Combined rapid urease test and histology for the diagnosis of helicobacter pylori infection

Significance@#Accurate detection of Helicobacter pylori (HP) is essential for the diagnosis of HP infection. The use of antibiotics and proton pump inhibitors (PPI) may result in false-negative rapid urease test (RUT) results. We aimed to determine the sensitivity and specificity of RUT compared with histology and assess the detection rate of combined RUT and histology for HP infection. @*Methodology@#Retrospective data collection was performed on 192 patients who were tested for both RUT and histology at the time of upper endoscopy from 2017 to 2018. At least two gastric biopsies (1 from corpus, 1 from antrum) were taken each for RUT and histology. The endoscopy was performed by a single gastroenterologist and a single pathologist was responsible for interpreting the histology with hematoxylin and eosin (H&E) and Giemsa stain. The gold standard test for the diagnosis of HP infection was histology. Demographic profile, RUT and histology results were reviewed. Tests for diagnostic accuracy were computed using SPSSv23. @*Results@#192 patients were tested for RUT and histology. 52(27.1%) were males and 140(72.9%) were females with a mean age of 54±17 years. Epigastric pain was the most common indication (42.7%). 24(12.5%) patients tested positive for HP infection. Among these; 16(8.3%) tested positive for both RUT and histology(true-positive), while 8(4.2%) tested negative for RUT but had positive histology(false-negative). 6 out of 8(75%) patients with false negative results had PPI use. The sensitivity and specificity of RUT for the diagnosis of HP infection were 66.7 and 98.2%, respectively. While the positive and negative likelihood ratio were 37.3 and 0.34, respectively with a diagnostic odds ratio of 110. @*Conclusion@#The HP detection rate of RUT combined with histology increased by 33% compared with RUT alone. RUT is a highly specific test for diagnosing HP infection. Given its modest sensitivity, histology plays an important role in the diagnosis of HP infection, especially in patients taking PPIs. We recommend doing histology when RUT is negative to increase the HP detection rate.

Efficacy of FNAC in early diagnosis of prostatic carcinoma

Background: Prostatic carcinoma is one of the most important causes of mortality in elderly men mainly because of the late detection despite of the fact that it is a potentially curable disease. Fine needle aspiration cytology (FNAC) is an easy to perform outpatient procedure requiring no expensive equipment or anesthesia. Objectives: The present study was carried out in an attempt to evaluate the fine needle aspiration cytology in the diagnosis of carcinoma prostate. Materials and methods: The present study was performed on 27 patients admitted in the surgical wards, with complaints suggestive of prostatic disease, in whom there was found to be a suspicion of malignancy of the prostate gland. Results: Among 27 patients, 14 patients were diagnosed as prostatic malignancy on per rectal FNAC whereas, 19 patients were confirmed with prostatic cancer histologically. Out of 19 histologically confirmed cancer cases, 16 were also positive on FNAC i.e. 84.21% accuracy of FNAC in detecting prostatic malignancy. Conclusion: Fine needle aspiration cytology is easily available and inexpensive procedure. It is a reliable method in the diagnosis of prostatic cancer. Its positive results are relatively more reliable than the negative ones. It is an effective method in follow up of the cancer cases.

To evaluate the role of sputum in the diagnosis of lung cancer in south Indian population.

Background: The main advantage of sputum cytology is its simplicity, non-invasiveness and minimal discomfort to the patient. Though, the sputum is evaluated in the diagnosis of lung cancer, the report on the same in the South Indian population was lacking. Therefore, the present study has been undertaken to evaluate the role of sputum in the diagnosis of lung cancer in South Indian population. Methods: The material consisted of sputum samples from 133 patients and was collected in clean wide mouthed disposable plastic containers. Patients were asked to collect sputum the next morning after washing the mouth properly. The sputum was immediately brought to the laboratory and poured into a watch glass. Four smears were prepared from each sample, out of which two smears were immediately fixed in methanol and the other two were air-dried. The methanol fixed smears were stained with Papanicolaou stain. Out of the two air dried smears, one was stained with May Grunwald Giemsa and the other with Gabbot's method for AFB. The smears were screened for malignant cells and a cytological diagnosis was made. The cytological diagnosis was correlated with the histopathological diagnosis. The data obtained were represented as mean percentages. Results: The observation of sputum smears showed numerous pleiomorphic keratinized squamous cells, keratinized squamous cell with hyper chromatic nucleus in well differentiated squamous cell carcinoma, pleiomorphic cells having vacuolated cytoplasm and vesicular nucleus with prominent nucleoli as in adenocarcinoma of the lung, cells arranged in small clusters and having scanty cytoplasm in small cell carcinoma and cells are slightly larger than lymphocyte with scanty cytoplasm and hyper chromatic, grooved nuclei in small cell carcinoma. Conclusion: Cytology of sputum is extremely useful and highly sensitive. The diagnostic accuracy is directly proportional to the number of samples. Sputum cytology is highly sensitive for the centrally located squamous cell carcinoma rather than the peripherally located adenocarcinoma. Properly collected, simple sputum examination alone can give results similar to other highly expensive methods like bronchoscopic material for the diagnosis of lung cancer.

