Parasitological versus molecular diagnosis of strongyloidiasis in serial stool samples: how many?

Strongyloidiasis is usually an asymptomatic disease in immunocompetent patients, caused by Strongyloides stercoralis. However, in immunocompromised patients it can produce a severe clinical profile. Therefore, a correct diagnosis is necessary in these cases and in those chronic asymptomatic patients. The low sensitivity of classical parasitological techniques requires the analysis of multiple serial stool samples. Molecular diagnostic techniques represent an improvement in the detection of the parasite. The objective of this study was to evaluate the minimum number of samples necessary to achieve maximum sensitivity by real-time polymerase chain reaction (PCR). A total of 116 stool samples from 39 patients were analysed by direct microscopic observation, agar culture, Harada-Mori and real-time PCR, in one, two, three and four or more consecutive samples. After two serial samples, 6 out of 39 patients were positive by parasitological and molecular techniques, while 16 of them were real-time PCR positive, and all the patients detected by parasitology were also detected by the molecular technique, reaching 100.00% sensitivity versus 83.00% when analysing a single sample. These data also reflect apparently low specificity (51.52%) and positive predictive value (PPV) (27.27 %) values, due to the high number of cases detected by real-time PCR and not by parasitological techniques. These cases were confirmed as true positives when analysing three, four or more samples from the same patient. In conclusion, the application of molecular techniques decreases the number of serial stool samples necessary to give a diagnosis with the maximum sensitivity.

Análise fitoquímica, potencial antioxidante e toxicidade do extrato bruto etanólico e das frações da espécie Senecio westermaniiDusén frente à Artemia salina / Phytochemical analysis, antioxidant potential and toxicity of crude ethanol extract and fractions of the species Senecio westermanii Dusén against Artemia salina

RESUMO A espécie Senecio westermanii Dusén pertencente à família Asteraceae é planta endêmica e nativa do Brasil encontrada na região da Floresta Atlântica, nos estados do Paraná e São Paulo. O objetivo deste trabalho foi avaliar a composição fitoquímica do extrato bruto etanólico e frações das partes aéreas (folha e caule) de S. westermanii utilizando-se a análise fitoquímica qualitativa e cromatografia líquida de alta eficiência com detector de arranjo de diodos (CLAE-DAD), avaliar in vitro a toxicidade preliminar utilizando Artemia salina e o potencial antioxidante. O estudo fitoquímico qualitativo revelou a presença de alcaloides, flavonoides, iridoides, esteroides/triterpenos, heterosídeos saponínicos e aminogrupos. Através da análise realizada por CLAE-DAD obteve-se o fingerprint característico de cada amostra. No ensaio frente à A. salina houve ausência de toxicidade das amostras, o resultado da DL50 para todas as amostras foi superior a 1000 μg/mL. Todas as amostras apresentaram atividade antioxidante pela redução do complexo fosfomolibdênio, com destaque para a fração clorofórmio que apresentou atividade antioxidante de 92,51% em relação ao padrão rutina. Com relação ao ensaio de redução do radical DPPH•, a fração acetato de etila apresentou IC50 de 26,98 μg/mL. Pelo ensaio do ácido tiobarbitúrico (TBARS) a fração hexano apresentou o melhor índice antioxidante em relação ao padrão rutina. Os resultados obtidos demonstram evidências de que a espécie é fonte potencial de antioxidantes naturais, estimulando assim novos estudos que viabilizam sua utilização no tratamento de patologias associadas aos radicais livres. Além disso, a espécie não apresentou atividade tóxica preliminar, assegurando sua aplicabilidade.

