scholarly journals APPLICABILITY OF kDNA-PCR FOR ROUTINE DIAGNOSIS OF AMERICAN TEGUMENTARY LEISHMANIASIS IN A TERTIARY REFERENCE HOSPITAL

2013 ◽  
Vol 55 (6) ◽  
pp. 393-399 ◽  
Author(s):  
Marcela M. Satow ◽  
Edite H. Yamashiro-Kanashiro ◽  
Mussya C. Rocha ◽  
Luiza K. Oyafuso ◽  
Rita C. Soler ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Molecular Method ◽  
Laboratory Methods ◽  
Tegumentary Leishmaniasis ◽  
Routine Diagnosis ◽  
Reference Hospital ◽  
American Tegumentary Leishmaniasis ◽  
Diagnosis Method ◽  
Pcr Method

SUMMARY This study evaluated the applicability of kDNA-PCR as a prospective routine diagnosis method for American tegumentary leishmaniasis (ATL) in patients from the Instituto de Infectologia Emílio Ribas (IIER), a reference center for infectious diseases in São Paulo - SP, Brazil. The kDNA-PCR method detected Leishmania DNA in 87.5% (112/128) of the clinically suspected ATL patients, while the traditional methods demonstrated the following percentages of positivity: 62.8% (49/78) for the Montenegro skin test, 61.8% (47/76) for direct investigation, and 19.3% (22/114) for in vitro culture. The molecular method was able to confirm the disease in samples considered negative or inconclusive by traditional laboratory methods, contributing to the final clinical diagnosis and therapy of ATL in this hospital. Thus, we strongly recommend the inclusion of kDNA-PCR amplification as an alternative diagnostic method for ATL, suggesting a new algorithm routine to be followed to help the diagnosis and treatment of ATL in IIER.

Morita-Baylis-Hillman adduct shows in vitro activity against Leishmania (Viannia) braziliensis associated with a reduction in IL-6 and IL-10 but independent of nitric oxide

Parasitology ◽  
2012 ◽  
Vol 140 (1) ◽  
pp. 29-38 ◽  
Author(s):  
F. M. AMORIM ◽  
Y. K. S. RODRIGUES ◽  
T. P. BARBOSA ◽  
P. L. N. NÉRIS ◽  
J. P. A. CALDAS ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cutaneous Leishmaniasis ◽  
Clinical Manifestations ◽  
Failure Rates ◽  
In Vitro Activity ◽  
Clinical Forms ◽  
Tegumentary Leishmaniasis ◽  
American Tegumentary Leishmaniasis ◽  
Human Infections

SUMMARYCurrent treatments for different clinical forms of leishmaniasis are unsatisfactory, highly toxic and associated with increasing failure rates resulting from the emergence of resistant parasites. Leishmania (Viannia) braziliensis is the main aetiological agent of different clinical forms of American tegumentary leishmaniasis, including the mucosal form for which treatment has high failure rates. The aim of this work was to investigate the activity of the Morita-Baylis-Hillman adduct, methyl 2-{2-[hydroxy(2-nitrophenyl)methyl])acryloyloxy} benzoate in vitro against isolates of L. (V.) braziliensis obtained from patients with different clinical manifestations of tegumentary leishmaniasis: localized cutaneous leishmaniasis, mucosal leishmaniasis and disseminated cutaneous leishmaniasis. The adduct effectively inhibited the growth of promastigotes of the different isolates of L. (V.) braziliensis (IC50 ⩽ 7·77 μg/ml), as well as reduced the infection rate of macrophages infected with these parasites (EC50 ⩽ 1·37 μg/ml). It is remarkable to state that the adduct was more effective against intracellular amastigotes (P ⩽ 0·0045). The anti-amastigote activity correlated with an immunomodulatory effect, since the adduct was able to decrease the production of IL-6 and IL-10 by the infected macrophages. However, its effect was independent of nitric oxide production. This work demonstrates the anti-leishmanial activity of methyl 2-{2-[hydroxy(2-nitrophenyl)methyl])acryloyloxy} benzoate and suggests its potential in the treatment of human infections caused by L. (V.) braziliensis.

scholarly journals Identification and Biological Characterization ofLeishmania (Viannia) guyanensisIsolated from a Patient with Tegumentary Leishmaniasis in Goiás, a Nonendemic Area for This Species in Brazil

