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1.
Surg Endosc ; 36(5): 2906-2913, 2022 05.
Artículo en Inglés | MEDLINE | ID: mdl-34231071

RESUMEN

BACKGROUND: Organ sparing by the transanal endoscopic microsurgery (TEM) procedure is a treatment for patients with locally advanced rectal cancer after chemoradiotherapy (CRT) and complete clinical response (cCR). AIMS: To assess the surgical and long-term oncological outcomes of TEM for the treatment in T2-3 rectal cancer after CRT and cCR. METHODS: This study was a retrospective review of a prospective database of patients with rectal cancer who underwent TEM after CRT and cCR from April 2011 to March 2020. RESULTS: 52 patients underwent TEM during a period of 9 years. This group of patients included 27 females and 25 males. The median age was 62 (32-86) years, lesion size was 2.5 (1-4) cm, and lesion distance from the anal verge 7.3 (4-10) cm. Median operative time was 79.5 (25-120) min and hospital stay was 1 day (14 h-4 days). Morbidity rate was 13.5% and reoperation rate due to major complications was 3.8%. Final histological findings confirmed 34 (65.4%) patients with ypT0, 7 (13.5%), 6 (11.5%), and 5 (9.6%) patients with carcinoma ypT1, ypT2, and ypT3, respectively. After a median follow-up period of 86 (5-107) months, 1 (2.4%) patient had local recurrences and 3 (7.3%) distant metastases. The 5-year disease-free survival was 91.7% and 5-year overall survival 89.5%. CONCLUSION: Our experience has shown significant rates of ypT0 and ypT1 associated with excellent long-term results. Performing TEM to treat T2-3N0 rectal cancer after CRT and cCR appears to be an oncologically safe and effective procedure.


Asunto(s)
Neoplasias del Recto , Microcirugía Endoscópica Transanal , Quimioradioterapia , Femenino , Humanos , Masculino , Microcirugia/métodos , Persona de Mediana Edad , Terapia Neoadyuvante , Recurrencia Local de Neoplasia/cirugía , Neoplasias del Recto/etiología , Neoplasias del Recto/cirugía , Estudios Retrospectivos , Microcirugía Endoscópica Transanal/efectos adversos , Resultado del Tratamiento
2.
Parasitol Res ; 120(8): 2897-2903, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34313806

RESUMEN

A cross-sectional study was carried out to determine the seroprevalence of Toxoplasma gondii and associated risk factors in pigs in the largest pork-producing region in Cuba. Serum samples from 420 pigs, including 210 sows and 210 post-weaning pigs, were tested for antibodies against T. gondii using a commercial indirect enzyme-linked immunosorbent assay. Anti-T. gondii antibodies were detected in 56 animals (13.3%, 95% CI: 10.1-16.6). A generalized estimating equations model revealed that the risk factors associated with higher seropositivity in pigs were altitude (higher in farm's location < 250 m above sea level (masl) versus ≥ 250 masl) and age (higher in sows compared to post-weaning pigs). The results indicated that this protozoan parasite is widely distributed on pig farms in the study area, which is a public health concern since the consumption of raw or undercooked pork meat products containing tissue cysts is considered one of the main routes of T. gondii transmission worldwide. Control measures should be implemented to reduce the risk of exposure to T. gondii in pigs in Cuba.


Asunto(s)
Sus scrofa/parasitología , Enfermedades de los Porcinos , Toxoplasma , Toxoplasmosis Animal , Animales , Anticuerpos Antiprotozoarios , Estudios Transversales , Cuba/epidemiología , Femenino , Masculino , Factores de Riesgo , Estudios Seroepidemiológicos , Porcinos , Enfermedades de los Porcinos/epidemiología , Toxoplasma/inmunología , Toxoplasmosis Animal/epidemiología
3.
Exp Parasitol ; 209: 107814, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31816280

