitin K; Wangsuphachart, Voranuch; Shipin, Oleg V

    2009-03-01

    Urban canals play a major socio-economic role in many tropical countries and, particularly, Thailand. One of the overlooked functions that they perform is a significant attenuation of waste-related pathogens posing considerable health risk, as well as pollution attenuation in general. The study dealt with a comparison of three canals receiving: (i) municipal, (ii) mainly industrial and (iii) mainly agricultural wastewater, listed in order of progressively decreasing organic loading. The occurrence and fate of waterborne Cryptosporidium parvum, Giardia lamblia and Escherichia coli were monitored in the canals by both real-time PCR and conventionally for 12 months. The pathogens are etiological agents of an estimated 38% and 47% of diarrhea cases worldwide and in Thailand, respectively. The geographic information system (GIS) was used to evaluate and map point and, particularly, non-point pollution sources which allowed differentiating the canal sections in terms of predominant pathogen sources. The flowthrough canals, which can be viewed as waste stabilization ponds, were found to be efficiently removing the pathogens at the following generalized specific rates: 0.3 (C. parvum), 1.2 (G. lamblia), 1.8 (E. coli) log10/km.d in the dry season. The rates decreased in the rainy season for E. coli and G. lamblia, but increased for C. parvum which indicated different removal mechanisms. Data suggest that E. coli and G. lamblia were mainly removed through sedimentation and sunlight (UV) irradiation, while the likely mechanism for C. parvum was predation. Overall, the specific pathogen removal rates positively correlated with the canal organic loading rates in the rainy season. As an important result, an estimate of the municipal pollution mitigation by over 2280 km canals in the Greater Bangkok suggests that concomitant to the pathogens at least 36-95 tons of BOD5 is being removed daily, thereby saving the receiving Chao Phraya River and Bight of Bangkok, by far exceeding

  18. In vitro susceptibility of Entamoeba histolytica and Giardia lamblia to plants used in Mexican traditional medicine for the treatment of gastrointestinal disorders.

    Science.gov (United States)

    Calzada, Fernando; Yépez-Mulia, Lilian; Aguilar, Abigaíl

    2006-12-06

    In our search for new antiprotozoal chemotherapy, we collected a selection of 26 plants used in Mexican traditional medicine for the treatment of gastrointestinal disorders. Methanolic extracts of these species were screened for their antiprotozoal activity against Entamoeba histolytica and Giardia lamblia trophozoites using in vitro tests. Among the tested extracts, the derivates of following species showed selectivity and significant antiprotozoal activity: Chiranthodendron pentadactylon, Annona cherimola and Punica granatum were the most active on Entamoeba histolytica with IC50 Mexican traditional medicine.

  19. Does the use of tubular digesters to treat livestock waste lower the risk of infection from Cryptosporidium parvum and Giardia lamblia?

    Science.gov (United States)

    Kinyua, Maureen N; Wald, Ileana; Camacho-Céspedes, Fabricio; Izurieta, Ricardo; Haas, Charles N; Ergas, Sarina J

    2016-10-01

    Worldwide, high incidences of cryptosporidiosis and giardiasis are attributed to livestock waste. Quantitative microbial risk assessment can be used to estimate the risk of livestock related infections from Cryptosporidium parvum and Giardia lamblia. The objective of this paper was to assess the occupational and public health risks associated with management of raw and anaerobically digested livestock waste in two rural communities in Costa Rica based on fomite, soil and crop contamination and livestock waste management exposure pathways. Risks related to cattle waste were greater than swine waste due to cattle shedding more (oo)cysts. Cryptosporidium parvum also posed a greater risk than Giardia lamblia in all exposure pathways due to livestock shedding high loads of Cryptosporidium parvum oocysts and oocysts' lower inactivation rates during anaerobic digestion compared with Giardia lamblia cysts. The risk of infection from exposure to contaminated soil and crops was significantly lower for a community using tubular anaerobic digesters to treat livestock waste compared to a community where the untreated waste was applied to soil. The results indicate that treatment of livestock waste in small-scale tubular anaerobic digesters has the potential to significantly decrease the risk of infection below the World Health Organization's acceptable individual annual risk of infection (10(-4)).

  20. Progress on Regulation of Gene Expression in Giardia lamblia%蓝氏贾第鞭毛虫基因表达调控的研究进展

    Institute of Scientific and Technical Information of China (English)

    王乙惠; 李雅杰

    2011-01-01

    蓝氏贾第鞭毛虫是一种重要的致病性寄生虫,人体感染后主要引起腹泻和营养不良等症状.作为一种源真核生物,其基因表达调控机制可能与其他真核生物有较大的区别.本文从蓝氏贾第虫鞭毛虫启动子、转录因子、转录后调节、翻译起始和表观遗传学等方面,对贾第虫基因表达调控的研究进展进行综述.%Giardia lamblia is an important human pathogen that causes diarrhea and malnutrition. As a late-branching eukaryote, G. lamblia may have special mechanisms for regulating gene expression which differ from other eukaryotes. In this paper, the mechanisms that governing regulation of G. lamblia gene expression, such as promoters, transcription factors, transcriptional regulation, translation initiation and epigenetic mechanisms are summarized.

  1. INHIBICIÓN DEL CRECIMIENTO DE Giardia lamblia POR ACCIÓN DEL EXTRACTO ACUOSO Y METANÓLICO DE SEMILLAS DE Cucurbita pepo

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    María Porfiria Barrón González

    2010-02-01

    Full Text Available Giardia lamblia es el protozoario parásito causante de la giardiasis, la cual se caracteriza por molestia abdominal, pérdida de peso y desnutrición. La droga de elección para su tratamiento es el metronidazol sin embargo, presenta diversos efectos secundarios adversos en el paciente. Por otra parte, en la herbolaria se conoce a la semilla de Cucurbita pepo por sus propiedades desparasitante, principalmente sobre Entamoeba histolytica y Taenia solium. Por lo anterior se planteó la hipótesis de que las semillas de C. pepo contienen principios activos capaces de inhibir el crecimiento de G. lamblia bajo condiciones axénicas in vitro. El objetivo de esta investigación fue evaluar la actividad biológica del extracto acuoso y metanólico de las semillas de C. pepo sobre el crecimiento de G. lamblia. Tanto el extracto metanólico como el acuoso se identificaron siete grupos funcionales y la actividad antioxidante no fue significativa

  2. Detection of a Giardia lamblia coproantigen by using a commercially available immunoenzymatic assay, in Belo Horizonte, Brazil Avaliação do ProSpeT Giardia ensaio em microplaca na detecção de coproantígenos de Giardia lamblia, em fezes de pacientes de Belo Horizonte, Brasil

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    Míriam Oliveira e ROCHA

    1999-05-01

    Full Text Available It is known that fecal examination to detect Giardia lamblia cysts or trophozoites produces a high percentage of false-negative results. A commercially available immunoenzymatic assay (ProSpecT Giardia Microplate Assay, Alexon, Inc., BIOBRÁS to detect G. lamblia specific coproantigen was evaluated for the first time in Brazil. A total of 90 specimens were tested. Each specimen was first tested as unpreserved stool, and then it was preserved in 10% Formalin to be tested 2 months later. The assay was able to identify all the 30 positive patients (sensitivity = 100.0% by visual or spectrophotometric examination in the unpreserved specimens and was negative in 57 of the 60 patients without G. lamblia (specificity = 95.0%. The assay identified 27 of the 30 positive patients (sensitivity = 90.0% and was negative in 59 of the 60 negatives (specificity = 98.3% in the preserved stools according to both readings. A marked difference was observed in the optical densities in both groups, preserved and unpreserved stools, when the G. lamblia-positive specimens were compared to the negative or positive for other intestinal parasites than G. lamblia. The assay seems a good alternative for giardiasis diagnosis, especially when the fecal examination was repeatedly negative and the patient presents giardiasislike symptoms.O diagnóstico da giardíase através da pesquisa de trofozoítos ou cistos do parasita nas fezes apresenta uma elevada percentagem de resultados falso-negativos. No presente trabalho foi feita, pela primeira vez no Brasil, uma avaliação do ProSpecT Giardia Ensaio em Microplaca (Alexon, Inc., BIOBRÁS, utilizando amostras fecais de pacientes de Belo Horizonte, Brasil. Um total de 90 amostras fecais foram testadas, primeiramente como fezes frescas e, então, preservadas em formalina 10%, para um novo teste 2 meses após. Com as amostras frescas, o ensaio imunoenzimático, através de leitura visual ou no espectrofotômetro, foi capaz de

  3. Giardia lamblia allergenic extract as diagnosis procedures for determining sensitization to this protozoa.

    Science.gov (United States)

    Crespo Guerrero, V; Alfonso Fernández, L A; Gómez Echevarría, A H

    1991-12-01

    We studied 200 patients assisting at the Allergy Department and the Gastroenterology Department in "Hermanos Ameijeiras" Clinical-Surgical Hospital. They were clinically and immunologically tested for giardiasis through duodenal fortis or gall bladder drainage and total IgE serum levels. All patients underwent intradermal and skin prick tests with Giardine allergenic extract. These skin tests showed high sensitivity and increased specificity. Thus, our procedure in diagnosis is accurate, accessible and economical.

  4. A unique mechanism of nuclear division in Giardia lamblia involves components of the ventral disk and the nuclear envelope.

    Science.gov (United States)

    Solari, Alberto J; Rahn, Monica I; Saura, Alicia; Lujan, Hugo D

    2003-12-01

    The fine structure of the binucleate, parasitic protist Giardia lamblia during interphase and divisional stages was studied by serial thin sectioning and three-dimensional reconstructions. The earlier sign of nuclear division is the development of a few peripheral areas of densely packed chromatin directly attached to the inner nuclear envelope. An intracytoplasmic sheet of ventral disk components grows from the cell periphery towards one of the nuclei, apparently constricting this nucleus, which becomes located at a ventral bulge. After the basal bodies become duplicated, a full nuclear division occurs in trophozoites, giving two pairs of parent-daughter nuclei. This full division occurs in a dorsal-ventral direction, with the resulting nuclear pairs located at the sides of the two sets of basal bodies. A new ventral disk is formed from the disk-derived sheets in the cell harboring the four nuclei. Cytokinesis is polymorphic, but at early stages is dorsal-to-dorsal. Encysting trophozoites show the development of Golgi cisternae stacks and dense, specific secretory granules. 3-D reconstructions show that cysts contain a single pair of incompletely strangled nuclei. The dividing Giardia lacks a typical, microtubular spindle either inside or outside the nuclei. The nuclear envelope seems to be the only structure involved in the final division of the parent-daughter nuclei.

  5. The transcriptional response to encystation stimuli in Giardia lamblia is restricted to a small set of genes.

    Science.gov (United States)

    Morf, Laura; Spycher, Cornelia; Rehrauer, Hubert; Fournier, Catharine Aquino; Morrison, Hilary G; Hehl, Adrian B

    2010-10-01

    The protozoan parasite Giardia lamblia undergoes stage differentiation in the small intestine of the host to an environmentally resistant and infectious cyst. Encystation involves the secretion of an extracellular matrix comprised of cyst wall proteins (CWPs) and a β(1-3)-GalNAc homopolymer. Upon the induction of encystation, genes coding for CWPs are switched on, and mRNAs coding for a Myb transcription factor and enzymes involved in cyst wall glycan synthesis are upregulated. Encystation in vitro is triggered by several protocols, which call for changes in bile concentrations or availability of lipids, and elevated pH. However, the conditions for induction are not standardized and we predicted significant protocol-specific side effects. This makes reliable identification of encystation factors difficult. Here, we exploited the possibility of inducing encystation with two different protocols, which we show to be equally effective, for a comparative mRNA profile analysis. The standard encystation protocol induced a bipartite transcriptional response with surprisingly minor involvement of stress genes. A comparative analysis revealed a core set of only 18 encystation genes and showed that a majority of genes was indeed upregulated as a side effect of inducing conditions. We also established a Myb binding sequence as a signature motif in encystation promoters, suggesting coordinated regulation of these factors.

  6. Identification of Giardia lamblia DHHC proteins and the role of protein S-palmitoylation in the encystation process.

    Science.gov (United States)

    Merino, María C; Zamponi, Nahuel; Vranych, Cecilia V; Touz, María C; Rópolo, Andrea S

    2014-07-01

    Protein S-palmitoylation, a hydrophobic post-translational modification, is performed by protein acyltransferases that have a common DHHC Cys-rich domain (DHHC proteins), and provides a regulatory switch for protein membrane association. In this work, we analyzed the presence of DHHC proteins in the protozoa parasite Giardia lamblia and the function of the reversible S-palmitoylation of proteins during parasite differentiation into cyst. Two specific events were observed: encysting cells displayed a larger amount of palmitoylated proteins, and parasites treated with palmitoylation inhibitors produced a reduced number of mature cysts. With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome. These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades. Although all Giardia DHHC proteins were found to be present in trophozoites and encysting cells, these proteins showed a different intracellular localization in trophozoites and seemed to be differently involved in the encystation process when they were overexpressed. dhhc transgenic parasites showed a different pattern of cyst wall protein expression and yielded different amounts of mature cysts when they were induced to encyst. Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.

  7. Engineering Giardia lamblia trimethylguanosine synthase (GlaTgs2) to transfer non-natural modifications to the RNA 5'-cap.

    Science.gov (United States)

    Holstein, Josephin Marie; Stummer, Daniela; Rentmeister, Andrea

    2015-06-01

    Trimethylguanosine synthase from Giardia lamblia (GlaTgs2) naturally catalyzes methyl transfer from S-adenosyl-L-methionine (AdoMet) to the exocyclic N(2) atom of the 5'-cap--a hallmark of eukaryotic mRNAs. The wild-type enzyme shows substrate promiscuity and can also use the AdoMet-analog AdoPropen for allyl transfer. Here we report on engineering GlaTgs2 to enhance the activity on AdoPropen. A mutational analysis, involving an alanine scan of 10 residues located around the active site, was performed. Positions V34 and S38 were identified as mutational hot spots and analyzed in greater detail by testing NNK libraries. Kinetic analysis and thermostability measurements revealed V34A as the best variant of GlaTgs2, with a ∼10-fold improved specificity for AdoPropen. Double mutants did not yield additional improvements due to low catalytic efficiencies and thermal destabilization. Homologous Tgs enzymes from Homo sapiens and G. intestinalis were also investigated regarding their catalytic activity on AdoPropen. While neither the human wild-type (WT) enzyme nor any of its variants showed activity on AdoPropen, the homologue from G. intestinalis (GinTgs) was remarkably active on AdoPropen. Introducing the best substitution at the homologous position led to variant T34A with ∼40-fold higher specificity for AdoPropen than the original GlaTgs2 WT.

  8. In vitro immunomodulatory effects of microemulsions with levamisole delivery systems on blood phagocytes interacting with Giardia lamblia.

    Science.gov (United States)

    Hernandes, Mara Rosa Gil; Moraes, Lucélia Campelo Albuquerque; Ribeiro, Elton Brito; Fagundes, Danny Laura Gomes; Honorio-França, Adenilda Cristina; França, Eduardo Luzía

    2017-06-01

    Giardiasis is one of the main parasites that infect the gastrointestinal tract of humans, affecting hundreds of millions of people worldwide, particularly in developing countries. Antiparasitics administered to treat giardiasis are inefficient in 20% of the cases, usually because of parasite resistance and side effects. In this scenario, microemulsions are a promising pharmaceutical alternative as carriers of molecules with therapeutic action that stimulate the immune system. The study evaluated the effects of a microemulsion delivery system with levamisole hydrochloride on the functional activity of MN phagocytes incubated with G. lamblia. The microemulsion formulated was incorporated with levamisole hydrochloride using distilled water, caprylic/capric triglyceride-Polymol 812®, Sorbitan Oleate-Span 80®, Polysorbate 80 - Tween 80® and 1-butanol. The activity of the microemulsion was analyzed by phagocytosis rate, microbicidal activity, apoptosis rate and intracellular calcium concentration. Phagocytosis rate, microbicidal activity and apoptosis index increased in the microemulsion treatment. The results suggest that the microemulsion improves the therapeutic efficacy of levamisole, increasing the functional activity of phagocytes. The microemulsion with a levamisole delivery system is therefore an efficient alternative for treating giardiasis, acting as an immunomodulator that probably causes fewer side effects than conventional drugs. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Incidence of Giardia lamblia Subspecies by PCR-RFLP in Stool Specimens of Hospitalized Children at Urmia Mutahhari Hospital, West Azerbaijan Province, Iran.

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    Khosro Hazrati Tappeh

    2014-12-01

    Full Text Available Giardia lamblia is one of the most prevalent intestinal flagellate protozoa that infects a wide range of vertebrate hosts causing severe intestinal disorder in children.This study was performed to determine subspecies of G.lamblia by the PCR-RFLP method, targeting the glutamate dehydrogenase(gdhlocus, in hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province,Iran and determining the infection transformational storages in this area.Overall, 720 stool specimens were collected from the hospitalized children, 34 samples were positive and Giardia cysts were detected under the microscope. Cysts were partially purified by the sucrose density gradient method and then washed with sterile distilled water to remove effectively the PCR inhibitors. Genomic DNA of G. lamblia isolates was extracted by freeze-thaw cycles followed by phenol/ chloroform/isoamyl alcohol method. The single step PCR-RFLP assay was used to differentiate the assemblages between A and B, which were found in humans. In this method, 432 bp expected size was amplified, and then for detection of subspecies, specific restriction RsaI and BspLI enzymes were used.Totally 34 samples were positive in terms of Giardia cyst out of 720 examined samples microscopically, so the parasite spread rate is reported 4.72%. Analysis PCR-RFLP on these samples revealed that 28 samples (93.3% have the genotype BIII and 2 samples (6.7% belong to the subgroup BIV.PCR-RFLP is a proper analytical method for determining the genotype among parasite types, using the glutamate dehydrogenizes zone's genes. Based on the results, an animal origin of infection cycle is suggested.

  10. An Improved PCR-RFLP Assay for Detection and Genotyping of Asymptomatic Giardia lamblia Infection in a Resource-Poor Setting.

    Science.gov (United States)

    Hawash, Yoursry; Ghonaim, M M; Al-Shehri, S S

    2016-02-01

    Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, RIDA(®) Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.

  11. Comparative analysis of the 5S rRNA and its associated proteins reveals unique primitive rather than parasitic features in Giardia lamblia.

    Science.gov (United States)

    Feng, Jin-Mei; Sun, Jun; Xin, De-Dong; Wen, Jian-Fan

    2012-01-01

    5S rRNA is a highly conserved ribosomal component. Eukaryotic 5S rRNA and its associated proteins (5S rRNA system) have become very well understood. Giardia lamblia was thought by some researchers to be the most primitive extant eukaryote while others considered it a highly evolved parasite. Previous reports have indicated that some aspects of its 5S rRNA system are simpler than that of common eukaryotes. We here explore whether this is true to its entire system, and whether this simplicity is a primitive or parasitic feature. By collecting and confirming pre-existing data and identifying new data, we obtained almost complete datasets of the system of three isolates of G. lamblia, two other parasitic excavates (Trichomonas vaginalis, Trypanosoma cruzi), and one free-living one (Naegleria gruberi). After comprehensively comparing each aspect of the system among these excavates and also with those of archaea and common eukaryotes, we found all the three Giardia isolates to harbor a same simplified 5S rRNA system, which is not only much simpler than that of common eukaryotes but also the simplest one among those of these excavates, and is surprisingly very similar to that of archaea; we also found among these excavates the system in parasitic species is not necessarily simpler than that in free-living species, conversely, the system of free-living species is even simpler in some respects than those of parasitic ones. The simplicity of Giardia 5S rRNA system should be considered a primitive rather than parasitically-degenerated feature. Therefore, Giardia 5S rRNA system might be a primitive system that is intermediate between that of archaea and the common eukaryotic model system, and it may reflect the evolutionary history of the eukaryotic 5S rRNA system from the archaeal form. Our results also imply G. lamblia might be a primitive eukaryote with secondary parasitically-degenerated features.

  12. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley

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    Díaz-Cinco Martha Elvia

    2002-01-01

    Full Text Available Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos. Es un estudio experimental efectuado entre 1995 y 1999, en el Centro de Investigación en Alimentación y Desarrollo, de la ciudad de Hermosillo, estado de Sonora, México, y cuya muestra estuvo integrada por dos grupos de 6 a 8 unidades experimentales, las cuales recibieron dos tipos de dieta y cinco tratamientos con diferente concentración de inóculo de quistes de Giardia lamblia. Durante la fase experimental se monitoreó la excreción de quistes, ganancia de peso, consumo de alimento, análisis del contenido intestinal y daños macro y microscópicos en mucosa intestinal. El tratamiento estadístico de los datos consistió en análisis de varianza, y de residuales cuando fue necesario. Resultados. En los animales alimentados con una dieta que cubre sus requerimientos nutricionales (DN se observó que es posible provocar la infección con 60 quistes, mientras que en los animales desnutridos (dieta DLN la concentración que se requiere es de únicamente seis quistes para establecer daños a nivel de mucosa intestinal. Conclusiones. La ganancia de peso monitoreada durante los 10 días de ensayo no resultó ser un buen indicador de la infección por Giardia lamblia. El porcentaje de infección depende de la concentración de quistes inoculados y del estado de nutrición de los animales evaluados.

  13. Evidence for Lateral Transfer of Genes Encoding Ferredoxins, Nitroreductases, NADH Oxidase, and Alcohol Dehydrogenase 3 from Anaerobic Prokaryotes to Giardia lamblia and Entamoeba histolytica

    Science.gov (United States)

    Nixon, Julie E. J.; Wang, Amy; Field, Jessica; Morrison, Hilary G.; McArthur, Andrew G.; Sogin, Mitchell L.; Loftus, Brendan J.; Samuelson, John

    2002-01-01

    Giardia lamblia and Entamoeba histolytica are amitochondriate, microaerophilic protists which use fermentation enzymes like those of bacteria to survive anaerobic conditions within the intestinal lumen. Genes encoding fermentation enzymes and related electron transport peptides (e.g., ferredoxins) in giardia organisms and amebae are hypothesized to be derived from either an ancient anaerobic eukaryote (amitochondriate fossil hypothesis), a mitochondrial endosymbiont (hydrogen hypothesis), or anaerobic bacteria (lateral transfer hypothesis). The goals here were to complete the molecular characterization of giardial and amebic fermentation enzymes and to determine the origins of the genes encoding them, when possible. A putative giardia [2Fe-2S]ferredoxin which had a hypothetical organelle-targeting sequence at its N terminus showed similarity to mitochondrial ferredoxins and the hydrogenosomal ferredoxin of Trichomonas vaginalis (another luminal protist). However, phylogenetic trees were star shaped, with weak bootstrap support, so we were unable to confirm or rule out the endosymbiotic origin of the giardia [2Fe-2S]ferredoxin gene. Putative giardial and amebic 6-kDa ferredoxins, ferredoxin-nitroreductase fusion proteins, and oxygen-insensitive nitroreductases each tentatively supported the lateral transfer hypothesis. Although there were not enough sequences to perform meaningful phylogenetic analyses, the unique common occurrence of these peptides and enzymes in giardia organisms, amebae, and the few anaerobic prokaryotes suggests the possibility of lateral transfer. In contrast, there was more robust phylogenetic evidence for the lateral transfer of G. lamblia genes encoding an NADH oxidase from a gram-positive coccus and a microbial group 3 alcohol dehydrogenase from thermoanaerobic prokaryotes. In further support of lateral transfer, the G. lamblia NADH oxidase and adh3 genes appeared to have an evolutionary history distinct from those of E. histolytica. PMID

  14. Comparison of four rapid diagnostic tests, ELISA, microscopy and PCR for the detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in feces.

    Science.gov (United States)

    Van den Bossche, Dorien; Cnops, Lieselotte; Verschueren, Jacob; Van Esbroeck, Marjan

    2015-03-01

    Microscopy is the diagnostic reference standard for the detection of parasites, but it is labor-intensive and requires experience. Rapid diagnostic tests (RDTs) can provide an alternative to microscopy. RDTs from four different manufacturers were compared to enzyme-linked immunosorbent assay (ELISA), microscopy and/or parasite-specific real-time PCR: ImmunoCardSTAT!®CGE (Meridian Bioscience Inc., Cincinnati, Ohio, USA) (A), Crypto/Giardia Duo-Strip (Coris Bioconcepts, Gembloux, Belgium) (B), RIDA®QUICK Cryptosporidium/Giardia/Entamoeba Combi (R-BioPharm, Darmstadt, Germany) (C) and Giardia/Cryptosporidium Quik Chek (Techlab Inc., Blacksburg, Virginia, USA) (D). Thirty frozen samples were analyzed retrospectively. For Giardia lamblia (n=12) and Cryptosporidium (n=12) sensitivities ranged from 58% (B), over 83% (A, C) to 100% (D) and from 92% (B) to 100% (A, C, D), respectively. Specificity for both G. lamblia and Cryptosporidium was 100% for all RDT brands. Sensitivity for Entamoeba histolytica (n=5) was 100%, while specificity reached 80% (A) to 88% (C). In a prospective study, fresh samples were tested. For G. lamblia (n=30), sensitivity ranged from 66% (B), over 79% (A) and 83% (C) to 100% (D) and specificity varied between 94% (D) and 100% (A, B, C). For Cryptosporidium (n=3), sensitivity was 100% for all brands except (B) (67%) and specificities were 95% (A, B), 98% (C) and 100% (D). E. histolytica (n=1) was detected by both (A) and (C), while specificity was 81% and 87% respectively. RDTs can be a valuable tool when microscopic expertise is poor and in remote and outbreak settings where other techniques are often not available and rapid diagnosis is required. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Oxidative stress-induced cell cycle blockage and a protease-independent programmed cell death in  microaerophilic Giardia lamblia

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    Esha Ghosh

    2009-03-01

    Full Text Available Esha Ghosh1, Arjun Ghosh1, Amar Nath Ghosh2, Tomoyoshi Nozaki3, Sandipan Ganguly11Division of Parasitology; 2Division of Electron Microscopy, National Institute of Cholera and Enteric Diseases, Beliaghata, Kolkata, West Bengal, India; 3Division of Parasitology, National Institute of Infectious Diseases, Tokyo, JapanAbstract: Giardia lamblia is a microaerophilic human gastrointestinal parasite and considered as an early-diverged eukaryote. In vitro oxidative stress generation plays a significant role in cell cycle progression and cell death of this parasite. In the present study hydrogen peroxide, metronidazole, and a modified growth medium without cysteine and ascorbic acid have been chosen as oxidative stress-inducing agents. Cell cycle progression has been found to be regulated by different types of oxidative stresses. Apoptosis is not an established pathway in Giardia, which is devoid of ideal mitochondria, but in the present investigation, apoptosis-like programmed cell death has been found by the experiments like AnnexinV-FITC assay, DNA fragmentation pattern, etc. On the contrary, Caspase-9 assay, which confirms the caspase-mediated apoptotic pathway, has been found to be negative in all the stress conditions. Protease inhibitor assay confirmed that, even in absence of any proteases, programmed cell death does occur in this primitive eukaryote. All these results signify a novel pathway of programmed suicidal death in Giardia lamblia under oxidative stress. This is the first demonstration of protease-independent programmed cell death regulation in Giardia exclusive for its own specialties.Keywords: Giardia lamblia, oxidative stress, reactive oxygen species, programmed cell death, apoptosis, early branching eukaryotes

  16. Simultaneous detection and differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii, Giardia lamblia and Cryptosporidium spp. in human fecal samples using multiplex PCR and qPCR-MCA.

    Science.gov (United States)

    Zebardast, Nozhat; Yeganeh, Farshid; Gharavi, Mohammad Javad; Abadi, Alireza; Seyyed Tabaei, Seyyed Javad; Haghighi, Ali

    2016-10-01

    Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp. are common causes of diarrheal and intestinal diseases all over the world. Microscopic methods are useful in the diagnosis of intestinal parasites (IPs), but their sensitivity was assessed approximately 60 percent. Recently, molecular techniques have been used increasingly for the identification and characterization of the parasites. Among those, in this study we have used multiplex PCR and Real-time PCR with melting curve analysis (qPCR-MCA) for simultaneous detection and differentiation of E. histolytica, E. dispar, E. moshkovskii, G. lamblia and Cryptosporidium spp. in human fecal samples. Twenty DNA samples from 12 E. histolytica and 8 E. dispar samples and twenty stool samples confirmed positive for G. lamblia and Cryptosporidium spp. were analyzed. After DNA extraction from the samples, multiplex PCR was done for detection and differentiation of above mentioned parasites. QPCR-MCA was also performed for the detection and differentiation of 11 isolates of above mentioned parasite in a cycle with a time and temperature. Multiplex PCR was able to simultaneous detect and differentiate of above mentioned parasite in a single reaction. QPCR-MCA was able to differentiate genus and species those five protozoa using melting temperature simultaneously at the same time and temperature programs. In total, qPCR-MCA diagnosed 7/11 isolation of E. histolytica, 6/8 isolation of E. dispar, 1/1 E. moshkovskii Laredo, 10/11 G. Lamblia and 6/11 Cryptosporidium spp. Application of multiplex PCR for detection of more than one species in a test in developing countries, at least in reference laboratories has accurate diagnosis and plays a critical role in differentiation of protozoan species. Multiplex PCR assay with a template and multi template had different results and it seems that using a set of primers with one template has higher diagnostic capability in compare with multi template. The results of this study

  17. Cyst-Wall-Protein-1 is fundamental for Golgi-like organelle neogenesis and cyst-wall biosynthesis in Giardia lamblia

    OpenAIRE

    2016-01-01

    The genome of the protozoan parasite Giardia lamblia is organized in two diploid nuclei, which has so far precluded complete analysis of gene function. Here we use a previously developed Cre/loxP-based knock-out and selection marker salvage strategy in the human-derived isolate WB-C6 to eliminate all four copies of the Cyst-Wall-Protein-1 locus (CWP1). Because these loci are silenced in proliferating trophozoites and highly expressed only in encysting cells, CWP1 ablation allows functional ch...

  18. vsp gene expression by Giardia lamblia clone GS/M-83-H7 during antigenic variation in vivo and in vitro.

    Science.gov (United States)

    Bienz, M; Siles-Lucas, M; Wittwer, P; Müller, N

    2001-09-01

    Giardia lamblia infections are associated with antigenic variation of the parasite, which is generated by a continuous change of the variant-specific surface proteins (VSPs). Many investigations on the process of antigenic variation were based on the use of G. lamblia clone GS/M-83-H7, which expresses VSP H7 as its major surface antigen. In the present study, mice were infected with the aforementioned clonal line to investigate vsp gene expression during the complex process of antigenic variation of the parasite. Trophozoites collected from the intestines of individual animals at different time points postinfection (p.i.) were analyzed directly for their vsp gene expression patterns, i.e., without cultivating the recovered parasites in vitro. Because few trophozoites were recovered at late time points p.i., a combined 5' rapid amplification of cDNA ends-reverse transcription-PCR approach was utilized. This allowed detection and subsequent sequence analysis of vsp gene transcripts upon generation of amplified cDNA analogues. The same PCR approach was applied for analysis of vsp gene expression in variants obtained after negative selection of axenic GS/M-83-H7 trophozoites by treatment with a cytotoxic, VSP H7-specific monoclonal antibody. In an overall view of the entire panel of in vivo- and in vitro-derived parasite populations, expression of 29 different vsp gene sequences could be demonstrated. In vivo antigenic variation of G. lamblia clone GS/M-83-H7 was shown to be a continuous process involving the consecutive appearance of relatively distinct sets of vsp transcripts. During the 42-day infection period investigated, this process activated at least 22 different vsp genes. Comparative molecular analyses of the amino acid level demonstrated that all cDNA segments identified encode structural elements typical of the terminal segment of Giardia VSP. The similarity of most of the GS/M-83-H7 VSP sequences identified in the present study supports previous

  19. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley

    OpenAIRE

    Díaz-Cinco Martha Elvia; Ballesteros-Vázquez Martha Nydia; Pérez-Morales Rosalba; Mata-Haro Verónica

    2002-01-01

    Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos. Es un estudio experimental efectuado entre 1995 y 1999, en el Centro de Investigación en Alimentación y Desarrollo, de la ciudad de Hermosillo, estado de Sonora, México, y cuya muestra estuvo integrada por dos grupos de 6 a 8 unidades experimentales, las cuales recibieron dos tipos d...

  20. The principal conductance in Giardia lamblia trophozoites possesses functional properties similar to the mammalian ClC-2 current.

    Science.gov (United States)

    Moreno-Galindo, Eloy G; Rodríguez-Elías, Julio C; Ramírez-Herrera, Mario A; Sánchez-Chapula, José A; Navarro-Polanco, Ricardo A

    2014-05-01

    The human intestinal pathogen Giardia lamblia is a flagellated unicellular protozoan with pronounced medical and biological relevance. However, the basic physiology of Giardia trophozoites has been sparsely studied, especially the electrical and ionic properties of their cellular membrane which are virtually unknown. In this study, we were able to record and characterize the macroscopic ionic currents of Giardia trophozoite membrane by electrophysiological methods of the patch clamp technique. Giardia trophozoites showed a high current density (∼600 pA/pF at -140 mV) that was activated upon hyperpolarization. This current was carried by a chloride-selective channel (I Cl-G) and it was the most important determinant of the membrane potential in Giardia trophozoites. Moreover, this conductance was able to carry other halide anions and the sequence of permeability was Br(-) > Cl(-) ≈ I(-) ≫ F(-). Besides the voltage-dependent inward-rectifying nature of I Cl-G, its activation and deactivation kinetics were comparable to those observed in ClC-2 channels. Extracellular pH modified the voltage-dependent properties of I Cl-G, shifting the activation curve from a V 1/2 = -79 ± 1 mV (pH 7.4) to -93 ± 2 mV (pH 8.4) and -112 ± 2 mV (pH 5.4). Furthermore, the maximal amplitude of I Cl-G measured at -100 mV showed dependence to external pH in a bell-shaped fashion reported only for ClC-2 channels. Therefore, our results suggest that I Cl-G possesses several functional properties similar to the mammalian ClC-2 channels.

  1. Rational Design Synthesis and Evaluation of First Generation Inhibitors of the Giardia Lamblia Fructose-1 6-biphosphate Aldolase

    Energy Technology Data Exchange (ETDEWEB)

    Z Li; Z Liu; D Cho; J Zou; M Gong; R Breece; A Galkin; L Li; H Zhao; et al.

    2011-12-31

    Inhibitors of the Giardia lamblia fructose 1,6-bisphosphate aldolase (GlFBPA), which transforms fructose 1,6-bisphosphate (FBP) to dihydroxyacetone phosphate and glyceraldehyde 3-phosphate, were designed based on 3-hydroxy-2-pyridone and 1,2-dihydroxypyridine scaffolds that position two negatively charged tetrahedral groups for interaction with substrate phosphate binding residues, a hydrogen bond donor to the catalytic Asp83, and a Zn{sup 2+} binding group. The inhibition activities for the GlFBPA catalyzed reaction of FBP of the prepared alkyl phosphonate/phosphate substituted 3-hydroxy-2-pyridinones and a dihydroxypyridine were determined. The 3-hydroxy-2-pyridone inhibitor 8 was found to bind to GlFBPA with an affinity (K{sub i} = 14 {micro}M) that is comparable to that of FBP (K{sub m} = 2 {micro}M) or its inert analog TBP (K{sub i} = 1 {micro}M). The X-ray structure of the GlFBPA-inhibitor 8 complex (2.3 {angstrom}) shows that 8 binds to the active site in the manner predicted by in silico docking with the exception of coordination with Zn{sup 2+}. The observed distances and orientation of the pyridone ring O=C-C-OH relative to Zn{sup 2+} are not consistent with a strong interaction. To determine if Zn{sup 2+} coordination occurs in the GlFBPA-inhibitor 8 complex in solution, EXAFS spectra were measured. A four coordinate geometry comprised of the three enzyme histidine ligands and an oxygen atom from the pyridone ring O=C-C-OH was indicated. Analysis of the Zn{sup 2+} coordination geometries in recently reported structures of class II FBPAs suggests that strong Zn{sup 2+} coordination is reserved for the enediolate-like transition state, accounting for minimal contribution of Zn{sup 2+} coordination to binding of 8 to GlFBPA.

  2. Identification of scaffold/Matrix Attachment (S/MAR like DNA element from the gastrointestinal protozoan parasite Giardia lamblia

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    Gautam Pennathur

    2010-06-01

    Full Text Available Abstract Background Chromatin in the nucleus of all eukaryotes is organized into a system of loops and domains. These loops remain fastened at their bases to the fundamental framework of the nucleus, the matrix or the scaffold. The DNA sequences which anchor the bases of the chromatin loops to the matrix are known as Scaffold/Matrix Attachment Regions or S/MARs. Though S/MARs have been studied in yeast and higher eukaryotes and they have been found to be associated with gene organization and regulation of gene expression, they have not been reported in protists like Giardia. Several tools have been discovered and formulated to predict S/MARs from a genome of a higher eukaryote which take into account a number of features. However, the lack of a definitive consensus sequence in S/MARs and the randomness of the protozoan genome in general, make it a challenge to predict and identify such sequences from protists. Results Here, we have analysed the Giardia genome for the probable S/MARs predicted by the available computational tools; and then shown these sequences to be physically associated with the nuclear matrix. Our study also reflects that while no single computational tool is competent to predict such complex elements from protist genomes, a combination of tools followed by experimental verification is the only way to confirm the presence of these elements from these organisms. Conclusion This is the first report of S/MAR elements from the protozoan parasite Giardia lamblia. This initial work is expected to lay a framework for future studies relating to genome organization as well as gene regulatory elements in this parasite.

  3. Structural and functional perturbation of Giardia lamblia triosephosphate isomerase by modification of a non-catalytic, non-conserved region.

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    Gloria Hernández-Alcántara

    Full Text Available BACKGROUND: We have previously proposed triosephosphate isomerase of Giardia lamblia (GlTIM as a target for rational drug design against giardiasis, one of the most common parasitic infections in humans. Since the enzyme exists in the parasite and the host, selective inhibition is a major challenge because essential regions that could be considered molecular targets are highly conserved. Previous biochemical evidence showed that chemical modification of the non-conserved non-catalytic cysteine 222 (C222 inactivates specifically GlTIM. The inactivation correlates with the physicochemical properties of the modifying agent: addition of a non-polar, small chemical group at C222 reduces the enzyme activity by one half, whereas negatively charged, large chemical groups cause full inactivation. RESULTS: In this work we used mutagenesis to extend our understanding of the functional and structural effects triggered by modification of C222. To this end, six GlTIM C222 mutants with side chains having diverse physicochemical characteristics were characterized. We found that the polarity, charge and volume of the side chain in the mutant amino acid differentially alter the activity, the affinity, the stability and the structure of the enzyme. The data show that mutagenesis of C222 mimics the effects of chemical modification. The crystallographic structure of C222D GlTIM shows the disruptive effects of introducing a negative charge at position 222: the mutation perturbs loop 7, a region of the enzyme whose interactions with the catalytic loop 6 are essential for TIM stability, ligand binding and catalysis. The amino acid sequence of TIM in phylogenetic diverse groups indicates that C222 and its surrounding residues are poorly conserved, supporting the proposal that this region is a good target for specific drug design. CONCLUSIONS: The results demonstrate that it is possible to inhibit species-specifically a ubiquitous, structurally highly conserved enzyme by

  4. Static Clathrin Assemblies at the Peripheral Vacuole-Plasma Membrane Interface of the Parasitic Protozoan Giardia lamblia.

    Science.gov (United States)

    Zumthor, Jon Paulin; Cernikova, Lenka; Rout, Samuel; Kaech, Andres; Faso, Carmen; Hehl, Adrian B

    2016-07-01

    Giardia lamblia is a parasitic protozoan that infects a wide range of vertebrate hosts including humans. Trophozoites are non-invasive but associate tightly with the enterocyte surface of the small intestine. This narrow ecological specialization entailed extensive morphological and functional adaptations during host-parasite co-evolution, including a distinctly polarized array of endocytic organelles termed peripheral vacuoles (PVs), which are confined to the dorsal cortical region exposed to the gut lumen and are in close proximity to the plasma membrane (PM). Here, we investigated the molecular consequences of these adaptations on the Giardia endocytic machinery and membrane coat complexes. Despite the absence of canonical clathrin coated vesicles in electron microscopy, Giardia possesses conserved PV-associated clathrin heavy chain (GlCHC), dynamin-related protein (GlDRP), and assembly polypeptide complex 2 (AP2) subunits, suggesting a novel function for GlCHC and its adaptors. We found that, in contrast to GFP-tagged AP2 subunits and DRP, CHC::GFP reporters have no detectable turnover in living cells, indicating fundamental differences in recruitment to the membrane and disassembly compared to previously characterized clathrin coats. Histochemical localization in electron tomography showed that these long-lived GlCHC assemblies localized at distinctive approximations between the plasma and PV membrane. A detailed protein interactome of GlCHC revealed all of the conserved factors in addition to novel or highly diverged proteins, including a putative clathrin light chain and lipid-binding proteins. Taken together, our data provide strong evidence for giardial CHC as a component of highly stable assemblies at PV-PM junctions that likely have a central role in organizing continuities between the PM and PV membranes for controlled sampling of the fluid environment. This suggests a novel function for CHC in Giardia and the extent of molecular remodeling of

  5. Modeling long-term host cell-Giardia lamblia interactions in an in vitro co-culture system.

    Science.gov (United States)

    Fisher, Bridget S; Estraño, Carlos E; Cole, Judith A

    2013-01-01

    Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite, Giardia lamblia, is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium, Giardia induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of Giardia pathology is the lack of a functionally relevant model for the long-term study of the parasite in vitro. Here we report on the development of an in vitro co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of Giardia trophozoites. Using the developed model, we show that Giardia trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and Giardia medium. Giardia induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by Giardia indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

  6. A microRNA derived from an apparent canonical biogenesis pathway regulates variant surface protein gene expression in Giardia lamblia

    Science.gov (United States)

    Saraiya, Ashesh A.; Li, Wei; Wang, Ching C.