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Alause da Silva Pires ◽  
Arissa Felipe Borges ◽  
Adriano Cappellazzo Coelho ◽  
Miriam Leandro Dorta ◽  
Ruy de Souza Lino Junior ◽  
...  
Keyword(s):  
Interferon Gamma ◽  
Murine Models ◽  
Biological Characterization ◽  
Field Isolates ◽  
Tegumentary Leishmaniasis ◽  
American Tegumentary Leishmaniasis ◽  
Growth Profiles

The aim of this study was to characterize clinical field isolates ofLeishmaniaspp. obtained from patients with American Tegumentary Leishmaniasis (ATL) who live in Goiás state, Brazil. The presumed areas of infection were in Goiás, Tocantins, and Pará states. Three isolates of parasites were identified asL. (Viannia) braziliensisand one asL. (V.) guyanensis. Thein vitrogrowth profiles were found to be similar for all parasites. Nevertheless, in C57BL/6 mice,L. (V.) guyanensisinfection was better controlled thanL. (V.) braziliensis. Yet in C57BL/6 mice deficient in interferon gamma,L. (V.) guyanensislesions developed faster than those caused byL. (V.) braziliensisisolates. In BALB/c mice, the development of lesions was similar for isolates from both species; however, on the 11th week of infection, amastigotes could not be observed in macrophages fromL. (V.) guyanensis-infected mice. Thus,L. (V.) guyanensiscan be circulating in Goiás, a state where autochthonous cases of this species had not yet been reported. Considering the difficulties to differentiateL. (V.) guyanensisfromL. (V.) braziliensisat the molecular, morphological, and clinical (human and murine models) levels, the presence ofL. (V.) guyanensisinfections is possibly underestimated in several regions of Brazil.

scholarly journals In vitro insights of Leishmania (Viannia) species related to their outcomes and virulence in American tegumentary leishmaniasis

2021 ◽  
Vol 1 (1) ◽  
pp. 1-15
Author(s):  
Daniele Stéfanie Sara Lopes Lera-Nonose ◽  
Áquila Carolina Fernandes Herculano Ramos-Milaré ◽  
Jully Oyama ◽  
Thaís Gomes Verzignassi Silveira ◽  
Izabel Galhardo Demarchi ◽  
...  
Keyword(s):  
Tegumentary Leishmaniasis ◽  
American Tegumentary Leishmaniasis

Clinical features and therapeutic response in adult and pediatric patients with American tegumentary leishmaniasis in Rio de Janeiro

2019 ◽  
Author(s):  
Tatiana C R Senna ◽  
Maria Inês F Pimentel ◽  
Liliane F A Oliveira ◽  
Marcelo R Lyra ◽  
Mauricio N Saheki ◽  
...  
Keyword(s):  
Clinical Features ◽  
Rio De Janeiro ◽  
Pediatric Patients ◽  
Therapeutic Response ◽  
Age Groups ◽  
Healing Time ◽  
The Body ◽  
Reference Centre ◽  
Tegumentary Leishmaniasis ◽  
American Tegumentary Leishmaniasis

Abstract Background American tegumentary leishmaniasis (ATL) is a neglected disease with wide territorial distribution. Knowledge is scarce in children and adolescents. This study aims to compare the clinical features and response to antimony treatment in pediatric and adult patients with cutaneous leishmaniasis. Methods A retrospective cohort study was performed with 659 patients who attended a reference centre in Rio de Janeiro, Brazil, from 2000 to 2015. The pediatric cohort consisted of 131 (20%) patients and the adult cohort consisted of 528 (80%) patients. Results The epidemiological profile, antimony therapeutic response and incidence of adverse events (AE) were different in the pediatric cohort compared with the adult cohort. Mucosal form was less frequent in the pediatric cohort (RR:0.49, p=0.011). Lesions in the head, neck and trunk were more frequent in the pediatric cohort (RR:1.49, p=0.043). The effectiveness of antimony treatment was superior in the pediatric cohort (88.3% vs 76.6%) with a shorter healing time (RR:0.49, p=0.009). Pediatric patients had lower proportions of moderate to severe AE compared with adults (RR:0.45, p=0.027). Clinical AE predominated in the adult cohort (RR:0.40, p=0.000) and laboratory AE in the pediatric cohort (RR:1.50, p=0.023). Conclusions This study adds to the body of knowledge on differences that exist between different age groups in ATL.

scholarly journals In Vitro Incorporation of Helicobacter pylori into Candida albicans Caused by Acidic pH Stress