Respuesta inflamatoria genital en la detección de alteraciones por virus del papiloma humano / The inflammatory genital reaction in the detection of alterations by human papilloma virus / Reagào inflamatoria genital na detecgào de alteragòes por Papilomavírus Humano

Disfunción vaginal (DV) (vaginosis/vaginitis) es el síndrome genérico de mayor prevalencia, alcanzando el 50% de todas las mujeres en edad fértil (sintomáticas y asintomáticas). El virus del Papiloma Humano (HPV) se detecta en 30 a 40% de mujeres en edad fértil (sintomáticas y asintomáticas) y se asocia a alteraciones pre-neoplásicas y a carcinoma invasor del cuello uterino. El diagnóstico sindrómico de DV y alteraciones inducidas por HPV es ineficiente y en la actualidad la morfología (macro y microscópica) es el gold standard, pero requiere ordenamiento. El Estudio del Contenido Vaginal es la prueba de laboratorio bacteriológico de mayor solicitud luego del urocultivo. BACOVA normatiza el diagnóstico de vaginosis/vaginitis y ERIGE aumenta el valor predictivo de células que alertan sobre alteraciones epiteliales. Desde 2007 al presente en los talleres BACOVA ERIGE (tinción de Giemsa) se evaluó la sensibilidad de la detección de células anormales exfoliadas. Un 99% de los participantes coincidió con la detección de koi-locitos. BACOVA/ERIGE no reemplaza al Papanicolaou de ninguna manera, pero puede y debe realizarse en laboratorios periféricos, con lo que además del diagnóstico de vaginosis/vaginitis con 100% de valor predictivo, aumentan la cobertura preventiva de estados proliferativos.

Vaginal dysfunction (DV) (vaginosis/vaginitis) is the generic syndrome of major prevalence, reaching 50% of all women in fertile age (symptomatic and asymptomatic). The Human Papillomavirus (HPV) is detected in 30-40% of women in fertile age (symptomatic and asymptomatic) and is associated to pre-neoplastic lesions and invading carcinoma of the uterine cervix. The diagnosis for the symptoms of DV and the alterations induced by HPV are inefficient and at present, the morphology (macroscopic and microscopic) is the standard gold, but it needs better classification. The Study of the Vaginal Content is the test of major request after urocultives in bacteriological laboratories. BACOVA establishes the procedure for the diagnosis of vaginosis/vaginitis and ERIGE increases the predictive value of cells that give the alarm on epithelial alterations. From 2007 to the present sensitivity in the detection of abnormal exfoliated cells from vagina and uterine cervix was evaluated during the BACOVA - ERIGE, (Giemsa's stain) workshops, 99% of the participants coincided with the detection of koilocytes. BACOVA/ERIGE does not replace the Papanicolaou by any means, but it can and must be performed in peripheral laboratories, where apart from the diagnosis of vaginosis/vaginitis with 100% of predictive value, it is possible to increase the detection of precocious proliferative changes of the squamous epithelium.