    2011-01-01

    We have previously shown that a snoRNA-derived microRNA, miR2, in Giardia lamblia potentially regulates the expression of 22 variant surface protein (VSP) genes. Here, we identified another miRNA, miR4, also capable of regulating the expression of several VSPs but derived from an unannotated open reading frame (ORF) rather than a snoRNA, suggesting a canonical miRNA biogenesis pathway in Giardia. miR4 represses expression of a reporter containing two miR4 antisense sequences at the 3′ UTR without causing a corresponding decrease in the mRNA level. This repression requires the presence of the Giardia Argonaute protein (GlAgo) and is reversed by 2′ O–methylated antisense oligo to miR4, suggesting an RNA-induced silencing complex (RISC)–mediated mechanism. Furthermore, in vivo and in vitro evidence suggested that the Giardia Dicer protein (GlDcr) is required for miR4 biogenesis. Coimmunoprecipitation of miR4 with GlAgo further verified miR4 as a miRNA. A total of 361 potential target sites for miR4 were bioinformatically identified in Giardia, out of which 69 (32.7%) were associated with VSP genes. miR4 reduces the expression of a reporter containing two copies of the target site from VSP (GL50803_36493) at the 3′ UTR. Sixteen of the 69 VSP genes were further found to contain partially overlapping miR2 and miR4 targeting sites. Expression of a reporter carrying the two overlapping sites was inhibited by either miR2 or miR4, but the inhibition was neither synergistic nor additive, suggesting a complex mechanism of miRNA regulation of VSP expression and the presence of a rich miRNAome in Giardia. PMID:22033329

  7. Modeling long-term host cell-Giardia lamblia interactions in an in vitro co-culture system.

    Directory of Open Access Journals (Sweden)

    Bridget S Fisher

    Full Text Available Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite, Giardia lamblia, is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium, Giardia induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of Giardia pathology is the lack of a functionally relevant model for the long-term study of the parasite in vitro. Here we report on the development of an in vitro co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of Giardia trophozoites. Using the developed model, we show that Giardia trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and Giardia medium. Giardia induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by Giardia indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

  8. Occurence of Giardia lamblia in children of municipal day-care centers from Botucatu, São Paulo State, Brazil Ocorrência de Giardia lamblia em crianças atendidas em creches municipais de Botucatu, Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Semíramis Guimarães

    1995-12-01

    Full Text Available Considering that the number of day-care centers for pre-school-age children has expanded rapidly in developing countries, and that these institutions presenting conditions that facilitate the transmission of many enteric agents, a parasitological survey was carried out in three municipal day-cares from Botucatu: two in the urban area (one in downtown area and the other one in the city periphery area and the third in the rural area. Three separate stool specimens were collected from 147 children ranging from 0 to 72 months old and 20 staff members. Each stool specimen was processed by Lutz and zinc sulfate flotation methods. The frequency of giardiasis observed among children of downtown, periphery and rural day-cares was 69.6%, 52.7% and 69.6%, respectively. Only one employee was positive for G. lamblia. The examination of three stool specimens increased the positivity for G. lamblia: from the ninety three final positive examinations, 24 (25.5% and 8 (8.5% were positives only after examination of the second and third samples, respectively. Others intestinal organisms like Ascaris lumbricoides (20.4%, Trichuris trichiura (19.0%. Hymenolepis nana (8.8%, Entamoeba coli (22.4% and Blastocystis hominis (32.0% were frequently found in the children. There was no significant association among localization of the day-cares, sex of the children and the levels of G. lamblia infection. According to the age, G. lamblia was found mainly in children between 12 to 47 months old.Foi realizado um levantamento parasitológico em três creches da rede municipal: duas na área urbana, uma no centro da cidade e a outra na periferia, e a terceira na zona rural. Três amostras de fezes, coletadas com intervalo de 7 dias, foram obtidas de 147 crianças com idade variando de 0 a 6 anos e de 20 funcionários. Cada amostra foi processada pelos métodos de sedimentação espontânea e centrífugo - flutuação com sulfato de zinco. Os resultados demonstraram que das 147

  9. Validation of housekeeping genes as an internal control for gene expression studies in Giardia lamblia using quantitative real-time PCR.

    Science.gov (United States)

    Marcial-Quino, Jaime; Fierro, Francisco; De la Mora-De la Mora, Ignacio; Enríquez-Flores, Sergio; Gómez-Manzo, Saúl; Vanoye-Carlo, America; Garcia-Torres, Itzhel; Sierra-Palacios, Edgar; Reyes-Vivas, Horacio

    2016-04-25

    The analysis of transcript levels of specific genes is important for understanding transcriptional regulation and for the characterization of gene function. Real-time quantitative reverse transcriptase PCR (RT-qPCR) has become a powerful tool to quantify gene expression. The objective of this study was to identify reliable housekeeping genes in Giardia lamblia. Twelve genes were selected for this purpose, and their expression was analyzed in the wild type WB strain and in two strains with resistance to nitazoxanide (NTZ) and metronidazole (MTZ), respectively. RefFinder software analysis showed that the expression of the genes is different in the three strains. The integrated data from the four analyses showed that the NADH oxidase (NADH) and aldolase (ALD) genes were the most steadily expressed genes, whereas the glyceraldehyde-3-phosphate dehydrogenase gene was the most unstable. Additionally, the relative expression of seven genes were quantified in the NTZ- and MTZ-resistant strains by RT-qPCR, using the aldolase gene as the internal control, and the results showed a consistent differential pattern of expression in both strains. The housekeeping genes found in this work will facilitate the analysis of mRNA expression levels of other genes of interest in G. lamblia.

  10. RESEARCH PROGRESS ON THE PATHOGENIC MECHANISM OF GIARDIA LAMBLIA%蓝氏贾第虫致病机制的研究进展

    Institute of Scientific and Technical Information of China (English)

    武省; 李国清

    2015-01-01

    蓝氏贾第虫(Giardialamblia)是一种重要的人畜共患寄生虫,可引起人和多种哺乳动物的腹泻。近年来人们对其致病机制进行了大量研究,包括贾第虫结构蛋白(贾第素)和排泄分泌物,表面抗原变异以及贾第虫对小肠的影响等,本文对此进行了综述。%Giardialambliais an important zoonotic parasite, and a leading cause of diarrhoeal disease in human beings and many kinds of mammals. In recent years, there have been an enormous amount of researches on the pathogenic mechanism of G.lamblia, including the cytoskeletal protein (giardin) and excretory secretory product, surface antigenic variation, and the effects of G.lamblia on small intestine of host including the enterocyte apoptosis, structural damage and dysfunction. This review summarizes the above aspects.

  11. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley The effect of malnourishment on the development of giardiosis in Sprague-Dawley rats

    National Research Council Canada - National Science Library

    Martha Elvia Díaz-Cinco; Martha Nydia Ballesteros-Vázquez; Rosalba Pérez-Morales; Verónica Mata-Haro

    2002-01-01

    Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos...

  12. Comparison of the Triage Micro Parasite Panel and Microscopy for the Detection of Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Cryptosporidium parvum in Stool Samples Collected in Kenya

    Directory of Open Access Journals (Sweden)

    Brett Swierczewski

    2012-01-01

    Full Text Available Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are three of the most important parasitic causes of acute diarrhea worldwide. Laboratory diagnosis of these parasites is usually done by ova and parasite examination (O&P examination via microscopy. The sensitivity and specificity of O&P examination varies among laboratories and can be labor intensive and time consuming. The Triage Micro Parasite Panel (BioSite, San Diego, California is an enzyme immunoassay kit that can detect E. histolytica/E. dispar, G. lamblia, and C. parvum simultaneously using fresh or frozen stool. The present study evaluated the Triage Micro Parasite Panel in detecting E. histolytica/E. dispar, G. lamblia, and C. parvum compared to O&P examination in 266 stool samples collected at medical facilities in Kenya. The sensitivity and specificity results for the Triage Micro Parasite Panel were: for E. histolytica/E. dispar: 100%, 100%, G. lamblia: 100%, 100% and C. parvum: 73%, 100%. There was no evidence of cross reactivity using the kit with other parasites identified in the stool specimens. These results indicate that the Triage Micro Parasite Panel is a highly sensitive kit that can be used for screening purposes in large scale studies or outbreak investigations or as a possible alternative to O&P examination.

  13. Different risk factors for infection with Giardia lamblia assemblages A and B in children attending day-care centres.

    Science.gov (United States)

    Pijnacker, R; Mughini-Gras, L; Heusinkveld, M; Roelfsema, J; van Pelt, W; Kortbeek, T

    2016-12-01

    Giardia lamblia is a major cause of diarrhoea in children, especially those attending day-care centres (DCCs). Only Giardia assemblages A and B infect humans. Given the lack of assemblage-specific epidemiological data, we aimed to identify risk factors for infection by assemblages A and B in DCC attendees. During 2010-2013, 5,015 faecal samples from ≤4-year-old children attending 40 DCCs participating in laboratory surveillance in the Netherlands were tested for Giardia using RT-PCR. Giardia-positive samples were typed for identification of assemblages A and B. We compared child- and DCC-level characteristics of Giardia-positive children with those of Giardia-negative children using mixed-effects logistic regression. Overall, 226 samples (4.5 %) tested positive for Giardia, and assemblages were determined for 138 of them: 62 (45 %) were assemblage A and 76 (55 %) were B. The only risk factor for assemblage A infection was attending DCCs with indoor sandpits and cats during spring/summer (odds ratio [OR] 13.5; 95% CI 1.8-101.3). For assemblage B, risk factors were attending DCCs with dedicated diaper-changing (OR 3.6; 95% CI 1.7-7.6) and laundry (OR 2.3; 95% CI 1.1-4.9) areas. Preventing sick children from attending day-care and having cloth-towels at the DCC decreased the risk of assemblage B infection (OR 0.0; 95% CI 0.0-0.5 and OR 0.3; 95% CI 0.1-0.6 respectively). Risk factors for assemblages A and B infection in DCC-attending children were different, with assemblage B being mainly related to anthroponotic transmission, and assemblage A being related to zoonotic transmission. Given these differences, interventions to reduce the burden of childhood giardiasis cannot ignore those assemblage-specific preferred reservoirs and transmission routes.

  14. The profile of snoRNA-derived microRNAs that regulate expression of variant surface proteins in Giardia lamblia.

    Science.gov (United States)

    Li, Wei; Saraiya, Ashesh A; Wang, Ching C

    2012-09-01

    In the current investigation, we analysed all the known small nucleolar RNAs (snoRNAs) in the deeply branching protozoan parasite Giardia lamblia for potential microRNAs (miRNAs) that might be derived from them. Two putative miRNAs have since been identified by Northern blot, primer extension, 3' RACE and co-immunoprecipitation with Giardia Argonaute (GlAgo), and designated miR6 and miR10. Giardia Dicer (GlDcr) is capable of processing the snoRNAs into the corresponding miRNAs in vitro. Potential miR6 and miR10 binding sites in Giardia genome were predicted bio-informatically. A miR6 binding site was found at the 3' untranslated regions (UTR) of 44 variant surface protein (vsp) genes, whereas a miR10 binding site was identified at the 3' end of 159 vsp open-reading frames. Thirty-three of these vsp genes turned out to contain binding sites for both miR6 and miR10. A reporter mRNA tagged with the 3' end of vsp1267, which contains the target sites for both miRNAs, was translationally repressed by both miRNAs in Giardia. Episomal expression of an N-terminal c-myc tagged VSP1267 was found significantly repressed by introducing either miR6 or miR10 into the cells and the repressive effects were additive. When the 2'-O-methyl antisense oligos (ASOs) of either miR6 or miR10 was introduced, however, there was an enhancement of tagged VSP1267 expression suggesting an inhibition of the repressive effects of endogenous miR6 or miR10 by the ASOs. Of the total 220 vsp genes in Giardia, we have now found 178 of them carrying putative binding sites for all the miRNAs that have been currently identified, suggesting that miRNAs are likely the regulators of VSP expression in Giardia.

  15. Rapid imaging, detection and quantification of Giardia lamblia cysts using mobile-phone based fluorescent microscopy and machine learning.

    Science.gov (United States)

    Koydemir, Hatice Ceylan; Gorocs, Zoltan; Tseng, Derek; Cortazar, Bingen; Feng, Steve; Chan, Raymond Yan Lok; Burbano, Jordi; McLeod, Euan; Ozcan, Aydogan

    2015-03-07

    Rapid and sensitive detection of waterborne pathogens in drinkable and recreational water sources is crucial for treating and preventing the spread of water related diseases, especially in resource-limited settings. Here we present a field-portable and cost-effective platform for detection and quantification of Giardia lamblia cysts, one of the most common waterborne parasites, which has a thick cell wall that makes it resistant to most water disinfection techniques including chlorination. The platform consists of a smartphone coupled with an opto-mechanical attachment weighing ~205 g, which utilizes a hand-held fluorescence microscope design aligned with the camera unit of the smartphone to image custom-designed disposable water sample cassettes. Each sample cassette is composed of absorbent pads and mechanical filter membranes; a membrane with 8 μm pore size is used as a porous spacing layer to prevent the backflow of particles to the upper membrane, while the top membrane with 5 μm pore size is used to capture the individual Giardia cysts that are fluorescently labeled. A fluorescence image of the filter surface (field-of-view: ~0.8 cm(2)) is captured and wirelessly transmitted via the mobile-phone to our servers for rapid processing using a machine learning algorithm that is trained on statistical features of Giardia cysts to automatically detect and count the cysts captured on the membrane. The results are then transmitted back to the mobile-phone in less than 2 minutes and are displayed through a smart application running on the phone. This mobile platform, along with our custom-developed sample preparation protocol, enables analysis of large volumes of water (e.g., 10-20 mL) for automated detection and enumeration of Giardia cysts in ~1 hour, including all the steps of sample preparation and analysis. We evaluated the performance of this approach using flow-cytometer-enumerated Giardia-contaminated water samples, demonstrating an average cyst capture

  16. Protein electrophoresis - serum

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003540.htm Protein electrophoresis - serum To use the sharing features on ... JavaScript. This lab test measures the types of protein in the fluid (serum) part of a blood ...

  17. Serum iron test

    Science.gov (United States)

    Fe+2; Ferric ion; Fe++; Ferrous ion; Iron - serum; Anemia - serum iron; Hemochromatosis - serum iron ... A blood sample is needed. Iron levels are highest in the morning. Your health care provider will likely have you do this test in the morning.

  18. Comparative analysis of the 5S rRNA and its associated proteins reveals unique primitive rather than parasitic features in Giardia lamblia.

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    Jin-Mei Feng

    Full Text Available BACKGROUND: 5S rRNA is a highly conserved ribosomal component. Eukaryotic 5S rRNA and its associated proteins (5S rRNA system have become very well understood. Giardia lamblia was thought by some researchers to be the most primitive extant eukaryote while others considered it a highly evolved parasite. Previous reports have indicated that some aspects of its 5S rRNA system are simpler than that of common eukaryotes. We here explore whether this is true to its entire system, and whether this simplicity is a primitive or parasitic feature. METHODOLOGY/PRINCIPAL FINDINGS: By collecting and confirming pre-existing data and identifying new data, we obtained almost complete datasets of the system of three isolates of G. lamblia, two other parasitic excavates (Trichomonas vaginalis, Trypanosoma cruzi, and one free-living one (Naegleria gruberi. After comprehensively comparing each aspect of the system among these excavates and also with those of archaea and common eukaryotes, we found all the three Giardia isolates to harbor a same simplified 5S rRNA system, which is not only much simpler than that of common eukaryotes but also the simplest one among those of these excavates, and is surprisingly very similar to that of archaea; we also found among these excavates the system in parasitic species is not necessarily simpler than that in free-living species, conversely, the system of free-living species is even simpler in some respects than those of parasitic ones. CONCLUSION/SIGNIFICANCE: The simplicity of Giardia 5S rRNA system should be considered a primitive rather than parasitically-degenerated feature. Therefore, Giardia 5S rRNA system might be a primitive system that is intermediate between that of archaea and the common eukaryotic model system, and it may reflect the evolutionary history of the eukaryotic 5S rRNA system from the archaeal form. Our results also imply G. lamblia might be a primitive eukaryote with secondary parasitically

  19. Comparative Analysis of the 5S rRNA and Its Associated Proteins Reveals Unique Primitive Rather Than Parasitic Features in Giardia lamblia

    Science.gov (United States)

    Xin, De-Dong; Wen, Jian-Fan

    2012-01-01

    Background 5S rRNA is a highly conserved ribosomal component. Eukaryotic 5S rRNA and its associated proteins (5S rRNA system) have become very well understood. Giardia lamblia was thought by some researchers to be the most primitive extant eukaryote while others considered it a highly evolved parasite. Previous reports have indicated that some aspects of its 5S rRNA system are simpler than that of common eukaryotes. We here explore whether this is true to its entire system, and whether this simplicity is a primitive or parasitic feature. Methodology/Principal Findings By collecting and confirming pre-existing data and identifying new data, we obtained almost complete datasets of the system of three isolates of G. lamblia, two other parasitic excavates (Trichomonas vaginalis, Trypanosoma cruzi), and one free-living one (Naegleria gruberi). After comprehensively comparing each aspect of the system among these excavates and also with those of archaea and common eukaryotes, we found all the three Giardia isolates to harbor a same simplified 5S rRNA system, which is not only much simpler than that of common eukaryotes but also the simplest one among those of these excavates, and is surprisingly very similar to that of archaea; we also found among these excavates the system in parasitic species is not necessarily simpler than that in free-living species, conversely, the system of free-living species is even simpler in some respects than those of parasitic ones. Conclusion/Significance The simplicity of Giardia 5S rRNA system should be considered a primitive rather than parasitically-degenerated feature. Therefore, Giardia 5S rRNA system might be a primitive system that is intermediate between that of archaea and the common eukaryotic model system, and it may reflect the evolutionary history of the eukaryotic 5S rRNA system from the archaeal form. Our results also imply G. lamblia might be a primitive eukaryote with secondary parasitically-degenerated features. PMID

  20. Genotyping of Giardia lamblia and methods of its detection%蓝氏贾第鞭毛虫基因型及其检测方法概述

    Institute of Scientific and Technical Information of China (English)

    郑佳

    2011-01-01

    蓝氏贾第鞭毛虫致病性与其基因型密切相关.最近发展的一项分子技术real-time PCR以其实时定量、灵敏度高、特异性强等优势,被广泛应用于病毒、细菌和寄生虫的临床检测.本文就蓝氏贾第鞭毛虫基因型及检测方法的研究进行了综述.%The pathogenicity of giardia is closely related to its genotype. Recent advances in a molecular technique, realtime PCR, have provided real-time quantification, a high degree of sensitivity, a high degree of specificity, and other advantages. This technique is widely used to clinically test for viruses, bacteria, and parasites. This paper summarizes the genotyping and detection of Giardia lamblia.

  1. The progress on detection methods of Giardia lamblia%蓝氏贾第鞭毛虫检测方法的研究进展

    Institute of Scientific and Technical Information of China (English)

    朱海波; 李国清

    2010-01-01

    Giardia lamblia is an important waterborne zoonotic parasitic protozoan. It can cause diarrhea in humans and animals. Traditional detection methods are mainly stool examinations. With the application of immunological and molecular biology techniques, its immunological and molecular detection methods have made considerable developments in recent years. This article reviewed the progress in this field.%蓝氏贾第鞭毛虫是一种重要的水源性人兽共患的寄生原虫,可致人和动物的腹泻.传统的检测方法主要为粪便检查,随着免疫学与分子生物学技术的应用,其免疫学检测和分子生物学检测近年来有了较大的发展.该文就上述研究进展进行综述.

  2. Iron-dependent hydrogenases of Entamoeba histolytica and Giardia lamblia: activity of the recombinant entamoebic enzyme and evidence for lateral gene transfer.

    Science.gov (United States)

    Nixon, Julie E J; Field, Jessica; McArthur, Andrew G; Sogin, Mitchell L; Yarlett, Nigel; Loftus, Brendan J; Samuelson, John

    2003-02-01

    Entamoeba histolytica and Spironucleus barkhanus have genes that encode short iron-dependent hydrogenases (Fe-hydrogenases), even though these protists lack hydrogenosomes. To understand better the biochemistry of the protist Fe-hydrogenases, we prepared a recombinant E. histolytica short Fe-hydrogenase and measured its activity in vitro. A Giardia lamblia gene encoding a short Fe-hydrogenase was identified from shotgun genomic sequences, and RT-PCR showed that cultured entamoebas and giardias transcribe short Fe-hydrogenase mRNAs. A second E. histolytica gene, which encoded a long Fe-hydrogenase, was identified from shotgun genomic sequences. Phylogenetic analyses suggested that the short Fe-hydrogenase genes of entamoeba and diplomonads share a common ancestor, while the long Fe-hydrogenase gene of entamoeba appears to have been laterally transferred from a bacterium. These results are discussed in the context of competing ideas for the origins of genes encoding fermentation enzymes of these protists.

  3. Detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in clinical stool samples by using multiplex real-time PCR after automated DNA isolation.

    Science.gov (United States)

    Van Lint, P; Rossen, J W; Vermeiren, S; Ver Elst, K; Weekx, S; Van Schaeren, J; Jeurissen, A

    2013-01-01

    Diagnosis of intestinal parasites in stool samples is generally still carried out by microscopy; however, this technique is known to suffer from a low sensitivity and is unable to discriminate between certain protozoa. In order to overcome these limitations, a real-time multiplex PCR was evaluated as an alternative approach for diagnosing Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in stool samples.Therefore, a total of 631 faecal samples were analysed both by microscopy as well as by real-time PCR following automated DNA extraction. Results showed that real-time PCR exhibited sensitivity and specificity of both 100%, whereas traditional microscopy exhibited sensitivity and specificity of 37.5% and 99.8% respectively. As real-time PCR provides simple, sensitive and specific detection of these three important pathogenic protozoan parasites, this technique, rather than microscopy, has become our diagnostic method of choice for the detection of enteric protozoan parasites for the majority of patients.

  4. Serum Levels of Zinc, Copper, Vitamin B12, Folate and Immunoglobulins in Individuals with Giardiasis

    Directory of Open Access Journals (Sweden)

    M Zarebavani

    2012-12-01

    Full Text Available Background: Giardia lamblia is one of the most important intestinal parasites. The aim of this study was to measure serum levels of IgA, IgE, zinc, copper, vitamin B12 and folate in individuals with giardiasis in comparison to normal subjects.Methods: The study was carried out among 49 Giardia positive and 39 age and sex matched healthy volunteers. Examination of stool samples was done by direct wet smear and formol-ether concentration method. Serum samples were obtained for further laboratory examination. IgA levels were measured by Single Radial Immune Diffusion (SRID. IgE levels were measured by ELISA kit. Zinc and copper levels was measured by Ziestchem Diagnostics Kit and colorimetric endpoint-method respectively. Vitamin B12 and folate levels were measured by DRG Diagnostics Kit and Enzyme Immunoassay method respectively. All data were analyzed using SPSS version 17.Results: There was a statistically significant difference in IgA, IgE, copper and zinc levels between positive and negative groups (P<0.05. There was no significant difference between vitamin B12 and folate levels between the two groups. Mean values of Giardia positive and negative groups for IgA were 309.26 and 216.89 mg/dl, IgE 167.34 and 35.49 IU/ml, copper 309.74 and 253.61 µg/dl and zinc 69.41 and 144.75 µg/dl respectively.Conclusion: The results showed levels of IgA may correlate more closely with giardiasis than IgE. Regarding trace elements, giardiasis elevated serum copper levels, while it decreased serum zinc. Finally, there was no significant difference in serum levels of vitamin B12 and folic acid between the two groups.

  5. 基于TPI基因的城市污水中蓝氏贾第鞭毛虫的分子鉴定%Molecular identification of Giardia lamblia isolates from raw urban wastewater using the TPI gene

    Institute of Scientific and Technical Information of China (English)

    张唯哲; 凌虹; 张晓丽; 李懿宏; 姬红; 杨凤坤; 沈玉娟; 刘爱芹

    2012-01-01

    目的 对哈尔滨市污水处理厂原水中的蓝氏贾第鞭毛虫进行分子鉴定.方法 收集哈尔滨市污水处理厂原水样,提取基因组DNA,巢式PCR扩增蓝氏贾第鞭毛虫TPI基因,通过序列分析确定蓝氏贾第鞭毛虫的集聚体类型和亚型,并进行同源性分析.结果 从哈尔滨市污水处理厂原水DNA提取物中扩增出蓝氏贾第鞭毛虫TPI基因,经分子鉴定为蓝氏贾第鞭毛虫集聚体AⅡ,其序列与文献报道人源蓝氏贾第鞭毛虫集聚体AII序列(AB516351,FJ560570,EF688021)的同源性为100%.结论 哈尔滨市污水处理厂原水中存在蓝氏贾第鞭毛虫集聚体AII,威胁当地居民健康.%Objective To molecularly identify Giardia lamblia isolates from raw urban wastewater at a wastewater treatment plant in Haerbin. Methods Genomic DNA of G. lamblia was extracted and the TPI gene was amplified by nested PCRj the assemblage/genotype and sub-assemblage/subtype of C lamblia were identified and sequenced. The ho-mology of the gene was analyzed with reference nucleotide sequences downloaded from GenBank. Results The TPI gene of G. lamblia was amplified from DNA extracts prepared from raw wastewater from a wastewater treatment plant in Harbin, and the isolate was identified as assemblage AH. The obtained sequence based on TPI had 100% homology with the previously reported sequences of human-derived G. lamblia assemblage All (AB516351 ;FJ560570;EF688021). Conclusion The presence of G. lamblia assemblage All in raw urban wastewater in Harbin poses a threat to the health of local inhabitants.

  6. Enquistación in vitro de Giardia lamblia: análisis por electroforesis bidimensional de proteínas expresadas diferencialmente.

    Directory of Open Access Journals (Sweden)

    Paula C. Hernández

    2002-09-01

    Full Text Available La reconstrucción in vitro del ciclo de vida de Giardia lamblia es un excelente instrumento para el estudio de la biología molecular del parásito. El presente trabajo pretende contribuir al desarrollo de algunas técnicas por las cuales se puede definir mejor un modelo de diferenciación celular del parásito. El estudio presenta un protocolo de enquistación in vitro y establece un método para el aislamiento y la purificación de los quistes producidos, cuyas características morfológicas, por microscopía de luz, coinciden con las de los quistes obtenidos in vivo. Se estudió el mapa de las proteínas de G. lamblia por medio de electroforesis bidimensional de alta resolución y de electroforesis en una dimensión. Estos estudios mostraron que la mayoría de las proteínas del parásito son de carácter ácido. Se amplió, con base en ese resultado, la resolución de la región ácida por medio de un isoelectroenfoque de pH 4-7 en la primera dimensión. Se encontraron diferencias en la expresión de las proteínas durante el proceso de enquistación. Se obtuvieron, además, imágenes por microscopía de luz y electrónica de transmisión que permitieron observar morfológica y ultraestructuralmente las células producidas durante el proceso de la enquistación.

  7. Both endo-siRNAs and tRNA-derived small RNAs are involved in the differentiation of primitive eukaryote Giardia lamblia.

    Science.gov (United States)

    Liao, Jian-You; Guo, Yan-Hua; Zheng, Ling-Ling; Li, Yan; Xu, Wen-Li; Zhang, Yu-Chan; Zhou, Hui; Lun, Zhao-Rong; Ayala, Francisco J; Qu, Liang-Hu

    2014-09-30

    Small RNAs (sRNAs), including microRNAs and endogenous siRNAs (endo-siRNAs), regulate most important biologic processes in eukaryotes, such as cell division and differentiation. Although sRNAs have been extensively studied in various eukaryotes, the role of sRNAs in the early emergence of eukaryotes is unclear. To address these questions, we deep sequenced the sRNA transcriptome of four different stages in the differentiation of Giardia lamblia, one of the most primitive eukaryotes. We identified a large number of endo-siRNAs in this fascinating parasitic protozoan and found that they were produced from live telomeric retrotransposons and three genomic regions (i.e., endo-siRNA generating regions [eSGRs]). eSGR-derived endo-siRNAs were proven to target mRNAs in trans. Gradual up-regulation of endo-siRNAs in the differentiation of Giardia suggested that they might be involved in the regulation of this process. This hypothesis was supported by the impairment of the differentiation ability of Giardia when GLDICER, essential for the biogenesis of endo-siRNAs, was knocked down. Endo-siRNAs are not the only sRNA regulators in Giardia differentiation, because a great number of tRNAs-derived sRNAs showed more dramatic expression changes than endo-siRNAs in this process. We totally identified five novel kinds of tRNAs-derived sRNAs and found that the biogenesis in four of them might be correlated with that of stress-induced tRNA-derived RNA (sitRNA), which was discovered in our previous studies. Our studies reveal an unexpected complex panorama of sRNA in G. lamblia and shed light on the origin and functional evolution of eukaryotic sRNAs.

  8. 蓝氏贾第鞭毛虫潜在致病机制初探%A preliminary exploration of the potential pathogenic mechanism of Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    余新刚; 胡伟; 李国清

    2015-01-01

    Giardia lamblia is a common zoonotic parasitic protozoa while giardiasis has been listed as one of the 10 major parasitic diseases endangering human health by World Health Organization.The harmfulness of giardiasis is attracting more and more attentions in recent years,however,the pathogenic mechanism of diarrhea caused by the parasite is still unclear.It may be related to many factors,such as cytoskeletal proteins,excretory/secretory products,L-arginine metabolism,surface antigen and antigenic variation of G.lamblia,and the immunological changes of the host.Researches concerning the pathogenic mechanism of the giardiasis were reviewed in this paper.%蓝氏贾第鞭毛虫(简称贾第虫)是一种常见的人兽共患寄生性原虫,由其导致的贾第虫病已被世界卫生组织列为危害人类健康的10种主要寄生虫病之一.近年来,贾第虫病的危害日益受到重视,然而贾第虫导致腹泻的致病机制尚不明确,可能与其细胞骨架蛋白、虫体分泌物、L精氨酸代谢以及表面抗原变异、免疫学改变等多种因素有关,该文对与贾第虫致病机理相关的研究作一综述.

  9. Putative SF2 helicases of the early-branching eukaryote Giardia lamblia are involved in antigenic variation and parasite differentiation into cysts.

    Science.gov (United States)

    Gargantini, Pablo R; Serradell, Marianela C; Torri, Alessandro; Lujan, Hugo D

    2012-11-28

    Regulation of surface antigenic variation in Giardia lamblia is controlled post-transcriptionally by an RNA-interference (RNAi) pathway that includes a Dicer-like bidentate RNase III (gDicer). This enzyme, however, lacks the RNA helicase domain present in Dicer enzymes from higher eukaryotes. The participation of several RNA helicases in practically all organisms in which RNAi was studied suggests that RNA helicases are potentially involved in antigenic variation, as well as during Giardia differentiation into cysts. An extensive in silico analysis of the Giardia genome identified 32 putative Super Family 2 RNA helicases that contain almost all the conserved RNA helicase motifs. Phylogenetic studies and sequence analysis separated them into 22 DEAD-box, 6 DEAH-box and 4 Ski2p-box RNA helicases, some of which are homologs of well-characterized helicases from higher organisms. No Giardia putative helicase was found to have significant homology to the RNA helicase domain of Dicer enzymes. Additionally a series of up- and down-regulated putative RNA helicases were found during encystation and antigenic variation by qPCR experiments. Finally, we were able to recognize 14 additional putative helicases from three different families (RecQ family, Swi2/Snf2 and Rad3 family) that could be considered DNA helicases. This is the first comprehensive analysis of the Super Family 2 helicases from the human intestinal parasite G. lamblia. The relative and variable expression of particular RNA helicases during both antigenic variation and encystation agrees with the proposed participation of these enzymes during both adaptive processes. The putatives RNA and DNA helicases identified in this early-branching eukaryote provide initial information regarding the biological role of these enzymes in cell adaptation and differentiation.

  10. ELISA y técnica de sedimentación espontánea para el diagnóstico de infección por Giardia lamblia en muestras fecales de niños de Perú ELISA and spontaneous sedimentation technique for the diagnosis of Giardia lamblia infection in stool samples of Peruvian children

    Directory of Open Access Journals (Sweden)

    Claudia Rodríguez-Ulloa

    2011-12-01

    Full Text Available OBJETIVO: Comparar un kit ELISA comercial para coproantígenos y la técnica de sedimentación espontánea en tubo (TSET para el diagnóstico de Giardia lamblia en muestras fecales de niños de una zona endémica peruana. MATERIAL Y MÉTODOS: Fueron analizadas 174 muestras mediante la TSET y el kit Giardia 2nd Generation ELISA. RESULTADOS: Fueron positivas 51 muestras por ELISA y 49 por TSET. CONCLUSIONES: El ELISA resultó ser altamente sensible y específico, sencillo y rápido; sin embargo, la muy buena concordancia, alta precisión, bajo costo y capacidad para detectar otros enteroparásitos hace que la TSET sea recomendable para el diagnóstico en zonas endémicas del Perú.OBJETIVE: To compare a commercial coproantigen ELISA kit and the technique of spontaneous sedimentation in tube (TSET for the diagnosis of Giardia lamblia in fecal specimens from children in a Peruvian endemic area. MATERIAL AND METHODS: 174 fecal samples were analyzed by TSET and 2nd Generation Giardia ELISA kit. RESULTS: 51 samples were positive by ELISA and 49 by TSET. CONCLUSIONS: The ELISA was highly sensitive and specific, simple and fast. However, the very good agreement, high precision, low cost and ability to detect other intestinal parasites makes use of TSET recommended for laboratory diagnosis in endemic areas of Peru.

  11. Research progress on multilocus genotyping for genetic diversity of Giardia lamblia%多位点基因分型用于蓝氏贾第鞭毛虫遗传多样性的研究进展

    Institute of Scientific and Technical Information of China (English)

    王斌; 沈玉娟; 刘华; 刘晖

    2015-01-01

    蓝氏贾第鞭毛虫(简称贾第虫)是一种重要的人兽共患肠道原虫,宿主广泛.目前,种内有8个有效聚集体型.随着分子生物学技术的发展,多位点基因分型(multi-locus genotyping,MLG)技术被广泛用于贾第虫的研究.该文就MLG技术在阐述贾第虫遗传多样性、评估人兽共患传播的可能性及分类学上的应用研究作一综述.%Giardia lamblia is an important zoonotic intestinal parasite with a wide range of hosts.Eight valid assemblages(A-H) have been identified within this parasite.With the development of molecular biological techniques,multi-locus genotyping(MLG) tools have been used increasingly to characterize G.lamblia isolates.This paper focused on reviewing the application of MLG in understanding genetic diversity and zoonotic potential of G.lamblia as well as its taxonomy.

  12. Serum pneumoproteins in firefighters

    NARCIS (Netherlands)

    Greven, Frans; Krop, Esmeralda; Burger, Nena; Kerstjens, Huib; Heederik, Dick

    2011-01-01

    Serum Clara cell protein (CC16) and surfactant-associated protein A (SP-A) were measured in a cross-sectional study in 402 firefighters. For the population as a whole, no associations were detected between serum pneumoproteins and smoke exposure. SP-A levels were increased in symptomatic subjects ex

  13. The intraspecific difference of the triose phosphate isomerase (tim) gene from Giardia lamblia%蓝氏贾第鞭毛虫磷酸丙糖异构酶基因种内差异研究

    Institute of Scientific and Technical Information of China (English)

    卢思奇; 李继红; 张亚平; 文建凡; 王凤云

    2002-01-01

    目的探讨蓝氏贾第鞭毛虫(Giardia lamblia)磷酸丙糖异构酶基因种内差异.方法提取虫体总DNA,对所有虫株磷酸丙糖异构酶(tim)基因部分片段进行PCR扩增.测定序列后,用简约法和NJ法构建系统树进行系统发育分析.结果共有124个位点存在变异(占所有测定序列中的23%),且大多数为发生在密码子的同义突变. 两种构树方法所得二树的分枝结构相似,均将受试的16株蓝氏贾第虫分为明显的两组.结论宿主及地理因素对蓝氏贾第虫群体的遗传多样性影响不大.在DNA分子进化水平上,自然选择的影响十分显著.可将tim基因作为蓝氏贾第虫群体遗传结构一个十分有效的遗传标记.%Objective To investigate the intraspecific difference of the triose phosphate isomerase(ti m) gene from Giardia lamblia (G. lamblia).Methods Total genomic DNA of G. lamblia was extracted and partial fragments of the triose phosphate isomerase (tim) gene were amplified by polymerase chain reactio n (PCR). All nucleotide sequences were analyzed by using a phylogenetic analysi s, which was constructed with parsimony and Neighbor-joining (N-J) methods. Results A total of 124 variable sites (23% of all sequences detected) was defined, most of which were found at the silent sites of codons. Two similar phylogenetic tre es were constructed, subdividing 16 Giardia isolates into two groups. Conclusion The genetic diversity of G. lamblia appeared to be little affected by facto rs of both host and geography, while natural-selection played an important role in DNA molecular evolution level of the tim gene. The tim gene may be consider ed a very useful genetic marker of the population genetic structure of G. lam blia.

  14. [Serum sickness in diphtheria].

    Science.gov (United States)

    Vozianova, Zh I; Chepilko, K I

    1999-01-01

    As many as 2247 patients with different clinical forms of diphtheria were examined. Antidiphtheric serum (ADS) was administered in 1556 children, the dosage being determined by condition of the patient. Serum sickness developed at day 7 to 9 in 24 (1.5%); 10 patients were found to run a mild course, 14--moderately severe. 6 patients had allergic reactions: 3--to antibiotic (penicillin), urticaria type, 1--to pertussoid-tetanic anatoxin, 2 had pollinosis-type reaction. Thus, serum sickness has practical value, which fact requires a detailed allergic history together with skin tests to be performed before the administration of ADS.

  15. Serum YKL-40

    DEFF Research Database (Denmark)

    Mylin, Anne K; Abildgaard, Niels; Johansen, Julia S

    2015-01-01

    In a time of increasing treatment options for multiple myeloma bone disease, risk factors predicting progression need to be elucidated. This study investigated the value of serum YKL-40, previously shown to be associated with radiographic progression of bone destruction, as a predictor for time...... for SRE and at 9 and 24 months for radiographic progression. Elevated serum YKL-40 was seen in 47% of patients and associated with high-risk disease (International Staging System stage III; p serum CTX/MMP; p ... to clinical progression, i.e. skeletal-related events (SREs), in 230 newly diagnosed patients with multiple myeloma receiving intravenous bisphosphonates. Serum concentrations of YKL-40 and biochemical bone markers (CTX-MMP, CTX-I, PINP) were measured at diagnosis. Patients were evaluated every third month...

  16. Prevalence and associated risk factors for Giardia lamblia infection among children hospitalized for diarrhea in Goiânia, Goiás State, Brazil Prevalência e fatores de risco associados à infecção por Giardia lamblia em crianças hospitalizadas com diarréia em Goiânia, Goiás, Brasil

    Directory of Open Access Journals (Sweden)

    Maria das Graças Cabral Pereira

    2007-06-01

    Full Text Available The objective of this study was to determine the prevalence and to identify risk factors associated with Giardia lamblia infection in diarrheic children hospitalized for diarrhea in Goiânia, State of Goiás, Brazil. A cross-sectional study was conducted and a comprehensive questionnaire was administered to the child's primary custodian. Fixed effects logistic regression was used to determine the association between infection status for G. lamblia and host, sociodemographic, environmental and zoonotic risk factors. A total of 445 fecal samples were collected and processed by the DFA methodology, and G. lamblia cysts were present in the feces of 44 diarrheic children (9.9%. A variety of factors were found to be associated with giardiasis in these population: age of children (OR, 1.18; 90% CI, 1.0 - 1.36; p = 0.052, number of children in the household (OR 1.45; 90% CI, 1.13 - 1.86; p = 0.015, number of cats in the household (OR, 1.26; 90% CI, 1.03 -1.53; p = 0.059, food hygiene (OR, 2.9; 90% CI, 1.34 - 6.43; p = 0.024, day-care centers attendance (OR, 2.3; 90% CI, 1.20 - 4.36; p = 0.034, living on a rural farm within the past six months prior hospitalization (OR, 5.4; CI 90%, 1.5 - 20.1; p = 0.03 and the number of household adults (OR, 0.59; 90% CI, 0.42 - 0.83; p = 0.012. Such factors appropriately managed may help to reduce the annual incidence of this protozoal infection in the studied population.O objetivo desse estudo foi estimar a prevalência e identificar fatores de risco associados à infecção por Giardia lamblia em crianças hospitalizadas com diarréia no município de Goiânia, Estado de Goiás, Brasil. Foi realizado um estudo transversal e um questionário estruturado foi administrado ao responsável pela criança. Análise multivariada foi realizada por meio de regressão logística para se determinar a associação entre infecção por G. lamblia e as variáveis sociodemográficas, ambientais e zoonóticas relacionadas ao

  17. Cyst-Wall-Protein-1 is fundamental for Golgi-like organelle neogenesis and cyst-wall biosynthesis in Giardia lamblia

    Science.gov (United States)

    Ebneter, Jacqueline A.; Heusser, Sally D.; Schraner, Elisabeth M.; Hehl, Adrian B.; Faso, Carmen

    2016-01-01

    The genome of the protozoan parasite Giardia lamblia is organized in two diploid nuclei, which has so far precluded complete analysis of gene function. Here we use a previously developed Cre/loxP-based knock-out and selection marker salvage strategy in the human-derived isolate WB-C6 to eliminate all four copies of the Cyst-Wall-Protein-1 locus (CWP1). Because these loci are silenced in proliferating trophozoites and highly expressed only in encysting cells, CWP1 ablation allows functional characterization of a conditional phenotype in parasites induced to encyst. We show that encysting Δcwp1 cells are unable to establish the stage-regulated trafficking machinery with Golgi-like encystation-specific vesicles required for cyst-wall formation but show morphological hallmarks of cyst development and karyokinesis. This ‘pseudocyst' phenotype is rescued by transfection of Δcwp1 cells with an episomally maintained CWP1 expression vector. Genome editing in genera Giardia and Trypanosoma are the only reported examples addressing questions on pathogen transmission within the Excavata supergroup. PMID:27976675

  18. PCR DETECTION OF GIARDIA LAMBLIA BY TIM GENE AMPLIFICATION%用PCR扩增tim基因检测蓝氏贾第鞭毛虫

    Institute of Scientific and Technical Information of China (English)

    丁慧萍; 卢思奇; 戎煜; 李凤舞; 王凤云

    2002-01-01

    采用聚合酶链反应(PCR)对蓝氏贾第鞭毛虫(Giardia lamblia)的磷酸丙糖异构酶(triose phosphate isomerase,缩写为tim)基因进行特异性扩增,结果扩增出1条683bp的DNA片段.此方法的特异性可高达100%,而其它DNA样本,如日本血吸虫(Schistosoma japonicum)、刚地弓形虫(Toxoplasma gondii)、微小隐孢子虫(Cryptosporidium parvum)、溶组织内阿米巴(Entamoeba histolytica)、旋毛虫(Trichinella spiralis)和阴道毛滴虫(Trichomonas vaginalis),以及人体血细胞等均未出现扩增反应.本法的敏感性也很高,可检测到0.4pg贾第虫包囊的DNA.13株来自不同地理位置和/或宿主的贾第虫DNA样本在PCR中均各产生1条长为683bp的目的片段.上述结果表明本实验建立的检测贾第虫的PCR方法有效.

