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.

Modified comet assay with Giemsa staining: a suitable method for assessing genotoxicity in rats / Ensaio cometa modificado com coloração de Giemsa: um método adequado para avaliar a genotoxicidade em ratos

The present study tested a comet assay that was modified for compatibility with Giemsa staining to assess the drug genotoxicity in the peripheral blood of rats. We analysed the peripheral blood of 16 female Wistar rats (N=8 rats/group) from a control group and from a group that was treated with an intraperitoneal injection of 50mg cyclophosphamide/kg. The comet assay was carried out with modifications of the blood volume and immersion time in the lysing solution and different combinations of electrophoresis conditions (running time, voltage and current), to Giemsa staining. The lysing time and electrophoresis conditions allowed for the expression of all classes of DNA damage during the electrophoresis run, and the comets were efficiently stained with Giemsa. The technique showed high reproducibility for the DNA classes. The results demonstrate that the modified comet assay with Giemsa staining can be standardized for routine laboratory procedures using a 20µL blood sample, 3h and 30min immersions in the lysing solution and electrophoresis runs with 23 to 25 V and 310 and 360mA of electrical current. The modified comet assay with Giemsa staining that was described in the present study was standardized to be applied in the laboratory routine.(AU)

O presente estudo testou um ensaio cometa modificado para a coloração de Giemsa para avaliar a genotoxicidade de fármacos no sangue periférico de ratos. Analisou-se o sangue periférico de 16 ratas Wistar (n=8 ratas/grupo) de um grupo controle e de um grupo que foi tratado com uma injeção intraperitoneal de 50mg/kg pv. de ciclofosfamida. O ensaio cometa foi realizado com modificações do volume sanguíneo e do tempo de imersão na solução de lise, bem como com diferentes combinações de condições de eletroforese (tempo de corrida, tensão e corrente), para coloração de Giemsa. O tempo de lise e as condições de eletroforese permitiram a expressão de todas as classes de danos no DNA durante a corrida de eletroforese, e os cometas foram eficientemente corados com Giemsa. A técnica mostrou alta reprodutibilidade para as classes de DNA. Os resultados demonstram que o ensaio cometa modificado com coloração de Giemsa foi padronizado para procedimentos laboratoriais de rotina usando-se uma amostra de sangue de 20µL, 3h30min de imersão na solução de lise e eletroforese com 23 a 25 V e 310 e 360mA. O ensaio cometa modificado com coloração de Giemsa descrito foi padronizado para ser aplicado na rotina laboratorial.(AU)

The diagnostic usefulness and correlation of nasal eosinophil count and percentage in children with rhinitis

PURPOSE: Allergic rhinitis is the most common atopic disease and the most common chronic disease of children. Eosinophil count and percentage in nasal smear are useful for differential diagnosis of allergic rhinitis. The aim of this study is to investigate the correlation between nasal eosinophil count and percentage.METHODS: Between January 2017 and August 2018, 221 children patients with a clinical history of rhinitis were tested at the outpatient respiratory and allergy unit of the Department of Pediatrics, School of Medicine, The Catholic University of Korea. Nasal secretion was collected by swabbing a children's nasal inferior turbinate 3–4 times with a cotton swab and then placed on to a glass slide. Later, the smear was stained by Giemsa stain.RESULTS: This is the first study to assess the comparison of nasal eosinophil count and percent. There is a positive correlation between nasal eosinophil count and percent Y=1.02 X+2.82 (Y=Eosinophil count, X=Eosinophil percentage). To determine the usefulness of nasal eosinophil count and percentage in the diagnosis of allergic rhinitis, we analyzed receiver operating characteristic curves. The cutoff value of the nasal eosinophil count was 6.5/high-power field, and that of the nasal eosinophil ratio was 3% for the diagnosis of allergic rhinitis.CONCLUSION: In patients with suspected rhinitis, one of the values of nasal eosinophil count or percentage can be used in clinical practice.