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 489 ◽  
Author(s):  
Kimberly Sánchez-Alonzo ◽  
Cristian Parra-Sepúlveda ◽  
Samuel Vega ◽  
Humberto Bernasconi ◽  
Víctor L. Campos ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Candida Albicans ◽  
Pcr Amplification ◽  
Growth Curves ◽  
Acidic Ph ◽  
Ph Values ◽  
16S Gene ◽  
Wide Range ◽  
H Pylori

Yeasts can adapt to a wide range of pH fluctuations (2 to 10), while Helicobacter pylori, a facultative intracellular bacterium, can adapt to a range from pH 6 to 8. This work analyzed if H. pylori J99 can protect itself from acidic pH by entering into Candida albicans ATCC 90028. Growth curves were determined for H. pylori and C. albicans at pH 3, 4, and 7. Both microorganisms were co-incubated at the same pH values, and the presence of intra-yeast bacteria was evaluated. Intra-yeast bacteria-like bodies were detected using wet mounting, and intra-yeast binding of anti-H. pylori antibodies was detected using immunofluorescence. The presence of the H. pylori rDNA 16S gene in total DNA from yeasts was demonstrated after PCR amplification. H. pylori showed larger death percentages at pH 3 and 4 than at pH 7. On the contrary, the viability of the yeast was not affected by any of the pHs evaluated. H. pylori entered into C. albicans at all the pH values assayed but to a greater extent at unfavorable pH values (pH 3 or 4, p = 0.014 and p = 0.001, respectively). In conclusion, it is possible to suggest that H. pylori can shelter itself within C. albicans under unfavorable pH conditions.

scholarly journals SBA1 Encodes a Yeast Hsp90 Cochaperone That Is Homologous to Vertebrate p23 Proteins

1998 ◽  
Vol 18 (7) ◽  
pp. 3727-3734 ◽  
Author(s):  
Yifang Fang ◽  
Albert E. Fliss ◽  
Jie Rao ◽  
Avrom J. Caplan
Keyword(s):  
Hormone Receptors ◽  
Pcr Amplification ◽  
Steroid Hormone Receptors ◽  
Loss Of Function ◽  
Vitro Assay ◽  
Growth Defects ◽  
Double Deletion ◽  
Affinity Isolation

ABSTRACT The Saccharomyces cerevisiae SBA1 gene was cloned by PCR amplification from yeast genomic DNA following its identification as encoding an ortholog of human p23, an Hsp90 cochaperone. TheSBA1 gene product is constitutively expressed and nonessential, although a disruption mutant grew more slowly than the wild type at both 18 and 37°C. A double deletion of SBA1and STI1, encoding an Hsp90 cochaperone, displayed synthetic growth defects. Affinity isolation of histidine-tagged Sba1p (Sba1His6) after expression in yeast led to coisolation of Hsp90 and the cyclophilin homolog Cpr6. Using an in vitro assembly assay, purified Sba1His6 bound to Hsp90 only in the presence of adenosine 5′-O-(3-thiotriphosphate) or adenyl-imidodiphosphate. Furthermore, interaction between purified Sba1His6 and Hsp90 in yeast extracts was inhibited by the benzoquinoid ansamycins geldanamycin and macbecin. The in vitro assay was also used to identify residues in Hsp90 that are important for complex formation with Sba1His6, and residues in both the N-terminal nucleotide binding domain and C-terminal half were characterized. In vivo analysis of known Hsp90 substrate proteins revealed that Sba1 loss of function had only a mild effect on the activity of the tyrosine kinase v-Src and steroid hormone receptors.

scholarly journals Establishment of Highly Efficient Agrobacterium Rhizogenes-mediated Transformation for Stevia Rebaudiana Bertoni Explants

2016 ◽  
Vol 58 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Żaneta Michalec-Warzecha ◽  
Laura Pistelli ◽  
Francesca D’Angiolillo ◽  
Marta Libik-Konieczny
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Hairy Roots ◽  
Molecular Level ◽  
Stevia Rebaudiana ◽  
Culture Conditions ◽  
Light Conditions ◽  
Inoculum Concentration ◽  
Stevia Rebaudiana Bertoni ◽  
Pcr Method

Abstract Leaves and internodes from Stevia rebaudiana Bertoni plants growing in different conditions were used for transformation with two strains of Agrobacterium rhizogenes: ATCC 15384 and LBA 9402. Hairy roots formation was observed and the percentage of the transformed explants depended on the type of explant, time of inoculation and inoculum concentration. Inoculation of explants from ex vitro and in vitro plants with LBA 9402 strain led to higher efficiency of transformation than inoculation with ATCC 15384 strain. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decreased significantly under light conditions. Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A. rhizogenes.

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