  19. Immune evasion mechanisms of Giardia lamblia%蓝氏贾第鞭毛虫的免疫逃避机制

    Institute of Scientific and Technical Information of China (English)

    王洋; 田喜凤

    2013-01-01

    蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)是一种世界性分布的机会性致病原虫,能够引起以腹泻为主要表现的贾第虫病,严重影响人类尤其是儿童的健康和发育.在贾第虫感染过程中,宿主的非特异性和特异性免疫系统均会产生强烈的抗贾第虫效应,但贾第虫通过抗原漂变、L-精氨酸饥饿等机制逃避和抑制宿主的免疫反应,引起宿主的长期或反复感染.本文对宿主的抗贾第虫免疫以及贾第虫的免疫逃避机制做一综述.

  20. Specific histone modifications play critical roles in the control of encystation and antigenic variation in the early-branching eukaryote Giardia lamblia.

    Science.gov (United States)

    Carranza, Pedro G; Gargantini, Pablo R; Prucca, César G; Torri, Alessandro; Saura, Alicia; Svärd, Staffan; Lujan, Hugo D

    2016-12-01

    During evolution, parasitic microorganisms have faced the challenges of adapting to different environments to colonize a variety of hosts. Giardia lamblia, a common cause of intestinal disease, has developed fascinating strategies to adapt both outside and inside its host's intestine, such as trophozoite differentiation into cyst and the switching of its major surface antigens. How gene expression is regulated during these adaptive processes remains undefined. Giardia lacks some typical eukaryotic features, like canonical transcription factors, linker histone H1, and complex promoter regions; suggesting that post-transcriptional and translational control of gene expression is essential for parasite survival. However, epigenetic factors may also play critical roles at the transcriptional level. Here, we describe the most common post-translational histone modifications; characterize enzymes involved in these reactions, and analyze their association with the Giardia's differentiation processes. We present evidence that NAD(+)-dependent and NAD(+)-independent histone deacetylases regulate encystation; however, a unique NAD(+)-independent histone deacetylase modulate antigenic switching. The rates of acetylation of H4K8 and H4K16 are critical for encystation, whereas a decrease in acetylation of H4K8 and methylation of H3K9 occur preferentially during antigenic variation. These results show the complexity of the mechanisms regulating gene expression in this minimalistic protozoan parasite. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Cross-modulation of pathogen-specific pathways enhances malnutrition during enteric co-infection with Giardia lamblia and enteroaggregative Escherichia coli

    Science.gov (United States)

    Bartelt, Luther A.; Bolick, David T.; Zaenker, Edna I.; Donowitz, Jeffery; Thomas-Beckett, Rose Viguna; Rogala, Allison; Carroll, Ian M.; Swann, Jonathan R.; Guerrant, Richard L.

    2017-01-01

    Diverse enteropathogen exposures associate with childhood malnutrition. To elucidate mechanistic pathways whereby enteric microbes interact during malnutrition, we used protein deficiency in mice to develop a new model of co-enteropathogen enteropathy. Focusing on common enteropathogens in malnourished children, Giardia lamblia and enteroaggregative Escherichia coli (EAEC), we provide new insights into intersecting pathogen-specific mechanisms that enhance malnutrition. We show for the first time that during protein malnutrition, the intestinal microbiota permits persistent Giardia colonization and simultaneously contributes to growth impairment. Despite signals of intestinal injury, such as IL1α, Giardia-infected mice lack pro-inflammatory intestinal responses, similar to endemic pediatric Giardia infections. Rather, Giardia perturbs microbial host co-metabolites of proteolysis during growth impairment, whereas host nicotinamide utilization adaptations that correspond with growth recovery increase. EAEC promotes intestinal inflammation and markers of myeloid cell activation. During co-infection, intestinal inflammatory signaling and cellular recruitment responses to EAEC are preserved together with a Giardia-mediated diminishment in myeloid cell activation. Conversely, EAEC extinguishes markers of host energy expenditure regulatory responses to Giardia, as host metabolic adaptations appear exhausted. Integrating immunologic and metabolic profiles during co-pathogen infection and malnutrition, we develop a working mechanistic model of how cumulative diet-induced and pathogen-triggered microbial perturbations result in an increasingly wasted host. PMID:28750066

  2. Comparison of the local immune response against Giardia lamblia cyst wall protein 2 induced by recombinant Lactococcus lactis and Streptococcus gordonii.

    Science.gov (United States)

    Lee, Peter; Abdul-Wahid, Aws; Faubert, Gaétan M

    2009-01-01

    Lactococcus lactis and Streptococcus gordonii are lactic acid bacteria (LAB) currently being advocated for use as live antigen delivery vehicles to mucosal sites. Since both vehicles differ in their capability to persist within the small intestine and in their mode of antigen delivery, we sought to compare them to determine which one was superior. In this study, we compared the efficacy of recombinant L. lactis and S. gordonii to stimulate intestinal immune responses against Giardia lamblia cyst wall protein-2 in BALB/c mice. Oral administration of either vector significantly increased the number of CD4(+) T helper and B-cells in the mesenteric lymph nodes (MLN) and Peyer's patches (PP) of immunized animals. Delivery of recombinant CWP2 (rCWP2) by L. lactis stimulated a balanced IFN-gamma/IL-4 response (MLN and PP cells) and a CWP2-specific intestinal IgA antibody response. Alternatively, delivery of rCWP2 by S. gordonii stimulated a higher frequency of IFN-gamma secreting MLN and PP cells, as well as doubling the amount of CWP2-specific intestinal IgA. In challenge studies, L. lactis and S. gordonii reduced cyst output by 71 and 90%, respectively. When compared to each other, S. gordonii-immunized animals shed 65% fewer cysts than their L. lactis-immunized counterparts. Based on these findings, we concluded that S. gordonii was superior to L. lactis as an intestinal vaccine delivery vehicle.

  3. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley The effect of malnourishment on the development of giardiosis in Sprague-Dawley rats

    Directory of Open Access Journals (Sweden)

    Martha Elvia Díaz-Cinco

    2002-07-01

    Full Text Available Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos. Es un estudio experimental efectuado entre 1995 y 1999, en el Centro de Investigación en Alimentación y Desarrollo, de la ciudad de Hermosillo, estado de Sonora, México, y cuya muestra estuvo integrada por dos grupos de 6 a 8 unidades experimentales, las cuales recibieron dos tipos de dieta y cinco tratamientos con diferente concentración de inóculo de quistes de Giardia lamblia. Durante la fase experimental se monitoreó la excreción de quistes, ganancia de peso, consumo de alimento, análisis del contenido intestinal y daños macro y microscópicos en mucosa intestinal. El tratamiento estadístico de los datos consistió en análisis de varianza, y de residuales cuando fue necesario. Resultados. En los animales alimentados con una dieta que cubre sus requerimientos nutricionales (DN se observó que es posible provocar la infección con 60 quistes, mientras que en los animales desnutridos (dieta DLN la concentración que se requiere es de únicamente seis quistes para establecer daños a nivel de mucosa intestinal. Conclusiones. La ganancia de peso monitoreada durante los 10 días de ensayo no resultó ser un buen indicador de la infección por Giardia lamblia. El porcentaje de infección depende de la concentración de quistes inoculados y del estado de nutrición de los animales evaluados.Objective. To assess the effect of malnutrition on the development of giardiosis in Sprague-Dawley rats, using different inoculum sizes of Giardia lamblia cysts. Material and Methods. An experimental study was conducted between 1995 and 1999 at Centro de Investigación, Alimentación y Desarrollo (Center for Research, Food, and Development, in Hermosillo, Sonora, Mexico. The study population consisted of two groups of six to eight

  4. Genome sequencing of Giardia lamblia genotypes A2 and B isolates (DH and GS) and comparative analysis with the genomes of genotypes A1 and E (WB and Pig).

    Science.gov (United States)

    Adam, Rodney D; Dahlstrom, Eric W; Martens, Craig A; Bruno, Daniel P; Barbian, Kent D; Ricklefs, Stacy M; Hernandez, Matthew M; Narla, Nirmala P; Patel, Rima B; Porcella, Stephen F; Nash, Theodore E

    2013-01-01

    Giardia lamblia (syn G. intestinalis, G. duodenalis) is the most common pathogenic intestinal parasite of humans worldwide and is a frequent cause of endemic and epidemic diarrhea. G. lamblia is divided into eight genotypes (A-H) which infect a wide range of mammals and humans, but human infections are caused by Genotypes A and B. To unambiguously determine the relationship among genotypes, we sequenced GS and DH (Genotypes B and A2) to high depth coverage and compared the assemblies with the nearly completed WB genome and draft sequencing surveys of Genotypes E (P15; pig isolate) and B (GS; human isolate). Our results identified DH as the smallest Giardia genome sequenced to date, while GS is the largest. Our open reading frame analyses and phylogenetic analyses showed that GS was more distant from the other three genomes than any of the other three were from each other. Whole-genome comparisons of DH_A2 and GS_B with the optically mapped WB_A1 demonstrated substantial synteny across all five chromosomes but also included a number of rearrangements, inversions, and chromosomal translocations that were more common toward the chromosome ends. However, the WB_A1/GS_B alignment demonstrated only about 70% sequence identity across the syntenic regions. Our findings add to information presented in previous reports suggesting that GS is a different species of Giardia as supported by the degree of genomic diversity, coding capacity, heterozygosity, phylogenetic distance, and known biological differences from WB_A1 and other G. lamblia genotypes.

  5. Giardia lamblia e alergia respiratória: estudo em uma amostra de crianças de área urbana com frequência elevada da protozoose

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    Valdênia M. O. Souza

    2012-06-01

    Full Text Available OBJETIVO: Uma elevada incidência de enteroparasitoses é encontrada em regiões urbanas do nordeste brasileiro. As infecções por Giardia lamblia têm sido relacionadas com aumento da prevalência de alergias cutâneas e gastrointestinais. Contudo, ainda existe pouca informação sobre a associação entre a giardíase e doenças alérgicas das vias aéreas. Diante disso, o presente estudo se propôs a verificar a relação entre a infecção por G. lamblia em crianças de área urbana e reatividade alérgica respiratória. MÉTODOS: O estudo incluiu 110 pacientes, de ambos os sexos e idades, entre 5 e 15 anos. Os pacientes foram submetidos a questionários de avaliação dos sintomas clínicos, testes cutâneos de leitura imediata e exames coproparasitológicos e sorológicos. RESULTADOS: Foi verificada uma frequência elevada de crianças infectadas por G. lamblia (45%, 50/110. A infecção pelo protozoário não foi associada com maior risco de alergias respiratórias (p = 0,075, elevação de IgE total (p = 0,701, IgE específica (p = 0,250 ou teste cutâneo positivo para diferentes alérgenos ambientais (p = 0,239. CONCLUSÃO: O estudo demonstrou que a presença dos sintomas de asma, atopia cutânea e marcadores sorológicos não foram associados com a presença de infecção pela G. lamblia nessa amostra de crianças.

  6. 犬源贾第虫β-贾第素基因的克隆及序列分析%Cloning and sequence analysis of bg gene of Giardia lamblia from dogs

    Institute of Scientific and Technical Information of China (English)

    李结; 王培园; 张萍; 朱海波; 郭建超; 李国清

    2011-01-01

    This study is to identify the genotype of Giardia lamblia from dogs in Foshan, Guangdong. Based on the published sequence in GenBank, two pairs of primers GF1/GR and GF2/GR were designed to amplify the bg gene of G. Lamblia from dogs by nested PCR. The am-plicons were purified and cloned into pMD-18T vector followed by being transformed into JM109. The positive clones were cultured and identified by PCR amplification and the insert was successfully sequenced. The results showed that the bg gene was 384 bp in length. The phylo-genetic tree revealed that the dog-derived Giardia strain was grouped into the D genotype of G. Lamblia, which was first reported in China. These results provided a foundation for further studies on molecular identification and molecular genetics of G. Lamblia.%为了鉴定犬源贾第虫佛山分离株的基因型,根据GenBank中登录的贾第虫β-贾第素(bg)基因序列(X85958),设计2对引物GF1/GR、GF2/GR,采用套式PCR方法从贾第虫基因组DNA中扩增出目的片段;将扩增产物纯化并连接到质粒载体pMD18-T,克隆转入感受态细胞JM109,挑选阳性克隆测序,然后进行序列分析.结果显示,犬源贾第虫bg序列长为384 bp,与预期目的片段一致;分子进化树表明该犬源贾第虫分离株为蓝氏贾第虫D型.该基因型在我国为首次报道,为贾第虫的分子鉴定以及分子遗传学研究奠定了基础.

  7. Impacto de la dieta sobre la inducción de infección con quistes de Giardia lamblia en ratas Sprague-Dawley The effect of malnourishment on the development of giardiosis in Sprague-Dawley rats

    OpenAIRE

    Martha Elvia Díaz-Cinco; Martha Nydia Ballesteros-Vázquez; Rosalba Pérez-Morales; Verónica Mata-Haro

    2002-01-01

    Objetivo. Evaluar el efecto de la desnutrición en la inducción de la giardiosis con diferentes concentraciones de quistes de Giardia lamblia utilizando como modelo animal ratas Sprague-Dawley. Material y métodos. Es un estudio experimental efectuado entre 1995 y 1999, en el Centro de Investigación en Alimentación y Desarrollo, de la ciudad de Hermosillo, estado de Sonora, México, y cuya muestra estuvo integrada por dos grupos de 6 a 8 unidades experimentales, las cuales recibieron dos tipos d...

  8. PREVALENCE OF GIARDIA LAMBLIA AND OTHER INTESTINAL PARASITES IN PRESCHOOL CHILDREN AND ITS RELATION TO RESIDENCE PLACE, SEX AND BLOOD GROUP IN ILAM COUNTY OF IRAN

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    Pourbabak

    1996-06-01

    Full Text Available 1711' object of this study was to determine the prevalence oj asymptomatic infestation with Giardia lamblia lind other intestinal parasites in children of urban anti rural communities oj /lam county and its relation with dwelling place, sex and blood grollp!.. 77w study designed as (l five-month pUTasitoulgica! .m,..£!' oj fecal ami blood specimens from humans anti performed in 10 urban hcalih-trcatmcru clinics of llam city, two urban health treatment clinics of Eyvun city, two rural health-treatment clinics oj Chavar and Sartaf villages, llam province west of fran, 17,e examined population was preschool {, to 7 year-old children without any 'gastrointestinal compliarus. Prevalence oj infestation in subject grOllp W(l."' 32.54% (n=3100. Among intestinal parasites' G. lambliu with 85.43o/c (27.8% oj all, n=JO(JI prevalence rate was the most common. Infestation with 11. nnrm with 1'/.93% and E. coli with 3.07';, were in the second and third ranks, respectively, Infestation shows a distinct relationship with gender (P<0.05 and dwelling place, but it lacks a significant relation with blood groups. This study ."lIOWS that the prevalence of intestinal infestation in 6 to 7 year old child oj llam county hi equivalent to the top oj tile line oj the statistical percentage all over the world. 17,e relation between the severity oj infestation and residence place may arouse the suspicion oj sever contamination oj imbibing water.

  9. Identification of Giardia lamblia and the human infectious-species of Cryptosporidium in drinking water resources in Western Saudi Arabia by nested-PCR assays.

    Science.gov (United States)

    Hawash, Y; Ghonaim, M; Hussein, Y; Alhazmi, A; Alturkistani, A

    2015-06-01

    The presence of Cryptosporidium and/or Giardia in drinking water represents a major public health problem. This study was the first report concerned with the occurrence of these protozoa in drinking water in Saudi Arabia. The study was undertaken in Al-Taif, a high altitude region, Western Saudi Arabia. Eight underground wells water, six desalinated water and five domestic brands of bottled water samples, 10 liter each, were monthly collected between May 2013 and April 2014. All samples (n = 228), were processed using an automated wash/elution station (IDEXX Laboratories, Inc.). Genomic DNA was directly isolated and purified from samples concentrates with QIAamp® Stool Mini Kit (Qiagen). The target protozoan DNA sequences were amplified using two previously published nested-PCR protocols. Of all the analyzed water, 31 samples (≈14%) were found contaminated with the target protozoa. Giardia lamblia was detected in ≈10% (7/72) of desalinated water and in ≈9% (9/96) of wells water. On the other hand, Cryptosporidium was identified in ≈8% (8/72) of desalinated water and in ≈7% (7/96) of wells water. All bottled water samples (n = 60) were (oo)cysts-free. Protozoan (oo)cysts were more frequently identified in water samples collected in the spring than in other seasons. The methodology established in our study proved sensitive, cost-effective and is amenable for future automation or semi-automation. For better understanding of the current situation that represent an important health threat to the local inhabitants, further studies concerned with (oo)cyst viability, infectivity, concentration and genotype identification are recommended.

  10. A Preliminary Study on the Lamin Gene of Giardia lamblia%蓝氏贾第虫核纤层蛋白基因的初步研究

    Institute of Scientific and Technical Information of China (English)

    陈万群; 文建凡; 卢思奇

    2004-01-01

    贾第虫一度被认为是迄今已知的最原始的真核细胞,但近来争议日盛.利用PCR和测序等技术,对蓝氏贾第虫(Giardia lamblia)的核纤层蛋白(lamin)基因进行了研究.结果表明:蓝氏贾第虫基因组中存在一个编码具有明显lamin特征的基因序列.如该基因序列的3′-端具有编码与核内膜亲和的特征性模体(motif)CaaX的序列;具有B型lamin基因所特有的高度保守的27 bp片段,该片段编码高度保守的位于α螺旋杆状区的9氨基酸片段等.同时,这些序列特征又与多细胞的后生动物存在一定差异.这些事实说明在贾第虫中已经进化产生了典型真核细胞的B 型lamin(基因)或至少是类似B 型的lamin(基因),该生物的进化地位可能并非过去所认为的那么原始.

  11. Factores de riesgo de la infección por Giardia lamblia en niños de guarderías infantiles de Ciudad de La Habana, Cuba Risk factors for Giardia lamblia in children in daycare centers in Havana, Cuba

    Directory of Open Access Journals (Sweden)

    Fidel Ángel Núñez

    2003-04-01

    Full Text Available En un estudio longitudinal sobre giardiasis desarrollado durante 18 meses en tres guarderías infantiles de Ciudad de La Habana, describimos un grupo de niños con tendencia o "predisposición" a la infección por Giardia lamblia; a los que se les encontró con síntomas clínicos más asociados a las diarreas. Este estudio de casos y controles se desarrolló para conocer si algunos factores socioeconómicos y hábitos higiénicos estaban asociados con este fenómeno. No se encontraron diferencias entre ambos grupos en las tasas de hacinamiento, el índice de personas por cama, en la carencia de determinados equipos eléctricos, en el nivel escolar de las madres, y en el promedio del ingreso familiar; sin embargo, predominó una frecuencia mayor de padres con nivel escolar menor de 12 grado en los casos que en los controles. No se encontraron diferencias entre ambos grupos en cuanto a la ausencia de lavado de manos antes de comer y después de defecar, pero se encontró un mayor porcentaje de lavado incorrecto de vegetales, y una menor frecuencia del hábito de hervir el agua de consumo en las familias de los casos. Estos resultados demuestran el papel del agua como vehículo de transmisión en la giardiasis y la importancia de algunos factores epidemiológicos.We conducted a longitudinal study on giardiasis in three daycare centers in Havana City for a period of 18 months and described a group of children with a "predisposition" or tendency towards re-infection with Giardia lamblia. This group was found to be more frequently associated with clinical symptoms such as diarrhea. A case-control study was designed to determine whether socioeconomic factors and hygiene were associated with this phenomenon. We found no differences between the groups with regard to overcrowding rates, number of persons per bed, absence of certain electric appliances, mother's schooling, or mean family income. However, there were proportionally more fathers with less

  12. The human serum metabolome.

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    Nikolaos Psychogios

    Full Text Available Continuing improvements in analytical technology along with an increased interest in performing comprehensive, quantitative metabolic profiling, is leading to increased interest pressures within the metabolomics community to develop centralized metabolite reference resources for certain clinically important biofluids, such as cerebrospinal fluid, urine and blood. As part of an ongoing effort to systematically characterize the human metabolome through the Human Metabolome Project, we have undertaken the task of characterizing the human serum metabolome. In doing so, we have combined targeted and non-targeted NMR, GC-MS and LC-MS methods with computer-aided literature mining to identify and quantify a comprehensive, if not absolutely complete, set of metabolites commonly detected and quantified (with today's technology in the human serum metabolome. Our use of multiple metabolomics platforms and technologies allowed us to substantially enhance the level of metabolome coverage while critically assessing the relative strengths and weaknesses of these platforms or technologies. Tables containing the complete set of 4229 confirmed and highly probable human serum compounds, their concentrations, related literature references and links to their known disease associations are freely available at http://www.serummetabolome.ca.

  13. Vaccination with Bivalent DNA Vaccine of α1-Giardin and CWP2 Delivered by Attenuated Salmonella typhimurium Reduces Trophozoites and Cysts in the Feces of Mice Infected with Giardia lamblia.

    Directory of Open Access Journals (Sweden)

    Xian-Min Feng

    Full Text Available Giardia lamblia is one of the most common infectious protozoans in human that may cause diarrhea in travelers. Searching for antigens that induced effectively protective immunity has become a key point in the development of vaccine against giardiasis.Mice vaccinated with G. lamblia trophozozite-specific α1-giardin DNA vaccine delivered orally by attenuated Salmonella typhimurium SL7027 elicited 74.2% trophozoite reduction, but only 28% reduction in cyst shedding compared with PBS buffer control. Oral vaccination with Salmonella-delivered cyst-specific CWP2 DNA produced 89% reduction in cysts shedding in feces of vaccinated mice. Significantly, the mice vaccinated with Salmonella-delivered bivalent α1-giardin and CWP2 DNA vaccines produced significant reduction in both trophozoite (79% and cyst (93% in feces of vaccinated mice. This parasite reduction is associated with the strong local mucosal IgA secretion and the IgG2a-dominant systemic immune responses in vaccinated mice.The results demonstrate that bivalent vaccines targeting α1-giardin and CWP2 can protect mice against the colonization of Giardia trophozoite and block the transformation of cyst in host at the same time, and can be used to prevent Giardia infection and block the transmission of giardiasis.

  14. 宠物犬蓝氏贾第鞭毛虫和隐孢子虫感染的分子检测%Molecular Detection of Giardia lamblia and Cryptosporidium Species in Pet Dogs

    Institute of Scientific and Technical Information of China (English)

    顾有方; 张衡; 张学亮; 凡自来; 李文超; 汪凯; 刘德义; 梅楠; 陈程; 陈涛; 韩敏敏; 周力; 曹佳彤

    2015-01-01

    Objective To determine the prevalence of Giardia lamblia and Cryptosporidium species infection in pet dogs, and identify the G. lamblia assemblages and Cryptosporidium species. Methods A total of 315 fresh fecal samples were collected from pet clinics in five counties of Anhui Province and in Hangzhou of Zhejiang Province. Hemi-nested-PCR targeting the GDH gene of G. lamblia and nested-PCR targeting the SSU rRNA gene of Cryptosporidium were performed in all the fecal samples. The PCR products were sequenced and analyzed using bioinformatics methods to identify the G. lamblia assemblages and Cryptosporidium species. Results The positive rates of G. lamblia and Cryptosporidium spp. infections in the 315 fecal samples were 3.2% (10/315) and 1.6%(5/315), respectively. Specifically, the two indicators were both significantly higher in dogs ≤12 months (17.8%and 11.1%, respectively) than in adult dogs (0.7% and 0.0%) (P<0.05). However, there was no significant difference in the two indicators between male and female dogs. In addition, two G. lamblia assemblages were identified, assemblages B (n=6) and D (n=4). Sequence analysis of PCR products of the SSU rRNA gene showed that the five Cryptosporidium isolates were C. canis (n=5). Conclusion The prevalences of G. lamblia and Cryptosporidium infection in pet dogs in Anhui and Zhejiang Provinces were 3.2 % and 1.6 %, respectively. The assemblages of G. lamblia in this study are of types B and D.%目的:了解皖浙两省部分城镇地区宠物犬蓝氏贾第鞭毛虫和隐孢子虫感染情况,及聚集体类型或虫种。方法在安徽多地和浙江杭州共采集315份新鲜宠物犬粪样,分别采用基于蓝氏贾第鞭毛虫谷氨酸脱氢酶( GDH )基因的半巢式 PCR 和隐孢子虫核糖体小亚基( SSU rRNA )基因的巢式 PCR 方法对所有粪样进行扩增,并对获得的阳性扩增产物进行测序和生物信息学分析,确定蓝氏贾第鞭毛虫的聚集体和隐孢

  15. Research Progress on the Mechanisms of Antigenic Variation in Giardia lamblia%蓝氏贾第鞭毛虫表面抗原变异机制研究进展

    Institute of Scientific and Technical Information of China (English)

    冯宪敏; 王月华; 鞠晓红

    2012-01-01

    蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)是引起腹泻的最常见的肠道寄生原虫之一.同大多数肠道寄生的病原体一样,表面抗原(variant-specific surface proteins,VSPs)变异是虫体逃避宿主免疫攻击的重要机制,从而导致疾病慢性化和/或重复感染的发生.本文将从基因水平的调控、转录和转录后调控、VSPs表达修饰调节和VSPs的加工、处理与转运等4个方面对贾第虫表面抗原变异机制的研究进展进行综述.%The intestinal protozoan parasite Giardia lamblia is one of the most common causes of diarrhoea and undergoes antigenic variation. In pathogenic microorganisms, antigenic variation is often described as a mechanism to evade the host immune system, resulting in chronic and/or recurrent infections. In the recent years, significant advances in the knowledge of the antigen switching process have been achieved. Here we review the principal knowledge on the mechanisms that regulate this process, including genomic organization, post-tanscriptional gene silencing, expressional modifications, and processing and turnover of VSPs.

  16. Vaccination with Bivalent DNA Vaccine of α1-Giardin and CWP2 Delivered by Attenuated Salmonella typhimurium Reduces Trophozoites and Cysts in the Feces of Mice Infected with Giardia lamblia.

    Science.gov (United States)

    Feng, Xian-Min; Zheng, Wen-Yu; Zhang, Hong-Mei; Shi, Wen-Yan; Li, Yao; Cui, Bai-Ji; Wang, Hui-Yan

    2016-01-01

    Giardia lamblia is one of the most common infectious protozoans in human that may cause diarrhea in travelers. Searching for antigens that induced effectively protective immunity has become a key point in the development of vaccine against giardiasis. Mice vaccinated with G. lamblia trophozozite-specific α1-giardin DNA vaccine delivered orally by attenuated Salmonella typhimurium SL7027 elicited 74.2% trophozoite reduction, but only 28% reduction in cyst shedding compared with PBS buffer control. Oral vaccination with Salmonella-delivered cyst-specific CWP2 DNA produced 89% reduction in cysts shedding in feces of vaccinated mice. Significantly, the mice vaccinated with Salmonella-delivered bivalent α1-giardin and CWP2 DNA vaccines produced significant reduction in both trophozoite (79%) and cyst (93%) in feces of vaccinated mice. This parasite reduction is associated with the strong local mucosal IgA secretion and the IgG2a-dominant systemic immune responses in vaccinated mice. The results demonstrate that bivalent vaccines targeting α1-giardin and CWP2 can protect mice against the colonization of Giardia trophozoite and block the transformation of cyst in host at the same time, and can be used to prevent Giardia infection and block the transmission of giardiasis.

  17. Apo And Calcium-Bound Crystal Structures of Alpha-11 Giardin, An Unusual Annexin From 'Giardia Lamblia'

    Energy Technology Data Exchange (ETDEWEB)

    Pathuri, P.; Nguyen, E.T.; Svard, S.G.; Luecke, H.; /UC, Irvine /Uppsala U. /Karolinska Inst.

    2007-07-12

    Alpha-11 giardin is a member of the multi-gene alpha-giardin family in the intestinal protozoan, Giardia lamblia. This gene family shares an ancestry with the annexin super family, whose common characteristic is calcium-dependent binding to membranes that contain acidic phospholipids. Several alpha giardins are highly expressed during parasite-induced diarrhea in humans. Despite being a member of a large family of proteins, little is known about the function and cellular localization of alpha-11 giardin, although giardins are often associated with the cytoskeleton. It has been shown that Giardia exhibits high levels of alpha-11 giardin mRNA transcript throughout its life cycle; however, constitutive over-expression of this protein is lethal to the parasite. Determining the three-dimensional structure of an alpha-giardin is essential to identifying functional domains shared in the alpha-giardin family. Here we report the crystal structures of the apo and Ca{sup 2+}-bound forms of alpha-11 giardin, the first alpha giardin to be characterized structurally. Crystals of apo and Ca{sup 2+}-bound alpha-11 giardin diffracted to 1.1 angstroms and 2.93 angstroms, respectively. The crystal structure of selenium-substituted apo alpha-11 giardin reveals a planar array of four tandem repeats of predominantly {alpha}-helical domains, reminiscent of previously determined annexin structures, making this the highest-resolution structure of an annexin to date. The apo alpha-11 giardin structure also reveals a hydrophobic core formed between repeats I/IV and II/III, a region typically hydrophilic in other annexins. Surprisingly, the Ca{sup 2+}-bound structure contains only a single calcium ion, located in the DE loop of repeat I and coordinated differently from the two types of calcium sites observed in previous annexin structures. The apo and Ca{sup 2+}-bound alpha-11 giardin structures assume overall similar conformations; however, Ca2+-bound alpha-11 giardin crystallized in a lower

  18. Apo and calcium-bound crystal structures of cytoskeletal protein alpha-14 giardin (annexin E1) from the intestinal protozoan parasite Giardia lamblia.

    Science.gov (United States)

    Pathuri, Puja; Nguyen, Emily Tam; Ozorowski, Gabriel; Svärd, Staffan G; Luecke, Hartmut

    2009-01-30

    Alpha-14 giardin (annexin E1), a member of the alpha giardin family of annexins, has been shown to localize to the flagella of the intestinal protozoan parasite Giardia lamblia. Alpha giardins show a common ancestry with the annexins, a family of proteins most of which bind to phospholipids and cellular membranes in a Ca(2+)-dependent manner and are implicated in numerous membrane-related processes including cytoskeletal rearrangements and membrane organization. It has been proposed that alpha-14 giardin may play a significant role during the cytoskeletal rearrangement during differentiation of Giardia. To gain a better understanding of alpha-14 giardin's mode of action and its biological role, we have determined the three-dimensional structure of alpha-14 giardin and its phospholipid-binding properties. Here, we report the apo crystal structure of alpha-14 giardin determined in two different crystal forms as well as the Ca(2+)-bound crystal structure of alpha-14 giardin, refined to 1.9, 1.6 and 1.65 A, respectively. Although the overall fold of alpha-14 giardin is similar to that of alpha-11 giardin, multiwavelength anomalous dispersion phasing was required to solve the alpha-14 giardin structure, indicating significant structural differences between these two members of the alpha giardin family. Unlike most annexin structures, which typically possess N-terminal domains, alpha-14 giardin is composed of only a core domain, followed by a C-terminal extension that may serve as a ligand for binding to cytoskeletal protein partners in Giardia. In the Ca(2+)-bound structure we detected five bound calcium ions, one of which is a novel, highly coordinated calcium-binding site not previously observed in annexin structures. This novel high-affinity calcium-binding site is composed of seven protein donor groups, a feature rarely observed in crystal structures. In addition, phospholipid-binding assays suggest that alpha-14 giardin exhibits calcium-dependent binding to

  19. Interpreting serum risperidone concentrations.

    Science.gov (United States)

    Boerth, Joel M; Caley, Charles F; Goethe, John W

    2005-02-01

    Risperidone is an atypical antipsychotic commonly used for treatment of schizophrenia and other psychotic disorders. Although therapeutic drug monitoring is not routine for any of the atypical antipsychotics, serum antipsychotic concentrations are measured routinely to assess treatment nonadherence. In humans, risperidone is metabolized by cytochrome P450 2D6 to 9-hydroxyrisperidone; together these constitute the active moiety. Dose-proportional increases in serum concentrations have not been reported for the parent drug, but have been reported for 9-hydroxyrisperidone and the active moiety (i.e., the combined concentrations of risperidone and 9-hydroxyrisperidone). We describe a 34-year-old Caucasian man of Sicilian descent with a history of schizophrenia, disorganized type. He was suspected to be noncompliant with his risperidone therapy. Initially, active moiety risperidone concentrations increased linearly with prescribed dosage increases. However, with continued increases, active moiety concentrations adjusted downward and remained 17-36% below anticipated levels. We propose a method for estimating target active moiety concentrations of risperidone based on dosage-a method that may be used to guide clinicians in assessing nonadherence to risperidone treatment.

  20. 实时荧光PCR检测蓝氏贾第鞭毛虫和隐孢子虫方法的建立%Detection of Giardia lamblia and Cryptosporidium by the real-time PCR method

    Institute of Scientific and Technical Information of China (English)

    杨晓华; 侯炎昌

    2013-01-01

    目的 建立贾第鞭毛虫和隐孢子虫的基因检测方法.方法 根据GenBank中贾第鞭毛虫和隐孢子虫相对保守序列,设计引物及其相应的Taqman探针.通过对引物和探针浓度、Taq酶以及反应条件等优化筛选后,建立检测贾第鞭毛虫和隐孢子虫的荧光PCR方法.结果 贾第鞭毛虫和隐孢子虫的检测结果为阳性,对其它DNA样本如日本血吸虫、刚地弓形虫、溶组织内阿米巴、旋毛虫和阴道毛滴虫的检测均为阴性.本法的敏感性高,可检测到10~102copies/μl的DNA浓度.结论 建立的基因检测方法,对贾第鞭毛虫和隐孢子虫的检测具有高度的特异性和敏感性,可用于饮用水和临床样本等的快速检测.%Objeetive The present study aims to establish a real-time PCR method for the detection of Giardia lamblia and Cryptosporidium.Methods Specific primers and the corresponding Taqman probes were designed according to the conserved sequences recorded in GenBank.Results Using the designed primers and probes,positive results were obtained from the samples of Giardia lamblia and Cryptosporidium.Non-specific detection of other DNA samples,such as Schistosoma japonicum,Toxoplasma gondii,Entamoeba histolytica,Trichinella spiralis and Trichomonas vaginalis was not observed.The sensitivity of this method is 10 ~ 102 copies/μl.Conclusion The established method is specific and sensitive for the detection of Giardia lamblia and Cryptosporidium.This method could be used in the rapid detection of drinking water and clinical samples.

  1. Giardia lamblia and other intestinal parasitic infections and their relationships with nutritional status in children in Brazilian Amazon Giardia lamblia e outros parasitas intestinais e sua relação com o status nutricional de crianças de uma área urbana na Amazônia Brasileira

    Directory of Open Access Journals (Sweden)

    Filipe Anibal Carvalho-Costa

    2007-06-01

    Full Text Available The objective of this survey was to assess the relationships between intestinal parasitism, nutritional status and hemoglobin level in children with Indian ascendancy living in an urban area in Brazilian Amazon. We carried out a cross-sectional survey obtaining anthropometric, parasitological and socioeconomic data, and hemoglobin measurements of children aged six to 84 months. Anthropometric data were expressed as z-scores for weight for age (WAZ, height for age (HAZ, weight for height (WHZ and mid upper circumference for age (MUACZ parameters. Parasitological examinations were performed through Ritchie (n = 307, Kato-Katz (n = 278, Baermann-Moraes (n = 238 and Safranin-methylene blue methods (n = 307. Hemoglobin measurements were obtained with a Hemocue® photometer (n = 282. Socioeconomic data were used in order to classify children in three family income strata (n = 242. Multiple linear regression analysis showed independent interactions between Giardia lamblia and WAZ (beta = -0.195, SE = 0.138, p = 0.003, WHZ (beta = -0.161, SE = 0.133, p = 0.018 and MUACZ (beta = -0.197, SE = 0.143, p = 0.011, controlling for age, sex, family income, Ascaris lumbricoides, and hookworm infection. Also, the multivariate model showed that the only variable associated with hemoglobin levels was age. Intestinal parasitism control should increase children's possibilities of full development in the studied area.O presente estudo objetivou avaliar a relação entre as parasitoses intestinais, o status nutricional e os níveis de hemoglobina em crianças vivendo em uma área urbana na Amazônia Brasileira. Foi realizado um estudo seccional, obtendo-se dados antropométricos, parasitológicos e socioeconômicos, além de dosagens de hemoglobina através do fotômetro Hemocue®, de crianças com idade entre seis e 84 meses. Os dados da antropometria foram expressos como escores de desvio-padrão (escores z para os parâmetros peso-idade (PI, altura-idade (AI, peso

  2. Presencia de Giardia lamblia y Cryptosporidium spp. en aguas residuales depuradas reutilizadas para riego agrícola en la isla de Tenerife, España. Efectos del transporte a larga distancia sobre la calidad del agua reutilizada

    Directory of Open Access Journals (Sweden)

    N. Abreu Acosta

    2002-06-01

    Full Text Available El empleo de aguas residuales depuradas para riego agrícola es la alternativa de reutilización con más relevancia hoy día, siempre y cuando se desarrolle con garantías sanitarias y medioambientales.Giardia lamblia y Cryptosporidium spp. son dos protozoos patógenos entéricos de amplia distribución ambiental, frecuentes en hábitats acuáticos. La investigación y detección de estos parásitos en aguas ha adquirido importancia en los últimos años debido a que poseen formas de dispersión resistentes a los tratamientos habituales, aplicados tanto en procesos de potabilización como de depuración, y a que han sido clasificados como patógenos emergentes causantes de importantes brotes de transmisión hídrica.En este trabajo se estudia la presencia de quistes de Giardia lamblia y ooquistes de Cryptosporidium spp. en el agua residual depurada de la ciudad de Santa Cruz de Tenerife que es transportada hasta el sur de la isla de Tenerife para su reutilización en agricultura. Asimismo se investiga el efecto del transporte, el almacenamiento y el tratamiento avanzado sobre la concentración de quistes y ooquistes a través del sistema, y la existencia de relaciones con otros parámetros bacteriológicos y físico-químicos.Los resultados obtenidos ponen de manifiesto comportamientos variables de los contenidos en quistes y ooquistes frente a los tratamientos aplicados, y el efecto depurador que el transporte a larga distancia, parece tener sobre el agua residual depurada.No se han encontrado relaciones entre las concentraciones de quistes y ooquistes en el agua residual depurada y el contenido de los indicadores tradicionales de contaminación fecal.

  3. Presencia de Giardia lamblia y Cryptosporidium spp. en aguas residuales depuradas reutilizadas para riego agrícola en la isla de Tenerife, España. Efectos del transporte a larga distancia sobre la calidad del agua reutilizada Giardia lamblia and Cryptosporidium spp. Presence in treated wastewater reutilised for irrigation in Tenerife island, Spain. Long-distance transport effects in the reutilised water quality

    Directory of Open Access Journals (Sweden)

    Escolástico Aguiar González

    2002-06-01

    Full Text Available El empleo de aguas residuales depuradas para riego agrícola es la alternativa de reutilización con más relevancia hoy día, siempre y cuando se desarrolle con garantías sanitarias y medioambientales.Giardia lamblia y Cryptosporidium spp. son dos protozoos patógenos entéricos de amplia distribución ambiental, frecuentes en hábitats acuáticos. La investigación y detección de estos parásitos en aguas ha adquirido importancia en los últimos años debido a que poseen formas de dispersión resistentes a los tratamientos habituales, aplicados tanto en procesos de potabilización como de depuración, y a que han sido clasificados como patógenos emergentes causantes de importantes brotes de transmisión hídrica.En este trabajo se estudia la presencia de quistes de Giardia lamblia y ooquistes de Cryptosporidium spp. en el agua residual depurada de la ciudad de Santa Cruz de Tenerife que es transportada hasta el sur de la isla de Tenerife para su reutilización en agricultura. Asimismo se investiga el efecto del transporte, el almacenamiento y el tratamiento avanzado sobre la concentración de quistes y ooquistes a través del sistema, y la existencia de relaciones con otros parámetros bacteriológicos y físico-químicos.Los resultados obtenidos ponen de manifiesto comportamientos variables de los contenidos en quistes y ooquistes frente a los tratamientos aplicados, y el efecto depurador que el transporte a larga distancia, parece tener sobre el agua residual depurada.No se han encontrado relaciones entre las concentraciones de quistes y ooquistes en el agua residual depurada y el contenido de los indicadores tradicionales de contaminación fecal.Treated wastewater use for irrigation is, nowadays, the more relevant reutilization alternative of wastewater, if it is developed with sanitary and environmental warranties.Giardia lamblia and Cryptosporidium spp. two wide distributed enteric pathogen protozoan, which are frequent in aquatic

  4. NL MIND-BEST: a web server for ligands and proteins discovery--theoretic-experimental study of proteins of Giardia lamblia and new compounds active against Plasmodium falciparum.

