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.

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.

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.

[The value of the parathyrin-related protein (PTH-RP) in the diagnosis of cancer-associated hypercalcemia]. / Valor de la proteína relacionada con la paratirina (PTH-RP) en el diagnóstico de la hipercalcemia asociada al cáncer.

BACKGROUND: The parathyrine related protein (PTH-RP) is very similar, both in structure and in function, to the PTH and is considered as a mediator in humoral hypercalcemia in cancer. The aim of this study was to know the clinical value of PTH-RP measurement. METHODS: Serum PTH-RP concentrations were studied in 22 healthy subjects, 13 patients with primary hyperparathyroidism, 9 patients with solid neoplasms and normocalcemia, 26 patients with solid neoplasms and hypercalcemia and 4 patients with hematologic neoplasms and hypercalcemia. The PTH-RP was quantified by a competitive radioimmunoassay technique using a specific antibody of the PTH-RP 1-40 fragment. Intact parathyrine (i-PTH) was quantified by an IRMA method using 2 polyclonal antibodies (INCSTAR). RESULTS: Fifteen (68%) of the healthy controls presented undetectable serum PTH-RP concentrations. The serum PTH-RP concentration was normal in all those patients with hyperparathyroidism. Elevated serum PTH-RP values were not found in patients with solid neoplasms and normocalcemia or in those with hematologic neoplasms and hypercalcemia. High values of PTH-RP were observed in 8 out of 9 (88%) of the patients with solid neoplasms and hypercalcemia with bone metastasis and in 7 out of 11 (63%) of the patients with bone involvement. CONCLUSIONS: Serum parathyrine-related protein was found to be high in a large proportion of patients with solid neoplasms and hypercalcemia. Serum PTH-RP determination is useful in the clinical investigation of patients with hypercalcemia. Even in patients with bone metastasis, hypercalcemia may have a humoral background.

[Hantavirus infection in Spain: seroepidemiologic study in the province of Soria]. / Infección por Hantavirus en España: estudio seroepidemiológico en la provincia de Soria.

BACKGROUND: The aim of the present was to study the seroprevalence of Hantavirus infection in the province of Soria, Spain. METHODS: Total specific antibodies against Hantavirus (Hantaan and Puumala virus) were studied by indirect immunofluorescence in 537 sera (251 males and 286 females) of a population from the province of Soria, distributed proportionally to the number of inhabitants from the health care areas. The age range of the population was from 1 to 95 years, the group mode ranging between 25 to 30 years and with a mean age of 36.4 years. The sera with antibody titers equal to or higher than 1/32 were considered as positive. RESULTS: Significant titers of antibodies against Hantaan or Puumala virus were detected in 2.2% of the total of sera studied (12 positive sera); 1.75% in women (5/286) and 2.8% in men (7/251). Of these 12 positive sera, 8 reacted against both virus (1.5%-8/537), 3 exclusively to Puumala and 1 only to Hantaan. Seroprevalence against Puumala was 2% (11/537) and 1.7% against Hantaan (9/537). The age of the subjects with positive sera ranged from 3 to 73 years with a mean of 33.8 years with higher prevalence in the group from 30 to 40 years of age (4.1%). CONCLUSIONS: The results of this study confirm the existence of infection by Hantavirus in Spain.

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.

[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.

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.

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.

[Variation of the incidence and antibiotic sensitivity of Neisseria gonorrhoeae in a 7-year period]. / Variación de la incidencia y sensibilidad antibiótica de Neisseria gonorrhoeae en un período de 7 años.

BACKGROUND: The aim of this study was to determine the incidence and antibiotic sensitivity of Neisseria gonorrhoeae versus penicillin, tetracycline, cefoxitin, ceftriaxone and spectinomycin from 1988 to 1994 in the province of Soria (Spain). METHODS: From January 1988 to December 1994, clinical samples of 57 strains of N. gonorrhoeae were isolated. Auxotype, serotype, and minimum inhibitory concentration (MIC) studies were performed versus 5 antimicrobials and analysis of plasmids in the penicillinase producer strains (PPNG), was carried out in the Bacteriology Department of the Instituto Carlos III in Madrid (Spain). RESULTS: The rate of incidence of the isolation of N. gonorrhoeae was similar from 1988 to 1990 (11.7 to 19.1 cases per 100,000 inhabitants) and decreased from 1991 to 1994 (6.4 to 0 cases per 100,000 inhabitants). The strains belonged to 30 different auxotypes/serotypes indicating a great heterogeneity among them. The proportion of penicillinase producing N. gonorrhoeae (PPNG) increased over the study period. The first strains with high resistance to tetracycline (TRNG) were first isolated in the authors' area in 1991 and the proportion increased up to the end of the study. All the strains were sensitive to ceftriaxone, cefoxitin and spectinomycin. CONCLUSIONS: A decrease was observed in the rate of incidence of N. gonorrhoeae since 1991 with an increase in the proportion of NGPP and TRNG strains. Ceftriaxone and spectinomycin present good activity versus all the strains studied with their empiric use as treatment being possible in the province of Soria (Spain).

[Evaluation of 4 media for the isolation of Helicobacter pylori from gastric biopsies]. / Evaluación de cuatro medios para el aislamiento de Helicobacter pylori en biopsias gástricas.