RESUMEN

Giardia duodenalis is one of the most important intestinal parasites globally, especially in children, and in Cuba is the leading cause of chronic paediatric diarrhoea in this population. G. duodenalis is composed of eight genetic groups (or assemblages), two of which (A and B) are apparently zoonotic, occurring in both humans and other animals. However, consensus on the most appropriate genotyping scheme for optimal characterization of G. duodenalis isolates is lacking. In this article we present the results of three descriptive observational studies conducted in Havana, Cuba between 2010 and 2013, with the aim of comparing the results from molecular (PCR) approaches targeting different genes in order to assign with confidence 224 isolates of G. duodenalis to the correct assemblages. In each sub-study, following DNA isolation by the phenol/chloroform/isoamyl alcohol extraction method, PCR targeting the triose phosphate isomerase (tpi) gene was used for molecular characterization, as well as one additional PCR-method targeting another gene or pair of genes. DNA amplification was obtained in 87%, 83%, and 80% in the three sub-studies. Although excellent agreement (kappa index = 1) was recorded between results from some pairs of genes, for other combinations only moderate or substantial agreement was achieved. These results highlight the importance of interpretation of genotyping data, especially when single genetic markers are used. From the results of our studies, PCR targeting a combination of the tpi gene and the intergenic spacer region of rDNA may be a useful approach for the molecular characterization of G. duodenalis isolates.


Asunto(s)
Técnicas de Genotipaje/normas , Giardia lamblia/clasificación , Giardiasis/parasitología , Reacción en Cadena de la Polimerasa/métodos , Animales , Niño , Preescolar , Cuba , ADN Protozoario/química , ADN Protozoario/aislamiento & purificación , ADN Espaciador Ribosómico/química , Heces/parasitología , Giardia lamblia/genética , Giardia lamblia/aislamiento & purificación , Glutamato Deshidrogenasa/genética , Humanos , Polimorfismo de Longitud del Fragmento de Restricción , Triosa-Fosfato Isomerasa/genética
4.
Parasitol Res ; 119(1): 345-349, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31776667

RESUMEN

Leishmaniasis is a parasitic disease of medical importance widely distributed around the world. Several methods are available for diagnosis but molecular approaches are highly recommended. To improve the sensitivity of an existing hsp20 gene based-PCR protocol to detect Leishmania parasites, primers were redesigned to amplify a shorter fragment using a new PCR variant (PCR-hsp20S). In this study, we aimed at characterizing the performance of the new method on cutaneous clinical samples and compare it with the former PCR-hsp20. The analytical sensitivity of the PCR-hsp20S was evaluated using DNA dilutions (100-0.1 pg) from Leishmania donovani and resulted in the detection of 10 fg of parasitic DNA, the equivalent to 0.05 parasite genome. For the diagnostic evaluation, a panel of 127 human clinical samples was used to calculate the parameters of sensitivity, specificity, accuracy, and positive and negative predictive values of the PCR-hsp20S. Diagnostic sensitivity was 94% (CI, 89.1-99.7%) and the specificity of 100% (CI, 98.6-100%). The same panel was also evaluated with the PCR-hsp20 to calculate the agreement between both molecular assays and to compare their performances. While both hsp20-based PCRs showed a good agreement coefficient (kappa index = 0.6), the performance of the novel variant, PCR-hsp20S, was significantly higher in terms of sensitivity (P = 0.0001) allowing the accurate detection of a higher number of Leishmania-positive clinical samples. We endorse the use of the PCR-hsp20S over the former protocol for the detection of Leishmania parasites from cutaneous clinical samples. In addition, as an improved sensitivity was achieved with the new method merely through the amplification of a shorter gene fragment, this investigation constitutes an experimental proof of this concept.


Asunto(s)
Proteínas del Choque Térmico HSP20/genética , Leishmania donovani/aislamiento & purificación , Leishmaniasis/parasitología , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Animales , Cartilla de ADN/química , ADN Protozoario/genética , Humanos , Leishmania donovani/genética , Proteínas Protozoarias/genética , Sensibilidad y Especificidad , Piel/parasitología
5.
Parasitol Res ; 116(7): 1843-1848, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28573463

RESUMEN

Leishmaniasis is highly prevalent in New World countries, where several methods are available for detection and identification of Leishmania spp. Two hsp70-based PCR protocols (PCR-N and PCR-F) and their corresponding restriction fragment length polymorphisms (RFLP) were applied for detection and identification of Leishmania spp. in clinical samples recruited in Colombia, Guatemala, and Honduras. A total of 93 cases were studied. The samples were classified into positive or suspected of leishmaniasis according to parasitological criteria. Molecular amplification of two different hsp70 gene fragments and further RFLP analysis for identification of Leishmania species was done. The detection in parasitologically positive samples was higher using PCR-N than PCR-F. In the total of samples studied, the main species identified were Leishmania panamensis, Leishmania braziliensis, and Leishmania infantum (chagasi). Although RFLP-N was more efficient for the identification, RFLP-F is necessary for discrimination between L. panamensis and Leishmania guyanesis, of great importance in Colombia. Unexpectedly, one sample from this country revealed an RFLP pattern corresponding to Leishmania naiffi. Both molecular variants are applicable for the study of clinical samples originated in Colombia, Honduras, and Guatemala. Choosing the better tool for each setting depends on the species circulating. More studies are needed to confirm the presence of L. naiffi in Colombian territory.