Comparison of different techniques and stains for direct diagnosis of Trichinella spiralis

Background: The aim of Trichinella inspection is to reliably detect larvae in meat at levels which are capable of causing human trichinellosis. Trichinoscopy is a rapid, cost effective but is less sensitive in detection of low muscle larvae (ML)numbers. Artificial digestion is more convenient, and flexible, but may destroy immature larvae. Baermann technique may enhance the efficiency of diagnosis. Giemsa and Leishman stains provide identical contrasting coloration as Hematoxylin and Eosin (H-E) with the both advantages of that they can be used for staining non-histological sectionsand rapid preparation.The primary aim of the study was to compare the efficiency of digestion and Baermann techniques for detection of ML in muscles. Secondary aim was detection of ML by Geimsa and Leishman stain and their comparison with routinely used stain H-E. Methods: Muscles from 36 mice orally infected with Trichinella spiralis encysted larvae were examined. Larvae from 1 g samples of different muscles were recovered by digestion and Baermann techniques after different dose of infection. Muscle samples were prepared for staining with Giemsa, Leishman and H-E. Results: Number of larvae recovered by Baermann was higher recovered by digestion method (P ˃0.5). Leishman stain was the best stain for rapid detection of ML after 30 min followed in validity by Geimsa then H-E. Conclusion: Baermann concentration technique showed higher sensitivity than digestion method. Leishman stain is superior to Giemsa stain as it takes less time. Both stains are considered good alternative to H-E for rapid and easy diagnosis of trichinellosis in post-mortem animals

Trisomía 9: reporte de un caso / Trisomy 9: a case report

La trisomía 9 es una enfermedad rara, que ha sido descrita por primera vez en 1970, a la fecha existen más de 150 casos reportados, caracterizados por dismorfias faciales, anomalías congénitas y retraso en el desarrollo psicomotor y/o discapacidad intelectual. Este es el primer caso reportado en nuestra población en un infante de sexo masculino con peso y talla bajos, fisura labiopalatina y retraso madurativo en varias áreas del desarrollo, en quien el cariotipo mostró un mosaico cromosómico con el 70% de sus células con la trisomía del cromosoma 9. El asesoramiento genético en estos casos es de vital importancia para orientar a los padres sobre posibles causas y explicar sobre la condición genética, su manejo y establecer pautas de seguimiento para hacer prevención terciaria

ABC Classification Is Less Useful for Older Koreans Born before 1960

BACKGROUND/AIMS: In the ABC classification system, group A consists of seronegative subjects without gastric corpus atrophy. This study aimed to determine the prevalence and characteristics of pseudo group A subjects. METHODS: Group A subjects were identified among consecutive Korean adults who underwent a serum anti-Helicobacter pylori immunoglobulin G (IgG) test and pepsinogen (PG) assay on the day of endoscopy. Past infection was defined as the presence of either eradication history or endoscopic findings suggesting past infection (i.e., gastric xanthoma, metaplastic gastritis, or advanced atrophy >closed-type 1). RESULTS: Among 2,620 group A subjects, 448 (17.1%) had eradication history, and 133 (5.1%) showed endoscopic findings suggesting past infection. Older age (odds ratio [OR], 1.148; 95% confidence interval [CI], 1.067 to 1.236) and earlier year of birth (OR, 1.086; 95% CI, 1.009 to 1.168) were independent risk factors for classification into pseudo group A, with cutoff points at 50.5 years and birth year of 1959.5, respectively. Positive H. pylori test findings were found in 22 subjects (3.1%) among the 715 subjects who underwent the urea breath test or Giemsa staining on the same day. Current infection was positively correlated with PG I and PG II levels (p<0.001) but not with age, anti-H. pylori IgG titer, or classification into pseudo group A. CONCLUSIONS: Among the group A subjects, 22.2% had past infection. The risk was higher in subjects older than 50 years, especially those born before 1960. Furthermore, current infection was found in 3.1% of the subjects and was correlated with increased gastric secretory ability.