    Science.gov (United States)

    González-Díaz, Humberto; Prado-Prado, Francisco; Sobarzo-Sánchez, Eduardo; Haddad, Mohamed; Maurel Chevalley, Séverine; Valentin, Alexis; Quetin-Leclercq, Joëlle; Dea-Ayuela, María A; Teresa Gomez-Muños, María; Munteanu, Cristian R; José Torres-Labandeira, Juan; García-Mera, Xerardo; Tapia, Ricardo A; Ubeira, Florencio M

    2011-05-07

    different experiments. In experiment 1, we report by first time Quantum QSAR study, synthesis, characterization, and experimental assay of antiplasmodial and cytotoxic activities of oxoisoaporphine alkaloids derivatives as well as NL MIND-BEST prediction of potential target proteins. In experiment 2, we report sampling, parasite culture, sample preparation, 2-DE, MALDI-TOF, and -TOF/TOF MS, MASCOT search, MM/MD 3D structure modeling, and NL MIND-BEST prediction for different peptides a new protein of the found in the proteome of the human parasite Giardia lamblia, which is promising for anti-parasite drug-targets discovery. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Serum albumin: touchstone or totem?

    Science.gov (United States)

    Margarson, M P; Soni, N

    1998-08-01

    A decrease in serum albumin concentrations is an almost inevitable finding in disease states, and is primarily mediated in the acute phase by alterations in vascular permeability and redistribution. This change is not disease specific but marked changes that persist are generally associated with a poorer prognosis. Critical appraisal of long-standing practices and the availability of alternative colloid solutions have led to a reduction in albumin replacement therapy, and a widespread tolerance of lower albumin concentrations in patients. The factors determining serum albumin concentrations, their measurement and the implications of hypoalbuminaemia are reviewed. The clinical value of serum albumin measurement is discussed.

  6. Study of Serum Amylase and Serum Cholinesterase in Organophosphorus Poisoning

    Directory of Open Access Journals (Sweden)

    Sharan Badiger

    2016-04-01

    Full Text Available Background: Poisoning due to organophosphorus compounds is most commonly seen. Earlier plasma cholinesterase level was used to assess the severity of poisoning. Presently serum amylase is being recommended as a better indicator of severity. Aims and Objectives: To study plasma cholinesterase and serum amylase levels in acute organophosphorus and to correlate serum amylase levels with clinical severity and outcome. Material and Methods: A total of 80 patients in the study admitted to a tertiary care centre within 24 hours with a history of organophosphorus poisoning were included in study. Estimation of plasma cholinesterase and serum rd amylase was done at the time of admission, and on 3 th day and on 5 day. Results: Occurrence of organophosphorus poisoning was more common among age group 21-30 years and among males (57.5%. They were 25 (31.2% farmers, 23 (28.8% st u d e n ts, a n d 2 2 ( 2 7 . 5% h o u s ewi v e s. Monocrotophos (45.0% was commonly used compound. Mean value of plasma cholinesterase and serum amylase at admission are 3693 U/L, and 185.4 U/L. There was significant inhibition of plasma cholinesterase and elevation of serum amylase at th admission with return to normal values on 5 day. Conclusion: Plasma cholinesterase inhibition 200 U/L has been associated with poor prognosis and proneness to respiratory failure.

  7. Second Trimester Maternal Serum Screening

    Science.gov (United States)

    ... page: Was this page helpful? Also known as: AFP Maternal; Maternal Serum AFP; MSAFP; msAFP; Triple Screen; Triple Test; Quad Screen; ... Free Fetal DNA Were you looking instead for AFP tumor markers , used to help diagnose and monitor ...

  8. SERUM LIPIDS IN ANXIETY NEUROSIS

    OpenAIRE

    Mishra, T.K.; Shankar, R.; Sharma, I.; Srivastava, P. K.

    1984-01-01

    SUMMARY Serum cholesterol, total triglycerides, HDL-cholesterol, LDL-cholesterol, VLDL-cholesterol, free cholesterol and total phospholipids were studied in 36 patients of anxiety neurosis and 24 control subjects. Serum triglycerides, VLDL-cholesterol and free-cholesterol were found to be significantly raised while esterified cholesterol WJS significantly lowered in anxiety neurosis. A significant negative correlation was observed between the anxiety score and free cholesterol in ferrule pati...

  9. Variations of serum copper values in pregnancy

    OpenAIRE

    Vukelić Jelka; Kapamadžija Aleksandra; Petrović Đorđe; Grujić Zorica; Novakov-Mikić Aleksandra; Kopitović Vesna; Bjelica Artur

    2012-01-01

    Introduction. Copper is essential micronutrient and has an important role in the human body. The serum copper increases during pregnancy and is doubled at full term. Lower levels of serum copper in pregnancy are connected with some pathological conditions. Objective. The aim of this study was to estimate the levels of serum copper in normal and pathological pregnancies, comparing them with values of serum copper in non-pregnant women, to determine if serum copper is lower in some pathol...

  10. Comparative Analysis of the 15.5kD Box C/D snoRNP Core Protein in the Primitive Eukaryote Giardia lamblia Reveals Unique Structural and Functional Features

    Energy Technology Data Exchange (ETDEWEB)

    Biswas, Shyamasri; Buhrman, Greg; Gagnon, Keith; Mattos, Carla; Brown, II, Bernard A.; Maxwell, E. Stuart (NCSU); (UTSMC)

    2012-07-11

    Box C/D ribonucleoproteins (RNP) guide the 2'-O-methylation of targeted nucleotides in archaeal and eukaryotic rRNAs. The archaeal L7Ae and eukaryotic 15.5kD box C/D RNP core protein homologues initiate RNP assembly by recognizing kink-turn (K-turn) motifs. The crystal structure of the 15.5kD core protein from the primitive eukaryote Giardia lamblia is described here to a resolution of 1.8 {angstrom}. The Giardia 15.5kD protein exhibits the typical {alpha}-{beta}-{alpha} sandwich fold exhibited by both archaeal L7Ae and eukaryotic 15.5kD proteins. Characteristic of eukaryotic homologues, the Giardia 15.5kD protein binds the K-turn motif but not the variant K-loop motif. The highly conserved residues of loop 9, critical for RNA binding, also exhibit conformations similar to those of the human 15.5kD protein when bound to the K-turn motif. However, comparative sequence analysis indicated a distinct evolutionary position between Archaea and Eukarya. Indeed, assessment of the Giardia 15.5kD protein in denaturing experiments demonstrated an intermediate stability in protein structure when compared with that of the eukaryotic mouse 15.5kD and archaeal Methanocaldococcus jannaschii L7Ae proteins. Most notable was the ability of the Giardia 15.5kD protein to assemble in vitro a catalytically active chimeric box C/D RNP utilizing the archaeal M. jannaschii Nop56/58 and fibrillarin core proteins. In contrast, a catalytically competent chimeric RNP could not be assembled using the mouse 15.5kD protein. Collectively, these analyses suggest that the G. lamblia 15.5kD protein occupies a unique position in the evolution of this box C/D RNP core protein retaining structural and functional features characteristic of both archaeal L7Ae and higher eukaryotic 15.5kD homologues.

  11. Blood serum mercury test report.

    Science.gov (United States)

    Vandenberge, J; Moodie, A S; Keller, R E

    1977-06-01

    A clinical blood serum mercury test of 111 dentists and auxiliaries revelaed that more than 50% had above normal serum mercury levels. This study showed that there may be a mercury health hazard in some dental environments. Acute mercury poisoning may be corrected simply by removing the cause, but long-term chronic effects are not known. Frequent screening of offices and personnel is advised. Experience reported here indicates that large amounts of mercury vapor are emitted when an amalgam carrier is heated over a flame ot dislodge particles, and also, that water-covered amalgam scrap relesases mercury vapor.

  12. Piezoelectric microcantilever serum protein detector

    Science.gov (United States)

    Capobianco, Joseph A.

    The development of a serum protein detector will provide opportunities for better screening of at-risk cancer patients, tighter surveillance of disease recurrence and better monitoring of treatment. An integrated system that can process clinical samples for a number of different types of biomarkers would be a useful tool in the early detection of cancer. Also, screening biomarkers such as antibodies in serum would provide clinicians with information regarding the patient's response to treatment. Therefore, the goal of this study is to develop a sensor which can be used for rapid, all-electrical, real-time, label-fee, in-situ, specific quantification of cancer markers, e.g., human epidermal receptor 2 (Her2) or antibodies, in serum. To achieve this end, piezoelectric microcantilever sensors (PEMS) were constructed using an 8 mum thick lead magnesium niobate-lead titanate (PMN-PT) freestanding film as the piezoelectric layer. The desired limit of detection is on the order of pg/mL. In order to achieve this goal the higher frequency lateral extension modes were used. Also, as the driving and sensing of the PEMS is electrical, the PEMS must be insulated in a manner that allows it to function in aqueous solutions. The insulation layer must also be compatible with standardized bioconjugation techniques. Finally, detection of both cancer antigens and antibodies in serum was carried out, and the results were compared to a standard commercialized protocol. PEMS have demonstrated the capability of detecting Her2 at a concentration of 5 pg/mL in diluted human serum (1:40) in less than 1 hour. The approach can be easily translated into the clinical setting because the sensitivity is more than sufficient for monitoring prognosis of breast cancer patients. In addition to Her2 detection, antibodies in serum were assayed in order to demonstrate the feasibility of monitoring the immune response for antibody-dependent cellular cytotoxicity (ADCC) in patients on antibody therapies

  13. Distribution of rDNA in the Nucleus of Giardia lamblia:Detection by Ag-Ⅰ Silver Stain%无核仁原生动物蓝氏贾第虫rDNA的分布

    Institute of Scientific and Technical Information of China (English)

    陈泳宏; 郭建; 李靖炎; 刘平; 张留宝

    2002-01-01

    过去的工作已表明,源真核生物(Archezoa)中的双滴虫类极其原始,核中尚无核仁发生,以蓝氏贾第虫(Giardia lamblia)作为双滴虫类的代表,用高度特异的核仁组织区银染法(改良的Ag-Ⅰ法,李靖炎,1985)在电镜下检视其rDNA在核中的分布.结果发现,代表rDNA之所在的银粒并不集中形成任何类似核仁组织区或核仁纤维区的结构;在作为对照的小眼虫(Euglena gracilis)体内,银粒则完全集中在核仁纤维区中,因此,作者以为贾第虫rDNA的这种分布代表着核仁组织区进化形成以前的一种原始状态.

  14. The study of urpan prefecture village primary students infection Giardia lamblia genotype%吐鲁番地区乡村小学生感染蓝氏贾第鞭毛虫基因型的研究

    Institute of Scientific and Technical Information of China (English)

    法蒂玛·木特力甫; 高剑; 刘君; 侯秋莲; 吾拉木·马木提

    2015-01-01

    目的 了解吐鲁番地区乡村小学生感染蓝氏贾第鞭毛虫(Giardia lamblia)的基因型及亚型,探索不同地域贾第虫分离虫株基因之间的蓝氏贾第鞭毛虫的相互关系.方法 对感染贾第鞭毛虫的小学生粪便内的蓝氏贾第鞭毛虫的磷酸丙糖异构酶(triose phosphate isomerase,简称为tin/tpi)基因进行基因检测和基因序列分析.结果 19个分离虫株为集聚体A型,11个分离虫株为集聚体B型.结论 人群感染蓝氏贾第鞭毛虫分离虫株种之间的进化关系不受不同地理区域分布的影响,而虫株亚种之间可能受不同地理区域分布的影响有差别.

  15. 人源蓝氏贾第虫(Giardia lamblia)病毒部分基因的克隆及序列分析%Cloning and sequence analysis of a partial gene of the Giardiavirus

    Institute of Scientific and Technical Information of China (English)

    田宗成; 张西臣; 李建华; 尹继刚; 杨举

    2002-01-01

    目的从我国蓝氏贾第虫(Giardia lamblia)中寻找贾第虫病毒.方法对人源贾第鞭毛虫北京株进行了体外纯培养,将其总核酸电泳,并用DNA酶和RNA酶处理.根据已发表的蓝氏贾第虫病毒基因序列合成一对引物并进行RT-PCR,将产物回收后连接到Pmd18-T载体上进行克隆并测序,通过BLAST对Gen Bank进行同源性搜索.结果在贾第虫总核酸电泳图谱上观察到一个分子量约7.0kb的片段.该核酸不能被DNA酶(100μg/ml)降解.但可被RNA酶(10μg/ml)降解.经RT-PCR扩增后得到1条预计856bp的片段,所得序列与蓝氏贾第虫病毒基因同源性为98%.结论在我国人源贾第鞭毛虫北京株中发现贾第虫病毒,该病毒属于蓝氏贾第虫病毒.

  16. Research progress on the molecular detection of waterborne Giardia lamblia%水源性蓝氏贾第鞭毛虫分子生物学检测方法研究进展

    Institute of Scientific and Technical Information of China (English)

    梁乐; 汪世龙; 王惠琴

    2009-01-01

    蓝氏贾第鞭毛虫具有一系列适于水源性传播的生物学特征,广泛存在于污水、原水、处理水等水体中,严重威胁公共卫牛安全.水源性贾第虫的检测主要是通过免疫磁珠分离后进行分子鉴别.运用单个基因作为分子标志的检测方法的可靠性值得商榷,而全基因组扩增技术和DNA微阵列技术具有重要的应用前景.%Giardia lamblia is waterborne transmission facilitated by a series of its biological traits.It exists in sewage water,raw water,treated water and other water systems extensively as a serious public health threatening pathogen.Waterborne Giardia is mainly identified by molecular methods after immunomagnetic sep aration.It is not reliable to use a single gene as molecular marker to detect this pathogen.Instead,whole ge nome amplification and DNA microarray are promising methods.

  17. Increased serum potassium affects renal outcomes

    DEFF Research Database (Denmark)

    Miao, Y; Dobre, D; Heerspink, H J Lambers;

    2011-01-01

    To assess the effect of an angiotensin receptor blocker (ARB) on serum potassium and the effect of a serum potassium change on renal outcomes in patients with type 2 diabetes and nephropathy.......To assess the effect of an angiotensin receptor blocker (ARB) on serum potassium and the effect of a serum potassium change on renal outcomes in patients with type 2 diabetes and nephropathy....

  18. Serum ferritin levels in hospital patients

    OpenAIRE

    竹田, 芳弘; 平木, 祥夫; 森本,節夫; 戸上, 泉; 白神, 敏明; 白石,則之; 木本, 真; 上者,郁夫; 橋本, 啓二; 青野, 要

    1985-01-01

    Serum ferritin levels were determined in 75 hematologic, 1205 malignant, and 538 benign diseases. In hematologic diseases the serum ferritin level was generally low in iron deficiency anemia, but high in hemochromatosis, aplastic anemia, and leukemia. In malignant diseases the serum ferritin concentration was increased remarkably in lung, liver, biliary, and urogential diseases and in malignant lymphoma. Among benign diseases hepatitis cases showed a tendency for high serum ferritin levels. S...

  19. Serum ferritin levels in hospital patients

    OpenAIRE

    竹田,芳弘; 平木,祥夫; 森本, 節夫; 戸上, 泉; 白神, 敏明; 白石, 則之; 木本, 真; 上者, 郁夫; 橋本, 啓二; 青野,要

    1985-01-01

    Serum ferritin levels were determined in 75 hematologic, 1205 malignant, and 538 benign diseases. In hematologic diseases the serum ferritin level was generally low in iron deficiency anemia, but high in hemochromatosis, aplastic anemia, and leukemia. In malignant diseases the serum ferritin concentration was increased remarkably in lung, liver, biliary, and urogential diseases and in malignant lymphoma. Among benign diseases hepatitis cases showed a tendency for high serum ferritin levels. S...

  20. Label-Free Proteomics of Serum

    NARCIS (Netherlands)

    Govorukhina, N.I; Horvatovich, P.; Bischoff, Rainer

    2008-01-01

    In this chapter we describe a method to analyze human serum with the goal of discovering disease-related changes in the serum proteome. The methodology is based on the removal of the six most abundant serum proteins by immunoaffinity chromatography. This step is followed by trypsin digestion and

  1. 1,6-二磷酸果糖对蓝氏贾第鞭毛虫滋养体形态学的影响%Effect of fructose 1,6-biphosphate on morphology of Giardia lamblia trophozoites

    Institute of Scientific and Technical Information of China (English)

    冯宪敏; 鞠晓红; 王月华; 朱枫; 藏秋雨; 时文艳; 卢思奇

    2012-01-01

    Objective To study the effect of fructose 1,6-biphosphate ( FBP ) on the morphology of Giarida lamblia trophozoites. Methods Fructose 1,6-diphosphate was added into each cultural tube with the final concentration of 30 ng/μ1 immediately for group A and 4 hours later for group B after the inoculation at concentration of 0. 5 × 10 /ml. The adherence, activity and morphology of trophozoites were detected under the inverted microscope at 24 h, 48 h, 72 h and 96 h after drug administration, respectively. Trophozoites in each group were collected at those time points. The concentrations of trophozoites were calculated and the growth curves were constructed. Meanwhile, a group cultured normally was set as a control( group C ). Results Compared with group C, morphology of the trophozoites changed significantly with activity and adherence of trophozoites decreasing at the time point of 48 h after exposing to FBP. No adherent trophozoites were found after 72 h. All the trophozoites died after 96 h. The growth curve showed that the growth of trophozoites was considerably depressed after 24 h exposing to FBP. Half of the trophozoites died after 48 h and no alive ones were detected after 96 h. The difference were statistically significant between group A, B and C ( P < 0. 01 ). Conclusions Over accumulation of fructose 1, 6-biphosphate can induce the morphology damage and death of Giardia lamblia trophozoites. But, further study of the mechanism should be performed.%目的 探讨1,6-二磷酸果糖对体外培养的蓝氏贾第鞭毛虫(贾第虫)滋养体形态学的影响.方法 按0.5 × 106/ml的贾第虫滋养体浓度接种于4 ml培养瓶,接种后即时(A组)或接种后4 h(B组)加入1,6-二磷酸果糖,终浓度30 ng/μl.分别于药物作用后24、48、72和96 h以倒置显微镜观察虫体的贴壁情况、活力和形态变化;收集各组、各时段虫体,计数虫体滋养体浓度并绘制虫体生长曲线;同时,设立正常培养

  2. Factores de riesgo de la infección por Giardia lamblia en niños de guarderías infantiles de Ciudad de La Habana, Cuba

    Directory of Open Access Journals (Sweden)

    Fidel Ángel Núñez

    Full Text Available En un estudio longitudinal sobre giardiasis desarrollado durante 18 meses en tres guarderías infantiles de Ciudad de La Habana, describimos un grupo de niños con tendencia o "predisposición" a la infección por Giardia lamblia; a los que se les encontró con síntomas clínicos más asociados a las diarreas. Este estudio de casos y controles se desarrolló para conocer si algunos factores socioeconómicos y hábitos higiénicos estaban asociados con este fenómeno. No se encontraron diferencias entre ambos grupos en las tasas de hacinamiento, el índice de personas por cama, en la carencia de determinados equipos eléctricos, en el nivel escolar de las madres, y en el promedio del ingreso familiar; sin embargo, predominó una frecuencia mayor de padres con nivel escolar menor de 12 grado en los casos que en los controles. No se encontraron diferencias entre ambos grupos en cuanto a la ausencia de lavado de manos antes de comer y después de defecar, pero se encontró un mayor porcentaje de lavado incorrecto de vegetales, y una menor frecuencia del hábito de hervir el agua de consumo en las familias de los casos. Estos resultados demuestran el papel del agua como vehículo de transmisión en la giardiasis y la importancia de algunos factores epidemiológicos.

  3. Factores de riesgo de la infección por Giardia lamblia en niños de guarderías infantiles de Ciudad de La Habana, Cuba

    Directory of Open Access Journals (Sweden)

    Núñez Fidel Ángel

    2003-01-01

    Full Text Available En un estudio longitudinal sobre giardiasis desarrollado durante 18 meses en tres guarderías infantiles de Ciudad de La Habana, describimos un grupo de niños con tendencia o "predisposición" a la infección por Giardia lamblia; a los que se les encontró con síntomas clínicos más asociados a las diarreas. Este estudio de casos y controles se desarrolló para conocer si algunos factores socioeconómicos y hábitos higiénicos estaban asociados con este fenómeno. No se encontraron diferencias entre ambos grupos en las tasas de hacinamiento, el índice de personas por cama, en la carencia de determinados equipos eléctricos, en el nivel escolar de las madres, y en el promedio del ingreso familiar; sin embargo, predominó una frecuencia mayor de padres con nivel escolar menor de 12 grado en los casos que en los controles. No se encontraron diferencias entre ambos grupos en cuanto a la ausencia de lavado de manos antes de comer y después de defecar, pero se encontró un mayor porcentaje de lavado incorrecto de vegetales, y una menor frecuencia del hábito de hervir el agua de consumo en las familias de los casos. Estos resultados demuestran el papel del agua como vehículo de transmisión en la giardiasis y la importancia de algunos factores epidemiológicos.

  4. C2株贾第虫SBDS基因的原核表达与生物信息学分析%Prokaryotic Expression and Bioinformatics Analysis of Giardia lamblia C2 Strain SBDS Gene

    Institute of Scientific and Technical Information of China (English)

    王沂; 余源; 田喜凤; 李冀; 周英斌; 李少东; 刘晓莉; 王洋

    2014-01-01

    目的:克隆、原核表达C2株蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)的Shwachman-Bodian-Diamond syndrome (SBDS),并对其进行生物信息学分析.方法:以C2株贾第虫基因组DNA为模板获得SBDS基因编码区,克隆入原核表达载体pET-28a(+),测序并进行生物信息学分析,将重组质粒pET-28a(+)-SBDS在大肠杆菌Rosetta(DE3)中诱导表达,SDS-PAGE及Western blot验证表达效果.结果:成功构建了C2株贾第虫SBDS原核表达载体并在在大肠杆菌中得到了高效表达,SDS-PAGE及Western blot显示,在相对分子量约29kDa的位置出现目的蛋白条带,与理论值相符;生物信息学分析显示贾第虫SBDS蛋白在空间上形成三个结构域,在进化上与其它现存真核生物亲缘关系较远.结论:原核表达并分析了贾第虫SBDS蛋白,为贾第虫SBDS的进一步研究提供了基础.

  5. Research progress on specific cytoskeletal proteins-giardins in Giardia lamblia%蓝氏贾第鞭毛虫特异性细胞骨架蛋白——贾第素研究进展

    Institute of Scientific and Technical Information of China (English)

    魏超君; 田喜凤; 卢思奇

    2014-01-01

    蓝氏贾第鞭毛虫具有由微管、微丝和骨架蛋白构成的高度发达的细胞骨架系统.近年来,贾第虫的细胞骨架与其致病力的密切关系已经得到证实.在参与细胞骨架构成的众多蛋白质中,特异性细胞骨架蛋白——贾第素是其中重要的成分之一.对贾第素的研究不仅可为贾第虫细胞骨架的研究增添新的知识和研究资料,还可为贾第虫病的致病机制及抗贾第虫药物靶点的选择提供实验依据.%The most significant features of Giardia lamblia are the highly developed cytoskeleton system which is composed of tubulin and a variety of microtubules.In recent years,the strong link between the cytoskeleton and virulence has been confirmed.Among a number of cytoskeletal proteins involved in composition,the specific cytoskeletal protein-giardin is one of the important ingredients.The study of the giardins will not only add new knowledge and research data to the cytoskeleton research but also provide an empirical basis for investigating the pathogenic mechanisms of giardiasis and searching for new drug targets.

  6. 蓝氏贾第鞭毛虫中国株组蛋白H2A基因的克隆与重组表达%Cloning and expression of H2A gene from Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    王云华; 郑国侠; 巨红妹; 张霞; 李雅杰

    2011-01-01

    Objective To obtain C1H2A gene from Giardia lamblia and relevant recombinant proteins. Methods The C1H2A gene of Giardia lamblia was amplified from genomic DNA by PCR. A His6-tag coding sequence and a Nco I site were included in upstream primer and a stop codon and a BamH I site were included in- downstream primer.The PCR products were subcloned into pMD-19T vector and the positive clones were selected by colony PCR.The recombinant plasmid was sequenced.The resulting sequence was queried in GenBank.The coding sequence of G1H2A was aligned with H2A sequences from the other organisms. The plasmid verified by sequencing was digested by Nco I and BamH I and then inserted into the expression vector pET28a, yielding pET28a-GlH2A.The E.coli BL21 (DE3) was transformed and induced with IPTG. The cells were collected and dispersed with sonication.The recombinant protein was purified by nickel affinity chromatography. Recombinant G1H2A was detected using Western blotting method. Results The gene sequence obtained was identical to G1H2A GL50803_14256 (Accession:NW_001844132),consisting of 375 bp and coding for 124 amino acids. The predicted molecular weight of G1H2A was 13 900 Mr, with PI value of 10.45. The amino acid sequence of H2A from different organisms was aligned and the result showed the divergent of G1H2A to the others H2A genes. The result also revealed conservation of a number of residues at some unique position such as H2A-H2A interface, H2A-H2B interface and DNA-binding site. The expression vector pET28a-GlH2A was constructed. rGlH2A was expressed in E.coli and accounted for about 20% of total protein.The recombinant protein was purified by nickel affinity chromatography and the purity reached 90%.The product was verified by Western blotting using anti-His6 tag antibody. Conclusions G1H2A gene of G.lamblia was cloned and sequenced.lt showed conservation at some positions and may play similar role as that of in other organisms. Recombinant G1H2A was expressed in E

  7. Leptin in first trimester pregnancy serum

    DEFF Research Database (Denmark)

    Hedley, Paula; Pihl, Kasper; Krebs, Lone

    2009-01-01

    Leptin is an adipocytokine that is also synthesized by the placenta. Leptin and its receptor, which is also expressed by the placenta, are believed to play an auto- and paracrine role in trophoblast invasion and placental development. The leptin concentration in first trimester maternal serum......, gestational age and body mass index (BMI). All were non-smokers. Leptin was measured in maternal serum in weeks 8-13 and was normalized for BMI with concentrations expressed as multiples of the median for the actual BMI. It was found that maternal serum leptin increased strongly (r = 0.7, P ... maternal BMI. There was no significant difference in maternal serum leptin concentrations between SGA and AGA pregnancies. In conclusion, SGA pregnancies are not associated with a lower maternal serum leptin concentration in first trimester. The maternal serum leptin concentration is largely determined...

  8. Variations of serum copper values in pregnancy

    Directory of Open Access Journals (Sweden)

    Vukelić Jelka

    2012-01-01

    Full Text Available Introduction. Copper is essential micronutrient and has an important role in the human body. The serum copper increases during pregnancy and is doubled at full term. Lower levels of serum copper in pregnancy are connected with some pathological conditions. Objective. The aim of this study was to estimate the levels of serum copper in normal and pathological pregnancies, comparing them with values of serum copper in non-pregnant women, to determine if serum copper is lower in some pathological pregnancies and if this is of some importance. Methods. A total of 2170 plasma samples for copper analyses were made in the following groups: healthy non-pregnant women; healthy pregnant women from the 5th-40th gestational week, during the first delivery stage and during the first three postpartum weeks, in pregnant women with habitual abortion, imminent abortion, abortion in progress, missed abortion (9th-24th weeks, missed labour and premature rupture of membranes (29th-40th weeks. Levels of serum copper were determined by colorimetric technique of bathocuproin with disulphate as a chromogen. Results. Serum copper values in non-pregnant women range from 11.6-25.8 μmol/L. In healthy pregnant women, there is a constant trend of the increase of serum copper. The mean serum copper values revealed three significant peaks at the 22nd, 27th and 35th gestational week. Serum copper values in the patients with some pathological pregnancies in relation to the serum copper values of the healthy pregnant women were significantly lower. Conclusion. Serum copper values can be used as an indicator of some pathological pregnancies.

  9. Variations of serum copper values in pregnancy.

    Science.gov (United States)

    Vukelić, Jelka; Kapamadzija, Aleksandra; Petrović, Djordje; Grujić, Zorica; Novakov-Mikić, Aleksandra; Kopitović, Vesna; Bjelica, Artur

    2012-01-01

    Copper is essential micronutrient and has an important role in the human body. The serum copper increases during pregnancy and is doubled at full term. Lower levels of serum copper in pregnancy are connected with some pathological conditions. The aim of this study was to estimate the levels of serum copper in normal and pathological pregnancies, comparing them with values of serum copper in non-pregnant women, to determine if serum copper is lower in some pathological pregnancies and if this is of some importance. A total of 2170 plasma samples for copper analyses were made in the following groups: healthy non-pregnant women; healthy pregnant women from the 5th-40th gestational week, during the first delivery stage and during the first three postpartum weeks, in pregnant women with habitual abortion, imminent abortion, abortion in progress, missed abortion (9th-24th weeks), missed labour and premature rupture of membranes (29th-40th weeks). Levels of serum copper were determined by colorimetric technique of bathocuproin with disulphate as a chromogen. Serum copper values in non-pregnant women range from 11.6-25.8 micromol/L. In healthy pregnant women, there is a constant trend of the increase of serum copper. The mean serum copper values revealed three significant peaks at the 22nd, 27th and 35th gestational week. Serum copper values in the patients with some pathological pregnancies in relation to the serum copper values of the healthy pregnant women were significantly lower. Serum copper values can be used as an indicator of some pathological pregnancies.

  10. Effects of dihydroartemisinin on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lamblia%双氢青蒿素对 C2株蓝氏贾第鞭毛虫Alpha-7.3 giardin 基因 mRNA 表达水平的影响

    Institute of Scientific and Technical Information of China (English)

    余源; 田喜凤; 陈阳; 葛爽; 王洋; 李巍伟; 赵丽娜; 刘阿倩; 林志强; 高雪

    2014-01-01

    目的:观察双氢青蒿素(dihydroartemisinin ,DHA)对C2株蓝氏贾第鞭毛虫(Giardia lamblia)Alpha-7.3 giar-din (α-贾第素)基因mRNA表达水平的影响,探讨其对蓝氏贾第鞭毛虫骨架蛋白的损伤作用。方法用双氢青蒿素浓度为100μg/mL、200μg/mL的改良TYI-S-33培养基分别培养C2株蓝氏贾第鞭毛虫2 h、4 h、8 h、12 h后,以不含药物组为对照,实时荧光定量RT-PCR检测药物作用后Alpha-7.3 giardin基因mRNA表达水平的变化。结果双氢青蒿素作用虫体后Al-pha-7.3 giardin基因mRNA表达水平明显低于对照组,二者有显著性差异。结论双氢青蒿素对C2株蓝氏贾第鞭毛虫Al-pha-7.3 giardin基因mRNA的表达具有明显的抑制作用,抑制效果与药物浓度和作用时间相关,提示双氢青蒿素对蓝氏贾第鞭毛虫骨架蛋白具有损伤作用。%Effects of dihydroartemisinin (DHA) on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lam-blia was investigated in this study to explore the damage to skeleton protein of C 2 Giardia lamblia .Giardia lamblia was culti-vated respectively for 2 ,4 ,8 ,and 12 hours with modified TYI-S-33 medium containing 100 μg/mL and 200 μg/mL DHA , while the control group performed in the same experimental conditions without DHA .The expressive quantity of Alpha-7 .3 gi-ardin mRNA was determined by using real-time reverse transcription PCR ,and then we found that the expressive quantities of Alpha-7 .3 giardin mRNA with DHA were significantly lower than those in the control group .It’s suggested that dihydroarte-misinin has obvious inhibitory effect on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lamblia .The actions of dihydroartemisinin on skeleton protein of C2 Giardia lamblia are effective .

  11. C2株蓝氏贾第鞭毛虫SUMO基因的克隆、原核表达和生物信息学分析%Cloning, prokaryotic expression and bioinformatics analysis of Giardia lamblia C2 strain SUMO gene

    Institute of Scientific and Technical Information of China (English)

    王洋; 王沂; LI Wei-wei; 李冀; 李思瑾; 楚晗; 田喜凤

    2013-01-01

    Small ubiquitin-like modifier (or SUMO) proteins are a family of small proteins that are covalently attached to and detached from other proteins in cells to modify their function.In order to express Giardia lamblia (C2 strain) SUMO prorein in E.coli,the full-length open reading frame of SUMO was amplified by PCR from Giardia lamblia genome DNA.The PCR product about 320 bp in length was cloned into prokaryotic expression vector pET 28a(+) with restriction enzymes Nco I and Xho I.Sequencing result showed the sequence of SUMO in C2 strain was identical with that in WB strain.Bioinformatics analysis showed that SUMO protein displayed an ubiquitin-fold structure in three dimensional space.Phylogenetic analysis showed a distant relationship of SUMO protein between Giardia lamblia and other eukaryotes.The recombinant vector pET-28a(+)-SUMO was transformed into E.coli Rosetta(DE3),then the recombinant SUMO protein was expressed by IPTG induction.SDS-PAGE and western blot using anti-His Tag antibody showed that the expression product of SUMO was a fusion protein about 12.7 kD.The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SUMO protein provide basis for antibody preparation and functional study of SUMO related protein.%目的 克隆、原核表达C2株蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)的小泛素相关修饰物(small ubiquit in-related modifier,SUMO)蛋白,并对其序列进行生物信息学分析.方法 提取C2株贾第虫基因组DNA,以基因组DNA为模板PCR获得SUMO基因片段,双酶切连入原核表达载体pET28a(+),经测序验证并进行生物信息学分析,将重组质粒pET 28a(+)-SUMO转化大肠杆菌Rosetta(DE3).IPTG诱导后收集菌体,裂解后进行SDS-PAGE及Western blot检测.结果 成功构建了C2株贾第虫SUMO基因原核表达载体pET 28a(+)-SUMO,经IPTG诱导后,在大肠杆菌中高效表达,SDSPAGE及Western blot分析显示,在相对分子量约12.7 kD的位置出现目的蛋

  12. Bone mineralisation in premature infants cannot be predicted from serum alkaline phosphatase or serum phosphate

    DEFF Research Database (Denmark)

    Faerk, J; Peitersen, Birgit; Petersen, S

    2002-01-01

    BACKGROUND: The bone mineral content of premature infants at term is lower than in mature infants at the same postconceptional age. Serum alkaline phosphatase and serum phosphate are often used as indicators of bone mineralisation. OBJECTIVE: To analyse the association between bone mineral content...... and serum alkaline phosphatase and serum phosphate. METHODS: Serum alkaline phosphatase and phosphate were measured at weekly intervals during admission in 108 premature infants of gestational age below 32 weeks (mean (SD) gestational age 29 (2) weeks; mean (SD) birth weight 1129 (279) g). Bone mineral...... content was measured at term (mean gestational age 41 weeks) by dual energy x ray absorptiometry and corrected for body size. RESULTS: Serum alkaline phosphatase was significantly negatively associated with serum phosphate (p serum alkaline...

  13. Peranan Kadar Feritin Serum terhadap Kejadian Preeklampsia

    Directory of Open Access Journals (Sweden)

    Mega Ulfah

    2015-09-01

    Full Text Available Abstrak Preeklampsia merupakan penyebab utama kematian maternal dan perinatal diseluruh dunia. Peningkatan kadar serum besi dan feritin memiliki potensi untuk digunakan secara diagnostik untuk memperingatkan preeklampsia tahap awal. Tujuan penelitian ini adalah untuk menentukan peranan kadar serum feritin terhadap kejadianpreeklampsia. Penelitian ini merupakan penelitian analitik observasional dengan desain case-control. Penelitian dilakukan dari Agustus 2013 hingga Juli 2014, bertempat di RS dr. M. Djamil, RS dr. Reksodiwiryo dan Laboratorium Biomedik UNAND Padang.  Jumlah sampel yang diteliti adalah 40 responden dimana sampel terdiri dari 2 kelompok,masing-masing terdiri dari 20 sampel. Pemeriksaan serum feritin dilakukan dengan metode ELISA. Perbedaan rerata kadar feritin serum antara kelompok preeklampsia dan kehamilan normal dianalisa dengan mengunakan independen ttest. Hasil penelitian diperoleh rerata kadar serum feritin pada kelompok preeklampsia dan kehamilan normal adalah50,46+4,37 ng/ml dan 17,64+1,6 ng/ml, dengan nilai p=0,004. Kadar feritin pada kedua kelompok masih dalam batas normal dan tidak ditemukan indikasi adanya kelebihan besi sebagai faktor resiko preeklampsia. Kesimpulan penelitian ini yaitu kadar serum feritin tidak memiliki peranan terhadap kejadian preeklampsia. Kata kunci: preeklampsia, hipertensi dalam kehamilan, serum feritin Abstract Preeclampsia is a major cause of worldwide maternal and prenatal mortality. The increase in iron serum and ferritin can be used as a diagnosis to warn of the early stage of preeclampsia. The objective of this study was to determine the impact of ferritin serum levels on preeclampsia. This study was an observational analytic study withcase-control design. It has been done from August 2013 to Juli 2014, in RS dr. M. Djamil, RS dr. Reksodiwiryo and Biomedical Laboratory of Andalas University Padang. Total sample evaluated was 40 samples. The sample consist of two groups, each group

  14. Characterization of feline serum-cobalt binding.

    Science.gov (United States)

    Schnelle, Amy N; Barger, Anne M; MacNeill, Amy L; Mitchell, Mark M; Solter, Philip

    2015-06-01

    Oxidative stress inhibits albumin's ability to complex with cobalt. Feline serum-cobalt binding has not been described. The objective was to develop a cobalt binding test for use with feline serum, and correlate the results with other biochemical and cellular constituents in blood, and with clinical diseases of cats. A colorimetric test of cobalt binding, based on the oxidation-reduction reaction of Co(+2) and dithiothreitol, was developed using feline serum. The test was used to measure cobalt binding in stored serum from 176 cats presented to the University of Illinois Veterinary Teaching Hospital for a variety of disease conditions. Time-matched hematology and biochemical data, and clinical information, were obtained from the medical record of each cat and correlated with the serum-cobalt binding results. Serial dilution of feline serum with phosphate-buffered saline resulted in a highly linear decrease in serum-cobalt binding (r(2)  = .9984). Serum-cobalt binding of the clinical samples also correlated with albumin concentrations in a stepwise linear regression model (r(2)  = .425), and both cobalt binding and albumin were significantly decreased in cases of inflammation. Albumin and cobalt binding also shared significant correlations with several erythron variables, and serum concentration of total calcium and bilirubin. The correlation of cobalt binding measured by a colorimetric test with albumin concentration in the clinical samples and with serum dilution is consistent with feline albumin-cobalt complex formation. Hypoalbuminemia is the likely cause of reduced serum-cobalt binding in inflammation and the correlations observed between cobalt binding and other variables. © 2015 American Society for Veterinary Clinical Pathology.

  15. Serum release boosts sweetness intensity in gels

    NARCIS (Netherlands)

    Sala, G.; Stieger, M.A.; Velde, van de F.

    2010-01-01

    This paper describes the effect of serum release on sweetness intensity in mixed whey protein isolate/gellan gum gels. The impact of gellan gum and sugar concentration on microstructure, permeability, serum release and large deformation properties of the gels was determined. With increasing gellan

  16. Serum biochemistries of Pacific black brant

    Science.gov (United States)

    Franson, J. Christian; Flint, Paul L.; Schmutz, Joel A.

    2017-01-01

    The data set contains results for nine serum biochemistries in molting Pacific black brant (Branta bernicla nigricans). These data were used to calculate reference intervals (sometimes referred to as normal values) for the nine serum biochemistries. All brant were after-hatch year. All samples were collected in 2006 and 2007 in the Teshekpuk Lake Special Area, Alaska.

  17. Serum Protein Profile Alterations in Hemodialysis Patients

    Energy Technology Data Exchange (ETDEWEB)

    Murphy, G A; Davies, R W; Choi, M W; Perkins, J; Turteltaub, K W; McCutchen-Maloney, S L; Langlois, R G; Curzi, M P; Trebes, J E; Fitch, J P; Dalmasso, E A; Colston, B W; Ying, Y; Chromy, B A

    2003-11-18

    Background: Serum protein profiling patterns can reflect the pathological state of a patient and therefore may be useful for clinical diagnostics. Here, we present results from a pilot study of proteomic expression patterns in hemodialysis patients designed to evaluate the range of serum proteomic alterations in this population. Methods: Surface-Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOFMS) was used to analyze serum obtained from patients on periodic hemodialysis treatment and healthy controls. Serum samples from patients and controls were first fractionated into six eluants on a strong anion exchange column, followed by application to four array chemistries representing cation exchange, anion exchange, metal affinity and hydrophobic surfaces. A total of 144 SELDI-TOF-MS spectra were obtained from each serum sample. Results: The overall profiles of the patient and control samples were consistent and reproducible. However, 30 well-defined protein differences were observed; 15 proteins were elevated and 15 were decreased in patients compared to controls. Serum from one patient exhibited novel protein peaks suggesting possible additional changes due to a secondary disease process. Conclusion: SELDI-TOF-MS demonstrated dramatic serum protein profile differences between patients and controls. Similarity in protein profiles among dialysis patients suggests that patient physiological responses to end-stage renal disease and/or dialysis therapy have a major effect on serum protein profiles.

  18. Serum paraoxonase 1 activity in dogs

    DEFF Research Database (Denmark)

    Rossi, Gabriele; Giordano, Alessia; Pezzia, Francesca

    2013-01-01

    Serum activity of paraoxonase (PON1) decreases during inflammation in many species. Little information is available on paraoxon-based tests and the possible role of PON1 in dogs.......Serum activity of paraoxonase (PON1) decreases during inflammation in many species. Little information is available on paraoxon-based tests and the possible role of PON1 in dogs....

  19. Serum triglycerides and risk of cardiovascular disease.

    NARCIS (Netherlands)

    Boullart, I.; Graaf, J. de; Stalenhoef, A.F.H.

    2012-01-01

    Dyslipidemia, especially elevated serum levels of cholesterol, is causally related to cardiovascular disease. The specific role of triglycerides has long been controversial. In this article we discuss the role of serum triglycerides in relation to the risk of cardiovascular disease. First, the

  20. Cloning, prokaryotic expression and sequencing of TCTP gene of Giardia lamblia C2 strain%C2株蓝氏贾第鞭毛虫TCTP基因的克隆、表达和序列分析

    Institute of Scientific and Technical Information of China (English)

    王沂; 余源; 田喜凤; 李冀; 楮晗; 王洋

    2015-01-01

    目的 克隆C2株蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)的翻译调控肿瘤蛋白(Translationally controlled tumor protein,TCTP)编码区,对其进行生物信息学分析和原核表达.方法 根据WB株贾第虫TCTP编码区序列设计引物,以C2株贾第虫基因组DNA为模板扩增获得TCTP编码区片段,双酶切连入原核表达载体pET-28a(+),将酶切和测序验证正确的重组质粒转化E.coli Rosetta(DE3),经IPTG诱导表达融合蛋白,SDS-PAGE及Western blot鉴定蛋白产物,采用Ni-NTA经亲和层析纯化重组蛋白.同时根据测序结果分析C2株贾第虫TCTP蛋白结构特征和进化关系.结果 克隆了包含酶切位点全长470 bp的C2株贾第虫TCTP编码区,构建了原核表达载体pET-28a(+)-TCTP,在Rosetta(DE3)中表达、纯化得到了相对分子量约18.5 kDa的融合蛋白,与预期相符;测序结果显示C2株贾第虫TCTP序列与WB株相同,生物信息学分析显示贾第虫TCTP蛋白虽然序列长度较其它物种更短但与裂殖酵母TCTP空间结构相似,均由4个β折叠和3个主要的α螺旋组成,位于序列中部的不规则环状结构更小,仅9个氨基酸残基组成,在进化上与其它真核生物亲缘关系较远.结论 成功克隆表达了贾第虫TCTP,为TCTP功能及贾第虫致病机制的研究提供了实验依据.