Asunto(s)
Proteínas HSP70 de Choque Térmico/genética , Leishmania/aislamiento & purificación , Leishmaniasis/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Animales , Colombia , Guatemala , Honduras , Humanos , Leishmania/genética , Leishmania braziliensis/genética , Leishmania braziliensis/aislamiento & purificación , Leishmania infantum/genética , Leishmania infantum/aislamiento & purificación , Leishmaniasis/diagnóstico , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y Especificidad
6.
Angew Chem Int Ed Engl ; 56(42): 12962-12966, 2017 10 09.
Artículo en Inglés | MEDLINE | ID: mdl-28816407

RESUMEN

Pyrrolidines are important heterocyclic compounds with endless applications in organic synthesis, metal catalysis, and organocatalysis. Their potential as ligands for first-row transition-metal catalysts inspired a new method to access complex poly-heterocyclic pyrrolidines in one step from available materials. This fundamental step forward is based on the discovery of an essential organoaluminum promoter that engages unactivated and electron-rich olefins in intermolecular [3+2] cycloadditions.

7.
Trop Med Int Health ; 19(2): 195-206, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24224809

RESUMEN

OBJECTIVE: Differential diagnosis of infection with Trypanosoma cruzi or T. rangeli is relevant for epidemiological studies and clinical practice as both species infect humans, but only T. cruzi causes Chagas' disease. Their common antigen determinants complicate the distinction between both species, while current PCR assays used for differentiation show some drawbacks. We developed and validated a generic PCR discriminating the species by restriction fragment length polymorphism (RFLP) analysis and a duplex PCR specifically amplifying a differently sized fragment of both species. METHODS: The assays are based upon a partial region of the heat-shock protein 70 gene (hsp70). The analytical sensitivity and specificity were determined for both PCRs. The assays were analytically evaluated on a panel of six T. cruzi, one T. cruzi marinkellei and four T. rangeli strains, various other infectious pathogens, a panel of spiked samples of T. cruzi, and artificially mixed infections of T. cruzi and T. rangeli. Finally, the tools were applied on 36 additional isolates of Trypanosoma species. RESULTS: The detection limit of the PCRs was between 0.05 and 0.5 parasite genomes, and 1-10 parasites spiked in 200 µl blood. In artificial mixtures, PCR-RFLP picked up both species in ratios up to 10(2) and duplex PCR up to 10(4) . In the 36 isolates tested, both single and mixed infections were identified. All assays were shown to be specific. CONCLUSION: Our PCRs show high potential for the differential diagnosis of T. cruzi and T. rangeli, which in view of their sensitivity can aid in the confirmation of infection with these parasites in vectors, reservoirs and clinical samples.


Asunto(s)
Proteínas HSP70 de Choque Térmico/genética , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Trypanosoma cruzi/genética , Trypanosoma rangeli/genética , Tripanosomiasis/parasitología , Enfermedad de Chagas/diagnóstico , Enfermedad de Chagas/parasitología , Diagnóstico Diferencial , Humanos , Reacción en Cadena de la Polimerasa/normas , Tripanosomiasis/diagnóstico
8.
Zoonoses Public Health ; 70(2): 125-133, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36416370