ABSTRACT The species Senecio westermanii Dusén, which belongs to the Asteraceae family, is an endemic and native plant from Brazil. It is found mainly in the Atlantic Forest region, in Paraná and São Paulo states. This study aimed to evaluate the phytochemical composition using a qualitative phytochemical analysis and high performance liquid chromatography with diode array detector (HPLC-DAD), to evaluate preliminary toxicity in vitro using Artemia salina and antioxidant potential. Qualitative phytochemical analysis revealed the presence of alkaloids, flavonoids, iridoids, steroids, triterpenes, saponinic glycosides and aminogroups. The analysis by HPLC-DAD provided the characteristic fingerprint of each sample. In the A. salina assay, the results of LD50 over 1000 μg/mL for all samples did not indicate toxicity of the evaluated extracts. All samples demonstrated antioxidant activity by reducing phosphomolybdenum complex. The highest activity was detected in the chloroform fraction, which presented activity of 92.51% compared to the standard rutin. Regarding the trial to reduce the radical DPPH•, the ethyl acetate fraction showed an IC50 of 26.98 µg/mL. In the thiobarbituric acid assay (TBARS), the hexane fraction showed the highest antioxidant activity compared to the standard rutin. These findings indicate that the Senecio westermanii is a potential source of natural antioxidants, stimulating new studies that enable their use in the treatment of disorders associated with free radicals. In addition, the species did not present toxicity in its evaluation, ensuring its applicability.

Molecular diagnosis of diphyllobothriasis in Spain, most presumably acquired via imported fish, or sojourn abroad.

Human diphyllobothriasis is sporadically detected in Spain. Diphyllobothrium latum and Diplogonoporus balaenopterae have been identified. In the study, four cases of presumably imported diphyllobothriasis in Spanish patients were appraised. Molecular diagnosis allowed us to identify 'exotic' fish tapeworms such as Diplogonoporus balaenopterae in one patient and Diphyllobothrium pacificum in the others.

Surveillance of Chagas disease in pregnant women in Madrid, Spain, from 2008 to 2010.

One of the most important modes of transmission of Trypanosoma cruzi infection in areas where it is not endemic is vertical transmission: from mother to child. The objective of this report is to assess the efficacy of different programmes of serological screening to monitor infection with T. cruzi in pregnant Latin American women living in Madrid (Spain). To achieve this, a retrospective study was undertaken from January 2008 to December 2010 in seven hospitals in the Autonomous Community of Madrid. Serological screening programmes were classified in two main strategies: a selective one (pregnant women from Bolivia) and a universal one (pregnant women from Latin America). A total of 3,839 pregnant women were tested and the overall prevalence was 3.96%. The rate of congenital transmission was 2.6%. The current monitoring programmes have variable coverage ranging between 26% (selective screening) and 100% (universal screening). Monitoring of pregnant women from Latin America only reaches full coverage if universal screening of pregnant women is carried out at any moment of pregnancy, including at delivery. A common national regulation is necessary in order to ensure homogenous implementation of screening.

Application of real-time PCR for the differentiation of Entamoeba histolytica and E. dispar in cyst-positive faecal samples from 130 immigrants living in Spain.

In an effort to improve the diagnosis of intestinal amoebiasis, a real-time PCR has been used for the detection and differentiation of Entamoeba histolytica and E. dispar infections in African or South American immigrants who live in Spain. Faecal samples from all of the 130 subjects had apparently been found to contain E. histolytica/E. dispar cysts by microscopical examination. Using the real-time PCR, E. histolytica DNA was detected in faecal samples from only 10 (7.7%) of the immigrants, with E. dispar DNA detected in the samples from another 117 (90.0%) of the subjects. The use of such PCR in the routine investigation of patients found positive for E. histolytica/E. dispar cysts (by microscopy) is recommended, especially in non-endemic areas.

Diarrea nosocomial por Clostridium difficile en mayores de 80 años / Nosocomial Clostridium difficile-associated diarrhoea in patients older than 80 years