BACKGROUND: Different culture mediums were used for the isolation of Helicobacter pylori. In this study three mediums commonly used for the culture of this microorganism were evaluated: chocolate agar, Martin-Lewis medium and CP of Dent medium as well as a new selective medium Pylori agar. METHODS AND RESULTS: Over a period of 7 months gastric biopsies were collected in 112 patients with clinical manifestations of chronic atrophic gastritis. H. pylori was isolated in some of these mediums on 73 occasions (65.1%). On 72 of the 73 occasions the microorganism grew in the Pylori agar medium (98.6%) being the medium achieving the greatest number of isolations, followed by the CP of Dent medium with 67 isolations (91.7%), 65 isolations with the Martin-Lewis medium (89.0%) and 57 isolations with chocolate agar (78.0%). The chocolate agar medium was that which most frequently presented contaminant flora (35 times: 61.4%) and the CP of Dent medium achieved the most rapid and widespread growth. The colonies were largest and therefore easier to recognize in the CP of Dent and Pylori agar mediums. CONCLUSIONS: The authors recommend the association of the CP of Dent and Pylori agar mediums in the isolation of Helicobacter pylori since these two mediums provide the greatest number of isolations with the greatest growth and ease for colony recognition.

[Seroprevalence of Coxiella burnetii and Rickettsia conorii infection in the province of Soria]. / Seroprevalencia de la infección por Coxiella burnetii y Rickettsia conorii en la provincia de Soria.

BACKGROUND: Determine the antibodies prevalence against Coxiella burnetii and Rickettsia conorii in Soria (Spain). METHODS: By an indirect immunofluorescence test we examined sera samples for antibodies (IgG, IgM and IgA) against C. burnetii (phase II, formolized antigen) and R. conorii (moroccan strain). The test has been applied to 298 sera (150 men and 148 women) from Soria (Spain). The age of the study population ranged between 2 and 91 years. The results were considered positive when the titres were greater than or equal to 1/80 for C. burnetii and 1/40 for R. conorii. RESULTS: Antibodies against C. burnetii were present in 20.8% of the examined sera, 8.8% for women (13/148) and 32.7% for men (49/150). We demonstrated titers of antibodies upper to 1/40 in all age groups, and a greater number of positive results in the persons older than 40 years old (p = 0.0001). In all areas we detected positive results and in many of these areas, the positivity was high (38%). We demonstrated positive results against R. conorii in 5% of the people (4.7% for women and 5.3% for men). More than 90% of the positive cases was present in the eastern part of Soria, and in an small area, the positivity was 20%. CONCLUSIONS: These results confirm a high prevalence of antibodies against C. burnetii in the Soria people, and the existence of R. conorii infection in this area of Spain with a particular distribution. We believe that it is very important to establish an epidemiological surveillance for the detection of clinic cases.

Evaluation of two commercial enzyme immunoassays for the diagnosis of Helicobacter pylori infection.

Sera from 65 patients with upper gastrointestinal tract symptoms and proven Helicobacter pylori infection, and from 42 negative controls were tested with two commercial EIAs (GAP test, BioRad; and ECP test, Biometra) and two non-commercial EIAs, one performed with whole sonicated cells and the other with acid extract of Helicobacter pylori as antigen. The GAP assay showed a sensitivity of 83.1% and a specificity of 47.6%. The ECP assay showed a sensitivity of 87.7% and a specificity of 61.9%. For both non-commercial EIAs these figures were 87.7% and 88.1%, respectively. Independent of the interpretive criteria established by the manufacturers, receiver operating characteristic curves were plotted for better evaluation of the four methods. Both commercial tests showed a lower probability of yielding a correct diagnosis than the non-commercial tests (p less than 0.05). Although commercial EIAs are convenient for the diagnosis of Helicobacter pylori infection, the accuracy of the two commercial tests evaluated in this study was lower compared to that of the two non-commercial EIAs.

[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.

[Lyme disease in the province of Soria: clinico-epidemiologic study]. / Enfermedad de Lyme en la provincia de Soria: estudio clinicoepidemiológico.

BACKGROUND: To determine the prevalence of the Borrelia burgdorferi infection in the Soria Province, and the detection of the human disease cases. METHODS: By an indirect immunofluorescence technique, we had studied total antibodies (IgG, IgM and IgA) against B. burgdorferi (B 31 strain) in the sera of 298 people (150 men y 148 women) of the Soria Province. The results were considered positive when the titers were same or upper to 1/256. All positive sera were tested for specific IgM antibodies by same method; IgM results were considered positive when the titers were upper to 1/16. RESULTS: 13.1% of the people had positive results (39 sera): 14 for women (9.5%) and 25 for men (16.7%). We detected 14 sera with positive IgM. We demonstrated titers same or upper to 1/256 in all age group, with a major prevalence in the age group between 41 and 50 years (22.6%). In all areas of the Province we detected positive results and in many of this areas the positivity was upper to 30% of the studied sera. 12 persons with positive IgM or high titers of total antibodies presented signs or symptoms of disease in relation with B. burgdorferi infection; 6 persons had alterations of the liver function do not explicable by other etiology. CONCLUSIONS: The B. burgdorferi infection present a high prevalence in this Province and a wide geographic distribution, with high endemic areas. We demonstrated patients with symptomatology compatible with Lyme borreliosis, and we wish to emphasize the frequency of alterations of the liver function in this group of people.