Comparative study of metachromatic staining methods in assessing the exfoliative cell types during oestrous cycle in Sprague-Dawley laboratory rats / Estudio comparativo de los métodos de tinción metacromática en la evaluación de los tipos de células exfoliativas durante el ciclo estral en ratas Sprague-Dawley de laboratorio

This study was aimed at comparing the commonly used metachromatic stains viz., Papanicolaou stain, WrightGiemsa, Toluidine blue and Methylene blue in the assessment of cell types of the oestrous cycle in rats. Eight female Sprague-Dawley rats aged 8-9 weeks were used for this assessment. Cotton Swabs were gently inserted in the animals vagina to obtain cells from which they were then transferred to glass slides for staining and evaluation under microscopy. The different cell types were compared for their morphological features and clarity of cellular detail under all four stains. The application, advantages and limitations of all stains were then discussed. It was concluded that the selection of the most effective stain in the assessment of vaginal cytology depends on their application to clinical or research which was based on the cellular detail of interest, time, cost and availability of each staining procedure.

El presente estudio tuvo como objetivo comparar las tinciones metacromáticas comúnmente utilizadas, Wright's-Giemsa, azul de toluidina, azul de metileno y tinción de Papanicolaou, en la evaluación de los tipos de células del ciclo estral en ratas. El estudio se realizó en ocho ratas hembras SpragueDawley, con edades entre 8 y 9 semanas, y se usaron hisopos vaginales de algodón para preparar portaobjetos. Los diferentes tipos de células se compararon por sus características morfológicas y claridad en las cuatro tinciones. La aplicación, ventajas y limitaciones de todas las tinciones fueron discutidas. Se concluye que la selección de la tinción más efectiva en la evaluación de la citología vaginal depende de su uso, es decir, clínico o de investigación, el detalle celular de interés, tiempo, costo y disponibilidad.

Identification of Nodular Gastritis among Patients Diagnosed with Lymphofollicular Gastritis on a Gastric Biopsied Specimen / 대한소화기학회지

BACKGROUND/AIMS: Nodular gastritis (NG) is a well-known endoscopic finding observed in patients with a Helicobacter pylori infection, which may lead to invasive gastric cancer. Lymphofollicular gastritis consists of lymphoid follicles or lymphoid cell aggregates, and is common in children. The aim of this study was to identify patients with NG from those in whom gastric biopsied specimens showed lymphoid follicles and lymphoid cell aggregates. METHODS: Subjects, whose gastric biopsy specimens showed lymphoid follicles or lymphoid cell aggregates, were included in this study. The inclusion criterion was that they underwent a serum pepsinogen assay on the day of upper gastrointestinal endoscopy. NG was diagnosed if the endoscopy findings revealed regular-sized, multiple, colorless subepithelial nodules. RESULTS: Among 108 subjects who showed lymphoid follicles or lymphoid cell aggregates, 13 (12.0%) revealed NG on endoscopy, and all these subjects showed positive Giemsa staining. Patients diagnosed with NG were younger (p=0.012) and showed a female predominance (p=0.001) compared to those without NG. The mean serum pepsinogen levels were higher (p=0.001) and lymphoid follicle-dominant subjects were more common (p<0.001) in the NG subjects than in those without NG. Logistic regression analysis revealed a younger age (p=0.041) and female gender (p=0.002) to be significant independent risk factors for NG. CONCLUSIONS: NG should be distinguished from lymphofollicular gastritis because only 12% of patients showing gastric biopsy findings of lymphoid follicles and lymphoid cell aggregates demonstrated NG on endoscopy. NG is an endoscopic finding that is more common in women and in the younger population, irrespective of the biopsy findings and gastric secretory ability.

Enhancement of quantum efficiency of a dye-sensitized electrochemical cell by using triturated zinc oxide mixed with two organic dyes, azure C and rose bengal

For efficiently utilising solar energy, when suitable nanoparticles are being engineered, triturated zinc oxide an eco-friendly, easily available, low-cost material has been used as an agent for solar energy conversion. Two organic dyes Azure C and Rose bengal having absorption bands in two different spectral regions at 545 nm and 610 nm respectively, were chosen in order to overcome the band absorption limits of each dye and utilise the broad spectrum of solar radiation. The material was mixed with these two dyes in a specially devised electrochemical cell and photovoltage with significant efficiency was generated. The energy conversion efficiency of the cell using three different potencies 6C, 30C and 200C of triturated zinc oxide with the same concentration of two dyes (0.5x10-5 M) in all cases are 0.39%, 0.43% and 0.35% respectively. The efficiency is only 0.15% for the mixed dye under similar conditions.