RESUMEN

Domestic pigs are considered as one of the main intermediate hosts in the zoonotic transmission of Toxoplasma gondii in many countries. Serological and molecular studies are warranted to better understand the epidemiology and transmission patterns of this parasite worldwide. To date, seroepidemiological information on T. gondii in domestic pigs in Cuba is very scarce and there are no reports of T. gondii genotypes circulating in this country. Here, we aimed to estimate the seroprevalence of T. gondii and provide genetic characterization of the strains circulating in slaughtered pigs intended for human consumption in Central Cuba. Seroprevalence was determined in 450 serum samples from slaughtered pigs in Villa Clara province using ELISA. Anti-Toxoplasma gondii IgG antibodies were detected in 100 animals (22.2%, 95% CI: 18.5-26.2). Conventional PCR of the 529-bp marker of T. gondii was performed in hearts and diaphragm tissues of all ELISA-seropositive pigs. Toxoplasma gondii DNA was detected in four animals. Further genetic characterization of the positive DNA samples was performed by multilocus PCR-RFLP and PCR-sequencing typing tools. Molecular analysis revealed four different genetic profiles that were combinations of type I, II, III and u-1 alleles, suggesting the circulation of non-clonal genotypes of T. gondii in domestic pigs in Cuba. Our results indicate that T. gondii is widely distributed in slaughtered pigs in this country, which might have important implications for public health. To the best of our knowledge, this is the first report on genetic characterization of T. gondii in Cuba. Although preliminary, the results suggest a high genetic diversity of T. gondii in the study region. Further studies based on parasite isolation are needed to definitively identify the genotypes circulating and characterize the virulence of strains detected in pigs in Cuba, and to assess the risk of zoonotic transmission from pork products in this country.


Asunto(s)
Enfermedades de los Porcinos , Toxoplasma , Toxoplasmosis Animal , Humanos , Porcinos , Animales , Sus scrofa , Estudios Seroepidemiológicos , Cuba/epidemiología , Toxoplasmosis Animal/epidemiología , Toxoplasmosis Animal/parasitología , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/parasitología , Anticuerpos Antiprotozoarios , Genotipo
9.
Arch Virol ; 157(9): 1789-95, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22653538

RESUMEN

The viral infection of the parasite with T. vaginalis virus (TVV) may have important implications for trichomonal virulence. In this study we identified the TVV species isolated from Cuban T. vaginalis, using specie specific Reverse Transcriptase-PCR. Of the 37 clinical isolates studied, 21 were infected with TVV, 6 contained TVV-1, 12, TVV- 2 and 3 were co-infected with TVV-1 and -2. The strains infected with TVV showing highest adhesion level in comparison to not infected strains, with high statistical significance. The strains infected only with TVV-2 showing highest adhesion level in comparison to strains infected with TVV-1, with high statistical significance. The parasites classified as mild symptomatic are infected only with TVV-1, however the severe only with TVV-2. According to our results, it seems that only two TVV species are infecting the Cuban isolates. Further studies using higher number of strains should be conducted in order to corroborate these results.


Asunto(s)
Adhesión Celular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Totiviridae/clasificación , Totiviridae/genética , Trichomonas vaginalis/patogenicidad , Trichomonas vaginalis/virología , Adolescente , Cuba , Genotipo , Humanos , Tipificación Molecular , Totiviridae/aislamiento & purificación , Tricomoniasis/parasitología , Trichomonas vaginalis/aislamiento & purificación , Virulencia
10.
Exp Parasitol ; 131(3): 393-8, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22584035

RESUMEN

Trichomonas vaginalis can be naturally infected with intracellular Mycoplasma hominis. This bacterial infection may have implications for trichomonal virulence and disease pathogenesis. The objective of the study was to report the presence of M. hominis in Cuban T. vaginalis isolates and to describe the association between the phenotype M. hominis infected with RAPD genetic polymorphism of T. vaginalis. The Random Amplified Polymorphic DNA (RAPD) technique was used to determine genetic differences among 40 isolates of T. vaginalis using a panel of 30 random primers and these genetic data were correlated with the infection of isolates with M. hominis. The trees drawn based on RAPD data showed no relations with metronidazole susceptibility and significantly association with the presence of M. hominis (P=0.043), which demonstrates the existence of concordance between the genetic relatedness and the presence of M. hominis in T. vaginalis isolates. This result could point to a predisposition of T. vaginalis for the bacterial enters and/or survival.


Asunto(s)
Mycoplasma hominis/aislamiento & purificación , Polimorfismo Genético , Trichomonas vaginalis/genética , Trichomonas vaginalis/microbiología , Cuba , ADN Bacteriano/análisis , ADN Bacteriano/química , Femenino , Humanos , Reacción en Cadena de la Polimerasa Multiplex , Mycoplasma hominis/genética , Filogenia , Técnica del ADN Polimorfo Amplificado Aleatorio , Tenericutes/clasificación , Tenericutes/genética , Tenericutes/aislamiento & purificación , Sistema Urogenital/microbiología , Sistema Urogenital/parasitología
11.
Mem Inst Oswaldo Cruz ; 107(8): 978-84, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23295746