Se realizó un estudio descriptivo entre abril de 2001 y marzo de 2002 en 381 pacientes ingresados en el servicio de Geriatría. Se investigó la adquisición nosocomial de diarrea asociada a Clostridium difficile, en pacientes mayores de 80 años. Se definió diarrea nosocomial asociada a C. difficile como la asociada al uso de antibióticos con estudio citotóxico positivo y aparición a las 72 h del ingreso. Se estudiaron los factores de riesgo asociados al desarrollo de la enfermedad. Se consideró «asociación de casos» cuando 1 o más se manifestaron durante el período sintomático o la fase de tratamiento de un caso previo. Veintiún pacientes tuvieron diarrea nosocomial por C. difficile (5,5%). Su edad media fue de 87 ± 4,1 años (un 57% de mujeres). Todos los pacientes tuvieron pluripatología crónica, desnutrición y dependencia funcional. Diecinueve pacientes tomaron 2 o más antibióticos. El tiempo medio de consumo de antibióticos fue de 21,6 días. Los más utilizados durante la hospitalización fueron amoxicilina-ácido clavulánico y ceftriaxona. La distribución de los casos se hizo en 3 fases. La estancia media de los pacientes fue superior a la media observada en el servicio. La sintomatología fue leve en 20 pacientes y remitió con el tratamiento en 19. La mayoría de los casos ocurrieron asociados en el tiempo. Esto puede indicar contaminación ambiental y transmisión nosocomial por C. difficile. En los servicios de Geriatría es necesario insistir en la importancia de los métodos de control de la infección nosocomial y de reducción de sus mecanismos de transmisión

We performed a descriptive study of a cohort of 381 patients admitted to a geriatric unit from April 2001 through March 2002. Nosocomial acquisition of Clostridium difficile-associated diarrhoea (CDAD) in elderly inpatients (> 80 years) was investigated. A patient with nosocomial CDAD was defined as an individual who had been in hospital for more than 72 hours, with antibiotic-associated diarrhoea and positive C. difficile cytotoxin studies. Risk factors for this disease were studied. Clustering of cases was defined as one or more cases of CDAD occurring during the symptomatic phase of the infection and/or treatment period of a previous case. Twenty-one patients had nosocomial CDAD (5.5%). The mean age of the patients was 87 years (SD 4.1) and 57% were women. All patients had severe concurrent illnesses, malnutrition and functional dependence. Nineteen patients had taken 2 or more antibiotics. The mean duration of antibiotic courses was 21.6 days. The most frequently used antibiotics were amoxicillin-clavulanic acid and ceftriaxone. Clusters of cases occurred in 3 periods. The mean length of hospital stay was greater than that of other patients admitted to the geriatric unit. The clinical course was mild in 20 patients and diarrhoea ceased with treatment in 19. Most cases of CDAD occurred in clusters, which could suggest environmental contamination and nosocomial transmission of C. difficile. The importance of nosocomial infection control and reduction of the transmission mechanisms of these infections should be emphasised in geriatric units

Detection of antibodies to tick salivary antigens among patients from a region of Spain.

With a view to determine which tick species bite humans most frequently in the province of Soria (Spain) and to know whether these species act as vectors of Rickettsia conorii and/or Borrelia burgdorferi, we analysed the presence of antibodies against salivary proteins of several tick species and of antibodies against R. conorii and B. burgdorferi sensu lato in 102 samples of serum. The sera were collected from 63 patients who were treated for tick bites at the health services of the Soria Health Area. Thirty-nine percent of the patients displayed antibodies against Dermacentor marginatus; 21% against Ixodes ricinus, and 6% against Rhipicephalus sanguineus. IgM antibodies against R. conorii were found in 15 patients, but none of the patients included in the study displayed IgG/IgM Abs against B. burgdorferi sensu lato. In 30% of the patients bitten by I. ricinus and in 24% bitten by D. marginatus, antibodies against R. conorii were found. None of the patients who had been bitten by R. sanguineus expressed antibodies against this pathogen. The results obtained here suggest that D. marginatus and I. ricinus are the main vectors of the spotted fever group rickettsiae in our environment and that the role of R. sanguineus in the transmission of these rickettsiae is debatable.

Donor age as a risk factor for patient survival in the liver transplant.