  1. Prokaryotic expression and characterization of Giardia lamblia extracellular nuclease%蓝氏贾第鞭毛虫胞外核酸酶的表达纯化和活性鉴定

    Institute of Scientific and Technical Information of China (English)

    王沂; 赵俊暕; 余源; 田喜凤; 李冀; 刘晓莉; 周英斌; 李少东; 王洋

    2016-01-01

    目的 克隆、原核表达蓝氏贾第鞭毛虫(Giardia lamblia,贾第虫)的胞外核酸酶编码区,并对其蛋白产物进行活性鉴定.方法 对贾第虫胞外核酸酶(GeNuc)蛋白进行生物信息学分析,根据分析结果以C2株贾第虫基因组DNA为模板扩增获得GeNuc去信号肽段编码区序列,双酶切连入原核表达载体pET-28a(+),将酶切和测序验证正确的重组质粒转化E.coli Rosetta(DE3),经IPTG诱导表达融合蛋白,SDS-PAGE及Western blot鉴定蛋白产物.Ni-NTA亲和层析纯化GeNuc蛋白,经复性后验证其对质粒DNA的水解能力.结果 成功克隆了长约800 bp的GeNuc编码区并构建了原核表达载体pET-28a(+)-GeNuc,测序结果显示C2株GeNuc序列与WB株相同;在大肠杆菌中诱导表达获得了相对分子量约30.8 kDa的融合蛋白;复性后的纯化GeNuc蛋白具有降解双链DNA的能力,但活性较商品化DNase Ⅰ低.结论 证明了GeNuc的存在,为GeNuc抗体的制备及贾第虫致病机制的研究提供了实验材料.

  2. Dihydroartemisinin inhibition on Delta giardin in C2 Giardia lamblia%双氢青蒿素对C2株蓝氏贾第鞭毛虫Delta giardin抑制作用的研究

    Institute of Scientific and Technical Information of China (English)

    刘阿倩; 王洋; 林志强; 张亚粉; 田喜凤; 余源

    2015-01-01

    目的 建立实时荧光定量RT-PCR(real-time quantitative,RT-PCR)检测C2株蓝氏贾第鞭毛虫(Giardia lamblia)Delta giardin基因mRNA表达量的方法,分析双氢青蒿素(Dihydroartemisinin,DHA)对体外C2株蓝氏贾第鞭毛虫Delta giardin基因mRNA表达水平的影响.方法 分别采用100 μg/mL、200 μg/mL的双氢青蒿素改良型TYI-S-33培养基培养C2株蓝氏贾第鞭毛虫,以不含药物组作为阴性对照,分别培养2h、4h、8h、12h,提取总RNA,逆转录合成cDNA,实时荧光定量PCR检测Delta giardin基因mRNA表达情况.结果 100 μg/mL双氢青蒿素培养2h、4h、8h、12h后,Delta giardin基因mRNA相对表达量分别为0.44、0.26、0.25、0.02;200 μg/mL双氢青蒿素培养2h、4h、8h、12h后,Delta giardin基因mRNA相对表达量分别为0.30,0.26,0.11,0.02.药物对照组中C2株蓝氏贾第鞭毛虫Delta giardin基因mRNA表达量明显低于阴性对照组.结论 双氢青蒿素对C2株蓝氏贾第鞭毛虫Delta giardin基因mRNA的表达具有明显的抑制作用,提示双氢青蒿素对蓝氏贾第鞭毛虫具有明显的防治效果.

  3. Serum Ferritin: Past, Present and Future

    Science.gov (United States)

    Wang, Wei; Knovich, Mary Ann; Coffman, Lan G.; Torti, Frank M.; Torti, Suzy V.

    2010-01-01

    Background Serum ferritin was discovered in the 1930’s, and was developed as a clinical test in the 1970’s. Many diseases are associated with iron overload or iron deficiency. Serum ferritin is widely used in diagnosing and monitoring these diseases. Scope of Review In this chapter, we discuss the role of serum ferritin in physiological and pathological processes and its use as a clinical tool. Major Conclusions Although many aspects of the fundamental biology of serum ferritin remain surprisingly unclear, a growing number of roles have been attributed to extracellular ferritin, including newly described roles in iron delivery, angiogenesis, inflammation, immunity, signaling and cancer. General Significance Serum ferritin remains a clinically useful tool. Further studies on the biology of this protein may provide new biological insights. PMID:20304033

  4. Novel Utilization of Serum in Tissue Decellularization

    Science.gov (United States)

    Gui, Liqiong; Chan, Stephen A.; Breuer, Christopher K.

    2010-01-01

    Decellularization of native tissues is a promising technique with numerous applications in tissue engineering and regenerative medicine. However, there are various limitations of currently available decellularization methods, such as alteration of extracellular matrix mechanics and restricted use on certain tissues. This study was conducted to explore the effect of serum on the decellularization of various types of tissues. Fetal bovine serum–containing cell culture medium endothelial growth media-2 removed DNA but not cellular β-actin from human umbilical artery after detergent treatment, without compromising the tissue mechanical strength assessed by burst pressure. In addition, the effect of serum-containing endothelial growth media-2 on DNA removal was replicated in other types of tissues such as tissue-engineered vessels and myocardium. Other types of serum, including human serum, were also shown to remove DNA from detergent-pretreated tissues. In conclusion, we describe a novel utilization of serum that may have broad applications in tissue decellularization. PMID:19419244

  5. Effectiveness of autologous serum as an alternative to fetal bovine serum in adipose-derived stem cell engineering.

    Science.gov (United States)

    Choi, Jaehoon; Chung, Jee-Hyeok; Kwon, Geun-Yong; Kim, Ki-Wan; Kim, Sukwha; Chang, Hak

    2013-09-01

    In cell culture, medium supplemented with fetal bovine serum is commonly used, and it is widely known that fetal bovine serum supplies an adequate environment for culture and differentiation of stem cells. Nevertheless, the use of xenogeneic serum can cause several problems. We compared the effects of four different concentrations of autologous serum (1, 2, 5, and 10%) on expansion and adipogenic differentiation of adipose-derived stem cells using 10% fetal bovine serum as a control. The stem cells were grafted on nude mice and the in vivo differentiation capacity was evaluated. The isolation of adipose-derived stem cells was successful irrespective of the culture medium. The proliferation potential was statistically significant at passage 2, as follows: 10% autologous serum > 10% fetal bovine serum = 5% autologous serum > 2% autologous serum = 1% autologous serum. The differentiation capacity appeared statistically significant at passage 4, as follows: 10% fetal bovine serum > 10% autologous serum = 5% autologous serum > 2% autologous serum = 1% autologous serum. Ten percent autologous serum and 10% fetal bovine serum had greater differentiation capacity than 1 and 2% autologous serum in vivo, and no significant difference was observed between the groups at ≥ 5% concentration at 14 weeks. In conclusion, 10% autologous serum was at least as effective as 10% fetal bovine serum with respect to the number of adipose-derived stem cells at the end of both isolation and expansion, whereas 1 and 2% autologous serum was inferior.

  6. [Serum hyaluronic acid in osteoarthritis].

    Science.gov (United States)

    Balblanc, J C; Hartmann, D; Noyer, D; Mathieu, P; Conrozier, T; Tron, A M; Piperno, M; Richard, M; Vignon, E

    1993-03-01

    In this prospective study, serum hyaluronate (SH) was assayed using a radiometric method (Pharmacia) in 73 osteoarthritis patients and 39 controls. All assays were performed between 8 h 00 and 9 h 00 a.m. because SH levels exhibit circadian variations. SH levels were significantly higher in patients with osteoarthritis than in controls (92 +/- 66 micrograms/l and 39 +/- 21 micrograms/l, respectively, p = 0.0001). Among 50 patients with osteoarthritis, including 29 with knee involvement and 21 with hip involvement, SH levels were not correlated with morning stiffness, duration of symptoms, Lequesne's algofunctional index, erythrocyte sedimentation rate, C-reactive protein, severity of roentgenographic changes in the affected knee or hip, disease extension, or severity. The lack of any relationship between changes in SH levels and Lequesne's is index values in 25 patients or between SH levels and joint space narrowing evaluated retrospectively in 16 patients, as well as the prompt return to high SH levels after arthroplasty and synovectomy in 14 patients with hip joint osteoarthritis, suggest that this potential marker is not useful for monitoring osteoarthritis in a single joint.

  7. [Serum iron and serum copper balance in the early diagnosis of metastases of breast cancer].

    Science.gov (United States)

    Wöllgens, P; Kuhne-Velte, H J; Franke-Lompa, C

    1980-02-01

    In a study made in the follow-up clinic on 684 patients with mammary carcinoma it was found that there were 64 cases of local recurrence and 244 cases of distant metastases. In both groups the serum iron and the serum copper balance in blood tests, in relation to the clinical proof of local recurrence and/or distant metastases, was investigated. It was found that there were 43.7% pathological serum iron and serum copper findings with local recurrences and 62.7% with distant metastases. The drift apart tendency of the blood serum values in patients with distant metastases could be proved in 78.7% of the cases and in 77.5% of the cases before any clinical proof. Thus, these observations allow the statement that the blood serum iron and serum copper imbalance in blood tests is of very real value in the early diagnosis of distant metastases.

  8. PRELIMINARY STUDIES ON THE EFFECTS OF A CHINESE HERB-RADIX SOPHORAE FLAVESCENTIS ON TROPHOZOITES OF GIARDIA LAMBLIA IN VITRO%苦参对蓝氏贾第鞭毛虫作用及其机理初步研究

    Institute of Scientific and Technical Information of China (English)

    朱艳红; 牛安欧; 卢思奇

    2005-01-01

    目的研究苦参对蓝氏贾第鞭毛虫的作用及其机理. 方法用苦参作用蓝氏贾第鞭毛虫滋养体,观察滋养体脱壁及死亡情况,并用光镜和透射电镜观察其形态和内部结构的变化. 结果在作用 6、8和24 h后,滋养体脱壁率分别为47.4%、70.9%和80.2%,死亡率为 24.5%、36.1%和54.2%;形态观察可见,苦参作用的滋养体收缩,胞质内容物减少,吸盘部分脱离胞体,有的滋养体部分胞膜内陷. 结论苦参在体外能引起滋养体脱壁、死亡,其抗虫作用是肯定的,吸盘脱离、质膜凹陷及胞质减少等,这可能是其对贾第虫作用机理所在.%Objective To investigate the effect and mechanism of radix Sophorae flavescentis on the trophozoites of Giardia lamblia in vitro. Methods Trophozoites of G. lamblia were exposed to aqueous extracts of radix S. flavescentis and then the detachment from the tube wall, the death rate and the morphological changes under light and transmission electron microscope of trophozoites were studied. Results After exposure to 1.6% aqueous extracts of radix S. flavescentis for 6, 8 and 24 h, the detachment rates were 47.4%, 70.9% and 80.2%, and the death rates were 24.5%, 36.1% and 54.2%, respectively. The radix S. flavescentis-treated trophozoites shrinked and became roundish. There were also changes in ultrastructure, such as depletion of the contents of cytoplasm, separation of adhesive disk from the cell body and appearance of pit in cell membrane of some treated trophozoites. Conclusion Radix S. flavescentis can kill trophozoites of G. lamblia and lead them to detach from the tube wall in vitro. The induced morphological changes may be partially responsible for the effects of radix S. flavescentis on giardiasis.

  9. Construction of Rapid Tagging Vector of Genes in Giardia lamblia%蓝氏贾第鞭毛虫基因快速标记载体的构建

    Institute of Scientific and Technical Information of China (English)

    巨红妹; 王乙惠; 张霞; 王云华; 李雅杰

    2012-01-01

    Objective To construct a recombinant vector for rapid gene bagging in Giardia lamblia. Methods To obtain the recombinant vector pGL gdh-Neo with the Neo selection marker, the Neo gene was put under the control of gdh promoter by overlap PCR and inserted into pGEM-5zf. A DNA fragment containing multiple cloning sites (MCS) followed by triple hemagglutinin(3HA) coding sequences was synthesized and cloned into the pGL gdh-Neo to construct a recombinant vector pCL MCS-3HA-gdh-Neo. Giardia H2A gene was selected as a tagging gene to validate the effectivity of the recombinant vector pGL MCS-3HA-gdh-Neo. The histone H2A coding sequence was amplified by PCR, digested with EcoR Ⅰ and Spe Ⅰ, and inserted into MCS of pGL MCS-3HA-gdh-Neo. The resulting plasmid was then linearized and transfected into Giardia trophozoites. The H2A recombinant strain selected by G418 was analyzed by PCR,Western blotting and immunofluorescence. Results A rapid tagging recombinant vector with multiple cloning sites and triple hemagglutinin (3HA) was constructed with a length of 4 260 bp. The H2A recombinant vector was transfected into Giardia trophozoites and integrate into the Giardia genome at the correct locus. The HA-tagged H2A protein was expressed with a molecular weight (Mr) of 16 900. Conclusion A rapid tagging recombinant vector of genes in Giardia lamblia, pGL MCS-3HA-gdh-Neo, has been constructed.%目的 构建可快速标记蓝氏贾第鞭毛虫(简称贾第虫)基因的重组载体.方法 采用重叠PCR法将新霉素(Neo)基因置于gdh启动子调控下,并插入pGEM-5zf载体,获得带有Neo筛选标记的重组载体pGL gdh-Neo;并将基因合成所得3'末端带有3×HA标签的多克隆位点区克隆至重组载体pGLgdh-Neo,构建可快速标记贾第虫基因的重组载体pGL MCS-3HA-gdh-Neo.以该载体标记贾第虫H2A基因验证其可用性.PCR扩增组蛋白H2A基因序列,用EcoR Ⅰ和Spe Ⅰ双酶切后克隆至重组载体pGL MCS-3HA-gdh-Neo多克隆位点区

  10. Serum adipokine profiles in Kawasaki disease.

    Science.gov (United States)

    Kemmotsu, Yasushi; Saji, Tsutomu; Kusunoki, Natsuko; Tanaka, Nahoko; Nishimura, Chiaki; Ishiguro, Akira; Kawai, Shinichi

    2012-02-01

    Adipokines are cytokines derived from adipose tissue. Recently it has been established that adipokines are closely linked to the pathophysiology of not only metabolic diseases, such as diabetes mellitus, obesity, and atherosclerosis, but also to inflammation and immune diseases. In this study we measured serum levels of adipokines in patients with acute Kawasaki disease to investigate the role of adipokines in the pathophysiology of Kawasaki disease. Serum resistin, high-molecular-weight (HMW) adiponectin, leptin, and visfatin levels were measured by enzyme-linked immunosorbent assay in a total of 117 subjects: 56 patients with acute Kawasaki disease, 30 healthy children, and 31 patients with acute infectious diseases. Serum resistin levels in patients with Kawasaki disease were significantly higher than those of healthy children and patients with acute infectious diseases. In contrast, mean serum HMW adiponectin, leptin, and visfatin levels in patients with Kawasaki disease exhibited no statistically significant differences compared with those in healthy children and patients with infectious diseases. Serum resistin levels decreased significantly after administration of intravenous immune globulin. Serum resistin levels on admission were significantly higher in nonresponders compared with responders to intravenous immune globulin therapy. A multivariate model revealed that C-reactive protein was a factor that was significantly related to elevated serum resistin level in patients with Kawasaki disease. In patients with Kawasaki disease, serum resistin levels were elevated, but decreased to nearly normal after intravenous administration of immune globulin. In contrast, serum HMW adiponectin, leptin, and visfatin levels showed no statistically significant changes. These findings suggest that resistin plays an important role, while other adipokines do not play a major role, in the pathogenesis of Kawasaki disease.

  11. Modeling Serum Creatinine in Septic ICU Patients

    DEFF Research Database (Denmark)

    De Gaetano, Andrea; Cortese, Giuliana; Pedersen, Morten Gram

    2004-01-01

    Serum creatinine is a metabolite assumed to be constantly produced by the normally functioning muscle mass and is a good measure for monitoring daily renal function in the intensive care unit (ICU). High serum creatinine levels or an abnormal departure from normal pre-disease basal levels....... The present work details the structure of a model describing observed creatinine serum concentration (CSC) variations, depending on the time-varying septic insult to renal function in ICU patients, as well as the estimation of its parameters. CSC determinations were routinely obtained from 12 patients...

  12. Serum gold concentrations during treatment with auranofin.

    Science.gov (United States)

    Van Riel, P L; Gribnau, F W; Van de Putte, L B; Arts, C W; Van Aernsbergen, A

    1987-03-01

    Serum gold concentrations were measured in rheumatoid arthritis patients during chronic treatment with the orally adsorbable gold compound auranofin. In agreement with data in the literature, the highest serum gold concentration was reached after 16 weeks of treatment with 6 mg auranofin daily. A striking finding in this study was that thereafter the serum gold concentrations did not appear to plateau but declined gradually. Statistically this resulted in a significantly lower concentration after one year as compared with week 16 (p less than 0.05, paired t-test). It is suggested that a shift from protein bound gold to cell-bound gold might be the explanation.

  13. Serum Lipids and Lipoproteins Levels and Selected Trace Metals In ...

    African Journals Online (AJOL)

    Serum Lipids and Lipoproteins Levels and Selected Trace Metals In Newly ... This study aim to determine the serum levels of trace metals and correlate same with serum levels of lipoproteins (an established marker of HBP) in ... Article Metrics.

  14. Serum obestatin and omentin levels in patients with diabetic ...

    African Journals Online (AJOL)

    2016-04-08

    Apr 8, 2016 ... Results: While serum omentin levels did not differ among two groups (P = 0.407), serum obestatin levels were .... creatine kinase (CK), serum iron, total iron binding capacity (TIBC), ferritin, complete blood count (CBC),.

  15. Radioimmunoassay of secretin in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Bonora, G.; Vezzadini, P.; Toni, R.; Labo, G. (Bologna Univ. (Italy))

    1984-06-27

    A sensitive radioimmunoassay for secretin has been developed. Antisera were raised against synthetic porcine secretin coupled to bovine serum albumin. N-..cap alpha..-desaminotyrosyl-..beta..-alanyl secretin was radioiodinated by a slight modification of the chloramine-T method. Pure synthetic porcine secretin was used as a standard. Free and bound hormone were separated by dextran-coated charcoal. No cross-reactivity was found with structurally and physiologically related peptides. The sensitivity of the assay was high enough to measure fasting secretin levels in human serum. Patients with acute or chronic pancreatitis had mean serum secretin concentration not significantly different from healthy subjects. In patients with pancreatic carcinoma the mean serum secretin concentration was significantly lower than in healthy subjects, although a wide overlap of the two groups was evident.

  16. Serum Prolactin in Diagnosis of Epileptic Seizures

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2005-09-01

    Full Text Available The results of studies in databases and references concerning serum prolactin levels (PRL in patients with suspected seizures were rated for quality and analyzed by members of the Therapeutics Subcommittee of the American Academy of Neurology.

  17. Ketoprofen analysis in serum by capillary electrophoresis.

    Science.gov (United States)

    Friedberg, M; Shihabi, Z K

    1997-07-18

    A method for the quantification of ketoprofen, a new non-prescription non-steroidal anti-inflammatory drug, in serum, by capillary zone electrophoresis for therapeutic monitoring and emergency toxicology is described. Serum is deproteinized with acetonitrile in the presence of an internal standard, to remove serum proteins and to induce sample stacking. The migration time was about 10 min. The assay was linear between 1-10 mg/l without any interferences. The method compared well to an HPLC assay. The HPLC afforded a better detection limit, but the CE was less expensive to operate. This method demonstrates that capillary electrophoresis is a simple and effective method for determination of ketoprofen as well as other drugs in human serum at levels close to 1 mg/l.

  18. Autologous serum therapy in chronic urticaria

    Directory of Open Access Journals (Sweden)

    Sharmila Patil

    2013-01-01

    Full Text Available Autologous serum therapy is a promising therapy for treatment resistant urticaria. This is useful in developing countries as this is economical option. Minimum instruments like centrifuge, syringe and needles are required for the procedure.

  19. Assessing possible hazards of reducing serum cholesterol.

    OpenAIRE

    Law, M. R.; Thompson, S. G.; Wald, N J

    1994-01-01

    OBJECTIVE--To assess whether low serum cholesterol concentration increases mortality from any cause. DESIGN--Systematic review of published data on mortality from causes other than ischaemic heart disease derived from the 10 largest cohort studies, two international studies, and 28 randomised trials, supplemented by unpublished data on causes of death obtained when necessary. MAIN OUTCOME MEASURES--Excess cause specific mortality associated with low or lowered serum cholesterol concentration....

  20. Revisiting the cardiometabolic relevance of serum amylase

    OpenAIRE

    Munakata Hiromi; Muneyuki Toshitaka; Nakajima Kei; Kakei Masafumi

    2011-01-01

    Abstract Background The pancreas has dual functions as a digestive organ and as an endocrine organ, by secreting digestive enzymes and endocrine hormones. Some early studies have revealed that serum amylase levels are lower in individuals with chronic pancreatitis, severe long-term type 2 diabetes or type 1 diabetes. Regarding this issue, we recently reported that low serum amylase levels were associated with metabolic syndrome and diabetes in asymptomatic adults. In the light of this, we fur...

  1. Revisiting the cardiometabolic relevance of serum amylase

    Directory of Open Access Journals (Sweden)

    Munakata Hiromi

    2011-10-01

    Full Text Available Abstract Background The pancreas has dual functions as a digestive organ and as an endocrine organ, by secreting digestive enzymes and endocrine hormones. Some early studies have revealed that serum amylase levels are lower in individuals with chronic pancreatitis, severe long-term type 2 diabetes or type 1 diabetes. Regarding this issue, we recently reported that low serum amylase levels were associated with metabolic syndrome and diabetes in asymptomatic adults. In the light of this, we further investigated the fundamental relationship between serum amylase and cardiometabolic aspects by reanalyzing previous data which comprised subjects without diabetes treatment with oral hypoglycemic drugs or insulin (n = 2,344. Findings Serum amylase was inversely correlated with body mass index independently of age. Higher serum amylase levels were noted in older subjects aged 55 years old or more (n = 1,114 than in younger subjects (P P Conclusions Revisiting the cardiometabolic relevance of serum amylase may yield novel insight not only into glucose homeostasis and metabolic abnormalities related to obesity, but also possibly carbohydrate absorption in the gut.

  2. Serum angiotensin converting enzyme in pemphigus vulgaris

    Directory of Open Access Journals (Sweden)

    Reza M Robati

    2014-01-01

    Full Text Available Background: Pemphigus vulgaris is an autoimmune blistering skin disease with unknown etiology. Drugs such as angiotensin-converting enzyme (ACE inhibitors may contribute in the pathogenesis of pemphigus. Objective: We plan this essay to evaluate the serum ACE level in pemphigus vulgaris patients in comparison with healthy controls to recognize its possible role in disease pathogenesis or activity. Methods: This study was planned and performed in the dermatology clinics of Shahid Beheshti University of MedicalSciences′ Hospitals between July 2010 and June 2011. Patients with new onset of pemphigus vulgaris were enrolled in our study. Control subjects were frequency-matched to cases by sex and age. Serum ACE was determined by the spectrophotometric method. Results: Thirty-four patients with pemphigus vulgaris and 35 healthy individuals were recruited in the study. No statistical significant difference was detected in the mean level of serum ACE of the two groups (t-test, P = 0.11. The mean ACE level was significantly lower in male patients compared with male controls (P = 0.04. Moreover, a significant higher serum ACE level of patients with cutaneous involvement was observed compared to patients with mucosal involvement (P = 0.02. Conclusions: Despite lack of any significant difference of serum ACE level between pemphigus and control group, the serum ACE level was considerably lower in male pemphigus vulgaris patients compared with male controls. Therefore, ACE might have some association with pemphigus vulgaris especially in male patients; however, further studies are required to confirm this association.

  3. Diarrea crónica refractaria y malabsorción secundaria a hipogammaglobulinemia común variable, infestación crónica por giardia lamblia y gastrectomía total por adenocarcinoma gástrico: un manejo nutricional complejo Chonic diarrhea and malabsorption due to common variable immunodeficiency, gastrectomy and giardiasis infection: a difficult nutritional management

    Directory of Open Access Journals (Sweden)

    M. E. Domínguez-López

    2011-08-01

    Full Text Available El adenocarcinoma gástrico es una de las causas más frecuentes de mortalidad en el mundo, siendo la cirugía el único tratamiento potencialmente curativo, aunque los efectos adversos digestivos y nutricionales son frecuentes y abundantes. La hipogammaglobulinemia variable común es causa de frecuentes manifestaciones digestivas, derivándose las más importantes en diarrea crónica causada por giardiasis, hiperplasia nodular linfoide o atrofia vellosa, siendo frecuente la malabsorción y la desnutrición. Los déficits nutricionales secundarios a la malabsorción (postgastrectomía y asociada a la atrofia vellosa y la giardiasis por hipogammaglobulinemia variable común son asimismo frecuentes. Presentamos el caso de un paciente gastrectomizado por adenocarcinoma gástrico y con hipogammaglobulinemia variable común e infestación crónica por giardiasis que presenta una importante diarrea crónica refractaria a tratamiento y malabsorción.Gastric cancer is a frequent cause of cancer-related mortality in the world. Surgery is the only potentially curative therapy, although the adverse effects of surgery are common and considerable. Common variable immunodeficiency is in many cases cause of gastrointestinal system problems such as chronic diarrhea caused by infestation with giardia lamblia, nodular lymphoid hiperplasia ad loss of villi leading frequently to malapsortion and malnutrition. Nutritional deficiencies due to malapsorption (postgastrectomy and secondary to loss of villi, giardiasis and common variable inmunodeficiency are common. We present the case of a patient with gastric cancer who underwent a gastrectomy with common variable hipogammaglobulinemia and chronic infestation by giardia lamblia, with serious diarrhea resistant to treatment and malabsorption.

  4. Serum lactic dehydrogenase isoenzymes and serum hydroxy butyric dehydrogenase in myocardial infarction

    Directory of Open Access Journals (Sweden)

    Kanekar D

    1979-01-01

    Full Text Available Total serum lactate dehydrogenase activity in cases of myocar-dial infarct is difficult to interpret as abnormal values can occur in diseases of liver, kidney and skeletal muscle. The estimation of its isoenzymes is of better diagnostic help because of its tissue specificity. Serum LDH isoenzymes were studied in patients o f myocardial infarction and results are quantitated by densitometry. As LDH 1 represents serum hydroxybutyric dehydrogenase when 2-oxylbutyrate is used as substrate, serum hydroxybutyric dehydro-genase was also estimated in above patients. Greater specificity in diagnosis is achieved with SHBDH because of its myocardial nature and lower incidence of false positive results.

  5. Treatment of serum with supernatants from cultures of Candida albicans reduces its serum-dependent phagocytosis

    Directory of Open Access Journals (Sweden)

    Santos Aderbal Antonio dos

    2002-01-01

    Full Text Available Candida albicans is a potent activator of the complement system, and heat labile opsonins produced by activation of C3 (C3b and iC3b enhance phagocytosis of C. albicans mediated by complement receptors. In this study we treated mouse serum with supernatants from cultures of a protease producer strain of C. albicans and evaluated the ability of this serum to enhance phagocytosis of C. albicans. Cell-free supernatants from cultures of C. albicans were concentrated 5 fold and added to mouse serum for 30 min at 37ºC, before using this serum for opsonization of glutaraldehyde-fixed yeast cells. We observed that normal mouse serum increased about 3 fold the phagocytosis of C. albicans by mice peritoneal macrophages, whereas supernatant-treated serum did not increase phagocytosis. This effect of supernatants on serum was prevented by addition of pepstatin (5 µg/ ml; an inhibitor of C. albicans acid proteases to the medium. Serum treated with supernatants from cultures of a protease-deficient mutant of C. albicans also increased about 3 fold phagocytosis of the yeast. These results suggest that a protease produced by C. albicans causes proteolysis of serum opsonins, thereby reducing the phagocytosis of the yeast.

  6. Effects of dihydroartemisinin on protein of Giardia lamblia trophozoites%双氢青蒿素对蓝氏贾第鞭毛虫滋养体蛋白质的损伤

    Institute of Scientific and Technical Information of China (English)

    余源; 陈阳; 王卫亮; 杨志宏; 田玉娜; 王玥; 田喜凤; 卢思奇; 赵永强

    2010-01-01

    目的 探讨双氢青蒿素 (dihydroartemisinin,DHA) 对体外培养的蓝氏贾第鞭毛虫(Giardia lamblia)滋养体蛋白质的损伤作用.方法 用改良TYI-S-33培养基培养蓝氏贾第鞭毛虫滋养体;设立实验组(DHA组)和正常对照组(C组).实验组用含双氢青蒿素的培养基(含LC50=200μg/mL的DHA)培养;对照组培养基不含药物.提取各组虫体总蛋白,经Bradford法进行蛋白质定量,样本经2D-电泳和硝酸银染色后, 扫描检测凝胶电泳图蛋白点的数量和种类,获取蛋白质点匹配信息,挑选匹配性高的蛋白点,进行质谱(MALDI-TOF MS)分析.结果 经双氢青蒿素作用后虫体总蛋白质含量(0.6993μg/mL)明显低于对照组(4.8241μg/mL),二者差异有显著性(P <0.001).蛋白质二维电泳凝胶图分析结果显示,正常对照组(C组)蛋白点密集,约1253个点,布满整块凝胶,蛋白表达量高(蛋白点颜色深且清晰、明亮);与此相反,实验组(DHA组)蛋白点显著减少,DHA作用6h后约617个点,作用12h后约326个点,蛋白表达量明显降低(蛋白点颜色浅,不清晰).其次,正常对照组与双氢青蒿素组的12kDa~30kDa、等电点pI 3.0~7.6范围内蛋白点表达量存在明显差异,即实验组该范围内大多数蛋白点完全消失, 45kD~60kDa的蛋白点仅剩6个点.质谱分析鉴定结果显示,在正常对照组胶图所选的80个匹配良好的蛋白点中,鉴定出6种骨架蛋白(14个点),而实验组(DHA组)蛋白胶图中相对应的蛋白点消失.结论 双氢青蒿素对蓝氏贾第鞭毛虫滋养体的蛋白质具有明显的损伤作用,其中包括细胞骨架蛋白.

  7. Fluorimetric determination of cholesterol in hypercholesterolemia serum

    Science.gov (United States)

    Lan, Xiufeng; Liu, Jiangang; Liu, Ying; Luo, Xiaosen; Lu, Jian; Ni, Xiaowu

    2005-01-01

    With the increase of people"s living standard and the changes of living form, the number of people who suffer from hypercholesterolemia is increasing. It is not only harmful to heart and blood vessel, but also leading to obstruction of cognition. The conventional blood detection technology has weakness such as complex operation, long detecting period, and bad visibility. In order to develop a new detection method that can checkout hypercholesterolemia conveniently, spectroscopy of cholesterol in hypercholesterolemia serum is obtained by the multifunctional grating spectrograph. The experiment results indicate that, under the excitation of light-emitting diode (LED) with the wavelength at 407 nm, the serum from normal human and the hypercholesterolemia serum emit different fluorescence spectra. The former can emit one fluorescence region with the peak locating at 516 nm while the latter can emit two more regions with peaks locating at 560 nm and 588 nm. Moreover, the fluorescence intensity of serum is non-linear increasing with the concentration of cholesterol increases when the concentration of cholesterol is lower than 13.8 mmol/L, and then, with the concentration of cholesterol increase, the fluorescence intensity decreases. However, the fluorescence intensity is still much higher than that of serum from normal human. Conclusions can be educed from the experiments: the intensity and the shape of fluorescence spectra of hypercholesterolemia serum are different of those of normal serum, from which the cholesterol abnormal in blood can be judged. The consequences in this paper may offer an experimental reference for the diagnosis of the hypercholesterolemia.

  8. Soluble serum Klotho levels in healthy subjects

    DEFF Research Database (Denmark)

    Pedersen, Lise Mariager; Pedersen, Susanne Møller; Brasen, Claus Lohman

    2013-01-01

    OBJECTIVE: Soluble serum Klotho, is a new biomarker linked to chronic kidney disease, cardiovascular disease and diabetes. This study describes the evaluation and comparison of two different immunoassays and establishment of assay specific reference intervals in adults. Design and methods Serum...... of serum Klotho. RESULTS: Serum Klotho levels were significantly higher in time-resolved fluorescence immunoassay (TRF) compared to an ELISA (IBL) and no correlation were found between the assays. No signal was obtained in either assay when the standard curve was switched between the two different...... immunoassays. The median serum Klotho concentration using TRF was 61ng/mL (2.5-97.5% reference limits; 11-181ng/mL) for males and 99ng/mL (2.5-97.5% reference limits; 19-316ng/mL) for females while the ELISA gave a mean value of 472pg/mL (2.5-97.5% reference limits; 204-741pg/mL) with no difference between...

  9. Peptidomic Identification of Serum Peptides Diagnosing Preeclampsia.

    Directory of Open Access Journals (Sweden)

    Qiaojun Wen

    Full Text Available We sought to identify serological markers capable of diagnosing preeclampsia (PE. We performed serum peptide analysis (liquid chromatography mass spectrometry of 62 unique samples from 31 PE patients and 31 healthy pregnant controls, with two-thirds used as a training set and the other third as a testing set. Differential serum peptide profiling identified 52 significant serum peptides, and a 19-peptide panel collectively discriminating PE in training sets (n = 21 PE, n = 21 control; specificity = 85.7% and sensitivity = 100% and testing sets (n = 10 PE, n = 10 control; specificity = 80% and sensitivity = 100%. The panel peptides were derived from 6 different protein precursors: 13 from fibrinogen alpha (FGA, 1 from alpha-1-antitrypsin (A1AT, 1 from apolipoprotein L1 (APO-L1, 1 from inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4, 2 from kininogen-1 (KNG1, and 1 from thymosin beta-4 (TMSB4. We concluded that serum peptides can accurately discriminate active PE. Measurement of a 19-peptide panel could be performed quickly and in a quantitative mass spectrometric platform available in clinical laboratories. This serum peptide panel quantification could provide clinical utility in predicting PE or differential diagnosis of PE from confounding chronic hypertension.

  10. Proteomic evaluation of sheep serum proteins

    Directory of Open Access Journals (Sweden)

    Chiaradia Elisabetta

    2012-05-01

    Full Text Available Abstract Background The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference map of sheep serum. The possible application of this approach was tested by analysing serum protein patterns in ewes with mild broncho-pulmonary disease, which is very common in sheep and in the peripartum period which is a stressful time, with a high incidence of infectious and parasitic diseases. Results This study generated the first reference 2-DE maps of sheep serum. Overall, 250 protein spots were analyzed, and 138 identified. Compared with healthy sheep, serum protein profiles of animals with rhino-tracheo-bronchitis showed a significant decrease in protein spots identified as transthyretin, apolipoprotein A1 and a significant increase in spots identified as haptoglobin, endopin 1b and alpha1B glycoprotein. In the peripartum period, haptoglobin, alpha-1-acid glycoprotein, apolipoprotein A1 levels rose, while transthyretin content dropped. Conclusions This study describes applications of proteomics in putative biomarker discovery for early diagnosis as well as for monitoring the physiological and metabolic situations critical for ovine welfare.

  11. Serum tryptase levels in adverse drug reactions.

    Science.gov (United States)

    Ordoqui, E; Zubeldia, J M; Aranzábal, A; Rubio, M; Herrero, T; Tornero, P; Rodríguez, V M; Prieto, A; Baeza, M L

    1997-11-01

    We evaluated the usefulness of individual tryptase levels and variations after adverse drug reactions in 64 patients. Our aim was to find a tool for the diagnosis of drug allergy. Thirty-seven subjects were confirmed to have drug allergy, 12 had nonsteroidal anti-inflammatory drug (NSAID) reactions, five had negative controlled drug challenges (NAAR), and 10 had symptoms after placebo intake (PLA). Serum tryptase levels greatly increased after anaphylactic shocks (2242%) and anaphylaxis (710.5%). Patients with allergic urticaria and those with idiosyncratic responses to acetylsalicylic acid (ASA) exhibited a small increase in serum tryptase (49.5% and 38.2%, respectively). In the other two groups (NAAR and PLA), no variation in this serum protease was observed. The time of appearance of the serum tryptase peak differed considerably among patients with similar clinical reactions (from 30 min to 6 h) and was independent of the latent period, severity of symptoms, or the amount of tryptase released. We conclude that serum tryptase determinations are helpful in the diagnosis of anaphylactic shock and anaphylaxis, but serial measurements may be needed to confirm mast-cell participation in milder reactions.

  12. Serum biochemical markers in carcinoma breast.

    Directory of Open Access Journals (Sweden)

    Seth R

    2003-08-01

    Full Text Available BACKGROUND: Despite the extensive research for many years throughout the world, the etiopathogenesis of cancer still remains obscure. For the early detection of carcinoma of various origins, a number of biochemical markers have been studied to evaluate the malignancy. AIM: To analyse serum gamma glutamyl transpeptidase (GGTP, lactate dehydrogenase (LDH and superoxide dismutase (SOD in carcinoma breast patients. SETTINGS & DESIGN: The serum biochemical markers were estimated in twenty five histopathologically confirmed patients with carcinoma breast and equal number of healthy age- matched individuals served as control. MATERIAL & METHODS: Serum gamma glutamyl transpeptidase (GGTP, lactate dehydrogenase (LDH and superoxide dismutase (SOD were estimated and their sensitivity determined. Statistics: Data was analysed with student′s ′t′-test and sensitivity score of these markers was determined. RESULTS & CONCLUSIONS: The mean serum GGTP, LDH and SOD activities in patients with carcinoma breast were tremendously increased as compared to controls, and a steady increase was observed in their activities from stage I through stage IV as well as following distant metastasis. Serum GGTP, LDH and SOD might prove to be most sensitive biomarkers in carcinoma breast in early detection of the disease.

  13. HIV and other predictors of serum folate, serum ferritin, and hemoglobin in pregnancy

    DEFF Research Database (Denmark)

    Friis, Henrik; Gomo, E; Kæstel, Pernille;

    2001-01-01

    BACKGROUND: Folate and iron status and hemoglobin concentrations are important to maternal and infant health. OBJECTIVE: Our goal was to identify predictors of serum folate, serum ferritin, and hemoglobin. DESIGN: This was a cross-sectional study of 1669 pregnant women (22-35 wk of gestation) in ...

  14. ALPHA-FETOPROTEIN IN FETAL SERUM, AMNIOTIC-FLUID, AND MATERNAL SERUM

    NARCIS (Netherlands)

    VANLITH, JMM; BEEKHUIS, [No Value; VANLOON, AJ; MANTINGH, A; DEWOLF, BTHM; BREED, ASPM

    1991-01-01

    In order to gain more insight into the association between alpha-fetoprotein (AFP) and fetal chromosomal disorders, especially Down's syndrome, we measured AFP in fetal serum, amniotic fluid, and maternal serum at cordocentesis. We compared the concentration and gradient of AFP in these three compar

  15. Serum bleomycin-detectable iron in patients with thalassemia major with normal range of serum iron.

    Directory of Open Access Journals (Sweden)

    Han,Khin Ei

    1995-06-01

    Full Text Available "Free" iron, a potentially radical-generating low mass iron, and not found in normal human blood, was increased in the serum of blood-transfused thalassemia major patients seen in the Yangon General Hospital, Yangon, Myanmar (Burma. The low mass iron was detected by the bleomycin assay. Fifty-one blood samples were analyzed (from 28 males and 23 females. High "free" iron was detected in 47 sera samples from thalassemia patients. Serum ferritin, which reflects the body store iron, was higher than the normal range (10-200 ng/ml in 49 patients. On the other hand, serum iron of 39 sera samples fell within the normal range (50-150 micrograms/dl. Four were less than 50 micrograms/dl and eight were more than 150 micrograms/dl. Almost all the patients' sera of normal or higher serum iron level contained "free" iron. Thus, almost all the sera from thalassemic patients from Myanmar contain bleomycin-detectable iron, even when serum iron is within the normal range. In developing countries where undernutrition is prevalent (serum albumin in these patients was 3.6 +/- 0.4 g/dl, P < 0.0001 vs. control value of 4.0 - 4.8 g/dl, normal serum iron does not preclude the presence of free iron in the serum.

  16. Serum cardiac troponin I in acute stroke is related to serum cortisol and TNF-alpha

    DEFF Research Database (Denmark)

    Christensen, Hanne Krarup; Johannesen, Helle Hjorth; Christensen, Anders Fogh

    2004-01-01

    Serum cardiac troponin I (cTnI) is a specific marker of myocardial injury related to in-patient fatality and cardiac injury in acute stroke. We investigated whether cTnI in acute stroke is related to serum cortisol, acute inflammatory response, and insular damage. We also investigated whether c...

  17. Prevalence of Entamoeba histolytica, Giardia lamblia, and ...

    African Journals Online (AJOL)

    2016-06-29

    Jun 29, 2016 ... Egyptian food handlers but not among the Libyan food handlers. .... Reassessment of the epidemiology of amebiasis: state of the art. Infect Genet Evol. ... Cambridge, United Kingdom: The Royal Society of Chemistry;. 2008. p.