Disfungào vaginal (DV) (vaginose / vaginite) é a síndrome mais prevalente genérica, atingindo 50% de todas as mulheres em idade fértil (sintomáticas e assintomáticas). O Papilomavírus Humano (HPV) é detectado em 30-40% das mulheres em idade fértil (sintomáticas e assintomáticas) e está associado a alteragòes pré-neoplásicas e a carcinoma invasivo do colo do útero. O diagnóstico sindrómico de DV e alteragòes induzidas pelo HPV é ineficiente e atualmente a morfologia (macroscópica e microscópica) é o padrào ouro, mas precisa de ordenamento. O Estudo do Conteúdo Vaginal é o exame de laboratòrio bacteriológico mais solicitado, seguido da urocultura. BACOVA normatiza o diagnòstico de vaginose/ vaginite e ERIGE aumenta o valor preditivo de células que alertam a respeito de alteragòes epiteliais. Desde 2007 até hoje, nos workshops BACOVA/ERIGE (coloragào de Giemsa), foi avaliada a sensibili-dade da detecgào de células anormais esfoliadas. 99% dos participantes coincidiram com a detecgào de coilócitos. Bacova/Erige nào substitui o Papanicolaou de forma alguma, mas pode e deve ser feito em laboratórios periféricos, com o qual além do diagnóstico de vaginose / vaginite com 100% de valor preditivo, aumentam a cobertura preventiva de estados proliferativos.

Evaluation of the effect of presence of blood in the stomach on endoscopic diagnostic tests for Helicobacter pylori infection.

Introduction: Presence of blood in the stomach has been thought to affect the performance of diagnostic tests used in detecting Helicobacter pylori (H. pylori) in the stomach. This study evaluated the effect of blood on the efficacy of rapid urease test (RUT) and microscopic appearance of the biopsy after staining with Giemsa stain. Materials and Methods: Patients with bleeding oesophageal varices who met the inclusion criteria were tested for H. pylori by RUT and microscopic examination of the biopsy. A repeat endoscopy, RUT and histology were done one month following initial presentation. The performance of the diagnostic tests was evaluated with and without the presence of intraluminal blood. A combined result of the two tests, RUT and histology, carried out in presence or absence of blood for the diagnosis of H. pylori, when considered together was considered as the gold standard. Results: Thirty six patients included in the study were in the ages ranging between 15-60 years (mean age = 44.14 years ±2.1). The combination of tests at both visits showed 20/36 (55.6%) patients were positive for H. pylori. The decrease in H. pylori positivity in the presence of blood was significant for RUT (8.3% vs. 38.9%; P=0.005) and combined test (19.4% vs. 47.2%; P=0.02) but the decrease in positivity for histology (11.1% vs 30.6%) was not significant (P=0.08). In the presence of blood, the sensitivity of RUT, histology and combined tests were 15%, 20% and 35%, respectively. In the absence of blood, the sensitivity of RUT, histology and combination of tests was 70%, 55% and 85%, respectively. Conclusion: Blood in the stomach significantly decreased the sensitivity of RUT, histology and the combination of both. Negative results of these tests in acute upper gastro intestinal (GI) bleeding should therefore be interpreted carefully.

Estudio de la infección de Leishmanias del complejo Viannia mediante citometría de flujo y coloración de Giemsa empleando líneas de macrófagos humanos y murinos (U-937 y J-774) / Study of Leishmania Viannia infection by means of flow cytometry and Giemsa stain using human and murine macrophage lines (U-937 and J-774)

Ante las dificultades frente al tratamiento de la leishmaniasis, es importante buscar alternativas terapéuticas que deben ser analizadas empleando modelos in vitro e in vivo adecuadamente estandarizados. Con este propósito, se implementó un modelo de infección in vitro de Leishmania conmacrófagos U-937 y J-774, para evaluar la internalización de promastigotes a distintos puntos tiempo (2 a 6 horas) y por dos técnicas: coloración de Giemsa (CG) y citometría de flujo (CF). En el análisis por CF, se evaluó la invasión teniendo en cuenta la emisión de fluorescencia de los parásitos transfectados con la proteína verde de fluorescencia (GFP) y el aumento de la densidad citoplasmática de los macrófagos debida a los parásitos internalizados, lo cual fue verificado con el recuento microscópico realizado con CG; se encontró que a una proporción 1:35 (células:parásitos) se pueden establecer cambios densitométricos asociados con la infección empleando cepas de parásitos no transfectadas. También se describe que las J774 internalizan más eficientemente promastigotes de Leishmania que las células U937 (P valor: 0,0006), y se observa a su vez para la línea murina un aumento del número de parásitos por célula, respecto a los macrófagos humanos empleados en este ensayo (P valor 0,0038). Este estudio nos permite concluir: (i) que los cambios en la densidad citoplasmática evidenciados por CF son suficientes para establecer el porcentaje de infección parasitaria, aun para aquellos parásitos no transfectados; (ii) que la CG es menos costosa que el uso de la CF para evaluar infección parasitaria, aunque por su carácter semicuantitativo, la variabilidad intra- e inter-observadores la hace menos precisa; (iii) que los resultados cuantitativos obtenidos por la CF se correlacionan con los observados en la CG, y permiten sugerir que estas dos técnicas resultan complementarias; y (iv) que el porcentaje de infección y el número de parásitos internalizados para la J-774 son mayores que lo encontrado para la U-937, lo cual puede deberse a que un mayor número de receptores de complemento sobre la línea murina favorece la internalización de patógenos intracelulares, proceso menos favorecido en la línea humana por los niveles reducidos de este tipo de receptores en su membrana celular.