OBJECTIVES: Our aims were to establish whether there is a relationship between donor age and patient and graft survival among liver transplant recipients and to determine the age at which this relationship emerges. PATIENTS AND METHODS: We reviewed 254 consecutive liver transplants performed at the Hospital Ramón y Cajal, Madrid in 206 patients over a 79-month period. Survival rates were determined using Kaplan-Meier curves analyzed by the log-rank method. RESULTS: The mean donors age was 42.08+/-17.89 years (range 8-79 years). The minimum and mean patient follow-up times were 6 months and 29.48+/-23.37 months. Mean patient and graft survival rates, along with their standard errors and 95% confidence intervals were 62.47+/-2.42(57.72-67.21) and 57.30+/-2.40(52.59-62.01) months, respectively. Mean survival was lower (P=.047) among patients who received a graft from a donor of 30 or more years (58.24+/-3.05[52.28-64.21] months) versus from a younger donor (66.19+/-3.55[59.23-73.15] months). Graft survival was also significantly different (P=.037) for donors older versus younger than 25 years (53.04+/-2.83[47.50-58.58] and 64.72+/-4.11[56.67-72.77] months, respectively). CONCLUSIONS: Patients undergoing liver transplant show lower survival when the donor is older than 30 and the survival of the implanted graft is also lower when the donor is over 25.

Impact of donor, recipient, and graft features on the development of primary dysfunction in liver transplants.

OBJECTIVES: To establish the utility of eight anatomic/pathologic suboptimal and 9 others graft features, versus in 20 donor versus 14 recipient characteristics to predict primary transplant dysfunction. PATIENTS AND METHODS: We reviewed 248 consecutive liver transplants performed at the Hospital Ramón y Cajal, Madrid, in 206 patients over a 79-month period. At least one biopsy specimen was obtained from 169 grafts (68.1%). Recipients were classified as showing primary function or dysfunction, the latter group being subdivided into primary failure and inadequate initial function. The primary function and inadequate initial function groups were defined in terms of transaminases less or more than 2000 IU and prothrombin activity over or under 50%, respectively during posttransplant days 2 to 7. RESULTS: The following graft-related rates were recorded: arteriopathy 6.5%, steatosis 29.4% (macrovesicular 26.4%, microvesicular 4.7%, or both 1.7%), hepatocyte vacuolization 14.2%, sinusoidal ectasia 12.4%, hepatocellular necrosis 44.7%, and neutrophilic infiltration 24.4%. The only significant factors in the multivariate analysis were cause of donor death other than cranioencephalic trauma (P=.032) and moderate steatosis (30%-60% affected hepatocytes); (P=.012). CONCLUSIONS: The only factors that seem to influence the development of primary liver dysfunction were a moderate degree of graft steatosis and a cause of brain death other than cranioencephalic trauma.

Survival and complication rates in liver transplant patients showing primary dysfunction versus normofunction.

AIMS: To determine rates of vascular and biliary duct complications, acute rejection, and graft and patient survival according to function status following liver transplantation. METHODS: We classified 248 consecutive liver transplants performed at the Hospital Ramón y Cajal, Madrid, over a 79-month period according to initial function as primary function (NP) versus dysfunction (PD). The latter group was subdivided into grafts showing primary failure (PF) or inadequate function (IPF). The classes NP and IPF were distinguished according to whether transaminase (GOT or GPT) levels and prothrombin activity were above or below 2000 IU and 50%, respectively. RESULTS: There were 23 (9.3%) patients with PD, of whom 12 (4.8%) showed PF. The incidence of vascular and biliary duct complications was similar in both groups, although acute rejection showed a significant difference (PD 3/23 versus NP 98/225; odds ratio =.18). In contrast, the mean survival rates of the grafts (NP 60.37 versus IPF 39.90 months) or patients (NP 63.02 versus PD 47.10 months) were not significantly different. Only 1- and 3-month graft survival rates significantly differed between the NP and IPF groups (NP 95% versus IPF 63%; P=.03 and NP 89% versus IPF 58%; P=.02, respectively). CONCLUSIONS: Recipients with PD or NP after liver transplant showed no differences in the incidence of vascular or biliary duct complications. These groups did vary, however, in terms of rates of acute rejection episodes. No differences in graft and patient survival rates were observed except a significantly lower graft survival at 1 and 3 months, among patients with inadequate primary function.

Primary cutaneous cryptococcosis in an immunocompetent host: case report and review of the literature.

We report a 46-year-old male who developed cellulitis of his third right finger after being injured with a metallic object. Cryptococcus neoformans var. neoformans was isolated from both the skin and the metallic object. Systemic cryptococcosis and immunosuppression was excluded by appropriate tests. Complete healing was achieved after 10 months of itraconazole 100 mg/12 h and surgical excision of the remaining lesion. Our literature review shows another 16 cases of primary cutaneous cryptococcosis in nonimmunocompromised hosts. The clinical, mycological and therapeutic characteristics of these patients are reviewed.