  18. Association of serum leptin with serum C-reactive protein in hemodialysis patients

    Directory of Open Access Journals (Sweden)

    Rastegari Ebrahim

    2012-04-01

    Full Text Available Introduction: Recent investigations have shown that leptin is cleared principally by the kidney. Objectives: To examine whether and how in patients on hemodialysis the level of C-reactive protein level correlate with serum leptin. Patients and Methods: The total patients were 36. The mean patients’ age were 46 (16 years. The median length of the time patients were on hemodialysis were 19 months. Results: The mean serum C-reactive protein was 8.7 (6.6 mg/l (median: 8 mg/l. The mean serum leptin was 9.4 (14 ng/ml (median: 5.75 ng/ml. In this study we found a significant inverse correlation of serum leptin with serum C-reactive protein (r= -0.57, p= 0.041 was seen. Conclusion: Our data supports, the positive effect of leptin on nutrition and support the theory of protective effects (reverse epidemiology of leptin in hemodialysis patients.

  19. Correlation between Serum T3 Resin Uptake and Serum Lipid Profile

    Directory of Open Access Journals (Sweden)

    Dinarvand, GH. ( MSc

    2015-05-01

    Full Text Available Background and Objective: T3 resin uptake test indicates the availability of thyroxin binding globulin (TBG, which is the carrier of a large amount of T3 and T4 in blood. We aimed to study the relationship between serum T3 Resin Uptake and serum lipids profile. Material and Methods: The participants aged 14.80 ± 38.29 were both males (59; 29.5% and females (141; 70.5 %. T3 Resin Uptake was measured by Immunoassay method and serum lipid profile components (cholesterol, triglycerides, HDL-C, LDL-C, and VLDL were measured by the kit. Results: Significant negative correlation was observed between serum T3 Resin Uptake and cholesterol, triglycerides and LDL-C (P≤ 0.05, but that was not the case for HDL-C and VLDL. Conclusion: Serum T3 Resin Uptake can be evaluated to assess the risk factors related to cardiovascular disorders

  20. Serum sialic acid and CEA concentrations in human breast cancer.

    OpenAIRE

    Hogan-Ryan, A.; Fennelly, J J; Jones, M.; Cantwell, B; Duffy, M J

    1980-01-01

    The concentration of bound sialic acid in the sera of 56 normal subjects and 65 subjects with breast cancer was measured, in order to determine (1) whether serum sialic acid concentrations are raised in breast cancer and (2) whether the concentration of sialic acid in serum reflects tumour stage. The amount of sialic acid in serum was compared to serum carcinoembryonic antigen (CEA) values. Urinary hydroxyproline and serum alkaline phosphatase concentrations were used as indicators of bone an...

  1. Serum fucose level in malignant diseases.

    Science.gov (United States)

    Sawke, N G; Sawke, G K

    2010-01-01

    We review the present knowledge of serum fucose with special attention to its relation with various malignant diseases. We summarize the role of serum fucose as a useful diagnostic and prognostic marker when used singly or in combination. The purpose of this review is to provide an expert opinion on the practical and applied aspect of serum fucose level in clinical practice and research settings. Our review is based on information from published research studies, library books, and electronic searches through Medline and PubMed. The available published data were used as the basis for recommendations. Each of the subsections concludes to provide information to assist the clinicians and the research scientists make informed decisions.

  2. How does whisky lower serum urate level?

    Science.gov (United States)

    Lu, Yang; Nakanishi, Takeo; Fukazawa, Miki; Tamai, Ikumi

    2014-05-01

    Clinical studies have shown that moderate whisky consumption increased renal excretion of urate into urine and decreased serum urate level, but the mechanism involved has not been established. Because renal reabsorption influences serum urate level, the effects of the whisky congeners on urate transporters, urate transporter 1 (URAT1), and voltage-driven urate transporter (URATv1) involved in reabsorptive transport of urate were examined. In transporter-expressing Xenopus oocytes, 12-year-old and 18-year-old whisky congeners inhibited urate uptake by URAT1 with IC50 values of 0.08 ± 0.01 and 0.04 ± 0.01 mg/mL, respectively, while urate uptake by URATv1 was inhibited only at 1 mg/mL. Decreased serum urate level after whisky consumption may be mainly due to inhibition of URAT1 by the congeners.

  3. Serum Calcium Concentration in Ethylene Glycol Poisoning.

    Science.gov (United States)

    Hodgman, Michael; Marraffa, Jeanna M; Wojcik, Susan; Grant, William

    2017-06-01

    The diagnosis of ethylene glycol intoxication can be challenging. Definitive testing for ethylene glycol is not readily available and clinical decisions are often based on clinical suspicion and the results of more readily available tests. One of these findings is hypocalcemia, presumable through complexation with the ethylene glycol metabolite oxalate. We performed a retrospective review of all patients admitted to a tertiary care hospital between 2005 and 2013 with laboratory confirmed ethylene glycol intoxication. Serum calcium on presentation was compared to blood gas pH on presentation as well as presentation serum bicarbonate. We did not find any relationship between calcium and serum pH either by linear regression or when dichotomized by pH ≥ or ethylene glycol poisoning, even in acidotic patients.

  4. Serum cholesterol in healthy postmenopausal women.

    Science.gov (United States)

    Samanta, B B

    1998-05-01

    Hypercholes erolaemia is a modifiable risk factor in atherosclerosis. Women lose their relative protection against coronory heart disease at menopause because of changed lipid profile due to oestrogen deficiency. Total serum cholesterol was estimated in 82 healthy postmenopausal women in the age group of 46-72 years (51.5 +/- 7.39). Thirty five healthy pre-menopausal women in the age group of 18-38 years (29.5 +/- 6.4) served as controls. The mean serum cholesterol concentration was significantly higher in the postmenopausal group compared to control group (178.5 +/- 39.8 Vs 155.4 +/- 24 mg/dl; P < 0.01). Serum cholesterol concentration in the study group was not related to social class, dietary habit and obesity.

  5. Imidazole binding to human serum albumin.

    Science.gov (United States)

    Rodrigo, M C; Ceballos, A; Mariño, E; Cachaza, J M; Domínguez-Gil, A; Kuemmerle, H P

    1988-06-01

    Imidazole is a substance released by the organism when a new salicylate derivative, imidazole salicylate is administered. A study was made of the binding of imidazole to human serum albumin by an in vitro assay employing an ultrafiltration technique. For the concentration range that imidazole was found in plasma following administration of the drug to healthy volunteers, the mean binding percentages were: 12.1 +/- 1.8 and 19.7 +/- 3.1 at 37 degrees C and 25 degrees C, respectively. The results obtained in the study follow a model entailing three equal and independent binding sites of imidazole to serum albumin and the values of the corresponding constants were determined. Apparently, the presence in the plasma samples of sodium salicylate at a concentration of 100 micrograms/ml does not affect the binding of imidazole to human serum albumin.

  6. Low serum zinc level in dpression

    Directory of Open Access Journals (Sweden)

    SA Mousavi

    2006-07-01

    Full Text Available BACKGROUND: Major Depressive Disorder (MDD is a common disorder, with a lifetime prevalence of about 15 percent, perhaps as high as 25 percent for women. The etiology of MDD is too complex to be explained totally by a single social, developmental, or biological theory. A variety of factors appear to work together to cause or precipitate depressive disorders. Various functions have been reported for trace elements such as zink in recovery or exacerbation of depression. METHODS: In this experimental study, we studied 46 patients with MDD based on DSM IV criteria, among the patients referred to mental disorders clininc of Noor Hospital. Twenty Patients were men and 26 were women. Thirty two volunteers of general population were evaluated for depression with Beck depression test who did not show any depressive symptoms with this test. A blood sample of 5cc was obtained from each person and the serum zinc concentration was measured. Data gathered and analyzed with SPSS, logistic regression and chi-squar tests. RESULTS: Serum zinc concentrations were 74 to 130 mg/dl in men and 60 to 128 mg/dl in women of control group. Serum zinc concentration was 30 to 60 mg/dl in depressive patients that it was lower in women than men. The difference between serum zinc concentrations of normal and depressive persons was meaningful (P = 0.02. CONCLUSION: In our study, the serum concentration of zinc was about half of normal value. This study replicates previous findings that major depressed subjects show significantly lowered serum zinc concentration. KEYWORDS: Depression, zinc.

  7. Serum adenosine deaminase activity in cutaneous anthrax.

    Science.gov (United States)

    Sunnetcioglu, Mahmut; Karadas, Sevdegul; Aslan, Mehmet; Ceylan, Mehmet Resat; Demir, Halit; Oncu, Mehmet Resit; Karahocagil, Mustafa Kasım; Sunnetcioglu, Aysel; Aypak, Cenk

    2014-07-06

    Adenosine deaminase (ADA) activity has been discovered in several inflammatory conditions; however, there are no data associated with cutaneous anthrax. The aim of this study was to investigate serum ADA activity in patients with cutaneous anthrax. Sixteen patients with cutaneous anthrax and 17 healthy controls were enrolled. We measured ADA activity; peripheral blood leukocyte, lymphocyte, neutrophil, and monocyte counts; erythrocyte sedimentation rate; and C reactive protein levels. Serum ADA activity was significantly higher in patients with cutaneous anthrax than in the controls (panthrax.

  8. I. Unbound serum gold: procedure for quantitation.

    Science.gov (United States)

    Lorber, A; Vibert, G J; Harralson, A F; Simon, T M

    1983-08-01

    The unbound fraction of many drugs appears to be the therapeutically active component. However, the major problem encountered in following unbound serum gold (UBSG) concentration during chrysotherapy has been the ability to quantitate such a small quantity of gold reliably without matrix interference. The methodology detailed here overcomes these difficulties and provides an effective means of monitoring the UBSG fraction during chrysotherapy. We have observed that the unbound fraction of gold dissipates quickly after gold sodium thiomalate administration and constitutes less than 2% of the total serum gold concentration.

  9. Accurate determination of serum ASAT isoenzymes.

    Science.gov (United States)

    Konttinen, A; Ojala, K

    1978-01-01

    An improved electrophoretic modification for measuring aspartate aminotransferase (ASAT) isoenzymes is presented. This method fulfils the clinical requirements for sensitivity and allows the detection of 1 U/l mitochondria ASAT activity at 25 degree C. The procedure is relatively simple, requiring about one hour for a series of 8 determinations. Mitochondrial ASAT activity was found in all patients suffering from acute myocardial infarction pathological activity was observed for several days longer than that of total serum ASAT enzyme. None of the 25 healthy people studied had mitochondrial ASAT in their serum.

  10. Serum YKL-40 and colorectal cancer

    DEFF Research Database (Denmark)

    Cintin, C; Johansen, J S; Christensen, Ib Jarle

    1999-01-01

    related to short survival. In the present study we analysed YKL-40 in preoperative sera from patients with colorectal cancer and evaluated its relation to survival. Serum YKL-40 was determined by RIA in 603 patients. Survival after operation was registered, and median follow-up time was 61 months. Three......YKL-40 is a mammalian member of the chitinase protein family. Although the function of YKL-40 is unknown, the pattern of its expression suggests a function in remodelling or degradation of extracellular matrix. High serum YKL-40 has been found in patients with recurrent breast cancer and has been...

  11. Relationship of serum uric acid, serum creatinine and serum cystatin C with maternal and fetal outcomes in rural Indian pregnant women

    Directory of Open Access Journals (Sweden)

    Padma Yalamati

    2015-10-01

    Conclusions: Serum creatinine and uric acid are independent risk factors for hypertensive disorders of pregnancy. High serum uric acid is associated with low birth weight and delivery by caesarian section whereas high serum creatinine with preterm delivery only before adjustment for confounding factors and not after adjustment. Serum cystatin C was not significantly associated with the maternal and fetal outcomes. [Int J Reprod Contracept Obstet Gynecol 2015; 4(5.000: 1505-1510

  12. Helicobacter pylori infection and serum ferritin

    DEFF Research Database (Denmark)

    Berg, Gabriele; Bode, G; Blettner, M

    2001-01-01

    OBJECTIVE: Helicobacter pylori may possibly affect the iron metabolism by occult bleeding, impaired absorption of non-hem iron, and by scavenging hem iron or ferritin, as some studies have suggested. The aim of this study was to analyze the association between H. pylori infection and serum ferrit...

  13. Serum Potassium Levels in Sigmoid Volvulus

    Science.gov (United States)

    Atamanalp, S. Selcuk; Keles, M. Sait; Aydinli, Bulent

    2009-01-01

    Objective: This study aimed to determine the serum potassium concentrations in patients with sigmoid volvulus (SV), which is a rare large bowel obstruction. Materials and Methods: The records of 86 patients with SV were reviewed retrospectively, while the records of 41 patients diagnosed with obstructive rectosigmoid cancer (ORC) were considered as the control group and as such, served as a source for comparison. Results: The analysis revealed a mean serum potassium concentration of 3.9 ± 0.6 mEq/L for the patients with SV, while the mean potassium concentration was 3.9 ± 0.5 mEq/L for the patients diagnosed with ORC (t:0.1, P>0.05). The number of hypokalemic and hyperkalemic patients identified in this study sample were 11 versus 5 patients and 1 versus 0 patients, respectively for the SV and ORC groups (x2 = 0.1 and 0.5, respectively with a P>0.05). Conclusions: No cause-and-effect relationship was observed between the serum potassium concentrations and SV. The serum potassium concentration is not pathognomonic for SV. PMID:25610090

  14. Pseudocholinesterase in the serum of allergic subjects

    NARCIS (Netherlands)

    Berrens, L.; Dijk, A.G. van

    1975-01-01

    There are no differences, between human normal and allergic sera, in the total activity of naphthylacetate-hydrolysing enzymes. No abnormal dibucaine-inhibited isoenzymes were detected. The Michaelis constants and the activation energies of serum pseudocholinesterases in the sera of patients with

  15. Serum sickness-like reaction with clarithromycin.

    Science.gov (United States)

    Sohail, Muhammad Adnan; Nasir, Junaid; Ikram, Umaira; Genese, Thomas

    2011-04-01

    Serum sickness-like reaction is a rare immunological condition which may develop following exposure to certain drugs such as penicillins, cephalosporins, and trimethoprim-sulfamethoxazole, among many others. It is described classically as a type III hypersensitivity response to heterologous proteins. Its true mechanism is still unclear. We present a case of serum sickness-like reaction to clarithromycin, a commonly prescribed drug for the treatment of respiratory tract infections. The patient had been taking this drug for 3 days when she experienced generalized body aches, rash, arthralgia, and shortness of breath, prompting presentation to the emergency department. Laboratory studies showed decreased C4 and total complement with a slightly elevated sedimentation rate. After exclusion of other possible causes, the diagnosis of serum sickness-like reaction was made. The patient responded well to nonsteroidal antiinflammatory medication, antihistamines, and a short, tapering dose of steroids. To our knowledge, serum sickness-like reaction to clarithromycin has never been reported previously. This case emphasizes the need for increased clinical awareness of such an adverse outcome to clarithromycin use.

  16. Abnormalities of serum calcium and magnesium

    Science.gov (United States)

    Neonatal hypocalcemia is defined as a total serum calcium concentration of <7 mg/dL or an ionized calcium concentration of <4 mg/dL (1mmol/L). In very low birth weight (VLBW) infants, ionized calcium values of 0.8 to 1 mmol/L are common and not usually associated with clinical symptoms. In larger in...

  17. Serum amyloid P inhibits dermal wound healing

    Science.gov (United States)

    The repair of open wounds depends on granulation tissue formation and contraction, which is primarily mediated by myofibroblasts. A subset of myofibroblasts originates from bone-marrow-derived monocytes which differentiate into fibroblast-like cells called fibrocytes. Serum amyloid P (SAP) inhibits ...

  18. Purification of human serum hyaluronidase using chromatofocusing

    DEFF Research Database (Denmark)

    Fenger, M

    1982-01-01

    A commercial chromatofocusing system was applied to Cohn's fraction III of human serum to purify hyaluronidase (E.C.3.2.1.3.5). The protein that eluted in the pH range 4.7-5.3 was pooled and precipitated by adding ammonium sulphate to 50% saturation. This sequence of fractionation purified...

  19. Serum proteomic profiling of major depressive disorder.

    Science.gov (United States)

    Bot, M; Chan, M K; Jansen, R; Lamers, F; Vogelzangs, N; Steiner, J; Leweke, F M; Rothermundt, M; Cooper, J; Bahn, S; Penninx, B W J H

    2015-07-14

    Much has still to be learned about the molecular mechanisms of depression. This study aims to gain insight into contributing mechanisms by identifying serum proteins related to major depressive disorder (MDD) in a large psychiatric cohort study. Our sample consisted of 1589 participants of the Netherlands Study of Depression and Anxiety, comprising 687 individuals with current MDD (cMDD), 482 individuals with remitted MDD (rMDD) and 420 controls. We studied the relationship between MDD status and the levels of 171 serum proteins detected on a multi-analyte profiling platform using adjusted linear regression models. Pooled analyses of two independent validation cohorts (totaling 78 MDD cases and 156 controls) was carried out to validate our top markers. Twenty-eight analytes differed significantly between cMDD cases and controls (P depression. Changes were more prominent in cMDD, suggesting that molecular alterations in serum are associated with acute depression symptomatology. These findings may help to establish serum-based biomarkers of depression and could improve our understanding of its pathophysiology.

  20. Pseudocholinesterase in the serum of allergic subjects

    NARCIS (Netherlands)

    Berrens, L.; Dijk, A.G. van

    1975-01-01

    There are no differences, between human normal and allergic sera, in the total activity of naphthylacetate-hydrolysing enzymes. No abnormal dibucaine-inhibited isoenzymes were detected. The Michaelis constants and the activation energies of serum pseudocholinesterases in the sera of patients with at

  1. Study of serum Helicobacter pylori soluble antigen

    Institute of Scientific and Technical Information of China (English)

    吴勤动; 朱永良

    2002-01-01

    Objective: to explore a new serological method for detecting Helicobac ter pylori ( H. pylori ) infection. Methods: Serum soluble antigen of H. p ylor i was detected by using avidin-biotin ELISA technique to evaluate the status of H. pylori infection and for comparison with rapid urease test ( RUT ), histo logi c examination and serology. Results: The sensitivity, specificity, positive pred ictive value and negative predictive value were 77.46%, 91.07%, 91.67% a nd 76.12 %, respectively. The prevalence rate of serum H. pylori soluble antigen in 138 patients undergoing endoscopy was similar to the rate obtained by 14 C-UBT met hods ( P>0.05 ). Conclusions: The detection of serum H. pylori solub le antigen( HpSAg) could be used as a new serological method which is accurate, and convenie nt, not affected by the memorizing reaction of serum antibody; is more sensitive , m ore specific and suitable for clinical diagnosis, and evaluation of eradication and for follow-up of H. pylori as well as for detection in children and pre gnant women.

  2. Dietary phosphorus, serum phosphorus, and cardiovascular disease.

    Science.gov (United States)

    Menon, Madhav C; Ix, Joachim H

    2013-10-01

    Recent epidemiologic studies have linked higher serum phosphorus concentrations to cardiovascular disease (CVD) events and mortality. This association has been identified in the general population and in those with chronic kidney disease (CKD). The risk of adverse outcomes appears to begin with phosphorus concentrations within the upper limit of the normal reference range. Multiple experimental studies have suggested pathogenetic mechanisms that involve direct and indirect effects of high phosphorus concentrations to explain these associations. Drawing from these observations, guideline-forming agencies have recommended that serum phosphorus concentrations be maintained within the normal reference range in patients with CKD and that dietary phosphorus restriction or use of intestinal phosphate binders should be considered to achieve this goal. However, outside the dialysis population, the links between dietary phosphorus intake and serum phosphorus concentrations, and dietary phosphorus intake and CVD events, are uncertain. With specific reference to the nondialysis populations, this review discusses the available data linking dietary phosphorus intake with serum phosphorus concentrations and CVD events.

  3. Comparison of Serum Calcium and Magnesium Between ...

    African Journals Online (AJOL)

    that serum magnesium seems to play a crucial role in the pathophysiology of preeclampsia in this environment. ... of the disease. Other contributory factors include obesity, ... association of blood levels of the elements and prevalence of preeclampsia ... Pregnant women with kidney disease, diabetes, endocrine disorder, or ...

  4. Serum selenium levels and prostate cancer risk

    Science.gov (United States)

    Cui, Zhigang; Liu, Dezhong; Liu, Chun; Liu, Gang

    2017-01-01

    Abstract Some observational studies have shown that elevated serum selenium levels are associated with reduced prostate cancer risk; however, not all published studies support these results. A literature search of PubMed, Embase, Medline, and the Cochrane Library up until September 2016 identified 17 studies suitable for further investigation. A meta-analysis was conducted on these studies to investigate the association between serum selenium levels and subsequent prostate cancer risk. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were used to evaluate the overall OR of prostate cancer for the highest versus the lowest levels of serum selenium. We found a pooled OR (95% CI) of 0.76 (0.64, 0.91; P selenium levels and prostate cancer risk was found in each of case–control studies, current and former smokers, high-grade cancer cases, advanced cancer cases, and different populations. Such correlations were not found for subgroups containing each of cohort studies, nonsmokers, low-grade cancer cases, and early stage cancer cases. In conclusion, our study suggests an inverse relationship between serum selenium levels and prostate cancer risk. However, further cohort studies and randomized control trials based on non-Western populations are required. PMID:28151881

  5. Serum chromium levels in gestational diabetes mellitus

    Directory of Open Access Journals (Sweden)

    P G Sundararaman

    2012-01-01

    Full Text Available Objective: To measure serum chromium level in women with gestational diabetes mellitus (GDM from Chennai, South India. Materials and Methods: Thirty women with gestational diabetes, 60 age matched controls. Inclusion criteria: Gestational age 22-28 weeks, age group 20-35 years. Exclusion Criteria: Gestational age beyond 28 weeks, malnutrition or presence of infection. Serum chromium was measured using inductive couple plasma emission spectrometer. Results: Serum chromium levels of women with GDM, 1.59+/-0.02 ng/ml (range: 0.16-4.0 ng/ml were lower than in controls (4.58+/-0.62 ng/ml; range 0.82-5.33 ng/ml (P < 0.001. However, there were no significant differences among cases and controls when subdivided by parity. Conclusions: Women with GDM from a South Indian city had lower levels of serum chromium compared to pregnant women without GDM. Studies may be done whether chromium supplementation is useful in this group of women.

  6. Serum hypercoagulability states in Coats’ disease

    Science.gov (United States)

    Ghassemi, Fariba; Shields, Carol L; Mohebbi, Masoumeh; Nili Ahmadabadi, Mehdi; Morsali, Fatemeh; Sabour, Siamak

    2017-01-01

    Purpose The purpose of this study was to investigate the serum hypercoagulability state and common viral and protozoan infections in Coats’ disease versus a normal control group. Materials and methods In this comparative case series, 22 consecutive patients with Coats’ disease and 19 non-Coats’ patients undergoing lensectomy for congenital, traumatic, or senile cataract between January 2011 and June 2014 were included. Laboratory data for hypercoagulability states and common viral and protozoan infections were investigated. Results The mean age for the Coats’ group was 14.5 years (median 8 years, range: 2 months to 59 years), and for the control group it was 30.6 years (median 17 years, range: 2–82 years). In patients aged 10 years or younger, anticytomegalovirus immunoglobulin G (IgG) (P≤0.01), homocysteine (P=0.03), and serum beta globulin (P<0.001) were associated with Coats’ disease. In those older than 10 years, higher serum protein S (P=0.04), beta globulin (P=0.05), and gamma globulin (P=0.04) were related to Coats’ diagnosis. After adjusting for sex and age as confounding factors, only beta globulin was found to be associated with Coats’ disease in logistic regression analysis (odds ratio: 1.8, 95% confidence interval: 1.0–3.1, P=0.02). Conclusion Serum beta globulin levels appear to be elevated in patients with Coats’ disease. PMID:28223774

  7. Aggregation and fibrillation of bovine serum albumin

    DEFF Research Database (Denmark)

    Holm, NK; Jespersen, SK; Thomassen, LV;

    2007-01-01

    The all-alpha helix multi-domain protein bovine serum albumin (BSA) aggregates at elevated temperatures. Here we show that these thermal aggregates have amyloid properties. They bind the fibril-specific dyes Thioflavin T and Congo Red, show elongated although somewhat worm-like morphology...

  8. Characteristics of serum endocan levels in infection.

    Directory of Open Access Journals (Sweden)

    Kazunori Seo

    Full Text Available Endocan is a newly recognized biomarker of sepsis. However, there have been no studies of the trends in endocan levels during infection and their associations with other clinical factors. The aim of this study was to assess the time course of endocan levels and the associations of endocan with clinical factors during infection by comparison with other biomarkers.Serum samples and blood cultures were obtained from patients who were diagnosed with infection from June 2013 to March 2014. Serum endocan, C-reactive protein (CRP, and procalcitonin (PCT levels during four periods during infection were measured (day 0, day 1-2, day 3-5, and day 6-10.A total of 78 patients were enrolled in this study. The median endocan level decreased by only 23% during infection, whereas both serum CRP and PCT levels decreased by more than 80%. Endocan levels were correlated to neither CRP levels nor PCT levels in each period. Endocan levels at day 0 in patients with bacteremia were higher than those without bacteremia (1.09 ng/mL vs 0.82 ng/mL, P=0.002, but neither CRP levels nor PCT levels at day 0 were different between the two groups. Areas under the receiver operator characteristic (ROC curves of endocan, CRP, and PCT at day 0 were 0.662, 0.343, and 0.563, respectively. Positive blood cultures tended to be related to high endocan levels, but not significantly (odds ratio: 4.24, 95% CI: 0.99-10.34, P=0.05.In bacteremic cases, serum endocan levels in bacteremia tended to be higher than in non-bacteremic cases. Although endocan level was not identified as a prognostic factor of bacteremia, further prospective study concerning the relationship between serum endocan level and bacteremia would be needed.

  9. EVALUATION OF SERUM CHOLESTEROL, AMINO TRANSFERASES

    Directory of Open Access Journals (Sweden)

    Anantha Babu

    2016-01-01

    Full Text Available BACKGROUND AND AIMS The purpose of this study was to determine the efficacy of red yeast rice (Monascus purpureus-fermented rice in lowering cholesterol in the blood. At the same time, alanine aminotranferase (ALT, aspartate aminotransferase (AST and gamma-glutamyl transferase (γ-GT were measured for notable side effects in the liver. Possible muscle damage was determined by measuring creatine kinase (CK. METHODS The cholesterol lowering effect in serum of red yeast rice-fed rats were studied over a 42-day feeding period. A total of 16 male Sprague-Dawley rats were randomised into 8 per group: control and treated. Treated rats were administered 1.35g/kg/day. Control rats were maintained on ordinary rat chow. RESULTS Serum cholesterol levels were significantly decreased by 19.13% in treated group compared to controls. This treatment also showed increase in serum ALT and AST activities by 41.90% and 21.53%, respectively. Mean CK activity in treated rats showed an increase by 32.32% when compared with control rats. γ-GT is the only enzyme that showed a decrease of 15.16% in sera of treated rats. Body weights of control and treated rats increased significantly by 10% end of feeding period but were not due to treatment. CONCLUSION Red yeast rice significantly decreased serum cholesterol level at a dosage of 1.35g/kg/day. However, the differences in serum enzyme activities between control and treated rats were not significant.

  10. Correlation of serum leptin with levels of hemoglobin in hemodialysis

    Directory of Open Access Journals (Sweden)

    Rafieian-Kopaei Mahmoud

    2012-04-01

    Full Text Available To examine the association of serum leptin level with anemia in hemodialysis, we investigated 36 patients (males: 21, diabetics: 11 under regular hemodialysis. For patients, complete blood counts, iron profile, serum leptin, and adequacy of hemodialysis were assessed. In this study a significant correlation of serum leptin with level of hemoglobin and body mass index was detected. An association between serum leptin and total iron binding capacity was observed. No correlation of serum ferritin with leptin level was seen. Our findings attest previous findings showing that greater serum leptin levels are associated with greater hemoglobin levels.

  11. The study of serum calcium and serum magnesium in pregnancy induced hypertension and normal pregnancy

    Directory of Open Access Journals (Sweden)

    Jagannath Pairu

    2015-02-01

    Full Text Available Background: Preeclampsia along with its complications is one of the major causes of maternal and fetal mortality and morbidity. Association of calcium and magnesium with pregnancy induced hypertension is known since decades. Evidence of decreased serum calcium and decreased serum magnesium has been observed in patients with pregnancy induced hypertension and has been implicated in the etiopathogenesis of preeclampsia. Methods: The present study was undertaken in 100 pregnant women. Data for the study was collected from 50 normotensive pregnant women with more than 20 weeks of gestational age (control group and 50 pregnancy induced hypertension patients (study group attending for the antenatal care in department of obstetrics and gynaecology in Vanivilas hospital, Bowring and Lady Curzon hospital attached to Bangalore medical college and research institute. Cases and controls were matched. Serum calcium and serum magnesium levels were estimated by spectrophotometry method. Results: The mean serum calcium is significantly lower in pregnancy induced hypertension group (8.15 +/- 0.37 mg/dl compared to normal pregnancy (9.16 +/- 0.82 mg/dl. The mean serum magnesium is lower in pregnancy induced hypertension group (1.78 +/- 0.70 mEq/L than normal pregnancy (2.08 +/- 0.46 mEq/L which is moderately significant. Conclusions: The serum calcium and serum magnesium levels are decreased in pregnancy induced hypertension patients compared to normotensive normal pregnant women, suggesting the possible role of calcium and magnesium in etiopathophysiology of pregnancy induced hypertension. [Int J Reprod Contracept Obstet Gynecol 2015; 4(1.000: 30-34

  12. 蓝氏贾第鞭毛虫alpha-8贾第素特异性锤头状核酶-GCV重组载体的构建%Construction of GCV-Specific Hammerhead Ribozyme Recombinant Vector of Alpha-8 Giardin in Giardia lamblia

    Institute of Scientific and Technical Information of China (English)

    魏超君; 卢思奇; 曹利静; 田喜凤

    2011-01-01

    Objective To construct a GCV- ribozyme recombinant vectors of a-8 giardin in Giardia lamblia. Methods The secondary structure of a-8 giardin mRNA (GenBank Accession No. AY781323) was analyzed with the RNA draw software. According to the proportion of G: C and principles of designing hammerhead ribozyme, suitable ribozyme cleavage points were chosen. A specific antisense-hammerhead ribozyme (H8)was designed and synthesized. The ribozyme was cloned into Giardia canis virus (GCV) vector to construct a recombinant viral vector-pGCV634/H8/1423. The vector was linearized and transcripted into the trophozoites of G. lamblia by electroporation method. The a-8 giardin mRNA level of the transfectants and normal trophozoites were analyzed 24 h after electroporation by RT-PCR. Results The recombinant vector of GCV-specific hammerhead ribozyme of a-8 giardin in Giardia lamblia (pGCV634/H8/ 1423) was constructed. RT-PCR assays showed the ribozyme (H8) mRNA can be detected 24h after transfection and a-8 giardin mRNA was cleaved effectively by ribozyme (H8) introcellularly. Conclusion pGCV634/H8/1423 can transfect Giardia trophozoites and cleave mRNA of a-8 giardin intracellularly.%目的 构建蓝氏贾第鞭毛虫alpha-8贾第素(α-8 giardin)特异性锤头状核酶-GCV重组载体. 方法 采用RNA draw软件对蓝氏贾第鞭毛虫α-8贾第素基因序列(GenBank登录号为AY781323)的二级结构进行模拟分析,按照G:C比例和锤头状核酶设计原则,选取合适的核酶切割靶点,设计特异性锤头状核酶(H8)序列,并将其与犬贾第虫病毒(GCV)连接,获得α-8贾第素特异性锤头状核酶-GCV重组载体(pGCV634/H8/1423).将载体线性化体外转录产物电击转染至贾第虫滋养体细胞内.提取转染后24 h的各组虫体总RNA,并以其为模板采用RT-PCR验证转染效果及对靶mRNA的切割效果. 结果 成功设计、合成了蓝氏贾第鞭毛虫α-8贾第素mRNA锤头状核酶序列(H8),将其与犬贾第虫病毒载

  13. 云南中缅边境居民贾第虫感染影响因素分析%Risk factors of Giardia Lamblia infection in residents at both sides of China-Myanmar border area of Nabang port,Yunnan province

    Institute of Scientific and Technical Information of China (English)

    孙晓东; 王剑; 梁桂亮; 杨捷; 邓艳; Lasi Ja-hkawn; 张再兴

    2012-01-01

    目的 了解云南那邦口岸中缅双边贾第虫感染危险因素.方法 2008年整群随机抽取中国盈江县那邦镇和缅甸拉咱市边境自然村,对调查点内居住时间>1年的边民进行流行病学问卷调查及新鲜粪便碘液涂片镜检,采用logistic回归方法对数据进行统计分析.结果 共调查6个自然村903人,平均年龄为(22.81±18.82)岁;总贾第虫感染率为13.51%,其中调查2个中国自然村286人,感染率为2.10%;调查4个缅甸自然村617人,感染率为18.80%;单因素分析结果表明,国籍、年龄、民族、用手抓饭、喝生水、饮用水源、近1月内腹泻、使用厕所类型和粪便带粘液为可能的危险因素;多因素分析结果表明,国籍、年龄、民族、喝生水和粪便带粘液为主要危险因素.结论 直接饮用被粪便污染的水源是缅甸边民贾第虫流行的主要原因.%Objective To examine the risk factors of Giardia Lamblia infection in the residents at both sides of China-Myanmar border area in Nabang port of Yunnan province. Methods An epidemiological survey was carried out with clustered random sampling in the border villages of Nabang township of China and Laiza city of Myanmar in 2008. The data and fecal samples were collected from the villagers who living in the sampled villages more than one year. The fecal samples with iodine staining were examined with microscopy. The data were analyzed with univariate and multivariate non-conditional logistic regression. Results Totally 903 residents from 6 villages were investigated and 13 kinds of risk factors were analyzed. The average age of the residents was 22. 81 ± 18. 82 years and the overall infection rate of Giardia Lamblia was 13.51%. Among the residents,there were 286 infected Chinese with a infection rate of 2.10% and 617 infected Burmese with a infection rate of 18. 8%. The results of univariate non-conditional logistic analysis showed that the risk factors of the infection

  14. C2株蓝氏贾第鞭毛虫 SU MO特异性蛋白酶基因的克隆、生物信息学分析及其催化活性区的原核表达%Cloning and bioinformatics analysis of Giardia lamblia C2 strain SENP gene and prokaryotic expression of SENP catalytic domain

    Institute of Scientific and Technical Information of China (English)

    李少东; 周英斌; 刘晓莉; 禇晗; 李思瑾; 田喜凤; 王洋

    2015-01-01

    目的:克隆C2株蓝氏贾第鞭毛虫(Giardia lamblia ,简称贾第虫)的SUMO‐Specific Protease(SENP)基因,并对其序列进行生物信息学分析,原核表达贾第虫SENP的催化活性区。方法提取C2株贾第虫基因组DNA ,以基因组DNA为模板获得SENP编码区全长片段,连入克隆载体pGM‐T ,测序后进行生物信息学分析;根据分析结果克隆SENP的催化活性区,构建其原核表达载体pET‐28a(+)‐SENPc ,在 E .coli Rosetta(DE3)中诱导表达,SDS‐PAGE及Western blot观察表达结果。结果成功克隆了C2株贾第虫SENP编码区全长序列,生物信息学分析显示C2株贾第虫SENP蛋白序列与WB株相同,二级结构以无规则卷曲为主,其催化活性区位于126‐497aa ,被一段插入序列分割成两个部分;构建了SENP催化活性区原核表达载体并在大肠杆菌中高效表达,在相对分子量约43 kD的位置出现目的蛋白条带,与理论值相符。结论成功克隆了贾第虫SENP基因并原核表达了其催化活性区,为贾第虫SENP蛋白功能的研究提供了基础。%SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a

  15. A low percentage of autologous serum can replace bovine serum to engineer human nasal cartilage

    Directory of Open Access Journals (Sweden)

    F Wolf

    2008-02-01

    Full Text Available For the generation of cell-based therapeutic products, it would be preferable to avoid the use of animal-derived components. Our study thus aimed at investigating the possibility to replace foetal bovine serum (FBS with autologous serum (AS for the engineering of cartilage grafts using expanded human nasal chondrocytes (HNC. HNC isolated from 7 donors were expanded in medium containing 10% FBS or AS at different concentrations (2%, 5% and 10% and cultured in pellets using serum-free medium or in Hyaff®-11 meshes using medium containing FBS or AS. Tissue forming capacity was assessed histologically (Safranin O, immunohistochemically (type II collagen and biochemically (glycosaminoglycans -GAG- and DNA. Differences among experimental groups were assessed by Mann Whitney tests. HNC expanded under the different serum conditions proliferated at comparable rates and generated cartilaginous pellets with similar histological appearance and amounts of GAG. Tissues generated by HNC from different donors cultured in Hyaff®-11 had variable quality, but the accumulated GAG amounts were comparable among the different serum conditions. Staining intensity for collagen type II was consistent with GAG deposition. Among the different serum conditions tested, the use of 2% AS resulted in the lowest variability in the GAG contents of generated tissues. In conclusion, a low percentage of AS can replace FBS both during the expansion and differentiation of HNC and reduce the variability in the quality of the resulting engineered cartilage tissues.

  16. Serum clusterin as a marker for diagnosing hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Ragaa A. Ramadan

    2014-09-01

    Conclusions: Although the diagnostic performance of serum AFP outperformed that of serum CLU, their combined parallel approach improved the sensitivity which is required in screening high risk populations such as CHC patients.

  17. Prognostic significance of serum immunoglobulin pareprotein in chronic lymphocytic leukemia

    Institute of Scientific and Technical Information of China (English)

    杨舒

    2012-01-01

    Objective To investigate the incidence of serum immunoglobulin (Ig) paraprotein in chronic lymphocytic leukemia(CLL) ,and to explore its clinical associated laboratory features and prognostic implication. Methods Serum protein electrophoresis and immunofixation

  18. Serum sialic acid and CEA concentrations in human breast cancer.

    Science.gov (United States)

    Hogan-Ryan, A; Fennelly, J J; Jones, M; Cantwell, B; Duffy, M J

    1980-04-01

    The concentration of bound sialic acid in the sera of 56 normal subjects and 65 subjects with breast cancer was measured, in order to determine (1) whether serum sialic acid concentrations are raised in breast cancer and (2) whether the concentration of sialic acid in serum reflects tumour stage. The amount of sialic acid in serum was compared to serum carcinoembryonic antigen (CEA) values. Urinary hydroxyproline and serum alkaline phosphatase concentrations were used as indicators of bone and liver involvement. Erythrocyte sedimentation rate (ESR) was also measured. Significantly elevated serum sialic acid concentrations were found in breast cancer, and showed correlation with tumour stage. Serum sialic acid values did not correlate with CEA values. The results suggest that measurement of serum sialic acid concentrations may be of adjunctive value in assessing tumour stage.

  19. Correlation between serum lipoproteins and abdominal fat pad in ...

    African Journals Online (AJOL)

    Correlation between serum lipoproteins and abdominal fat pad in broiler chickens. ... The high density lipoprotein (HDL) concentration in birds fed fish oil diet was higher than other treatments, but the serum low density ... Article Metrics.

  20. Survey of serum procalcitonin in cirrhotic patients.

    Science.gov (United States)

    Rahimkhani, Monireh; Einollahi, Nahid; Khavari Daneshvar, Hossein; Dashti, Nasrin

    2013-04-06

    Procalcitonin (PCT) is a prohormone that has been used as a marker for the diagnosis of bacterial infections. The aim of this study was to survey PCT levels in patients with cirrhosis. Sixty-four patients with hepatic cirrhosis and 32 healthy blood donors were enrolled in this study. Serum PCT levels was detected using immunoluminometric assay. The rate of positive PCT was higher in patients with hepatitis C cirrhosis (92.8%) than the other groups. Among other cirrhotic patients, positive PCT levels were 77% for hepatitis B, 70% for cancer and 53.3% for unknown groups respectively. Serum procalcitonin levels were significantly higher in cirrhotic patients with bacterial infection (2.65±1.11 ng/ml) than those without infection (0.59±0.16 ng/ml, P=0.0001). PCT assay in cirrhotic patients may help diagnosis of sepsis and reduce unnecessary antibiotic use.

  1. Survey of Serum Procalcitonin in Cirrhotic Patients

    Directory of Open Access Journals (Sweden)

    Monireh Rahimkhani

    2013-03-01

    Full Text Available Procalcitonin (PCT is a prohormone that has been used as a marker for the diagnosis of bacterial infections. The aim of this study was to survey PCT levels in patients with cirrhosis. Sixty-four patients with hepatic cirrhosis and 32 healthy blood donors were enrolled in this study. Serum PCT levels was detected using immunoluminometric assay. The rate of positive PCT was higher in patients with hepatitis C cirrhosis (92.8% than the other groups. Among other cirrhotic patients, positive PCT levels were 77% for hepatitis B, 70% for cancer and 53.3% for unknown groups respectively. Serum procalcitonin levels were significantly higher in cirrhotic patients with bacterial infection (2.65±1.11 ng/ml than those without infection (0.59±0.16 ng/ml, P=0.0001. PCT assay in cirrhotic patients may help diagnosis of sepsis and reduce unnecessary antibiotic use

  2. Purification of NAD(+) glycohydrolase from human serum.

    Science.gov (United States)

    Coşkun, Ozlem; Nurten, Rüstem

    2013-07-01

    In the present study, NAD(+) glycohydrolase was purified from serum samples collected from healthy individuals using ammonium sulfate fractionation, Affi-Gel blue (Cibacron Blue F3GA) affinity chromatography, Sephadex G-100 column chromatography and isoelectric focusing. The final step was followed by a second Sephadex G-100 column chromatography assay in order to remove the ampholytes from the isoelectric focusing step. In terms of enhancement of specific activity, the NAD(+) glycohydrolase protein was purified ∼480-fold, with a yield of 1% compared with the initial serum fraction. The purified fraction appeared to be homogeneous, with a molecular weight of 39 kDa, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, and also corresponded to the soluble (monomeric) form of surface antigen CD38.

