The conventional diagnosis challenge: Real-time PCR and nested PCR correlation with the scoring system for individuals at high-risk of Pneumocystis jirovecii pneumonia

Introduction. Pneumocystis jirovecii is an opportunistic fungus that affects mainly people living with HIV (CD4 cell count lower than 200 cells/ml) and other immunosuppressed patients. Since P. jirovecii does not grow on routine mycological media, diagnosis of P. jirovecii pneumonia relies on indirect evidence of its presence in respiratory samples. Objectives. To associate the results of direct immunofluorescence and two molecular methods with a score to predict P. jirovecii pneumonia in patients with AIDS. Materials and methods. A prospective study was conducted with 40 patients. A respiratory sample collected before treatment was subjected to direct immunofluorescence using the Merifluor kit, to nested PCR targeting the mitochondrial large subunit ribosomal RNA, and to the VIASURE real-time PCR kit. Results. These three techniques revealed P. jirovecii in 6, 12, and 15 samples, respectively. All positive samples by direct immunofluorescence were positive by nested PCR, and all positive samples by nested PCR amplified by real-time PCR. There was a statistically significant association between the P. jirovecii pneumonia score and the molecular methods. Two patients were early diagnosed and responded well to treatment. Conclusion. Molecular methods, especially real-time PCR, are recommended for early diagnosis of P. jirovecii pneumonia in AIDS patients.

Introducción. Pneumocystis jirovecii es un hongo oportunista que afecta principalmente a personas con HIV (recuento de CD4 menor de 200 células/ml) y a otros pacientes inmunosuprimidos. Como P. jirovecii no crece en los medios micológicos de rutina, el diagnóstico de neumonía por P. jirovecii se basa en la evidencia presente en muestras respiratorias. Objetivos. Asociar los resultados de la inmunofluorescencia directa y los de dos métodos moleculares con un puntaje para predecir la neumonía causada por P. jirovecii en pacientes con sida. Materiales y métodos. Se realizó un estudio prospectivo de 40 pacientes. Se recolectó una muestra respiratoria antes del inicio de tratamiento y se sometió a una prueba de inmunofluorescencia directa con el kit Merifluor, una PCR anidada para la amplificación de la subunidad larga del ribosoma mitocondrial y una PCR en tiempo real usando el kit VIASURE. Resultados. Estas tres técnicas evidenciaron la presencia de P. jirovecii en 6, 12 y 15 muestras, respectivamente. Todas las muestras positivas por inmunofluorescencia directa fueron positivas en la PCR anidada y todas las muestras positivas en la PCR anidada amplificaron por PCR en tiempo real. Se encontró una asociación estadística entre los valores de la neumonía causada por P. jirovecii y los métodos moleculares. Dos pacientes con diagnóstico temprano respondieron satisfactoriamente al tratamiento. Conclusión. Se recomiendan los métodos moleculares, especialmente la PCR en tiempo real, para el diagnóstico temprano de neumonía causada por P. jirovecii en pacientes con sida.

Immunohistochemical detection of Treponema pallidum in skin samples with clinical and histopathological correlations and Warthin-Starry staining critical analysis

Abstract Background: Syphilis in its different phases may be a difficult diagnosis in clinical and histopathological grounds. Objectives: The present study objectives were to evaluate the detection and tissue distribution of Treponema pallidum in skin lesions of syphilis. Methods: A blinded diagnostic accuracy study was performed with immunohistochemistry and Warthin-Starry silver staining in skin samples from patients with syphilis and other diseases. Patients attended two tertiary hospitals between 2000 and 2019. Prevalence ratios (PR) and 95% confidence intervals (95% CI) were calculated for the association between immunohistochemistry positivity and clinical-histopathological variables. Results: Thirty-eight patients with syphilis and their 40 biopsy specimens were included in the study. Thirty-six skin samples were used as non-syphilis controls. The Warthin-Starry technique was unable to accurately demonstrate bacteria in all samples. Immunohistochemistry showed spirochetes only in skin samples from patients with syphilis (24/40) with 60% sensitivity (95% CI 44.8-75.2). Specificity was 100% and accuracy, 78.9% (95% CI 69.8-88.1). Most cases had spirochetes in both dermis and epidermis and there was a high bacterial load. Study limitations: Correlation between immunohistochemistry and clinical or histopathological characteristics was observed but was limited statistically due to the small sample size. Conclusions: Spirochetes were promptly seen in an immunohistochemistry protocol, which can contribute to the diagnosis of syphilis in skin biopsy samples. On the other hand, the Warthin-Starry technique showed to be of no practical value.