The treatment for Leishmaniasis has presented some difficulties related with adverse effects and resistance. For these reasons it is important to search therapeutic alternatives which must be analyzed using adequately standardized in vitro and in vivo models. With this purpose, we implemented an in vitro model for leishmania infection using U-937 and J-774macrophages, and evaluating promastigote internalization at consecutive time spans (from 2 to 6 hours). The first approximation assayed involves the flow cytometric (CF) analysis for invasion quantification by measuring the fluorescence emitted by parasites previously transfected with green fluorescence protein (GFP). In the alternative strategy, parasitized cells were subjected to Giemsa stain and CF was applied to measure the increase of macrophages cytoplasm density owed to internalized parasites. Giemsa stain also allowed us to estimate the number of parasites within each cell. We report that the presence of 35 parasites per macrophage produces an increase in cytoplasmic density enough to be detected by CF so that infection can be clearly reported. We also found that J-774 macrophages internalize Leishmania promastigotes more efficiently than U-937 cells (P value: 0.0006). Cells of the murine line were infected by a higher number of parasites than the human counterparts used in this study (P value 0.0038). From the resultas, we conclude: (i) the change in macrophage cytoplasm density demonstrated by CF after Giemsa stain are sufficient to estimate the percentage of infection. (ii) Giemsa stain provides a less expensive strategy to evaluate Leishmania infection than the fluorescence based option, although the intra and inter observer variability (semi-quantitative procedure) makes it less precise; (iii) quantitative results obtained by both techniques correlate to each other, suggesting that these two tools can be considered complementary; and (iv) both the percentage of infection and the number of internalized parasites are higher for J-774 than for U-937 macrophages. This probably suggests the presence of more receptor molecules (binding targets) for Leishmania ligands on the murine cell membrane determining a more efficient internalization rate than in human macrophages.

Comparación de las coloraciones de Giemsa y Grocott en el diagnóstico de la histoplasmosis / Comparison of Giemsa and Grocott stains in the diagnosis of histoplasmosis

El diagnóstico de histoplasmosis se realiza tradicionalmente mediante el reconocimiento de típicas levaduras intracelulares de Histoplasma capsulatum en preparaciones microscópicas teñidas con Giemsa. Se comparó la eficacia de una modificación rápida de la técnica de Grocott (MRG) y la tradicional de Giemsa para el diagnóstico de la histoplasmosis, a partir de la aplicación de ambas a 10 secreciones respiratorias, 8 escarificaciones de lesiones cutáneas y una biopsia ganglionar, pertenecientes todas a pacientes con sospecha clínica de esta micosis. En 15 de las 19 muestras no se encontraron diferencias significativas en la capacidad y rapidez para arribar al diagnóstico, mientras que en las 4 restantes, fueron reconocidas con la MRG estructuras que pasaron desapercibidas con la coloración de Giemsa. La modificación rápida permitió un reconocimiento más rápido del H. capsulatum en materiales donde este hongo se observó en escaso número y permitió además identificar con seguridad otros patógenos fúngicos diferentes de H. capsulatum, como Pneumocystis jiroveci, Paracoccidioides brasiliensis y Cryptococcus neoformans, difíciles de observar con la coloración de Giemsa. Se propone la técnica de Grocott o su modificación rápida para el diagnóstico de la histoplasmosis, especialmente cuando el empleo de la coloración de Giemsa da lugar a resultados negativos o dudosos.