  3. Macrophage serum markers in pneumococcal bacteremia

    DEFF Research Database (Denmark)

    Møller, Holger Jon; Moestrup, Søren K; Weis, Nina

    2006-01-01

    OBJECTIVE: Soluble CD163 (sCD163) is a new macrophage-specific serum marker. This study investigated sCD163 and other markers of macrophage activation (neopterin, ferritin, transcobalamin, and soluble urokinase plasminogen activator receptor [suPAR]) as prognostic factors in patients with pneumoc......OBJECTIVE: Soluble CD163 (sCD163) is a new macrophage-specific serum marker. This study investigated sCD163 and other markers of macrophage activation (neopterin, ferritin, transcobalamin, and soluble urokinase plasminogen activator receptor [suPAR]) as prognostic factors in patients...... on the probability of survival when sCD163 and CRP were known (p = .25). CONCLUSIONS: Macrophage marker response in pneumococcal bacteremia was compromised in old age. In patients disease outcome....

  4. Hypoxemia increases serum interleukin-6 in humans

    DEFF Research Database (Denmark)

    Klausen, T; Olsen, Niels Vidiendal; Poulsen, T D

    1997-01-01

    Serum concentrations of interleukin (IL) 1 beta, IL-1 receptor antagonist (IL-1ra), IL-6, tumor necrosis factor (TNF) alpha, and C-reactive protein (CRP) were determined in ten healthy men at sea level and during four days of altitude hypoxia (4350m above sea level). The mean (SD) arterial blood...... oxygen saturations were 78.6 (7.3)%, 82.4 (4.9)%, and 83.4 (5.3)% in the first, second, and third days at altitude, respectively. A symptom score of acute mountain sickness (AMS) revealed that the subjects had mostly light symptoms of AMS. Mean serum IL-6 increased from 1.36 (1.04) pg x ml(-1) at sea...

  5. Serum triglycerides and risk of cardiovascular disease.

    Science.gov (United States)

    Boullart, A C I; de Graaf, J; Stalenhoef, A F

    2012-05-01

    Dyslipidemia, especially elevated serum levels of cholesterol, is causally related to cardiovascular disease. The specific role of triglycerides has long been controversial. In this article we discuss the role of serum triglycerides in relation to the risk of cardiovascular disease. First, the (patho)physiology of triglycerides is described, including the definition and a short summary of the primary and secondary causes of hypertriglyceridemia. Furthermore, we will give an overview of the published epidemiological studies concerning hypertriglyceridemia and cardiovascular disease to support the view that triglyceride-rich lipoproteins are an independently associated risk factor. Finally, treatment strategies and treatment targets are discussed. This article is part of a Special Issue entitled Triglyceride Metabolism and Disease. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. Comparison of fortified calf serum, serum substitutes and fetal calf serum with or without extenders for propagation of cell cultures for virus plaque assays.

    Science.gov (United States)

    Dahling, D R; Wright, B A

    1990-03-01

    Two studies comparing sewage-isolated and laboratory stock viruses were conducted to determine if alternative forms of serum or serum extenders could be used in place of fetal bovine serum without a significant loss of viral titer. In the first study, BGM cells were grown in standard MEM-L15 medium which was supplemented with Nuserum, Sigma serum replacement (CPSR-1), HyClone defined iron supplemented calf bovine serum, fetal bovine serum (FBS) or FBS supplemented with either SerXtend or Mito serum extenders. Comparison of virus titers showed that CPSR-1 gave the best overall results and was comparable to FBS. Of the serum extenders, only SerXtend improved virus recovery from sewage samples. In the second study, all sera were tested with and without SerXtend. In these experiments, SerXtend enhanced virus sensitivity of the BGM cell line grown in the HyClone serum but reduced the sensitivity of those cultured in Sigma serum. In both series, the growth of BGM cells was monitored for 12 weeks and all test products were shown to support long-term cell growth.

  7. Serum sickness-like illness associated with ciprofloxacin.

    Science.gov (United States)

    Slama, T G

    1990-05-01

    Serum sickness is a systemic hypersensitivity reaction initially reported in children receiving horse serum. Drugs such as penicillins, cephalosporins, and sulfonamides are now noted to be the most common etiologic agents of serum sickness-like reactions. This case report describes a serum sickness-like reaction temporally related to ciprofloxacin, a previously unreported adverse effect of this drug or any of the other quinolones.

  8. Photodynamically generated bovine serum albumin radicals

    DEFF Research Database (Denmark)

    Silvester, J A; Timmins, G S; Davies, Michael Jonathan

    1998-01-01

    Porphyrin-sensitized photoxidation of bovine serum albumin (BSA) results in oxidation of the protein at (at least) two different, specific sites: the Cys-34 residue giving rise to a thiyl radical (RS.); and one or both of the tryptophan residues (Trp-134 and Trp-214) resulting in the formation of...... of proteases. The generation of protein-derived radicals also results in an enhancement of photobleaching of the porphyrin, suggesting that protein radical generation is linked to porphyrin photooxidation....

  9. Group specific component in serum and otosclerosis

    DEFF Research Database (Denmark)

    Kjeldsen, A D; Vase, P; Thymann, M

    1994-01-01

    An earlier Swedish study suggested a positive association between otosclerosis and the group-specific component GC*1F marker. We investigated the distribution of GC subtypes in 101 Danish patients with otosclerosis who all had surgery performed in the county of Funen. Compared to 1674 Danish...... controls we found no evidence of any association between markers from the GC serum protein system and otosclerosis....

  10. Mammographic Breast Density and Serum Phytoestrogen Levels

    OpenAIRE

    Lowry, Sarah J.; Sprague, Brian L; Bowles, Erin J. Aiello; Hedman, Curtis J.; Hemming, Jocelyn; Hampton, John M.; Burnside, Elizabeth S.; Sisney, Gale A.; Buist, Diana S. M.; Trentham-Dietz, Amy

    2012-01-01

    Some forms of estrogen are associated with breast cancer risk as well as with mammographic density (MD), a strong marker of breast cancer risk. Whether phytoestrogen intake affects breast density, however, remains unclear. We evaluated the association between serum levels of phytoestrogens and MD in postmenopausal women. We enrolled 269 women, ages 55–70 yr, who received a screening mammogram and had no history of postmenopausal hormone use. Subjects completed a survey on diet and factors rel...

  11. Serum ferritin levels in hemoglobin H disease.

    Science.gov (United States)

    Galanello, R; Melis, M A; Paglietti, E; Cornacchia, G; de Virgiliis, S; Cao, A

    1983-01-01

    This study shows that hemoglobin H disease patients aged between 0.5 and 44 years, usually (27 out of 30) have normal serum ferritin levels according to age. This reconfirms that in this disease there are usually normal iron stores. However, in a few patients (3 out of 30) increased levels were found. This may be due to inappropriate iron medication, transfusions or associated idiopathic hereditary hemocromatosis gene.

  12. Proteomic evaluation of sheep serum proteins

    OpenAIRE

    Chiaradia Elisabetta; Avellini Luca; Tartaglia Micaela; Gaiti Alberto; Just Ingo; Scoppetta Fausto; Czentnar Zoltan; Pich Andreas

    2012-01-01

    Abstract Background The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference...

  13. Comparative serum protein binding of anthracycline derivatives.

    Science.gov (United States)

    Chassany, O; Urien, S; Claudepierre, P; Bastian, G; Tillement, J P

    1996-01-01

    The binding of doxorubicin, iododoxorubicin, daunorubicin, epirubicin, pirarubicin, zorubicin, aclarubicin, and mitoxantrone to 600 microM human serum albumin and 50 microM alpha 1-acid glycoprotein was studied by ultrafiltration at 37 degrees C and pH 7.4. Anthracycline concentrations (total and free) were determined by high-performance liquid chromatography (HPLC) with fluorometric detection. Binding to albumin (600 microM) varied from 61% (daunorubicin) to 94% (iododoxorubicin). The binding to alpha 1-acid glycoprotein (50 microM) was more variable, ranging from 31% (epirubicin) to 64% (zorubicin), and was essentially related to the hydrophobicity of the derivatives. Simulations showed that the total serum binding varied over a broad range from 71% (doxorubicin) to 96% (iododoxorubicin). We recently reported that the binding to lipoproteins of a series of eight anthracycline analogues could be ascribed to chemicophysical determinants of lipophilicity [2]. The present study was conducted to evaluate in vitro the contribution of albumin and alpha 1-acid glycoprotein to the total serum binding of these drugs.

  14. Serum Adiponectin in Women with Gestational Diabetes

    Directory of Open Access Journals (Sweden)

    2014-03-01

    Full Text Available Background &Objective: Adiponectin is an adipose tissue adipokin that may contribute to obesity and insulin resistantance. The aim of this study was to evaluate the associations between serum concentrations of adiponectin and insulin resistance in gestational diabetes (GDM.Materials & Methods: Serum adiponectin levels, fasting blood sugar (FBS, glycated hemoglobin (HbA1C, insulin levels and blood lipids were measured in 66 women with GDM and 70 pregnant women without GDM. The associations between serum concentrations of adiponectin and insulin resistance were evaluated using the homeostasis model assessment of insulin resistance (HOMA–IR and quantitative insulin sensitivity check index (QUICKI.Results: There were statistically significant between-group differences in FBS, HbA1C and HOMA–IR. Adiponectin concentrations were not significantly different in GDM women in comparison with the control group. However, GDM women above the age of 30 have significantly lower adiponectin concentrations than those without GDM. Adiponectin was positively associated with QUICKI (r = 0.268, P < 0.03 and inversely related to HOMA–IR (r = 0.238, P < 0.05.Conclusion: Adiponectin is significantly decreased in older women with GDM. Deficiency of adiponectin may correlate with insulin resistance in GDM.

  15. Correlations Between Macular, Skin, and Serum Carotenoids

    Science.gov (United States)

    Conrady, Christopher D.; Bell, James P.; Besch, Brian M.; Gorusupudi, Aruna; Farnsworth, Kelliann; Ermakov, Igor; Sharifzadeh, Mohsen; Ermakova, Maia; Gellermann, Werner; Bernstein, Paul S.

    2017-01-01

    Purpose Ocular and systemic measurement and imaging of the macular carotenoids lutein and zeaxanthin have been employed extensively as potential biomarkers of AMD risk. In this study, we systematically compare dual wavelength retinal autofluorescence imaging (AFI) of macular pigment with skin resonance Raman spectroscopy (RRS) and serum carotenoid levels in a clinic-based population. Methods Eighty-eight patients were recruited from retina and general ophthalmology practices from a tertiary referral center and excluded only if they did not have all three modalities tested, had a diagnosis of macular telangiectasia (MacTel) or Stargardt disease, or had poor AFI image quality. Skin, macular, and serum carotenoid levels were measured by RRS, AFI, and HPLC, respectively. Results Skin RRS measurements and serum zeaxanthin concentrations correlated most strongly with AFI macular pigment volume under the curve (MPVUC) measurements up to 9° eccentricity relative to MPVUC or rotationally averaged macular pigment optical density (MPOD) measurements at smaller eccentricities. These measurements were reproducible and not significantly affected by cataracts. We also found that these techniques could readily identify subjects taking oral carotenoid-containing supplements. Conclusions Larger macular pigment volume AFI and skin RRS measurements are noninvasive, objective, and reliable methods to assess ocular and systemic carotenoid levels. They are an attractive alternative to psychophysical and optical methods that measure MPOD at a limited number of eccentricities. Consequently, skin RRS and MPVUC at 9° are both reasonable biomarkers of macular carotenoid status that could be readily adapted to research and clinical settings. PMID:28728169

  16. Childhood serum sickness: a case report.

    Science.gov (United States)

    Chao, Y K; Shyur, S D; Wu, C Y; Wang, C Y

    2001-09-01

    Childhood serum sickness is a rare allergic disease that follows the administration of a foreign antigenic material, most commonly caused by injecting a protein or haptenic drug. The disease is a type III hypersensitivity reaction mediated by deposits of circulating immune complexes in small vessels, which leads to complement activation and subsequent inflammation. The clinical features are fever, cutaneous eruptions, lymphadenopathy, arthralgias, albuminuria, and nephritis. Serum sickness is an acute self-limited disease. We report a 3-year-old child who presented with fever and a rash; an invasive bacterial infection was strongly suspected. He was therefore given penicillin and gentamicin and responded well. At day 4 after admission, he developed a serum sickness reaction and showed symptoms of arthralgias, generalized edema, purpura, and gross hematuria. The white blood cell count was 12 190/mm3 with 7% eosinophils. Urinalysis revealed red blood cell above 100 per high power field, white blood cell 10 to 15 per high power field, and proteinuria. The antibiotics were discontinued and hydrocortisone (20 mg/kg/d), diphenhydramine HCl (4 mg/kg/d), aspirin (66 mg/kg/d) was administered, plus 1 dose of epinephrine (0.01 mL/kg) administered intramuscularly. On day 7, the 3rd day after withholding antibiotics, his condition dramatically improved. The clinical symptoms resolved progressively and his urinalysis returned to normal.

  17. Serum copeptin and pregnancy outcome in preeclampsia

    Directory of Open Access Journals (Sweden)

    Kehinde Sola Akinlade

    2015-01-01

    Full Text Available Background: A number of biochemical predictors of preeclampsia have been reported, but little is known about their possible relationship with maternal and fetal outcomes. This study determined serum copeptin in pregnant women with preeclampsia and assessed its relationship with pregnancy outcomes. Materials and Methods: Thirty women with severe preeclampsia (SP, 30 with mild preeclampsia (MP, and 30 with uncomplicated pregnancy were enrolled into this study. Serum copeptin, creatinine, and liver function were determined using enzyme-linked immunosorbent assay and colorimetry as appropriate. Pregnancy outcomes, both maternal and fetal, were taken using standard methods. Results: Copeptin was significantly elevated in preeclampsia subjects compared with controls and in SP compared with MP. Assessing the diagnostic property of copeptin for preeclampsia, the area under the curve for copeptin was 0.99. Nine (30% and 3 (10% of SP and MP, respectively had abruptio placenta while 6 (20%, 2 (6.7%, and 1 (3.3% still births were recorded in SP, MP, and controls, respectively. Neonates of mothers with preeclampsia had significantly lower birth weight, infant length, ponderal index, and head circumference compared with neonates of the controls. Copeptin had a significant inverse relationship with birth weight, ponderal index, head circumference, Apgar score, and infant length in neonates of mothers with preeclampsia. Conclusion: Serum copeptin level in the third trimester could predict preeclampsia and its elevation is associated with adverse perinatal outcome.

  18. Study of serum Helicobacter pylori soluble antigen

    Institute of Scientific and Technical Information of China (English)

    吴勤动; 朱永良

    2002-01-01

    Objective:to explore a new serological method for detecting Helicobacter pylori(H.pylori) infection.Methods:Serum soluble antigen of H.pylori was detected by using avidin-biotin ELISA technique to evaluate the status of H.pylori infection and for comparison with rapid urease test(RUT).histologic examination and serology,Results:The sensitivity,specificity,positive predictive value and negative predictive value were 77.46% ,91.07%,91.67% and 76.12%,respectively.The prevalence rate of werum H. pylori soluble antigen in 138 patients undergong endoscopy was similar to the rate obtained by 14 C-UBT methods(P>0.05).Conclusions:The detection of serum H.pylori soluble antigen(HpSAg) could be used as a new serological method which is accurate,and convenient,not affected by the memorizing raction of serum antibody;is more sensitive,more specific and suitable for dinical diagriosis,and evaluation of eradication and for follow-up of H.pylori as well as for detection in children and pregnant women.

  19. Serum Antibody Repertoire Profiling Using In Silico Antigen Screen.

    Directory of Open Access Journals (Sweden)

    Xinyue Liu

    Full Text Available Serum antibodies are valuable source of information on the health state of an organism. The profiles of serum antibody reactivity can be generated by using a high throughput sequencing of peptide-coding DNA from combinatorial random peptide phage display libraries selected for binding to serum antibodies. Here we demonstrate that the targets of immune response, which are recognized by serum antibodies directed against sequential epitopes, can be identified using the serum antibody repertoire profiles generated by high throughput sequencing. We developed an algorithm to filter the results of the protein database BLAST search for selected peptides to distinguish real antigens recognized by serum antibodies from irrelevant proteins retrieved randomly. When we used this algorithm to analyze serum antibodies from mice immunized with human protein, we were able to identify the protein used for immunizations among the top candidate antigens. When we analyzed human serum sample from the metastatic melanoma patient, the recombinant protein, corresponding to the top candidate from the list generated using the algorithm, was recognized by antibodies from metastatic melanoma serum on the western blot, thus confirming that the method can identify autoantigens recognized by serum antibodies. We demonstrated also that our unbiased method of looking at the repertoire of serum antibodies reveals quantitative information on the epitope composition of the targets of immune response. A method for deciphering information contained in the serum antibody repertoire profiles may help to identify autoantibodies that can be used for diagnosing and monitoring autoimmune diseases or malignancies.

  20. Iron and ADHD: Time to Move beyond Serum Ferritin Levels

    Science.gov (United States)

    Donfrancesco, Renato; Parisi, Pasquale; Vanacore, Nicola; Martines, Francesca; Sargentini, Vittorio; Cortese, Samuele

    2013-01-01

    Objective: (a) To compare serum ferritin levels in a sample of stimulant-naive children with ADHD and matched controls and (b) to assess the association of serum ferritin to ADHD symptoms severity, ADHD subtypes, and IQ. Method: The ADHD and the control groups included 101 and 93 children, respectively. Serum ferritin levels were determined with…

  1. Binding of disodium cromoglycate to human serum albumin

    Science.gov (United States)

    Ochoa de Aspuru, Eduardo; Zatón, Ana M. L.

    1998-07-01

    The binding of several benzopiranone derivatives to human serum albumin was determined. The antiallergic drug disodium cromoglycate binds weakly to serum albumin. However, its precursors, chromones of smaller size, were able to bind in a hydrophobic pocket in the protein, and are carried by serum albumin in blood.

  2. Iron and ADHD: Time to Move beyond Serum Ferritin Levels

    Science.gov (United States)

    Donfrancesco, Renato; Parisi, Pasquale; Vanacore, Nicola; Martines, Francesca; Sargentini, Vittorio; Cortese, Samuele

    2013-01-01

    Objective: (a) To compare serum ferritin levels in a sample of stimulant-naive children with ADHD and matched controls and (b) to assess the association of serum ferritin to ADHD symptoms severity, ADHD subtypes, and IQ. Method: The ADHD and the control groups included 101 and 93 children, respectively. Serum ferritin levels were determined with…

  3. Albendazole increases serum retinol concentration in primary schoolchildren with low serum retinol

    Directory of Open Access Journals (Sweden)

    Jane Florida Kalumpiu

    2015-08-01

    Full Text Available Background Soil-transmitted helminth (STH infection is still a health problem in Indonesia. Vitamin A deficiency (VAD is often found together with STH. The effect of the deworming program on the vitamin A status of primary school children is unknown. The objective of this study was to evaluate the effect of albendazole on serum retinol concentrations in primary school children. Methods An experimental study of pre-post test design without control was carried out on 118 children from grades 3-5 at one primary school in North Jakarta, from July to September 2014. Flotac examination was used to determine STH infections and high performance liquid chromatography was performed to measure serum retinol concentration. Stool and blood samples were taken twice: before and three weeks after treatment with albendazole 400 mg single dose for three consecutive days given to all children either infected or uninfected. Paired t-test was used to analyze the data. Results Of the 118 children, the prevalence of STH was 71.2%. The prevalence of low retinol serum (<1.05 mmol/L was 17 (17.2% and of mild VAD was 2 (2.0%. Albendazole given in 3 consecutive days could not increase serum retinol concentrations (p=0.055. Serum retinol concentration significantly increased after albendazole treatment in children with low retinol serum at baseline (p=0.001. Conclusions In children with low serum retinol concentration, treatment with albendazole increased retinol concentration. Integrated deworming and vitamin A supplementation strategies should be evaluated in this population.

  4. SERUM PARAOXONASE ACTIVITY AND ITS RELATION TO SERUM LIPIDS IN CORONARY ARTERY DISEASE

    Directory of Open Access Journals (Sweden)

    Rakhi S

    2015-10-01

    Full Text Available Human serum paraoxonase1 (PON1 is a calcium dependent es terase enzyme that hydrolyses lipid peroxides accumulating on low density lipoproteins. In the serum, PON1 enzyme is almost exclusively located on the high density lipoprotein (HDL. It hydrolyses oxidised lipids in low density lipoprotein (LDL and could therefore retard the development of atherosclerosis. As PON1 activity has a role in preventing atherosclerosis and coronary heart disease is common in Kerala, this study was conducted to assess the activity of the enzyme and its corelation to serum lipids. MATERIALS AND METHODS : In this case - control study, one hundred patients with coronary artery disease and one hundred healthy controls were included. Serum paraoxonase activity was meausured using phenyl acetate as substrate and lipid profile was done in a uto analyzer . RESULTS : The mean PON 1 activity {(70.21  27.62 in cases against (135.86  33.48 in controls (p value .000} was significantly low in CAD group compared to the control group. The CHD group had a significantly lower mean total cholesterol level (202.82  57.77 against 219.45  46.08 and lower mean HDL level (40.88  10.08 against 56.79  17.24. Correlation of the activity of PON1 with other variables for the combined group ( C ases and controls taken together showed that age is negatively correlated an d HDL is positively correlated with PON 1 activity. CONCLUSIONS : The activity of serum paraoxonase enzyme (PON1 is low in patients with coronary artery disease compared to healthy controls. Thus low serum PON1 enzyme activity is a risk factor for CAD. The activity of the serum PON1 enzyme increases with increase in HDL level and decreases with increase in age for the combined group.

  5. Relationship between the Serum Uric Acid Level, Visceral Fat Accumulation and Serum Adiponectin Concentration in Japanese Men

    National Research Council Canada - National Science Library

    Tamba, Sachiko; Nishizawa, Hitoshi; Funahashi, Tohru; Okauchi, Yukiyoshi; Ogawa, Tomoko; Noguchi, Midori; Fujita, Koichi; Ryo, Miwa; Kihara, Shinji; Iwahashi, Hiromi; Yamagata, Kazuya; Nakamura, Tadashi; Shimomura, Iichiro; Matsuzawa, Yuji

    2008-01-01

    ...). Hypoadiponectinemia is one of the key molecules of the MetS. In the present study, we investigated the relationship between the serum uric acid level, visceral fat accumulation and serum adiponectin concentration in Japanese men...

  6. Serum- and substratum-dependent modulation of neuritic growth.

    Science.gov (United States)

    Skaper, S D; Selak, I; Varon, S

    1983-01-01

    Explants of embryonic day 8 (E8) chicken dorsal root ganglia (DRG) have been cultured with medium containing serum or the serum-free supplement N1 on one of three substrata: collagen, polyornithine (PORN), or PORN exposed to a polyornithine-binding neurite-promoting factor (PNPF-PORN). Replicate cultures were maintained with or without nerve growth factor (NGF). NGF elicited its classical neuritic outgrowth on all three substrata in serum-containing or serum-free medium. In the absence of NGF, however, a gradation of increasing neurite growth was seen with: PNPF-PORN greater than PORN greater than collagen. This response occurred in both media. In addition, the neuritic halo in each instance was markedly more developed in the absence of serum, especially on PNPF-PORN. Nonneuronal behaviors reflected both serum and substratum influences: thus, nonneuronal outgrowth consisted mainly of flat cells with serum and collagen, was nonexistent with serum and PORN or PNPF-PORN, and involved mostly Schwann-like scattered cells in the absence of serum on any one substratum. The serum-dependent behaviors of ganglionic neurites were examined further with explants from chicken E11 sympathetic ganglia. A single substratum was used (PORN), without exogenous trophic factor. Neurite outgrowth was depressed by the presence of fetal calf serum, thus supporting the generality of this phenomenon. Lastly, PC12 cells, a clonal line of rat pheochromocytoma, will grow neurites in the presence of NGF after 48 hr in serum-free, but not serum-containing media. Addition of serum to serum-free cultures at this time results in the rapid and complete retraction of neurites.

  7. Serotriflin, a CRISP family protein with binding affinity for small serum protein-2 in snake serum.

    Science.gov (United States)

    Aoki, Narumi; Sakiyama, Akie; Kuroki, Kimiko; Maenaka, Katsumi; Kohda, Daisuke; Deshimaru, Masanobu; Terada, Shigeyuki

    2008-04-01

    Habu (Trimeresurus flavoviridis) serum contains 3 small serum proteins (SSP-1, SSP-2, and SSP-3) with molecular masses of 6.5 to 10 kDa. Gel filtration analysis showed that all the SSPs exist in high molecular mass forms of approximately 60 kDa in the serum. Ultrafiltration of Habu serum showed that SSPs dissociated from the complex below a pH of 4. An SSP-binding protein was purified from Habu serum by gel filtration, ion exchange, and reverse-phase HPLC. N-terminal sequencing yielded a 39-amino acid sequence, similar to the N-terminal region of triflin, which is a snake venom-derived Ca2+ channel blocker that suppresses smooth muscle contraction. The amino acid sequence of this protein, termed serotriflin, was established by peptide analysis and cDNA cloning. Serotriflin is a glycosylated protein and consists of 221 amino acids. Among the 3 SSPs, only SSP-2 formed a noncovalent complex with serotriflin. It was bound to triflin and serotriflin with high affinity, as evidenced by surface plasmon resonance. SSP-2 is considered to be a protein that prevents self injury by accidental leaking of venom into the blood.

  8. Serum fibroblast growth factor 23, serum iron and bone mineral density in premenopausal women.

    Science.gov (United States)

    Imel, Erik A; Liu, Ziyue; McQueen, Amie K; Acton, Dena; Acton, Anthony; Padgett, Leah R; Peacock, Munro; Econs, Michael J

    2016-05-01

    Fibroblast growth factor 23 (FGF23) circulates as active protein and inactive fragments. Low iron status increases FGF23 gene expression, and iron deficiency is common. We hypothesized that in healthy premenopausal women, serum iron influences C-terminal and intact FGF23 concentrations, and that iron and FGF23 associate with bone mineral density (BMD). Serum iron, iron binding capacity, percent iron saturation, phosphorus, and other biochemistries were measured in stored fasting samples from healthy premenopausal white (n=1898) and black women (n=994), age 20-55years. Serum C-terminal and intact FGF23 were measured in a subset (1631 white and 296 black women). BMD was measured at the lumbar spine and femur neck. Serum phosphorus, calcium, alkaline phosphatase and creatinine were lower in white women than black women (piron (piron. C-terminal FGF23 correlated inversely with iron (white women r=-0.134, piron iron predicted changes in C-terminal FGF23. Spine BMD correlated with iron negatively (r=-0.076, piron negatively (r=-0.119, piron did not relate to intact FGF23, but was inversely related to C-terminal FGF23. Intact FGF23 correlated with serum phosphorus. In weight-adjusted models, BMD was not related to intact FGF23, C-terminal FGF23 or iron. The influence of iron on FGF23 gene expression is not important in determining bone density in healthy premenopausal women.

  9. Predicting brain metastases of breast cancer based on serum S100B and serum HER2

    DEFF Research Database (Denmark)

    Bechmann, Troels; Madsen, Jonna Skov; Brandslund, Ivan

    2013-01-01

    Brain metastases are a major cause of morbidity and mortality in breast cancer. The aim of the current study was to evaluate the prediction of brain metastases based on serum S100B and human epidermal growth factor receptor 2 (HER2). A total of 107 breast cancer patients were included in the curr......Brain metastases are a major cause of morbidity and mortality in breast cancer. The aim of the current study was to evaluate the prediction of brain metastases based on serum S100B and human epidermal growth factor receptor 2 (HER2). A total of 107 breast cancer patients were included...... in the current study from two prospective cohort studies with either elevated serum HER2 levels >15 ng/ml or brain metastases verified by magnetic resonance imaging (MRI) or computer tomography (CT). Following the exclusion of six patients, the remaining 101 patients were divided into two groups: Group 0 (n=55......), patients with normal MRI results; and group 1 (n=46), patients with brain metastases. The levels of serum S100B and HER2 in the two groups were analyzed prior to MRI or CT of the brain, and no significant differences were identified in the serum HER2 (P=0.060) or S100B levels (P=0.623) between the groups...

  10. Serum transferrin receptor, serum ferritin and serum transferrin receptor-ferritin index in adults with iron deficiency anaemia.

    Science.gov (United States)

    Nadeem, Seema; Shah, Shahida; Iqbal, Touqueer; Iqbal, Zafar; Hanif, Ejaz

    2011-01-01

    Serum Ferritin (SF) and iron both show acute phase responses to inflammation, so iron may fall and ferritin rise independent of the marrow iron store. Bone marrow iron store has been considered the gold standard, but is invasive, painful and expensive and not suitable for everyone. Serum transferrin receptor (sTfR) which is the concentration of the soluble fragment of transferrin receptor in serum, is an important new haematological parameter. The ratio of sTfR to log SF is known as sTfR-SF index. This study was conducted to evaluate sTfR, Ferritin and sTfR-F Index in diagnosing and differentiating iron deficiency anaemia (IDA) from anaemia of chronic disease (ACD). One hundred and sixteen (116) adult subjects (80 anaemic and 36 controls) who already had their bone marrow examination done for various reasons were included in the study. sTfR, SF, and their index were measured and compared with bone marrow iron stores. Absence of iron stores denoted IDA whereas increased macrophage iron with decreased siderocytes and sideroblasts was diagnostic of ACD. Out of 80 anaemic patients, 47 were diagnosed as IDA while 33 were diagnosed as ACD. In case of IDA the diagnostic accuracy of index was 91.57%, sTfR had accuracy of 85.54% while SF had accuracy of 75.90%. In case of ACD, the diagnostic accuracy of sTfR was 91.30%, index 89.86%, while SF had accuracy of 79.71%. sTfR-SF index is a better parameter than sTfR or ferritin alone but should only be used when the results of these parameters seem altered or a bone marrow aspiration is mandatory for diagnosis of ACD. The estimation of sTfR or index may offer a simple non invasive method that may enable more accurate assessments of iron status in such patients.

  11. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sorensen, G.L.; Hjelmborg, J.V.; Kyvik, K.O.

    2006-01-01

    defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located...... in the NH(2)-terminal region (Met11Thr) of the mature protein is significantly associated with the serum SP-D levels. A classic twin study was performed on a twin population including 1,476 self-reported healthy adults. The serum SP-D levels increased with male sex, age, and smoking status. The intraclass...... correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum...

  12. Low serum alkaline phosphatase activity in Wilson's disease.

    Science.gov (United States)

    Shaver, W A; Bhatt, H; Combes, B

    1986-01-01

    Low values for serum alkaline phosphatase activity were observed early in the course of two patients with Wilson's disease presenting with the combination of severe liver disease and Coombs' negative acute hemolytic anemia. A review of other cases of Wilson's disease revealed that 11 of 12 patients presenting with hemolytic anemia had values for serum alkaline phosphatase less than their respective sex- and age-adjusted mean values; in eight, serum alkaline phosphatase activity was less than the lower value for the normal range of the test. Low values for serum alkaline phosphatase were much less common in Wilson's disease patients with more chronic forms of presentation. Copper added in high concentration to serum in vitro did not have an important effect on serum alkaline phosphatase activity. The mechanism responsible for the decrease in serum alkaline phosphatase activity in patients is uncertain.

  13. EXPLORE ON DIAGNOSTIC AND PROGNOSTIC VALUES OF EXTRAHEPATIC CHOLANGIOCARCINOMA:UTILITY OF SERUM CA19-9 AND SERUM CEA

    Institute of Scientific and Technical Information of China (English)

    秦兴雷; 王作仁; 鲁敏; 王林; 何铨儒

    2004-01-01

    Objective To evaluate the validity of a serum carbohydrate antigen 19-9(CA19-9) determination in the diagnosis of extrahepatic cholangiocarcinoma(EHCC). Methods Serum CA19-9 concentration and serum carcinoembryonic antigen(CEA) concentration were prospectively measured by an immunoradiometric assay without knowledge of the clinical diagnosis in patients with EHCC (n=51), benign biliary diseases (n=42), and healthy individuals (n=15). Using a receiver operating characteristic (ROC) curve define a new strategy for interpreting CA19-9 and CEA in EHCC. Results The sensitivity of CA19-9 and CEA in diagnosing EHCC were 86.3% (44/51) and 25.5%(13/51), respectively. When compared with the benign biliary diseases group, the true negative rates of serum CA19-9 and serum CEA were 85.7% (36/42) and 95.2%(40/42), respectively. The false positive rates of serum CA19-9 and serum CEA were 14.3 %( 6/42) and 4.8 %( 2/42), whereas the accuracy of serum CA19-9 and serum CEA were 57.0%(53/93) and 86.0%(80/93), respectively. Serum CA19-9 concentration and serum CEA concentration were significantly elevated (P0.05), respectively. In 22 patients undergoing curative resection of EHCC, the mean serum CA19-9 concentration decreased from preoperative level of (456.6±120.4) kU·L-1 to postoperative level (62.8±17.3) kU·L-1(P<0.001). The outcomes showed that serum CA19-9 had greater diagnosis performances than serum CEA. Conclusion Serum CA19-9 is an effective tumor marker in diagnosing of cholangiocarcinoma, deciding whether the tumor has been radically resected and monitoring effect of treatment

  14. A study of serum magnesium and serum calcium in major depressive disorder

    Directory of Open Access Journals (Sweden)

    Niladri Deb

    2016-01-01

    Full Text Available Background: Depression is arguably the epidemic of our time. In one of the reports of the World Health Organization, it was projected that depression and heart disease will be the most common diseases on Earth by 2020. The paper attempts to determine the relationship between the pathophysiology of major depressive disorder (MDD and serum magnesium and serum calcium levels. Methodology: The study is based on a sample of 60 subjects–30 healthy normal individuals and 30 indoor and outdoor patients of the Department of Psychiatry of Gauhati Medical College and Hospital, Guwahati, who were identified by psychiatrist as having MDD as per the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV criteria. Estimation of the parameters of the study was done by using spectrophotometre (Spectra scan UV 2600. The result values of both the groups were compared by using independent ‘t’ test. Further, the correlation between serum magnesium and serum calcium among individuals of control as well as case groups were carried out by using Pearson’s correlation test. Results: Analysis of data reveals the mean value of serum magnesium among subjects of control group was higher than that of the subjects of the case (before treatment group while in case of serum calcium, it was the vice versa. Moreover, in both the cases, the differences in mean values were found to be statistically significant. Conclusion: The present study shows that alterations in the concentrations of magnesium and calcium may play a role in depressive illnesses and thus, may have a possible role in causing various mood disorders like MDD.

  15. Serum Albumin Domain Structures in Human Blood Serum by Mass Spectrometry and Computational Biology.

    Science.gov (United States)

    Belsom, Adam; Schneider, Michael; Fischer, Lutz; Brock, Oliver; Rappsilber, Juri

    2016-03-01

    Chemical cross-linking combined with mass spectrometry has proven useful for studying protein-protein interactions and protein structure, however the low density of cross-link data has so far precluded its use in determining structures de novo. Cross-linking density has been typically limited by the chemical selectivity of the standard cross-linking reagents that are commonly used for protein cross-linking. We have implemented the use of a heterobifunctional cross-linking reagent, sulfosuccinimidyl 4,4'-azipentanoate (sulfo-SDA), combining a traditional sulfo-N-hydroxysuccinimide (sulfo-NHS) ester and a UV photoactivatable diazirine group. This diazirine yields a highly reactive and promiscuous carbene species, the net result being a greatly increased number of cross-links compared with homobifunctional, NHS-based cross-linkers. We present a novel methodology that combines the use of this high density photo-cross-linking data with conformational space search to investigate the structure of human serum albumin domains, from purified samples, and in its native environment, human blood serum. Our approach is able to determine human serum albumin domain structures with good accuracy: root-mean-square deviation to crystal structure are 2.8/5.6/2.9 Å (purified samples) and 4.5/5.9/4.8Å (serum samples) for domains A/B/C for the first selected structure; 2.5/4.9/2.9 Å (purified samples) and 3.5/5.2/3.8 Å (serum samples) for the best out of top five selected structures. Our proof-of-concept study on human serum albumin demonstrates initial potential of our approach for determining the structures of more proteins in the complex biological contexts in which they function and which they may require for correct folding. Data are available via ProteomeXchange with identifier PXD001692.

  16. Reference Interval and Status for Serum Folate and Serum Vitamin B12 in a Norwegian Population.

    Science.gov (United States)

    Schwettmann, Lutz; Berbu, Siw

    2015-01-01

    Deficiencies of folate and vitamin B12 lead to an elevated serum concentration of homocysteine which has been associated with many diseases including cardiovascular disease. Laboratory algorithms often include initial testing of serum folate and vitamin B12. Reference intervals for these vitamins can vary significantly among populations for which dietary intakes may be different. The aim of this study was to establish reference intervals in a Norwegian population and to assess the folate and vitamin B12 status related to reference intervals. Blood samples were taken from 144 healthy volunteers aged 18 - 65 years. A questionnaire provided data of medication, medical history, vitamin supplementation, alcohol consumption, and use of oral contraceptives and others. Serum folate and vitamin B12 concentrations were measured on the Abbott Architect i2000. Reference values were calculated using the bootstrap method. Results of serum folate, vitamin B12, and homocysteine from 1190 individuals from regional primary health care centers were evaluated related to reference values and the proportion of individuals with deficiency was estimated. Mean serum concentrations of folate and vitamin B12 were 11.9 nmol/L and 328 pmol/L, respectively. Men were found to have statistically significant higher vitamin B12 concentrations than women. 95%-reference intervals were calculated to 5.2 - 29.2 nmol/L for folate and 133 - 595 pmol/L for vitamin B12. 1.1% of the study population has serum vitamin B12-concentrations vitamin B12 for a healthy, not vitamin-supplemented adult population were determined from 144 subjects. The application of these intervals will assist in the evaluation of folate and vitamin status.

  17. Molecular sex differences in human serum.

    Directory of Open Access Journals (Sweden)

    Jordan M Ramsey

    Full Text Available BACKGROUND: Sex is an important factor in the prevalence, incidence, progression, and response to treatment of many medical conditions, including autoimmune and cardiovascular diseases and psychiatric conditions. Identification of molecular differences between typical males and females can provide a valuable basis for exploring conditions differentially affected by sex. METHODOLOGY/PRINCIPAL FINDINGS: Using multiplexed immunoassays, we analyzed 174 serum molecules in 9 independent cohorts of typical individuals, comprising 196 males and 196 females. Sex differences in analyte levels were quantified using a meta-analysis approach and put into biological context using k-means to generate clusters of analytes with distinct biological functions. Natural sex differences were established in these analyte groups and these were applied to illustrate sexually dimorphic analyte expression in a cohort of 22 males and 22 females with Asperger syndrome. Reproducible sex differences were found in the levels of 77 analytes in serum of typical controls, and these comprised clusters of molecules enriched with distinct biological functions. Analytes involved in fatty acid oxidation/hormone regulation, immune cell growth and activation, and cell death were found at higher levels in females, and analytes involved in immune cell chemotaxis and other indistinct functions were higher in males. Comparison of these naturally occurring sex differences against a cohort of people with Asperger syndrome indicated that a cluster of analytes that had functions related to fatty acid oxidation/hormone regulation was associated with sex and the occurrence of this condition. CONCLUSIONS/SIGNIFICANCE: Sex-specific molecular differences were detected in serum of typical controls and these were reproducible across independent cohorts. This study extends current knowledge of sex differences in biological functions involved in metabolism and immune function. Deviations from typical

  18. Serum amyloid a in clinical practice

    Directory of Open Access Journals (Sweden)

    Jovanović Dijana B.

    2004-01-01

    Full Text Available Serum amyloid A (SAA is an acute phase first class protein discovered a quarter of the century ago. Its concentration depends on clinical findings of the patient, illness activity and the therapy applied. SAA increases moderately to markedly (100-1000 mg/l in bacterial and fungal infections, invasive malignant diseases, tissue injuries in the acute myocardial infarction and autoimmune diseases such as rheumatoid arthritis and vasculitis. Mild elevation (10-100 mg/l is often seen in viral infections, systemic lupus erythematosus and localized inflammation or tissue injuries in cystitis and cerebral infarction. SAA as sensitive, non-invasive parameter is used in organ transplantation where early and correct diagnosis is needed as well as where prompt therapy is required. Besides acute kidney allograft rejection, SAA is used in the diagnosis of rejection after liver transplantation, simultaneous pancreas and kidney transplantation and also in bone marrow transplantation (acute „graft vs. host disease". Simultaneous determination of C-reactive protein (CRP and SAA may point to acute kidney allograft rejection. Standard immunosuppressive therapy with cyclosporine A and prednisolone significantly suppresses the acute phase CRP reaction both in operation itself and acute rejection, but not in infection. On the other hand, SAA rejection in operation, acute allograft rejection and infection is present in spite of cyclosporine A and steroids therapy. Different reaction of SAA and CRP in transplant patients to cyclosporine A therapy helps in differentiation between the infection and rejection. Although CRP and SAA are sensitive and acute phase reactants, their serum concentrations cannot be valued as prognostic and diagnostic criteria without creatinine serum concentration and clinical findings. In addition, they offer important information for clinical diagnosis as well as the kind of therapy.

  19. Serum fructosamine and retinopathy of prematurity.

    Science.gov (United States)

    Bozdag, Senol; Oguz, Serife Suna; Gokmen, Tulin; Tunay, Zuhal; Tok, Levent; Uras, Nurdan; Erdeve, Omer; Dilmen, Ugur

    2011-12-01

    To determine whether serum fructosamine which is a good marker for detecting hyperglycemia during the previous 2 to 3 wk in infants could predict the development of retinopathy of prematurity in very low birth weight infants. One hundred sixty seven premature infants who had a birth weight of < 1500 g and a gestational age of less than 32 wk were investigated in the present study. Blood glucose was measured at the bedside and infants were recorded as hyperglycemic if their mean blood glucose levels were higher than 150 mg/dL. Serum corrected fructosamine level was obtained from the cord blood at birth and after the first month of life. The infants' eyes were examined by ophthalmologists to detect retinopathy of prematurity at the gestational age of 32 wk or at four wk after birth, whichever came first. Corrected fructosamine was 319.6 ± 59.6 and 272.8 ± 50.6 mmol/l for group 1 on 1(st) and 30(th) day respectively; 320 ± 61.7 and 268.2 ± 47.3 mmol/l for groups 2 + 3 on 1(st) and 30(th) day respectively which did not differ between groups (p = 0.766 and p = 0.665), whereas duration of hyperglycemia was 1.69 ± 1.1 day in group 1 compared with 3.05 ± 2.4 day in groups 2 + 3 which was significantly different (p = 0.019). The multivariate regression analysis indicated that the duration of hyperglycemia in days was significantly correlated with the development of retinopathy of prematurity (OR 3.26; 95% CI 1.09-9.80; p = 0.035). Although the duration of hyperglycemia may contribute to the development of retinopathy of prematurity, serum corrected fructosamine does not have a good predictive value in developing retinopathy of prematurity in very-low-birth-weight (VLBW) infants.