Frequency of helicobacter pylori in histologically proven gastritis cases - a study of 100 cases

Objective: To determine the frequency of Helicobacter pylori [H. pylori] based on endoscopic of special stains in gastritis cases

Study Design: Prospective descriptive study

Place and Duration of Study: Histopathology department, Army Medical College Rawalpindi, from Oct 2016 to Mar 2017

Material and Methods: One hundred [100] cases were included in the study. Gastric biopsies of the patients histologically diagnosed as gastritis were included in the study, which were evaluated for the presence of H. pylori with the help of special stain [Modified Giemsa stain]

Results: Gastric biopsies of 100 patients, who were diagnosed as gastritis on histopathological examination were analyzed with the help of Giemsa stain for the presence or absence of H. pylori. Out of these 100 cases, 60 were males and 35 were females. Most patients were between the age group of 30-40 years. Histological examination and special stain analysis revealed presence of H. pylori in 30 cases [30%], while rest of the 70 cases [70%] showed no H. pylori. Out of 30 cases positive for H. pylori, 28 cases [93%] had chronic active gastritis, while 2 cases [7%] had no element of activity

Conclusion: Among the cases of chronic gastritis, H. pylori negative gastritis was more common than H. pylori associated gastritis. A significantly decreased frequency of H. pylori in histologically gastritis cases in our population may be due to more frequent use of complete or incomplete therapies against H. pylori by general practitioners at some stage of disease

Usefulness of Dual Priming Oligonucleotide-Polymerase Chain Reaction for Diagnosis and Treatment of Helicobacter pylori

BACKGROUND/AIMS: The eradication rate of triple therapy for Helicobacter pylori is decreasing and one of the main causes is increased clarithromycin resistance. Recently, new methods have been introduced for the diagnosis of clarithromycin resistance. The aim of this study was to investigate the diagnostic rate of dual priming oligonucleotide-polymerase chain reaction (DPO-PCR) compared with histology and the eradication rates of triple therapy for clarithromycin susceptible H. pylori. MATERIALS AND METHODS: We retrospectively reviewed patients who underwent DPO-PCR exam and Giemsa stain for diagnosis of H. pylori between January, 2015 and March, 2016 at Incheon St. Mary's Hospital. Clarithromycin resistance of H. pylori was determined by DPO-PCR and the diagnostic accuracy of DPO-PCR was compared with histology. We also examined the eradication rates of triple therapy for clarithromycin susceptible strains. RESULTS: A total of 928 patients underwent DPO-PCR exam and Giemsa stain for diagnosis of H. pylori. The resistance rate for clarithromycin was 39%. The sensitivity and specificity of PCR exam compared with histology were 96.2% and 96.9%. The positive predictive values, negative predictive values, and accuracy were 90.54%, 98.87%, 96.88%, each. A total of 53 patients received triple therapy, and 39 patients completed ¹³C-urea breath test. The overall eradication rate was 97.4%. CONCLUSIONS: DPO-PPR showed high accuracy compared with biopsy and the eradication rates of triple therapy for clarithromycin susceptible H. pylori was 97.4%. DPO-PCR may be effective in determining treatment regimens in areas of high clarithromycin resistance.

Hemozoin Pigment: An Important Tool for Low Parasitemic Malarial Diagnosis

Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.