RESUMEN

Propolis is a resinous mixture of different plant exudates collected by honeybees. Currently, propolis is widely used as a food supplement and in folk medicine. We have evaluated 20 Cuban propolis extracts of different chemical types, brown (BCP), red and yellow (YCP), with respect to their in vitro antibacterial, antifungal and antiprotozoal properties. The extracts inhibited the growth of Staphylococcus aureus and Trichophyton rubrum at low µg/mL concentrations, whereas they were not active against Escherichia coli and Candida albicans. The major activity of the extracts was found against the protozoa Leishmania, Trypanosoma and Plasmodium, although cytotoxicity against MRC-5 cells was also observed. The BCP-3, YCP-39 and YCP-60 extracts showed the highest activity against P. falciparum, with 50% of microbial growth (IC50) values of 0.2 µg/mL. A positive correlation between the biological activity and the chemical composition was observed for YCP extracts. The most promising antimicrobial activity corresponds to YCP subtype B, which contains acetyl triterpenes as the main constituents. The present in vitro study highlights the potential of propolis against protozoa, but further research is needed to increase selectivity towards the parasite. The observed chemical composition-activity relationship of propolis can contribute to the identification of the active principles and standardisation of this bee product.


Asunto(s)
Antibacterianos/farmacología , Antifúngicos/farmacología , Antiprotozoarios/farmacología , Própolis/farmacología , Animales , Candida albicans/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Concentración 50 Inhibidora , Leishmania/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Pruebas de Sensibilidad Parasitaria , Plasmodium/efectos de los fármacos , Própolis/química , Staphylococcus aureus/efectos de los fármacos , Trichophyton/efectos de los fármacos , Trypanosoma/efectos de los fármacos
12.
Trop Dis Travel Med Vaccines ; 8(1): 15, 2022 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-35773710

RESUMEN

BACKGROUND: Leishmaniasis is a vector-borne disease caused by several species from genus Leishmania. An increase in the number of cases related to human movement has been informed in the last years. Due to the increase of suspicious leishmaniasis cases arriving in Cuba during 2017, a general analysis is presented herein. METHODS: Clinical samples were collected from 5 patients suspicious of leishmaniasis, received from January to December 2017 at the Institute of Tropical Medicine Pedro Kourí, Cuba. Skin lesion samples were analyzed using different diagnostic assays: direct smear, histological examination, and molecular analysis for species identification. Epidemiological and demographic data were requested from each case and analyzed. Treatment and follow up of patient was also performed. RESULTS: Five cases were confirmed as Leishmania infection according to microscopic observation and molecular methods results. PCR-18S, PCR-N/RFLP and PCR-F/RFLP identified the following species: L. panamensis (2 cases), L. braziliensis (1 case), L.panamensis/L.guyanensis (1 case), L. mexicana complex (1 case). In treated patients, drugs were well tolerated, cure were documented and no relapse have been currently reported (3 years later). CONCLUSIONS: Clinical characteristics, demographic data, and epidemiological features of infection for each case evidence the potential risk related with travel to endemic areas of leishmaniasis. KEYWORKS: Cutaneous leishmaniasis, Epidemiology, Imported cases.

13.
Exp Parasitol ; 127(2): 593-9, 2011 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-20875411

RESUMEN

Trichomonas vaginalis can be infected with double-stranded RNA (dsRNA) viruses known as T. vaginalis virus (TVV). This viral infection may have important implications for trichomonal virulence and disease pathogenesis. The objective of this study was to determine the possible correlation between the T. vaginalis genetic polymorphism and the isolate infection with TVV. The Random Amplified Polymorphic DNA (RAPD) technique was used to determine genetic differences among 37 isolates of T. vaginalis using a panel of 30 random primers and these genetic data were correlated with the infection of isolates with TVV. The trees drawn based on RAPD data showed significantly association with the presence of TVV (P = 0.028) demonstrating the existence of concordance between the genetic relatedness and the presence of TVV in T. vaginalis isolates. This result could point to a predisposition of T. vaginalis for the viral enters and/or survival.


