Effect of all-trans retinoic acid on maturation and differentiation of mouse hepatic progenitor cells induced by bone morphogenetic protein 9 and its mechanism / 解剖学报

Objective To investigate the effect of 1 μmol/L and 10 μmol/L all trans retinoic acid（ATRA） on bone morphogenetic protein 9（BMP9）-induced maturation and differentiation of hepatic progenitor cells. Methods BMP9, BMP9 + 1 μmol/L ATRA and BMP9 + 10 μmol/L ATRA acted on HP14-19, respectively. The expression of albumin-drive gussid（LAB-Glus） was detected by luciferase reporter gene. The mRNA levels of ALB, cytokeratin 18（CK18）, tyrosine aminotransferase（TAT）, apolipoprotein B（ApoB） were detected by Real-time PCR. The expressions of ALB and uridine diphosphate glucuronosyltransferase 1 A（UGT1 A） were detected by immunofluorescence. Periodic acid-schiff（PAS） staining and indocyanine green（ICG） uptake assay were used to detect the metabolism and glycogen synthesis of hepatocytes. Real-time PCR and Western blotting were used to detect the expression of retinoic acid receptor（RAR）α, RARβ、RARγ and BMP9 signal related molecules Samd1, Samd 5 and Samd 8. Ad-siRARα、Ad-siRARβ、Ad-siRARγ infected cells were treated with BMP9+10 μmol/L ATRA, the cell morphology and PAS staining result were observed, the mRNA levels of ALB, CK18, TAT and ApoB were detected by Real-time PCR. Results BMP9 could significantly induce the maturation and differentiation of HP14-19 cells. The morphology of HP14-19 cells looked like polygonal paving stone. The expressions of ALB, CK18, ApoB and UGT1 A were significantly up-regulated. Some cells had the function of metabolic detoxification and glycogen synthesis. Compared with the BMP9 group, BMP9+1 μmol/L ATRA group had more mature morphology and larger volume. The expressions of Alb, CK18, ApoB and UGT1 A were up-regulated significantly（P＜0.05）. The number of ICG and PAS positive cells increased. Compared with the BMP9+1 μmol/L ATRA group, BMP9 + 10 μmol/L ATRA group showed long spindle, spindle and polygonal shapes, and the expression of hepatocyte related markers decreased, and the number of ICG and PAS positive cells decreased. ATRA（1 μmol/L） significantly increased the expression of RARα, RARβ and RARγ. Compared with the 1 μmol/L ATRA group, 10 μmol/L ATRA group only increased the expression of RARα. BMP9 did not affect the expression levels of Samd1, Samd5 and Samd8, but up-regulated their phosphorylation. Ad-siRARα could improve cell morphology and PAS staining induced by 10 μmol/L ATRA, while increased the expression of Alb and CK18（P＜0.05）. Conclusion ATRA（1 μmol/L） can promote the maturation and differentiation of hepatic progenitor cells（HPCs） induced by BMP9, while 10 μmol/L ATRA can weaken the differentiation of hepatic progenitor cells. Excessive ATRA may over activate RARα signal to affect the differentiation of hepatic progenitor cells.

Epidemiología de la Onicomicosis Pedis en adultos hospitalizados en un servicio de medicina en un hospital de tercer nivel en Chile / Epidemiology of Onychomycosis pedisinhospitalized adults at the internal medicine ward of a high complexity hospital in Chile

La prevalencia global de la onicomicosis pedis es de 4,3%, y en hospitalizados puede llegar hasta 8,9%. Aun así, se propone que está ampliamente subdiagnosticada. Personas añosas con comorbilidades presentan mayor riesgo de onicomicosis pedis y de sus complicaciones. Se examinaron aleatoriamente a 64 pacientes hospitalizados en el Servicio de Medicina del Hospital San José. A aquellos con signos clínicos de onicomicosis pedis se les realizó un examen micológico directo (MD) y estudio histopatológico de un corte de uña teñido con PAS (Bp/PAS). Muestra de 64 pacientes, un 78,1% presentó onicomicosis pedis clínica y en un 70,3% se confirmó el diagnóstico con MD y/o Bp/PAS positivo. De los pacientes con onicomicosis confirmada, el promedio de edad fue de 67,8 +/- 12,3 años. Un 44% correspondió al sexo femenino y un 56% al sexo masculino. La onicomicosis pedis en el servicio de medicina interna del Hospital San José es una condición frecuente. El conjunto de MD y Bp/PAS podría ser considerado como una buena alternativa diagnóstica. (AU)

Onychomycosis of the toenails has a global prevalence of 4,3% and can reach up to 8,9% in hospitalized patients. It has been hypothesized that it is widely under diagnosed. Aged patients with multiple diseases have an increased risk of Onychomycosis and its complications. 64 patients of the internal medicine ward were randomly selected. Those who had clinical signs of onychomycosis of the toenails were tested with direct microscopy and histological study of the nail plate with PAS staining. Of the 64 patients, 78,1% (50) had clinical signs of onychomycosis of the toenails and in 70,3% (45) the diagnosis was confirmed either by direct microscopy and/or by histological study of the nail plate with PAS staining. The mean age for the group with onychomycosis was 67,8 +/- 12,3 ages. 44% were female and 56% were male. Onychomycosis of the toenails is a frequent condition at the internal medicine ward of the San José Hospital. The direct microscopy together with the histological study of the nail plate with PAS staining seem to be a good diagnosis alternative. (AU)