Validation of an algorithm based on clinical, histopathological and immunohistochemical data for the diagnosis of early-stage mycosis fungoides

Abstract Background: Diagnosis of mycosis fungoides is challenging due to the non-specificity of clinical and histopathological findings. The literature indicates an average delay of 4-6 years for a conclusive diagnosis. Refinement of the histopathological criteria for the diagnosis of patients in early stages of the disease is considered of interest. Objectives: To study the histopathological aspects of early-stage mycosis fungoides and the applicability, in a retrospective form, of the diagnostic algorithm proposed by Pimpinelli et al. Methods: Observational, retrospective, transversal study based on revision of histopathological exams of patients with suspected mycosis fungoides. Medical records were reviewed, and complementary immunohistochemistry performed. Results: Sixty-seven patients were included. The most frequent histopathological features were superficial perivascular lymphoid infiltrate (71.6%), epidermotropism (68.7%), lymphocytic atypia (63.8%), hyperkeratosis (62.7%) and acanthosis (62.7%). Forty-three patients scored 4 points at the algorithm, by clinical and histological evaluation. Immunohistochemistry was performed on 23 of the 24 patients with less than 4 points. Of those 23, 22 scored 1 point, allowing a total of 61 patients (91%) with the diagnosis of early-stage mycosis fungoides. Study limitations: Its retrospective character, reduced sample size and incomplete application of the algorithm. Conclusions: Application of the Pimpinelli et al. algorithm, even in an incomplete form, increased the percentage of cases diagnosed as mycosis fungoides. Routine application of the algorithm may contribute to earlier and specific management and improvement of the patients' outcome.

Gait variability and symmetry assessment with inertial sensors for quantitative discrimination of Trendelenburg sign in total hip arthroplasty patients: a pilot study based on convenience sampling

Abstract Introduction The aim of this pilot study based on convenience sampling was to analyze the feasibility to quantitatively discriminate Trendelenburg sign (TS), a characteristic drop in pelvic position during gait in hip disfunctions, in patients with total hip arthroplasty (THA), by assessing gait variability and symmetry using inertial sensors. Methods Thirteen patients with right THA, divided into two groups with (GTS, n=4) and without TS (GnTS, n=9) assessed by experienced physician, were enrolled in the study. Harris Hip Score was applied for specific evaluation of THA. The protocol consisted in walking on a level treadmill during 3 minutes with two inertial sensors attached at anterior superior iliac spine of both sides. For each left and right step, features were extracted from the Y-axis gyroscope signals: peak value, mean absolute value, standard deviation and range. For each feature, a symmetry ratio was calculated as the ratio between left and right side. Results No significant differences were found in Harris Hip Score between groups. The variability assessed by standard deviation for left step, contralateral to the replaced side, was significantly larger for GTS group (p<0.001). Significant differences in the symmetry ratios were found between GTS and GnTS for all features extracted from gyroscopes Y-axis (W=144, p<0.001). The symmetry ratios for GnTS group were approximately equal one (except for range), whereas for the GTS group they exceed the 10% criterion. Conclusion The variability and symmetry ratios of gait features extracted from inertial sensors were successful to discriminate TS in THA patients.

Gastrointestinal parasites of swine raised in different management systems in the State of Rio de Janeiro, Brazil / Parasitos gastrintestinais de suínos criados sob diferentes sistemas de manejo no Estado do Rio de Janeiro

From 2012 to 2013 were surveyed gastrointestinal parasites from pig farms located in different municipalities in the state of Rio de Janeiro. Fecal samples from 790 pigs were collected from the rectum on 88 family farms and 702 farms with industrial production. The samples were subjected to Faust et al., Sheather, Ritchie, Lutz and direct examination faecal techniques. The estimated parasite prevalence was 93.1% in family farms and 59.1% in industrial farms. Balantidium coli, coccidia and Entamoeba sp. were the parasites with the highest frequencies, and the male and female reproductive categories and fatteners pigs the most infected (p<0.05). Trophozoites of B. coli were most evident in stool samples from semi-solid followed by solid and diarrheal consistencies. Strongyles eggs and Trichuris suis have been detected exclusively in family farms. Ascaris suum eggs and Strongyloides ransomi showed low frequency. The high degree of parasitism, especially protozoa, indicates the need to reassess the management of pigs in both types of production...