A Case of Solitary Bullous Mastocytoma Occurring at a Site of Spinal Tap / 대한피부과학회지

Mastocytosis is characterized by an accumulation of mast cells in various organs, most frequently in the skin. A solitary mastocytoma is a clinical variant of cutaneous mastocytosis. It is defined as a localized collection of mast cells in the skin without evidence of extracutaneous organ involvement. Here we report on a 2-year-old female patient presenting with Solitary erythematous bulla on her lower back. The patient had a history of spinal tap on the lower back for evaluation of meningitis at 5 months of age, which resulted in trauma at the site. Histopathology showed mast cells infiltrating the papillary and reticular dermis and metachromatic purple cytoplasmic granules seen with Giemsa staining. As a result, the patient was diagnosed with a solitary bullous mastocytoma and administered antihistamine. The patient showed complete remission at 3 months. Herein, we report a rare case of solitary bullous mastocytoma occurring at a trauma site.

Comparison of Serum-Free Media in RBC Differentiation from Human Hematopoietic Stem Cells / 대한수혈학회지

BACKGROUND: Research on RBC production from hematopoietic stem cells has been conducted competitively in many countries. However those were in vitro successes and many hurdles still remain for large scale transfusable RBC production from stem cells. A need for large volume of culture media is a crucial factor for culture condition which researchers must overcome. In this study, we evaluated the efficiency of two commercial serum-free media, StemPro(R)-34 SFM and Stemline II hematopoietic stem cell expansion medium, in RBC differentiation from cord derived stem cells. METHODS: We cultured cord derived CD34+ cells in vitro and evaluated over the periods of 7 days, 14 days, 17 days and 21 days in culture for expanded cell count, cell morphology and differential count using the Wright Giemsa stain. RESULTS: Cell expansion and RBC differentiation developed rapidly in Stemline media compared to StemPro media. Enucleated RBCs were observed at 10~14 culture days and orthochromatic erythroblasts were shown up to 50% among culture cells at 17 days in Stemline media. The enucleated RBCs were observed at 17 days in StemPro Media. Although the erythroblasts in StemPro media are slow at differentiation, they maintain continuous expansion up to 21 days. CONCLUSION: In Stemline media, the expansion and differentiation to mature RBCs are processed much faster, but the cell condition slows down after 17 days. In the RBC production aspects, Stemline media is better than StemPro media as a rapid differentiation because it reduces the cost due to in vitro short culture duration.

case of urticaria pigmentosa

Urticaria pigmentosa is the most common variant of cutaneous mastocytosis. A three and a half-month-old female child presented with generalized eruption of multiple brownish macular and papular lesions on the trunk, limbs, face and neck of two months duration. On stroking the individual lesion, there was formation of wheal and erythema [Darier's sign positive]. Histopathological examination of lesional skin and staining with Giemsa stain and hematoxylin eosin showed infiltration of numerous spindle shaped mast cells in perivascular and periappendageal location in the upper dermis. These findings were suggestive of mastocytosis. There was no systemic involvement. We present a typical case of urticaria pigmentosa

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.

Cultivo de la línea celular HEp-2: doblaje poblacional y coloración con Giemsa Perspectivas para el estudio de la infección con Chlamydia trachomatis / Cultivation of the cell line hep-2: dubbing of population and coloring with Giemsa

Los cultivos celulares se han convertido en herramientas esenciales para la investigación básica. Se aplican en inmunología, virología, biología molecular, ingeniería genética y farmacología, entre otras áreas. Se usan también en procesos industriales farmacéuticos, en técnicas de diagnóstico clínico y para estudio de trasplante de tejidos. En bacteriología, estos cultivos permiten confirmar una infección, evaluar la eficiencia de antimicrobianos, realizar estudios de infectividad, investigar sobre nuevas especies, obtener gran cantidad de microorganismos no cultivables para optimizar técnicas y examinar las relaciones entre la célula huésped y los microorganismos intracelulares (virus, bacterias y parásitos). La línea celular HEp-2 (Human Epidermoid Cancer Cells) es utilizada en estudios de infección con diferentes bacterias, entre ellas Chlamydia trachomatis (CT), con el fin de determinar los mecanismos por los cuales este patógeno sobrevive en la célula huésped. También se emplea para observar la acción de péptidos antimicrobianos y de extractos para combatir la infección causada por dicha bacteria. Para este estudio se realizaron curvas de crecimiento en la línea celular HEp-2 con medios DMEM-F12 y MEM. Se estandarizó, además, la coloración con Giemsa y se calculó el doblaje poblacional con diferentes inóculos para evaluar el desarrollo de la línea celular en cultivo y seleccionar las condiciones óptimas para realizar futuros ensayos de infección con parásitos intracelulares, en particular con CT serovar L2.