Asunto(s)
Polimorfismo Genético , Totiviridae/fisiología , Trichomonas vaginalis/genética , Trichomonas vaginalis/virología , Adolescente , Femenino , Humanos , Filogenia , Técnica del ADN Polimorfo Amplificado Aleatorio , Vaginitis por Trichomonas/parasitología , Trichomonas vaginalis/clasificación
14.
Cerebrospinal Fluid Res ; 6: 2, 2009 Mar 06.
Artículo en Inglés | MEDLINE | ID: mdl-19267913

RESUMEN

BACKGROUND: Toxoplasmic encephalitis (TE) is one of the most common opportunistic infections in immunocompromised patients. In Cuba, despite the highly active antiretroviral therapy, TE is still the most important cause of cerebral mass lesions in patients infected with the human immunodeficiency virus (HIV). The detection of Toxoplasma gondii by PCR may facilitate the diagnosis and follow-up of TE in acquired immunodeficiency syndrome (AIDS) patients by direct identification of parasite DNA in clinical samples. The aim of the present study was to evaluate a rapid PCR method using the B1 gene to detect T. gondii in cerebrospinal fluid (CSF) samples from patients with suspected TE. METHODS: CSF samples from AIDS and HIV-negative patients were analyzed. Patients were divided into two groups according to the Centre for Disease Control and Prevention (CDC) criteria for AIDS-related TE: AIDS patients with suspected neurotoxoplasmosis and AIDS and HIV-negative patients with other confirmed neurological diseases but no suspicions of TE. Predictive values, diagnostic accuracy, sensitivity and specificity of the PCR B1 method were calculated. RESULTS: The results obtained from 190 patients showed that this assay has a good sensitivity and specificity (83.3% and 95.7%, respectively) for the diagnosis of TE in AIDS patients. CONCLUSION: PCR using the B1 gene and B22/B23 set of primers is a single, rapid and reliable method that may be valuable for discrimination between toxoplasmosis and other central nervous system (CNS) diseases.

15.
Med Sci Monit ; 15(2): BR47-54, 2009 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19179961

RESUMEN

BACKGROUND: Bone marrow-derived stem cell transplantation is a potentially viable therapeutic option for the treatment of neurodegenerative disease. MATERIAL/METHODS: We have isolated bone marrow stem cells by standard method. We then evaluated the survival of rats' bone marrow mononuclear cells implanted in rats' brain. The cells were extracted from rats' femurs, and marked for monitoring purposes by adenoviral transduction with Green Fluorescent Protein (GFP). Labeled cells were implanted within the area of rats' striatum lesions that were induced a month earlier employing quinolinic acid-based method. The implants were phenotyped by monitoring CD34; CD38; CD45 and CD90 expression. Bone marrow stromal cells were extracted from rats' femurs and cultivated until monolayer bone marrow stromal cells were obtained. The ability of bone marrow stromal cells to express NGF and GDNF was evaluated by RT-PCR. RESULTS: Implanted cells survived for at least one month after transplantation and dispersed from the area of injection towards corpus callosum and brain cortex. Interestingly, passaged rat bone marrow stromal cells expressed NGF and GDNF mRNA. CONCLUSIONS: The bone marrow cells could be successfully transplanted to the brain either for the purpose of trans-differentiation, or for the expression of desired growth factors.


Asunto(s)
Células de la Médula Ósea/citología , Trasplante de Tejido Encefálico , Encéfalo/patología , Supervivencia de Injerto , Neuronas/metabolismo , Trasplante de Células Madre , Adenoviridae , Animales , Antígenos CD/metabolismo , Biomarcadores/metabolismo , Regulación de la Expresión Génica , Leucocitos Mononucleares/virología , Masculino , Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/metabolismo , Ácido Quinolínico , Ratas , Ratas Sprague-Dawley , Coloración y Etiquetado , Factores de Tiempo
16.
Exp Parasitol ; 122(3): 203-7, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19318095

RESUMEN

Highly active antiretroviral therapy (HAART) has decreased the incidence of opportunistic infections in the central nervous system (CNS) in AIDS patients. However, toxoplasmic encephalitis (TE) still represents the most common cerebral mass lesion in patients infected with human immunodeficiency virus (HIV). The aim of this study was to evaluate nested PCR-B1 using cerebrospinal fluid (CSF) to detect Toxoplasma gondii DNA for the diagnosis of TE. A total of 114 samples were evaluated, and 33/44 samples from patients with TE were positive by PCR (sensitivity 75%), demonstrating the diagnostic usefulness of PCR technique. PCR-B1 products were analyzed by restriction fragment length polymorphism (RFLP) in 30 samples. Only type I allele at B1 was identified in these samples according banding patterns. This is the first report of evaluation of S1-AS1/S2-AS2 set of primers in more than 100 clinical samples as well as the first genotyping study of T. gondii in Cuba.


Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/diagnóstico , Líquido Cefalorraquídeo/parasitología , ADN Protozoario/líquido cefalorraquídeo , Encefalitis/diagnóstico , Toxoplasma/aislamiento & purificación , Toxoplasmosis Cerebral/diagnóstico , Infecciones Oportunistas Relacionadas con el SIDA/líquido cefalorraquídeo , Infecciones Oportunistas Relacionadas con el SIDA/parasitología , Animales , Encefalitis/líquido cefalorraquídeo , Encefalitis/parasitología , Genotipo , Humanos , Ratones , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y Especificidad , Toxoplasma/clasificación , Toxoplasma/genética , Toxoplasmosis Cerebral/líquido cefalorraquídeo , Toxoplasmosis Cerebral/parasitología
17.
Acta Parasitol ; 64(2): 300-307, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31165993

RESUMEN

INTRODUCCTION: Trichomonas vaginalis is a highly prevalent parasitic that causes the sexually transmitted disease trichomoniasis with some serious health complications. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, epidemiology of the infection and drug susceptibility. For this end, we conducted analysis of a fragment (23 kDa) of the p60 of T. vaginalis gene. MATERIAL AND METHODS: The restriction fragment length polymorphism (RFLP) methods was used. RESULT AND DISCUSSION: RFLP analysis showed the difference between T. vaginalis isolates from symptomatic and asymptomatic patients, suggesting a relation between the genetic identity of the isolates and their clinical manifestations.


Asunto(s)
Variación Genética , Péptido Hidrolasas/genética , Proteínas Protozoarias/genética , Trichomonas vaginalis/genética , Adolescente , ADN Protozoario/genética , Femenino , Humanos , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción , Tricomoniasis/parasitología , Trichomonas vaginalis/enzimología
18.
Biomedica ; 28(4): 597-606, 2008 Dec.
Artículo en Español | MEDLINE | ID: mdl-19462565

RESUMEN

INTRODUCTION: The analysis of the PCR-restriction fragment length polymorphism and random amplified polymorphic DNA have been useful tools for Leishmania identification. OBJECTIVES: Molecular procedures were demonstrated for identification and typing of reference strains of New World Leishmania and their applicability was validated for clinical samples. MATERIALS AND METHODS: DNA was extracted from 16 reference strains of Latin American Leishmania as well as from clinical samples of leishmaniasis patients. A sequence coding for cysteine proteinase B was amplified by PCR and subjected to restriction fragment length polymorphism analysis. The enzyme used was Taq1. For eight of the reference strains, the random amplified polymorphic desoxyribonucleic acid technique (RAPD) was applied. Band patterns for Leishmania species differentiation were established each each method. The sample size of the clinical sample was of 5. RESULTS: PCR products of the cysteine proteinase B gene were obtained for L. braziliensis, L. peruviana, L. panamensis and L. guyanensis. For the other species, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, no amplification occurred. The patterns of restriction fragments revealed band patterns in common for L. peruviana, L. guyanensis and L. panamensis, whereas L. braziliensis had a distinctive pattern. When human samples were examined, amplification occurred for all cases, and the profiles corresponded to the common profile of L. peruviana, L. guyanensis and L. panamensis. The RAPD technique demonstrated reproducible and distinctive patterns for each of the 8 reference strains, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, making possible to differentiate all them. The advantages and limitations of each procedure are discussed. CONCLUSIONS: The combination of RFP and RAPD methodologies provide useful tools to identify medical important species of Leishmania by recognizing DNA sequences characteristic of each species.


Asunto(s)
Leishmania , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Técnica del ADN Polimorfo Amplificado Aleatorio , Animales , Humanos , Leishmania/clasificación , Leishmania/genética , Leishmaniasis/etiología , Leishmaniasis/fisiopatología , Clima Tropical
19.
Artículo en Inglés | MEDLINE | ID: mdl-30094054