De 2012 a 2013 foram pesquisados parasitos gastrintestinais de suínos de granjas localizadas em diferentes municípios do estado do Rio de Janeiro. Amostras fecais de 790 suínos foram coletadas da ampola retal, sendo 88 de propriedades familiares e 702 de granjas com produção industrial. As amostras foram submetidas às técnicas de Faust et al., Sheather, Ritchie, Lutz e exame direto. A prevalência estimada foi de 93,1% nas granjas familiares e 59,1% nas granjas industriais. Balantidium coli, coccídios e Entamoeba sp. foram os parasitos que apresentaram as maiores frequências, sendo as categorias machos e fêmeas reprodutoras e leitões de terminação as mais infectadas (p<0,05). Trofozoítas de B. coli foram mais evidenciados nas amostras fecais de consistências semi-sólida, seguida pelas fezes com consistências sólida e diarreica. Ovos de estrongilídeos e Trichuris suis foram detectados exclusivamente em criações familiares. Ovos de Ascaris suum e de Strongyloides ransomi apresentaram baixa frequência. O alto grau de parasitismo, principalmente de protozoários, indica a necessidade de reavaliação do manejo dos suínos em ambos os tipos de produção...

Occurrence of anti-Toxoplasma gondii antibodies in cattle and pigs slaughtered, State of Rio de Janeiro / Ocorrência de anticorpos anti-Toxoplasma gondii em bovinos e suínos abatidos, Estado do Rio de Janeiro

This study aimed to estimate the occurrence of anti-Toxoplasma gondii antibodies by means of the Indirect Immunufluorescent Antibody Assay (IFAT) in cattle and pigs from slaughterhouses in two municipalities in the State of Rio de Janeiro. IgG anti-T. gondii antibodies was found in 1.96 percent (9/459) of the cattle and 7.64 percent (31/406) of the pigs, without any positive association (p > 0.05) with risk factors for transmission and seropositivity. Taking the cutoff point of IFAT ³ 1:64, titers of 64 and 256 were detected, suggesting chronic infection. Despite the low prevalence rates found for cattle and pigs, these indicated environmental contamination in the municipalities investigated, biosafety failures at the technified pig farm evaluated and the possibility of human and animal infection through eating raw or undercooked meat from these animals.

Este trabalho objetivou estimar a ocorrência de anticorpos anti-Toxoplasma gondii por meio da Reação de Imunofluorescência Indireta (RIFI) em bovinos e suínos de abatedouros de dois municípios do Estado do Rio de Janeiro. Anticorpos IgG anti-T. gondii foram encontrados em 1,96 por cento (9/459) dos bovinos e em 7,64 por cento (31/406) dos suínos, sem associação positiva (p > 0,05) em relação aos fatores de risco de transmissão e a soropositividade. Considerando-se o ponto de corte (RIFI ³ 1:64), foram detectados os títulos de 64 e 256, sugestivos de infecção crônica. Apesar de baixas, as prevalências encontradas para bovinos e suínos indicaram contaminação ambiental nos municípios avaliados, falhas na biossegurança da granja tecnificada de suínos e a possibilidade de infecção humana e animal pela ingestão da carne crua ou mal passada desses animais.

Cisto dentígero seqüencial: relato de caso / Sequential dentigerous cyst: case report

O cisto dentígero (CD) é definido como um cisto que se origina pela separação do folículo pericoronáriode um dente incluso devendo estar associado à coroa do mesmo e unido à junção cemento-esmalte.CD seqüencial (CDS) pode ser definido como o desenvolvimento de maneira seqüencial de mais de um cistodentígero em diferentes momentos da vida do paciente. Paciente de 16 anos, sexo masculino, queixando-sede aumento de volume do lado esquerdo da mandíbula com evolução lenta. Após exame físico, radiográficoe histopatológico, foi diagnosticado cisto dentígero. Após 12 anos de controle do paciente, houve aparecimentode imagem radiolucida associada ao dente 48, que teve o diagnóstico de cisto dentígero. Esta descriçãorepresenta um caso raro de CDS. O paciente encontra-se em acompanhamento e após de 16 anos daprimeira ocorrência do CD, não apresenta sinais clínicos e radiográficos de recidivas.

Dentigerous cyst (DC) is originate from separation of dental follicle of the crown of an enclosed tooth,must be associated to the crown of exactly joined to the cementoenamel junction. Sequential DC (SDC)represents the development of DC in sequential way at different moments of one determined period of life ofthe patient. Patient of 16 years, male, complaining of increase of volume of the left side of the jaw with slowevolution. After clinical, radiographic and histopathology exam, the diagnosis of DC was performed. After 12-years follow-up, it was observed a radioluscence, well-defined area associated to the 48 tooth. Also, it wasdiagnosed as DC. This case report represents a rare case of SDC. The patient is free-disease after 16 yearsof follow-up.