Cell cultures have become essential tools for basic research. They are applied in immunology, virology, molecular biology, genetic engineering and pharmacology, among other areas. They are also used in pharmaceutical industrial processes, in techniques of clinical diagnostic, and to study tissue transplantation. In bacteriology, these crops allow us to confirm an infection, assess the efficiency of antimicrobials, carry out studies of infectivity, investigate on new species, obtaining a large number of microorganisms non-arable to optimize techniques, and to examine the relationship between the host cell and intracellular microorganisms (bacteria, viruses, and parasites). The HEp-2 cell line (Human epidermoid cancer cells) is used in studies of infection with different bacteria, including Chlamydia trachomatis (CT), in order to determine the mechanisms by which the pathogen survives in the host cell. It is also used to observe the action of antimicrobial peptides and extracts to combat the infection caused by the bacterium. For this study, growth curves of the HEp-2 cell line were carried out with DMEM-F12 and MEM media. In addition, the staining with Giemsa was standardized, and the population dubbing was calculated with different inocula for assessing the development of the cultured cell line and select the optimal conditions for future tests of infection with intracellular parasites, in particular with CT serovar L2.

The application of an in situ karyotyping technique for mesenchymal stromal cells: a validation and comparison study with classical G-banding

The cytogenetic analysis of mesenchymal stromal cells (MSCs) is essential for verifying the safety and stability of MSCs. An in situ technique, which uses cells grown on coverslips for karyotyping and minimizes cell manipulation, is the standard protocol for the chromosome analysis of amniotic fluids. Therefore, we applied the in situ karyotyping technique in MSCs and compared the quality of metaphases and karyotyping results with classical G-banding and chromosomal abnormalities with fluorescence in situ hybridization (FISH). Human adipose- and umbilical cord-derived MSC cell lines (American Type Culture Collection PCS-500-011, PCS-500-010) were used for evaluation. The quality of metaphases was assessed by analyzing the chromosome numbers in each metaphase, the overlaps of chromosomes and the mean length of chromosome 1. FISH was performed in the interphase nuclei of MSCs for 6q, 7q and 17q abnormalities and for the enumeration of chromosomes via oligo-FISH in adipose-derived MSCs. The number of chromosomes in each metaphase was more variable in classical G-banding. The overlap of chromosomes and the mean length of chromosome 1 as observed via in situ karyotyping were comparable to those of classical G-banding (P=0.218 and 0.674, respectively). Classical G-banding and in situ karyotyping by two personnel showed normal karyotypes for both cell lines in five passages. No numerical or structural chromosomal abnormalities were found by the interphase-FISH. In situ karyotyping showed equivalent karyotype results, and the quality of the metaphases was not inferior to classical G-banding. Thus, in situ karyotyping with minimized cell manipulation and the use of less cells would be useful for karyotyping MSCs.

A Case of Plasmodium ovale wallikeri Infection in a Chinese Worker Returning from West Africa

In contrast to the gradual reduction in the number of locally transmitted malaria cases in China, the number of imported malaria cases has been increasing since 2008. Here, we report a case of a 39-year-old Chinese man who acquired Plasmodium ovale wallikeri infection while staying in Ghana, West Africa for 6 months in 2012. Microscopic examinations of Giemsa-stained thin and thick blood smears indicated Plasmodium vivax infection. However, the results of rapid diagnostic tests, which were conducted 3 times, were not in agreement with P. vivax. To further check the diagnosis, standard PCR analysis of the small-subunit rRNA gene was conducted, based on which a phylogeny tree was constructed. The results of gene sequencing indicated that this malaria is a variant of P. ovale (P. ovale wallikeri). The infection in this patient was not a new infection, but a relapse of the infection from the one that he had contracted in West Africa.