RESUMEN

BACKGROUND: Leishmaniasis is a neglected parasitic disease caused by Leishmania spp., which is not endemic in Cuba. However, several factors (such as human activities, climate changes, and tourism) have led to an increase in the number of leishmaniasis cases in all regions, raising diagnosis and surveillance issues. We aim to present the retrospective analysis of 16 human cases suspicious of leishmaniasis, which were received during 2006-2016 for diagnosis at the Department of Parasitology from the Institute of Tropical Medicine Pedro Kourí, Cuba. METHODS: Clinical samples were collected and analyzed via different diagnostic assays, including direct smear, cultivation, histological analysis, and molecular analysis. Epidemiology and background of infection, clinical features, sex and age from each patient was recorded. RESULTS: From the 16 suspicious cases, 5 cases were confirmed for Leishmania infection, based on at least two positive results using different methods: PCR-based diagnosis [18S rRNA (5/5), hsp20 gene (4/5), hsp70 gene (3/5)], histopathology evaluation (2/3), parasite cultivation (2/3), or direct smears (2/3). L. braziliensis and L. mexicana were identified as the involving species in two cases, according to hsp70 PCR-RFLP protocols. Demographic and clinical features, as well as treatment and follow up, are described for every case. CONCLUSIONS: The combination of parasitological and molecular methods allowed proper diagnosis of imported leishmaniasis cases in Cuba. The utility and advantages of molecular diagnosis assays in non-endemic countries like Cuba are discussed.

20.
Rev. cuba. cir ; 62(3)sept. 2023.
Artículo en Español | LILACS, CUMED | ID: biblio-1550825

RESUMEN

Introducción: Las indicaciones de la Microcirugía Transanal Endoscópica han evolucionado desde la cirugía de tumoraciones rectales hasta otras enfermedades pélvicas. La asociación de esta y la escisión total del mesorrecto transanal ofrece una serie de ventajas. Objetivo: Determinar las indicaciones, describir la técnica quirúrgica y mostrar los resultados a largo plazo obtenidos en la realización de la escisión total del mesorrecto transanal en el tratamiento del cáncer del recto medio y bajo. Métodos: Se realizó un estudio observacional descriptivo y prospectivo de los pacientes con cáncer del recto medio y bajo sometidos a esta técnica quirúrgica en el período comprendido entre febrero de 2017 y febrero de 2022 en el Centro Nacional de Cirugía de Mínimo Acceso. Resultados: Se operaron 13 pacientes, 9 con cáncer del recto bajo y 4 con cáncer del recto medio y un promedio de edad de 56,2 años (rango 28-76). El promedio de tiempo quirúrgico fue de 183 minutos (rango 120-270) y las pérdidas hemáticas estimadas de 68 mililitros. La incidencia de morbilidad mayor fue de 15,4 por ciento y la media de estadía hospitalaria de 5,4 días. La media del período de seguimiento fue de 35 (rango 9-69) meses con una recidiva local de 7,7 por ciento y una supervivencia global a los 5 años de 100 por ciento. Conclusiones: La escisión total del mesorrecto transanal combinado con cirugía laparoscópica es una técnica factible y segura. La introducción de la variante técnica utilizando el instrumental de la Microcirugía Transanal Endoscópica es más ergonómica y disminuye los costos(AU)


Introduction: The indications for transanal endoscopic microsurgery have evolved from surgery of rectal tumors to other pelvic diseases. The association between this and total excision of the transanal mesorectum offers a series of advantages. Objective: To determine the indications, to describe the surgical technique and to show the long-term outcomes obtained in the performance of total excision of the transanal mesorectum for treating cancer of the middle and lower rectum. Methods: A descriptive and prospective observational study was carried out of patients with cancer of the middle and lower rectum who underwent this surgical technique in the period from February 2017 to February 2022 at Centro Nacional de Cirugía de Mínimo Acceso. Results: Thirteen patients were operated on, 9 with cancer of the lower rectum and 4 with cancer of the middle rectum, as well as an average age of 56.2 years (range 28-76). The average surgical time was 183 minutes (range 120-270) and estimated blood loss was 68 milliliters. The incidence of highest morbidity was 15.4 percent and mean hospital stay was 5.4 days. The median follow-up period was 35 (range 9-69) months, with a local recurrence of 7.7 percent and an overall 5-year survival of 100 percent . Conclusions: Total excision of the transanal mesorectum combined with laparoscopic surgery is a feasible and safe technique. The introduction of the variant technique using the instruments of endoscopic transanal microsurgery is more ergonomic and reduces costs(AU)


Asunto(s)
Humanos , Persona de Mediana Edad , Microcirugía Endoscópica Transanal/métodos , Epidemiología Descriptiva , Estudios Prospectivos , Estudios Observacionales como Asunto
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