The diagnosis of histoplasmosis is traditionally achieved by recognizing the typical intracellular yeasts of Histoplasma capsulatum, in smears stained with Giemsa stain. The usefulness of a rapid modification of Grocott and of traditional Giemsa stains for the diagnosis of histoplasmosis was compared applying both techniques in 10 respiratory secretions, 8 cutaneous lesions scrapings and 1 adenomegaly biopsy, all of them belonging to patients with clinically suspected histoplasmosis. In 15 out of the 19 evaluated samples, no significant differences were found in the ability or speed to reach the diagnosis with the applied techniques; while in the remaining 4 samples, structures that had not been observed with Giemsa stain were recognized with the rapid modification. The modification enabled quicker recognition of H. capsulatum than Giemsa stain in those clinical samples where the number of these fungal pathogens was scant. Additionally, the rapid modification also enabled the recognition of fungal pathogens other than H. capsulatum, as Pneumocystis jiroveci, Paracoccidioides brasiliensis and Cryptococcus neoformans, difficult to observe with the Giemsa stain. Use of Grocott technique or rapid modification stain is proposed for the diagnosis of histoplasmosis, when the result obtained with the Giemsa stain is doubtful or negative.

Application of Giemsa stain for easy detection of Trichinella spiralis muscle larvae

The application of Giemsa technique to stain compressed diaphragm samples obtained from rodents experimentally infected with Trichinella spiralis is described. Diaphragm samples from rats heavily infected with 20 muscle larvae per gram of body weight (20 ML/gbw) were cut into several pieces and stained with Giemsa; on the other hand, whole diaphragms from slightly infected mice (1 ML/gbw) were also stained with Giemsa. Besides, muscle samples were also stained with Giemsa. Observation at 10 x magnification revealed that both ML and nurse cells (NC) look as bluish structures clearly contrasting with the pinkish color of the non-infected muscle fibers. NC in the diaphragms of mice could be easily observed at naked eye as blue points contrasting with the pink surrounding areas formed by the non-infected muscle fibers. Among NC observed in the diaphragms of rats infected with 20 ML/gbw, 4.4% was multiple infection. These findings were confirmed in sectioned and hematoxylin-eosin stained specimens. This data could be usefulness for a rapid diagnosis of trichinellosis in post-mortem mammals without magnification procedures.

Application of Giemsa stain for easy detection of Trichinella spiralis muscle larvae

The application of Giemsa technique to stain compressed diaphragm samples obtained from rodents experimentally infected with Trichinella spiralis is described. Diaphragm samples from rats heavily infected with 20 muscle larvae per gram of body weight (20 ML/gbw) were cut into several pieces and stained with Giemsa; on the other hand, whole diaphragms from slightly infected mice (1 ML/gbw) were also stained with Giemsa. Besides, muscle samples were also stained with Giemsa. Observation at 10 x magnification revealed that both ML and nurse cells (NC) look as bluish structures clearly contrasting with the pinkish color of the non-infected muscle fibers. NC in the diaphragms of mice could be easily observed at naked eye as blue points contrasting with the pink surrounding areas formed by the non-infected muscle fibers. Among NC observed in the diaphragms of rats infected with 20 ML/gbw, 4.4% was multiple infection. These findings were confirmed in sectioned and hematoxylin-eosin stained specimens. This data could be usefulness for a rapid diagnosis of trichinellosis in post-mortem mammals without magnification procedures.

An attempt to determine contributory factors in the appearance of the purplish sheath of W. bancrofti microfilariae when stained with dilute giemsa

The staining experiment was an attempt to study and possibly identify some factors which may have influence the unusual appearance of purplish sheath of W. bancrofti when stained with dilute Giemsa. The results showed, subject to the limitations cited in connection with the basic data, that no significant differences in the appearance of the purplish sheath could be ascribed to the variety of Giemsa staining solutions use (American, German and Japanese). The use of tap water both as stain diluent and for washing excess stain appears to be the least favorable cause for the appearance of this unusual staining reaction. On the other hand, the likelihood of defecting microfilariae with purplish sheaths is highest at the thin sections of smears that were stained with Giemsa diluted with buffer solution and subsequently washed with tap water to remove excess stain. (Summary and conclusions)