  20. Serum dioxin concentration and age at Menarche

    Energy Technology Data Exchange (ETDEWEB)

    Warner, M.; Samuels, S.; Eskenazi, B. [Univ. of California at Berkeley (United States); Mocarelli, P.; Gerthoux, P.M. [Univ. of Milano-Biococca (Italy); Needham, L.; Patterson, D. Jr. [Centers for Disease Control and Prevention (United States)

    2004-09-15

    To date, no epidemiologic studies have examined the association of TCDD exposure and age at menarche. Three studies, however, have examined the relation of dioxin-like compounds to pubertal development, with inconsistent conclusions. A study of daughters of Michigan women who had consumed polybrominated biphenyls (PBB) in food in 1973, found an earlier age at menarche among daughters whose mothers had higher serum PBB levels 8. No differences were found in age at menarche of Taiwanese women who were exposed postnatally (but premenarche) to PCBs and PCDFs via consumption of contaminated rice oil (Yu-Cheng) compared to unexposed 9. In Flemish adolescents, there was no relation of age at menarche with current serum levels of dioxin-like compounds as measured by Chemical Activated LUciferase gene eXpression bioassay toxic equivalents (CALUX-TEQ) or individual PCB congeners 118, 153, and 180 10. On July 10, 1976, as a result of a chemical explosion, residents of Seveso, Italy experienced the highest levels of TCDD exposure in a human population. Twenty years later (1996-1998), the Seveso Women's Health Study (SWHS), a retrospective cohort study, was initiated to determine whether the women were at higher risk for reproductive disease. Among participants in SWHS, we have observed that TCDD levels are associated with an increase in menstrual cycle length among those who were premenarcheal at exposure, but not in those who were postmenarcheal at exposure 11. Consistent with animal studies 12, this suggests that females may be particularly susceptible to the effects of TCDD during early stages of development, e.g. in utero or pre-pubertal. Thus, here we examine the association of individual serum TCDD and age of menarche among women who were premenarcheal in 1976, at the time of explosion.

  1. Serum potassium concentrations: Importance of normokalaemia.

    Science.gov (United States)

    Heras, Manuel; Fernández-Reyes, María José

    2017-06-21

    Abnormalities in potassium concentrations are associated with morbidity and mortality. In recent years it has been considered that small variations in serum potassium concentrations within normal intervals may also be associated with mortality. Strategies for achieving normokalaemia include dietary measures, limiting the use of potassium retaining drugs, and use of conventional cation exchange resins (calcium/sodium polystyrene sulfonate) and/or the new non-absorbed cation exchange polymer (patiromer). Copyright © 2017 Elsevier España, S.L.U. All rights reserved.

  2. Interaction of Nicotine and Bovine Serum Albumin

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The binding of nicotine to bovine serum albumin (BSA) was studied by UV absorption, fluorescence, and 1H NMR methods. With the addition of nicotine, the absorption band of BSA at about 210 nm decreased gradually, moved to longer wavelengths, and narrowed. BSA fluorescence of tryptophan residue was quenched by nicotine. The 1H NMR peaks of nicotine moved to downfield by the addition of BSA. The experimental results showed that nicotine was capable of binding with BSA to form a 1:1 complex. BSA's high selectivity for nicotine binding suggests a unique role for this protein in the detoxification and/or transport of nicotine.

  3. Osteoprotegerin Serum Levels in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Kolarz G

    2003-01-01

    Full Text Available Osteoprotegerin (OPG wird in der entzündeten Synovialis bei chronischer Polyarthritis von Makrophagen, dendritischen Zellen und B-Zellen produziert. OPG hemmt die Differenzierung von Osteoklasten und ihre Aktivierung durch die Bindung an RANKL (receptor activator of NFkappaBbeta ligand. OPG könnte daher eine wichtige Rolle in der Verhinderung des Auftretens von Erosionen bei chronischer Polyarthritis spielen. In der vorliegenden Studie wurde untersucht, ob die Serumspiegel von Osteoprotegerin mit Entzündungsparametern oder der Gelenksdestruktion bei chronischer Polyarthritiskorrelieren. Patienten und Methoden: In 85 Sera von 68 Patienten mit chronischer Polyarthritis wurden die Serumspiegel von OPG untersucht und mit klinischen Parametern, Laborbefunden und dem Larsen-Score verglichen. Zur OPG-Bestimmung wurde ein Sandwich-ELISA (enzyme-linked-im-munosorbent-assay mit zwei spezifischen OPG-Antikörpern verwendet. Mit dem bindenden Antikörper wurde eine Mikrotiterplatte beladen, der zweite Biotin-konjugierte Antikörper wurde durch Streptavidin-Peroxidase und TMB gemessen. Der Immunoassay mißt sowohl freies als auch an RANKL gebundenes OPG. Zur statistischen Auswertung wurde ein SAS-Programm verwendet, als Tests wurden der Mann-Whitney-Test, die Spearman-Korrelation und die Varianzanalyse (ANOVA angewandt. Ergebnisse: Verglichen mit einer gesunden Kontrollgruppe (Alter 56-76 Jahre lagen die Mittelwerte von Serum-OPG bei chronischer Polyarthritis höher. Wenn man den Mittelwert der Gesunden für die Einteilung in höhere oder niedrigere Serum-OPG-Werte bei Patienten mit chronischer Polyarthritis verwendet, zeigt sich im Mann-Whitney-Test eine signifikante Korrelation nicht nur mit BSG und Rheumafaktor, sondern auch mit dem Larsen-Score. Die OPG-Spiegel korrelierten mit dem Rheumafaktor, der BSG und dem Serum-CRP (C-reaktives Protein signifikant positiv, der Larsen-Score zeigte nur einen statistischen Trend. Diskussion: OPG blockiert die

  4. Serum zinc levels in gestational diabetes

    Directory of Open Access Journals (Sweden)

    Rahimi Sharbaf F

    2008-12-01

    Full Text Available "nBackground: Maternal zinc deficiency during pregnancy has been related to adverse pregnancy outcomes. Most studies in which pregnant women have been supplemented with zinc to examine its effects on the outcome of the pregnancy have been carried out in industrialized countries and the results have been inconclusive. It has been shown that women with gestational diabetes (GDM have lower serum zinc levels than healthy pregnant women, and higher rates of macrosomia. Zinc is required for normal glucose metabolism, and strengthens the insulin-induced transportation of glucose into cells by its effect on the insulin signaling pathway. The purpose of this study was to assess the serum zinc levels of GDM patients and evaluate the effect of zinc supplementation. "nMethods: In the first stage of this prospective controlled study, we enrolled 70 women who were 24-28 weeks pregnant at the Prenatal Care Center of Mirza Kochak Khan Hospital, Tehran, Iran. The serum zinc level of each subject was determined. In the second stage, among these 70 subjects, the diabetics receiving insulin were divided into two groups, only one of which received a zinc supplement and the other group was the control group. Birth weight of neonates and insulin dosages were recorded. "nResults: The mean serum zinc level in the GDM group was lower than that of the control group (94.83 vs. 103.49mg/dl, respectively and the mean birth weight of neonates from the GDM women who received the zinc supplement was lower than that of the control group (3849g vs. 4136g. The rate of macrosomia was lower in the zinc supplemented group (20% vs. 53%. The mean of increase of insulin after receiving the zinc supplement was lower (8.4u vs. 13.53. "nConclusion: Maternal insulin resistance is associated with the accumulation of maternal fat tissue during early stages of pregnancy and greater fetoplacental nutrient availability in later stages, when 70% of fetal growth occurs, resulting in macrosomia. In

  5. Purification of human serum hyaluronidase using chromatofocusing

    DEFF Research Database (Denmark)

    Fenger, M

    1982-01-01

    A commercial chromatofocusing system was applied to Cohn's fraction III of human serum to purify hyaluronidase (E.C.3.2.1.3.5). The protein that eluted in the pH range 4.7-5.3 was pooled and precipitated by adding ammonium sulphate to 50% saturation. This sequence of fractionation purified...... hyaluronidase extensively by immunological criteria. It is shown that hyaluronidase is a population of enzymes displaying microheterogeneity. The commercial chromatofocusing system behaved as theoretically expected. The capacity of the gel is 3 mg per ml gel. Any overload will be trapped or precipitated...

  6. Serum Antibodies to Glycans in Peripheral Neuropathies.

    Science.gov (United States)

    Sonnino, Sandro; Chiricozzi, Elena; Ciampa, Maria Grazia; Mauri, Laura; Prinetti, Alessandro; Toffano, Gino; Aureli, Massimo

    2017-03-01

    In peripheral neuropathies, such as sensorimotor neuropathies, motor neuron diseases, or the Guillain-Barré syndrome, serum antibodies recognizing saccharide units, portion of oligosaccharides, or oligosaccharide chains, have been found. These antibodies are called anti-glycosphingolipid (GSL) or anti-ganglioside antibodies. However, the information on the aglycone carrying the hydrophilic oligosaccharide remains elusive. The absolute and unique association of GSL to the onset, development and symptomatology of the peripheral neuropathies could be misleading. Here, we report some thoughts on the matter.

  7. Total serum calcium level may have adverse effects on serum cholesterol and triglycerides among female university faculty and staffs.

    Science.gov (United States)

    He, Lianping; Qian, Yifan; Ren, Xiaohua; Jin, Yuelong; Chang, Weiwei; Li, Jie; Chen, Yan; Song, Xiuli; Tang, Hui; Ding, Lingling; Guo, Daoxia; Yao, Yingshui

    2014-03-01

    Our previous studies showed that serum calcium level may have influence in the blood pressure to older male subjects, but the relationship between serum calcium level and blood lipids is unclear. The aim of this study was to evaluate the relationship between total serum calcium level and blood lipids. In our study, total serum calcium level and blood lipids were measured among 1,075 subjects, with age range of 30-60 years, who were recruited for the routine health screening in 2006. The results showed that serum calcium level was positively correlated with triglyceride and total cholesterol weight, but not HDL-cholesterol and LDL-cholesterol in female subjects (P 0.05). These findings suggest that a higher total serum calcium level may have a adverse effects on serum cholesterol and triglycerides among female subjects.

  8. Serum adiponectin levels in gestational diabetes mellitus

    Science.gov (United States)

    Bhograj, Abhijit; Suryanarayana, K. M.; Nayak, Ashwini; Murthy, N. S.; Dharmalingam, Mala; Kalra, Pramila

    2016-01-01

    Introduction: Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance with onset or first recognition during pregnancy.[1] Pregnancy is a unique situation in which there is a physiological temporary increase in insulin resistance (IR). The mechanisms responsible for the gestational-induced IR are not completely understood. The current study was undertaken to compare adiponectin levels during 24–28 weeks period of gestation in drug-naive newly diagnosed GDM women with a cohort of normoglycemic pregnant women. Subjects and Methods: A total of 47 pregnant women in the age group of 18–40 years were included in this cross-sectional study, of which 13 were GDM cases and 34 were normoglycemic controls. Serum adiponectin level was analyzed by enzyme-linked immunosorbent assay. Results: The mean adiponectin level was 16.92 ng/ml (standard deviation [SD] = 2.78) and 19.38 ng/ml (SD = 2.71) in case and control groups, respectively, and the difference was found to be statistically significant (P = 0.008). Conclusion: Our study demonstrated decreased serum adiponectin levels in women with GDM when compared with age- and body mass index-matched euglycemic pregnant women. PMID:27867874

  9. Serum prolactin concentrations are elevated after syncope.

    Science.gov (United States)

    Oribe, E; Amini, R; Nissenbaum, E; Boal, B

    1996-07-01

    The distinction between syncope and epileptic seizures is a common clinical diagnostic problem. Elevated serum prolactin (PRL) concentrations are used to help differentiate epileptic from nonepileptic attacks such as pseudoseizures. Reports of PRL concentrations following syncope have been variable. To determine whether PRL rises after syncope, we measured serum PRL concentrations during a 45-minute passive 60-degree head-up tilt in 21 patients with a history of near-fainting or syncope. Head-up tilt triggered hypotension (mean arterial pressure 51 mm Hg, 95% CI = 45-57) with syncope in 11 patients. PRL concentrations were elevated ( > 19 ng/mL) and reached a maximum within the first 30 minutes after tilt-induced syncope in nine patients (PRL supine: 11 ng/mL, 95% CI = 7-15, vs. PRL after syncope: 52 ng/mL, 95% CI = 36-67; a greater than fourfold rise), while they remained unchanged in 10 patients who had a normal response to head-up tilt (PRL supine: 6 ng/mL, 95% CI = 5-8, vs. maximum PRL while upright: 8 ng/mL, 95% CI = 6-10). The findings indicate that elevated PRL concentrations are present after hypotensive syncope and are of little use in differentiating such syncope from epileptic seizures.

  10. Immune functions of serum amyloid A.

    Science.gov (United States)

    Eklund, Kari K; Niemi, K; Kovanen, P T

    2012-01-01

    Serum amyloid A (SAA) is a highly conserved, acute-phase protein synthesized predominantly by the liver. After secretion into the circulation, it associates with high-density lipoprotein (HDL) particles. During acute inflammation, serum SAA levels may rise up to 1000-fold, and under these conditions, SAA displaces apolipoprotein A-I from HDL, thus becoming the major apolipoprotein of circulating HDL3. SAA exhibits significant immunological activity by, for example, inducing the synthesis of several cytokines and by being chemotactic for neutrophils and mast cells. It exerts many of its immunological activities by binding and activating cell-surface receptors, including Toll-like receptor (TLR) 2 and TLR4, formyl peptide receptor-like 1 (FPRL1), class B scavenger receptor CD36, and the ATP receptor P2X7. SAA also recently has been shown to activate the inflammasome cascade, which has a key role in immune activation, thus further stressing the unique role of SAA in immunomodulation. Traditionally, SAA has been considered to have a key role in the pathogenesis of amyloid A-type amyloidosis, but we now understand that it may also participate in the pathogenesis of chronic inflammatory diseases, such as rheumatoid arthritis and atherosclerosis. Thus, SAA is one potential target in the treatment of diseases associated with chronic inflammation. The purpose of this review is to shed light on SAA as an immunologically active protein. We also focus on the recent findings implicating SAA in the regulation of the inflammasome cascade.

  11. Interaction of Citrinin with Human Serum Albumin

    Directory of Open Access Journals (Sweden)

    Miklós Poór

    2015-12-01

    Full Text Available Citrinin (CIT is a mycotoxin produced by several Aspergillus, Penicillium, and Monascus species. CIT occurs worldwide in different foods and drinks and causes health problems for humans and animals. Human serum albumin (HSA is the most abundant plasma protein in human circulation. Albumin forms stable complexes with many drugs and xenobiotics; therefore, HSA commonly plays important role in the pharmacokinetics or toxicokinetics of numerous compounds. However, the interaction of CIT with HSA is poorly characterized yet. In this study, the complex formation of CIT with HSA was investigated using fluorescence spectroscopy and ultrafiltration techniques. For the deeper understanding of the interaction, thermodynamic, and molecular modeling studies were performed as well. Our results suggest that CIT forms stable complex with HSA (logK ~ 5.3 and its primary binding site is located in subdomain IIA (Sudlow’s Site I. In vitro cell experiments also recommend that CIT-HSA interaction may have biological relevance. Finally, the complex formations of CIT with bovine, porcine, and rat serum albumin were investigated, in order to test the potential species differences of CIT-albumin interactions.

  12. Lower Serum Paraoxonase-1 Activity Is Related to Higher Serum Amyloid A Levels in Metabolic Syndrome

    NARCIS (Netherlands)

    Kappelle, Paul Jan Willem Herman; Bijzet, Johan; Hazenberg, Bouke Pier; Dullaart, Robin Pieter Frank

    Background and Aims. High-density lipoproteins (HDL) contain the anti-oxidative enzyme, paraoxonase-1 (PON-1), which is important for atheroprotection. The acute phase reactant, serum amyloid A (SAA), an HDL-associated apolipoprotein, may impair PON-1 activity, whereas SAA and PON-1 are reciprocally

  13. Discovery and identification of Serum Amyloid A protein elevated in lung cancer serum

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two hundred and eighteen serum samples from 175 lung cancer patients and 43 healthy individuals were analyzed by using Surface Enhaced Laser Desorption/Ionization Time of Flight Mass Spectrome- try (SELDI-TOF-MS). The data analyzed by both Biomarker Wizard? and Biomarker Patterns? software showed that a protein peak with the molecular weight of 11.6 kDa significantly increased in lung cancer. Meanwhile,the level of this biomarker was progressively increased with the clinical stages of lung cancer. The candidate biomarker was then obtained from tricine one-dimensional sodium dodecyl sul- fate-polyacrylamide gel electrophoresis by matching the molecular weight with peaks on WCX2 chips and was identified as Serum Amyloid A protein (SAA) by MALDI/MS-MS and database searching. It was further validated in the same serum samples by immunoprecipitation with commercial SAA antibody. To confirm the SAA differential expression in lung cancer patients, the same set of serum samples was measured by ELISA assay. The result showed that at the cutoff point 0.446(OD value)on the Receiver Operating Characteristic (ROC) curve, SAA could better discriminate lung cancer from healthy indi- viduals with sensitivity of 84.1% and specificity of 80%. These findings demonstrated that SAA could be characterized as a biomarker related to pathological stages of lung cancer.

  14. Lower Serum Paraoxonase-1 Activity Is Related to Higher Serum Amyloid A Levels in Metabolic Syndrome

    NARCIS (Netherlands)

    Kappelle, Paul Jan Willem Herman; Bijzet, Johan; Hazenberg, Bouke Pier; Dullaart, Robin Pieter Frank

    2011-01-01

    Background and Aims. High-density lipoproteins (HDL) contain the anti-oxidative enzyme, paraoxonase-1 (PON-1), which is important for atheroprotection. The acute phase reactant, serum amyloid A (SAA), an HDL-associated apolipoprotein, may impair PON-1 activity, whereas SAA and PON-1 are reciprocally

  15. Blood serum components and serum protein test of Hybro-PG broilers of different ages

    Directory of Open Access Journals (Sweden)

    PRL Silva

    2007-12-01

    Full Text Available Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1, 30 from 35-day-old birds (G2, and 30 from 42-day-old birds (G3, with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT, aspartate aminotransferase (AST, creatine kinase (CK, alkaline phosphatase (ALP, and lactate dehydrogenase (LDH, serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

  16. Reference intervals for serum cystatin C and serum creatinine in adults.

    Science.gov (United States)

    Erlandsen, E J; Randers, E; Kristensen, J H

    1998-06-01

    The aim of this study was to establish reference intervals for cerum cystatin C and serum creatinine in adults. Blood samples were collected from 270 healthy blood donors (135 men and 135 women between 20 and 65 years old with 15 men and 15 women in each five-year-interval). Serum cystatin C was analyzed using an automated particle-enhanced immunoassay (DAKO Cystatin C PET kit) on the Cobas Mira S analyzer. Serum creatinine was analyzed using the Vitros Creatinine Slide, an enzymatic method on the Vitros 950 chemistry analyzer. The calculated reference intervals for serum cystatin C were 0.62-1.15 mg/l in women (median 0.84 mg/l, range 0.56-1.29 mg/l) and 0.51-1.25 mg/l in men (median 0.87 mg/l, range 0.42-1.39 mg/l). The Mann-Whithey U-test revealed no gender-related difference for cystatin C (p = 0.48). A common reference interval in women and men was calculated to be 0.54-1.21 mg/l (median 0.85 mg/l, range 0.42-1.39 mg/l). The non-parametric reference interval for serum creatinine was 57-95 mumol/l in women (median 72 mumol/l, range 44-105 mumol/l) and 69-111 mumol/l in men (median 89 mumol/l, range 58-123 mumol/l).

  17. Serum Iron, Haemoglobin and Serum Lipid Peroxidation in Neonates with Respiratory Disorders

    Directory of Open Access Journals (Sweden)

    Sushama P. Dhonde

    2010-01-01

    Full Text Available Respiratory disorders are the most common cause for higher morbidity and mortality rate in India. Higher oxygen concentration of extrauterine existence causes increased erythrocyte lysis lead to release of iron in neonates. Iron is known to catalyze the formation of Reactive oxygen species. Involvement of hemoglobin and iron in oxygen-mediated reactions stimulate us to study the role of these in neonates.Objectives :iTo estimate the concentration of Haemoglobin, serum iron, serum lipid peroxidation in neonates with respiratory disorders; and compare those with that of healthy controls. iiTo study the role of these parameters in neonates with respiratory disorders.Materials and Methods: Present study includes 50 neonates suffering from respiratory disorders and 50 healthy neonates as controls. Samples collected from these were used for the estimation of haemoglobin, serum iron, and serum lipid peroxidation.Observations: Significantly (p<0.001 increased levels of serum iron and lipid peroxidation were observed in neonates with respiratory disorders when compared those with control values. These levels were found significantly (p<0.001 higher in preterm than full-term neonates. Concentration of haemoglobin showed no significant difference in both groups.Conclusion: Exacerbation of oxidative stress in neonates with respiratory disorders may be due to hypoxia induced free radical generation, higher oxidative tendency of HbF and elevated iron. Premature neonates are probably unprepared for extra uterine life in an oxygen rich environment and due to this they are more prone to oxidative insult. Thus this study reveals the pro-oxidant role of HbF and iron, which enhances the oxidative stress in respiratory disorder.

  18. Superovulation in the cow with pregnant mare serum gonadotrophin: effects of dose and antipregnant mare serum gonadotrophin serum.

    Science.gov (United States)

    Gonzalez, A; Wang, H; Carruthers, T D; Murphy, B D; Mapletoft, R J

    1994-01-01

    The effects of pregnant mare serum gonadotrophin (PMSG) dose and PMSG antiserum on superovulation in crossbred beef cows were studied. In experiment I, three groups were treated with 1200, 2400 or 3600 IU of PMSG and 48 h later with prostaglandin (PGF). The mean numbers of corpora lutea (CL), unovulated follicles, and total ova/embryos collected increased as the PMSG dose increased. The percent of fertilized ova and transferable embryos was lowest in the highest dose group (p superovulation of beef cows with PMSG and treatment with PMSG antiserum will induce a higher superovulatory response and will result in higher CL numbers and fewer unovulated follicles. Further, the variability in the superovulatory response to PMSG treatment was still evident when PMSG antiserum was administered. PMID:8055430

  19. Severe serum sickness reaction to oral and intramuscular penicillin.

    Science.gov (United States)

    Clark, Brychan M; Kotti, George H; Shah, Anand D; Conger, Nicholas G

    2006-05-01

    Serum sickness is a type III hypersensitivity reaction mediated by immune complex deposition with subsequent complement activation, small-vessel vasculitis, and tissue inflammation. Although the overall incidence of serum sickness is declining because of decreased use of heterologous sera and improved vaccinations, rare sporadic cases of serum sickness from nonprotein drugs such as penicillins continue to occur. Drug-induced serum sickness is usually self-limited, with symptoms lasting only 1-2 weeks before resolving. We report an unusual case of a severe and prolonged serum sickness reaction that occurred after exposure to an intramuscular penicillin depot injection (probable relationship by Naranjo score) and discuss how pharmacokinetics may have played a role. Clinicians should be familiar with serum sickness reactions particularly as they relate to long-acting penicillin preparations. Accurate diagnosis in conjunction with cessation of drug exposure and prompt initiation of antiinflammatory treatment with corticosteroids can produce complete recovery

  20. Relation of serum uric acid to cardiovascular disease.

    Science.gov (United States)

    Wu, Audrey H; Gladden, James D; Ahmed, Mustafa; Ahmed, Ali; Filippatos, Gerasimos

    2016-06-15

    This review summarizes recent published literature on the association between serum uric acid and cardiovascular disease, a relationship which is complex and not fully elucidated. Uric acid may be a marker for risk, a causative agent in cardiovascular disease, or both. Various biologic factors can influence serum uric acid levels, and serum uric acid level itself is closely related to conditions such as hypertension, dyslipidemia, obesity, and impaired glucose metabolism, that contribute to cardiovascular disease pathophysiology. Serum uric acid levels have been found to be associated with adverse outcomes, including mortality, in the general population. In addition, serum uric acid is associated with increased risk for incident coronary heart disease, heart failure, and atrial fibrillation. In the setting of established systolic heart failure, serum uric acid is positively associated with disease severity and mortality risk. Whether targeting treatment based on uric acid levels might affect clinical outcomes is still being studied.

  1. Genetic and environmental influences of surfactant protein D serum levels

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Hjelmborg, Jacob v. B.; Kyvik, Kirsten Ohm

    2006-01-01

    defining the constitutional serum level of SP-D and determine the magnitude of the genetic contribution to serum SP-D in the adult population. Recent studies have demonstrated that serum SP-D concentrations in children are genetically determined and that a single nucleotide polymorphism (SNP) located...... correlation was significantly higher for monozygotic (MZ) twin pairs than for dizygotic (DZ) twin pairs. Serum SP-D variance was influenced by nonshared environmental effects and additive genetic effects. Multivariate analysis of MZ and DZ covariance matrixes showed significant genetic correlation among serum...... SP-D and metabolic variables. The Met11Thr variant explained a significant part of the heritability indicating that serum SP-D variance could be decomposed into non-shared environmental effects (e(2) = 0.19), additive genetic effects (h(2) = 0.42), and the effect of the Met11Thr variations (q(2) = 0.39)....

  2. Immunoassay of serum muramidase (lysozyme) in ocular diseases.

    Science.gov (United States)

    Sen, D K; Sarin, G S

    1987-01-01

    Serum levels of muramidase activity were measured in 162 patients with different ocular diseases and 84 healthy subjects by electro-immuno-diffusion technique. We demonstrated for the first time that electro-immuno-diffusion technique could be successfully applied for the estimation of serum muramidase concentrations. Serum muramidase was found to be high in significant number of cases with granulomatous uveitis, tuberculous keratitis, central serous retinopathy and Eales' disease. Tuberculosis was presumed to be the cause in them by the process of exlusion. Patients with high serum muramidase activity were subjected to anti-tubercular treatment with a marked clinical improvement. It is suggested that high serum muramidase could be an useful parameter in deciding the line of treatment in patients with ocular diseases of uncertain etiology. Serum muramidase concentrations showed return to normal levels with the clinical improvement of the diseases with treatment. It increased again with the re-appearance of the activity of the diseases.

  3. The determination of ultrafiltrable calcium and magnesium in serum.

    Science.gov (United States)

    Danielson, B G; Pallin, E; Sohtell, M

    1982-01-01

    Ultrafiltrate of human serum was investigated in order to evaluate the serum content of calcium and magnesium. The acid and base concentrations and pH of the serum was altered through titration with HCl- or NaOH-solutions. The Pco2 was varied in the titrated serum using different carbon dioxide tensions. This was performed when serum was filtered in a recycling system. It is shown that the analysis of calcium and magnesium have to be done under anaerobic conditions or at standardized pH and Pco2 situations, as the concentrations vary with both pH and Pco2. The concentration ratio between ultrafiltrate and serum for calcium and magnesium was found to be 0.56 and 0.74 respectively at pH=7.41 and Pco2=40 mmHg.

  4. Serum testosterone concentrations in men with alcoholic cirrhosis

    DEFF Research Database (Denmark)

    Gluud, C

    1987-01-01

    Median serum testosterone concentration of men with alcoholic cirrhosis (n = 216) did not differ significantly from normal controls (n = 51), but serum testosterone concentrations varied by a factor 43.9 in patients compared to 3.2 in controls (P less than .001). Nineteen percent of the patients...... had serum testosterone concentrations above 30 nmol/L. Serum concentrations of sex-hormone-binding globulin (SHBG) were significantly (P less than .001) raised, and serum concentrations of calculated nonprotein-bound and non-SHBG-bound testosterone were significantly (P less than .001) decreased...... in patients compared to normal control values. A number of background variables were analyzed with reference to serum testosterone concentrations by means of multiple regression techniques after having divided the patients into groups (A, B, C) with decreasing liver function by a modification of the Child...

  5. Serum beta2-microglobulin in cadmium exposed workers.

    Science.gov (United States)

    Piscator, M

    1978-09-01

    In cadmium exposed workers with renal tubular dysfunction the determination of beta2m in urine is an important diagnostic test. Cadmium exposure's influence on serum beta2m levels and its relationship to urinary excretion of beta2m were studied in 24 cadmium exposed workers with normal serum creatinine levels (less than 10 mg/l)) and no obvious tubular dysfunction. With increasing blood levels of cadmium beta2m was found to increase in serum. There was no concomitant increase in the urinary excretion of beta2m. Serum beta2m was not dependent on serum creatinine within the range studied. The results suggest that for evaluating renal glomerular function in cadmium exposed workers, it might be better to use the serum creatinine level, creatinine clearance or inulin clearance since beta2m might give some false positive results.

  6. Potential Synthetic Biolubricant as an Alternative to Bovine Serum

    OpenAIRE

    Scholes, Susan C.; Colledge, Coral J.; Naylor, Andrew; Mahdi, Mohammed H.; Smith, Alan M.; Joyce, Thomas

    2016-01-01

    It is crucial that orthopaedic implant materials are tested in conditions that replicate the natural body’s environment as closely as possible. Bovine serum is currently recommended for use by the International Organisation for Standardisation (ISO) for the wear testing of these implant materials, however, the rheological properties of bovine serum do not match fully those of the body’s natural lubricant, synovial fluid. This study looks at a potential alternative to bovine serum for the test...

  7. Antibacterial activities of serum from the Komodo Dragon (Varanus komodoensis

    Directory of Open Access Journals (Sweden)

    Mark Merchant

    2013-07-01

    Full Text Available Komodo dragons (Varanus komodoensis are able to feed on large prey items by injecting a dose of toxic bacteria with their bite that, over time, kills the prey by systemic infection. Dragons also suffer bites from other members of their own species during territorial disputes and feeding frenzies. However, they do not suffer the same fate as their prey, suggesting that they have developed a strong immunity to bacterial infections. This study was undertaken to determine the antibacterial activities of serum from the Komodo dragon. Bacterial cultures were treated with different volumes serum from Varanus komodoensis and the growth was monitored by optical density at 430 nm. In addition, the serum was treated with protease, chelators of divalent metal ions, or with mild heat to determine the mechanism of antibacterial activities. Treatment of bacterial cultures with serum from Komodo dragons (Varanus komodoensis resulted in a volume-dependent decrease in bacterial growth. Cultures of Escherichia coli, Staphylococcus aureus, and Klebsiella oxytoca exhibited moderate-strong growth inhibition by V. komodoensis serum, while cultures of Streptococcus epidermitis, Salmonella typhimurium, Providencia stuartii, and Shigella flexneri were nearly completely obliterated for 24 h by only 10% (v/v serum. The antibacterial activity of V. komodensis serum occurred very rapidly, as 18% of E. coli growth was inhibited by a five min exposure to serum. Furthermore, 10- and 20-min incubations of E. coli with serum from V. komodoensis resulted in 43 and 68% inhibition of bacterial growth, respectively. The bactericidal capacity of the serum against E. coli was 2,075,000 bacteria/μL serum, and was inhibited by mild heat treatment, pronase, EDTA, and phosphate, indicating that the anti-bacterial action is most probably due to the presence of a potent serum complement protein system.

  8. Serum IgG subclasses in autoimmune diseases.

    Science.gov (United States)

    Zhang, Haoze; Li, Ping; Wu, Di; Xu, Dong; Hou, Yong; Wang, Qian; Li, Mengtao; Li, Yongzhe; Zeng, Xiaofeng; Zhang, Fengchun; Shi, Qun

    2015-01-01

    To characterize serum IgG subclass levels in several autoimmune diseases, including primary Sjogren syndrome (pSS), systemic sclerosis (SSc), systemic lupus erythematosus (SLE), and primary biliary cirrhosis (PBC). We aimed to analyze serum IgG subclass distribution and to test whether serum IgG4 levels are elevated in these diseases. Serum IgG subclass levels from 102 pSS, 102 SSc, 100 SLE, and 59 PBC patients, as well as 40 healthy controls (HCs), were measured using the immunonephelometric assay. The distribution of IgG subclasses among these autoimmune diseases was analyzed. In this cross-sectional study, serum IgG1 (IgG1/IgG) and/or IgG3 (IgG3/IgG) were significantly increased, compared with those in HCs. Only 6.34% of patients had levels of serum IgG4 >135 mg/dL. There were no significant differences in the frequency of elevated serum IgG4 levels between patients and HC. In pSS, serum IgG1 levels were much higher than those in other disease groups, whereas serum IgG2 and IgG3 levels were most prominently increased in PBC. A strikingly different serum IgG subclass distribution was detected in patients with autoimmune diseases compared with HCs. Serum IgG subclass levels also showed distinct characteristics among different autoimmune diseases. Serum IgG4 levels in these patients were lower or not much higher than those in HCs, which differed from IgG4-related diseases.

  9. Insulin reduces the requirement for serum in Plasmodium falciparum culture

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Tosta

    1984-03-01

    Full Text Available Insulin added to Plasmodium falciparum cultures (0.2 IU/ml reduced the requirement for human serum from ten to five percent. This represents an obvious advantage by its serum-sparing effect and by reducing the chances of using contaminated serum in cultures. The growth-promoting ability of insulin was observed eitherin culture- adapted P. falciparum or in newly-isolated samples.

  10. Studies on serum and hepatic ferritin in alcoholic liver diseases

    OpenAIRE

    杉山, 明

    1987-01-01

    The kinetics of serum ferritin were studied in 42 male patients with alcoholic liver diseases (17 with alcoholic fibrosis, 10 with alcoholic hepatitis and 15 with alcoholic cirrhosis). In addition, the chromatic reaction of biopsied liver tissue to ferritin staining (PAP method) was examined and compared with serum ferritin levels and the iron-staining data. Serum ferritin levels in patients with alcoholic liver diseases, determined immediately after abstinence, were significantly higher than...

  11. Farm Animal Serum Proteomics and Impact on Human Health

    Directory of Open Access Journals (Sweden)

    Francesco Di Girolamo

    2014-09-01

    Full Text Available Due to the incompleteness of animal genome sequencing, the analysis and characterization of serum proteomes of most farm animals are still in their infancy, compared to the already well-documented human serum proteome. This review focuses on the implications of the farm animal serum proteomics in order to identify novel biomarkers for animal welfare, early diagnosis, prognosis and monitoring of infectious disease treatment, and develop new vaccines, aiming at determining the reciprocal benefits for humans and animals.

  12. Phenotype of asthma related with high serum periostin levels

    Directory of Open Access Journals (Sweden)

    Masako Matsusaka

    2015-04-01

    Conclusion: Elevated periostin concentrations in serum were correlated with a specific phenotype of eosinophilic asthma, late-onset and often complicated by obstructive pulmonary dysfunction and nasal disorders.

  13. Klotho is a serum factor related to human aging

    Institute of Scientific and Technical Information of China (English)

    肖能明; 张焱明; 郑权; 顾军

    2004-01-01

    Background Does klotho (KL) protein exist in human serum, and is there any correlation between KL protein in serum with human aging? In order to answer those questions, we identified KL protein in human serum and established the correlation between KL protein in human serum and aging.Methods We prepared a polyclonal antibody against human KL protein that was able to recognize the C-terminal of human secreted KL protein. Western blot and enzyme-linked immunosorbent assay (ELISA) were used to identify KL protein in human serum.Results In Western blot, the antibody specifically recognized a 60-kD KL protein in both human and mice serum. The population aged from 0 to 91 years screened by ELISA revealed that the level of serum KL declined while age increased, though each individual level was variable and that the trend of decreasing in serum KL had no difference in sex.Conclusion Our data suggest that KL is a serum factor related to human aging.

  14. Alterations of serum cholinesterase in patients with gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Shan-Zhi Gu; Xin-Han Zhao; Ping Quan; Sheng-Bin Li; Bo-Rong Pan

    2005-01-01

    AIM: To understand the correlation of serum cholinesterase (CHE) activity with gastric cancer and to assess their clinical significance.METHODS: The velocity method was adopted to detect the activity of serum CHE in patients with gastric cancer and in patients with non-malignant tumor as controls.RESULTS: The serum CHE activity in the treatment group was significantly lower than that in the control group with a very significant difference between the two groups (83.3:113.1,P = 0.0003). Age was significantly associated with the incidence of gastric caner.CONCLUSION: Serum CHE activity has a close relation with the incidence of gastric cancer.

  15. Inhibin B as a serum marker of spermatogenesis

    DEFF Research Database (Denmark)

    Jensen, T K; Andersson, A M; Hjollund, N H

    1997-01-01

    Recent studies have focused on reproductive health of men in the general population. However, semen samples are difficult to obtain within sampling frames that allow comparisons. Blood samples are easier to obtain than ejaculates. Therefore, serum biomarkers of spermatogenesis are of major interest...... for population studies. FSH has previously been used as a marker of spermatogenesis, although it is also influenced by the hypothalamus. Serum inhibin B was recently suggested as a possible, more direct serum marker of spermatogenesis in men with testicular disorders. In a Danish nationwide collaborative study...... reproductive health, may serve as a new marker of spermatogenesis, in addition to sperm concentration and serum FSH....

  16. Relationship between Serum Ferritin Levels and Dyslipidemia in Korean Adolescents

    National Research Council Canada - National Science Library

    Kim, Young-Eun; Kim, Do-Hoon; Roh, Yong-Kyun; Ju, Sang-Yhun; Yoon, Yeo-Joon; Nam, Ga-Eun; Nam, Hyo-Yun; Choi, Jun-Seok; Lee, Jong-Eun; Sang, Jung-Eun; Han, Kyungdo; Park, Yong-Gyu

    2016-01-01

    .... We aimed to study the association between serum ferritin levels and dyslipidemia in adolescents, because dyslipidemia is considered an important modifiable cardiovascular risk factor in the young...

  17. PPARalpha regulates the production of serum Vanin-1 by liver.

    Science.gov (United States)

    Rommelaere, Samuel; Millet, Virginie; Gensollen, Thomas; Bourges, Christophe; Eeckhoute, Jérôme; Hennuyer, Nathalie; Baugé, Eric; Chasson, Lionel; Cacciatore, Ivana; Staels, Bart; Pitari, Giuseppina; Galland, Franck; Naquet, Philippe

    2013-11-15

    The membrane-bound Vanin-1 pantetheinase regulates tissue adaptation to stress. We investigated Vnn1 expression and its regulation in liver. Vnn1 is expressed by centrolobular hepatocytes. Using novel tools, we identify a soluble form of Vnn1 in mouse and human serum and show the contribution of a cysteine to its catalytic activity. We show that liver contributes to Vanin-1 secretion in serum and that PPARalpha is a limiting factor in serum Vnn1 production. Functional PPRE sites are identified in the Vnn1 promoter. These results indicate that serum Vnn1 might be a reliable reporter of PPARalpha activity in liver.

  18. Maternal serum copeptin as a marker for fetal growth restriction

    Directory of Open Access Journals (Sweden)

    Ashraf A. Foda

    2013-09-01

    Conclusion: Maternal serum copeptin level can differentiate between the normal sized and small for gestational age fetuses. Also, it can differentiate between constitutionally small and growth restricted fetuses.

  19. Serum endocan levels in children with febrile neutropenia

    Directory of Open Access Journals (Sweden)

    Eylem Kiral

    2016-03-01

    Full Text Available Endocan is an endotelial cell specific molecule; previous studies have shown that serum endocan levels increased in cancer and sepsis and are also related to the severity of sepsis. There are no clinical study about serum endocan levels in children with febrile neutropenia. The aim of this study was to evaluate serum endocan levels in pediatric leukemia patients with febrile neutropenia (n=33 and compare them with children with leukemia without fever (n=33 and also with healthy children (n=24. The median serum endocan level in the first group (children with febrile neutropenia was statistically significantly higher compared to the leukemic children without febrile neutropenia and also control group (P<0.01 for both. No difference was determined between the serum endocan levels of the leukaemia patients without febrile neutropenia and the healthy control group (P>0.05. Serum endocan levels were also similar with febrile neutropenia due to bacterial causes comparing with the idiopathic febril neutropenia. The results of this study showed increased serum endocan in children with leukemia during the febrile neutropenia episode, and no changes of serum endocan levels in children without leukemia without infection/fever. The monitoring of a series of serum endocan levels would be helpful for the course of febrile neutropenia.

  20. Hyperphenylalaninemia and pterin metabolism in serum and erythrocytes.

    Science.gov (United States)

    Ponzone, A; Guardamagna, O; Spada, M; Ponzone, R; Sartore, M; Kierat, L; Heizmann, C W; Blau, N

    1993-07-16

    The relationship between blood phenylalanine concentrations and serum and erythrocyte biopterin and neopterin concentrations was investigated in 20 phenylketonuric patients with different dietary compliance. At serum phenylalanine concentrations ranging from 43 to 1004 mumol/l, a good correlation was found with serum biopterin (r = 0.76, P < 0.001) and with red blood cell biopterin (r = 0.62, P < 0.001). A similar correlation was found between serum neopterin and phenylalanine (r = 0.60, P < 0.001). The correlation between red blood cell neopterin and serum phenylalanine was less evident, however (r = 0.47, P < 0.005). After oral loading with phenylalanine (100 mg/kg body weight), serum and red blood cell biopterin concentrations increased in patients with classical phenylketonuria as well as in one patient with dihydropteridine reductase deficiency in response to the induced acute hyperphenylalaninemia. One patient suffering from 6-pyruvoyl tetrahydropterin synthase deficiency was loaded orally with tetrahydrobiopterin (20 mg/kg body weight). The kinetics of administered cofactor confirmed its rapid absorption, with early increase of serum concentrations followed by its transport into the red blood cells. The half-life of biopterin was approximately 7 h in serum and 15 h in red blood cells. Because both values are less than the half-life of phenylalanine (20-30 h) in serum, biopterin measurement offers no advantage in monitoring dietary control in hyperphenylalaninemic patients.