Epidemiological characteristics of dogs with Lyme borreliosis in the province of Soria (Spain).

From the point of view of the human disease, dogs are the most important animal reservoir of Lyme borreliosis; therefore, they are used as 'sentinel animals'. In order to know the epidemiological characteristics of dogs with antibodies against Borrelia burgdorferi, 146 canine sera samples collected during 1993-94 have been studied. The antibody detection was made by an indirect immunofluorescence method and confirmed by a haemagglutination test. Seventeen dogs (11.6%) were seropositive, of which seven were hunting dogs, three were shepherd dogs, five were domestic pets, one was a watchdog and one was a stray dog. Seven dogs had longhair phenotype, 2 medium length type, 4 hard-hair and the other 4 were short-haired. Twelve seropositive dogs were males and five were females. All seropositive dogs were more than 1 year old. The dogs with greater seroprevalence were those that harboured ticks more often than the rest. Spring and summer were the seasons when more ticks were observed. The seroprevalence found in dogs was similar to that previously obtained in humans in this area. This finding gave evidence to an intimate association between human and canine seroprevalence.

Resection of colorectal liver metastases: analysis of risk factors.

OBJECTIVE: to analyze the survival after surgery to resect liver metastases from colorectal cancer, and to investigate the influence of 15 potentially prognostic factors on survival. METHODS: this retrospective study included all patients diagnosed as having colorectal cancer who were operated on with curative intent (without no evidence of tumor after surgery) during the 10-year period from 1988 to 1998. Survival analysis was done using the Kaplan-Meier method, and the prognostic value of each factor was determined with the log-rank test. Of the 60 surgical patients we studied, 58 survived the postoperative period (< 30 days). We analyzed their epidemiological, histological and chronopathological characteristics, tumor markers, and type of resection. RESULTS: mean survival was 33. 06 +/- 3.2 months, and ranged from 2 to 108 months. Survival at 1, 3 and 5 years was 90.1%, 48.9% and 28.9% respectively. None of the 15 prognostic factors studied was significantly related with survival (p < 0.05). CONCLUSIONS: surgical treatment of liver metastases from colorectal cancer is a safe option and offers good survival rates. The potentially prognostic factors studied here do not allow us to identify those patients who might benefit most from this procedure.

[Cutaneous myiasis by Sarcophaga sp]. / Miasis cutánea por Sarcophaga sp.

BACKGROUND: Myasis are due to the invasion of tissue or cavities of animal organism by dipterous larvae. PATIENTS AND METHODS: Two cases of semi-specific myasis caused by Sarcophaga larvae are described. RESULTS: Case 1: A 77-year-old woman with Kaposi sarcoma in the left leg developed cutaneous radionecrosis secondary to radiotherapy. In June, 1998 five fly larvae were observed moving freeing within the wound. These were removed with forceps and local dressing of the wound was performed with povidone-iodine. Case 2: A 87-year-old man with moderate dementia, progressive immobilization syndrome prostasic neoplasm and gastric ulcer reported. In the posterior part of the right outer ear the presence of three fly larvae were observed with some dermic orifices made. The larvae were removed with forceps and local dressing was carried out with povidone-iodine. In the laboratory an adult form was obtained from one of the larva. CONCLUSIONS: Myasis in infrequent in Spain and appears particularly in people with some predisposing factor. Treatment consists in the elimination of the larvae in the infected tissue and disinfection of the wound.

[Pelvic lipomatosis: a case report with diagnostic and therapeutic approach]. / Lipomatosis pélvica: un caso con enfoque diagnóstico y terapéutico.

Pelvic lipomatosis is a rare disease of unknown etiology characterized by benign proliferation of fat in the pelvis. We describe a 27-year-old man with pelvic and retroperitoneal lipomatosis causing a severe urinary and fecal obstruction. The diagnosis was evaluated by barium enema, intravenous urogram, computerized tomography and magnetic resonance imaging. The therapeutic approach is described and discussed.