Value of fluorescence staining in histopathological diagnosis of Acanthamoeba keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the application value of fluorescent staining technique in the detection of amoebic pathogens in corneal tissue biopsy, and to apply fluorescent staining technique in the histopathological diagnosis of Acanthamoeba keratitis(AK), comparing the results with those of hemotoxyiln-eosin staining(HE staining)and periodic acid-schiff staining(PAS staining), and analyzing the sensitivity and specificity of these three staining methods.<p>METHODS:Specimens of infected corneal tissue were collected from 74 cases(75 eyes), and then they were divided into an AK group and a non-Acanthamoeba keratitis(NAK)group based on the results of corneal scraping, culture and histopathological diagnosis. The tissues of consecutive sections were stained with HE staining, PAS staining and fluorescence respectively, and the sensitivity and specificity of the three staining methods for the diagnosis of AK were analyzed. Area under the curve(AUC)was calculated using the receiver operating characteristic(ROC)curve. Further analysis was performed to count the number of Acanthamoeba pathogens found by the three staining methods under the same magnification field of view at the same site, and to clarify the diagnostic value of fluorescent staining technique for AK.<p>RESULTS: The sensitivity of HE staining was 69%(27/39)with a specificity of 92%; the sensitivity of PAS staining was 62%(24/39)with a specificity of 97%, and the sensitivity of fluorescent staining was 95%(37/39)with a specificity of 97%. There were differences in the sensitivity of the three staining methods for the diagnosis of AK(χ2=19.857, <i>P</i><0.001), and pairwise comparison revealed that the differences between HE staining and fluorescent staining, PAS staining and fluorescent staining for the diagnosis of AK were statistically significant(<i>P</i>=0.003,<0.001), while the difference in sensitivity between HE staining and PAS staining for the diagnosis of AK was not statistically significant(<i>P</i>=0.978). The maximum AUC was 0.960 for fluorescence staining, followed by 0.804 for HE staining and 0.794 for PAS staining, respectively. The median number of amoeba cysts detected by HE staining, PAS staining and fluorescent staining at the same site under the same magnification field of view was 4(0, 11), 2(0, 9)and 12(3, 33), respectively(χ2=56.561, <i>P</i><0.001). Pairwise comparison revealed that the differences in the number of amoeba cysts found by HE staining and fluorescence staining, PAS staining and fluorescence staining were statistically significant(<i>P</i><0.001), while the difference in the number of amoeba cysts found by HE staining and PAS staining was not statistically significant(<i>P</i>=0.210). Fluorescently stained histopathological sections make it easier to identify amoebic pathogens.<p>CONCLUSION:Fluorescent staining technique is more sensitive to histopathological diagnosis of AK than HE staining and PAS staining, which can significantly improve the positive rate of detection of amoebic pathogens.

Fungal infection in surgical pathology – Our institutional experience

Background: Fungal infection has a worldwide prevalence and it affects almost all age groups and both the sexes. Due to neoteric treatment modalities nowadays, people are enjoying longevity of life. However, on the other hand, immune status is compromised that has resulted in increased prevalence of fungal infection in society. Fungi reported by surgical pathologist, either incidentally or in clinically suspicious cases, greatly improve morbidity and outcome of the patients. Objective: The objective of this study is to study the prevalence of fungal infection in surgical pathology tissue and frequency and spectrum-based analysis in relation to age, gender, and systemic involvement. Materials and Methods: Ethically approved retrospective observational secondary data analysis is done based on histomorphological and cytomorphological evaluation of 17 fungal cases diagnosed at the Department of Pathology, New Civil Hospital, Surat, from January 2015 to January 2017. Results: Out of 17 cases, most common fungus seen was Candida albicans mainly affecting gastrointestinal tract. Common age group affected was 41–60 years and few rare fungi, i.e., mucormycosis and tinea corporis were also reported. Conclusion: Distribution of fungi in tissue section in relation to frequency, age, sex, various organs, and its correlation with clinical symptoms is analyzed in our study and results are found to be yielding that provided outlook in the prevalence of infectious agent.

Cytological intracellular glycogen evaluation using PAS and PAS-D stains to correlate plasma glucose in diabetics

Background: Oral cytology studies have claimed that cytoplasmic Periodic Acid Schiff (PAS) positivity in type-2 diabetics is due to glycogen content. But, it can also be due to mucin and glycoconjugates. Aim: 1. To confirm that cytoplasmic PAS positivity in type-2 diabetics is due to glycogen using diastase. 2. To know the effect of diabetes by determining the number of glycogen-containing cells in the smear. 3. To assess the impact of duration of diabetes based on PAS staining of cells. 4. To correlate between random blood glucose level and the number of PAS-positive cells. Materials and Methods: Study population comprised 45 individuals with 30 type-2 diabetics as case group (Group I < 5 years duration; Group II > 5 years duration) and 15 healthy volunteers (age and gender-matched) as control. For all subjects, random blood glucose was estimated and two cytosmears were obtained. The smears were stained with PAS and PAS-diastase stains (PAS-D). Staining intensity was documented as score 1 (mild-to-moderate) and score 2 (moderate-to-intense) and data obtained were statistically analyzed in SPSS version 16.0. Results: Mann-Whitney U test revealed that in diabetics cytoplasmic PAS positivity is because of glycogen (P < 0.05). There is an increase in the number of glycogen-containing cells (P < 0.05) in diabetics. The duration of diabetes had less impact on intracellular glycogen accumulation (P > 0.05). Spearman's correlation test revealed no significant correlation (P > 0.05) between random blood glucose and a number of PAS-positive cells. Conclusion: PAS positivity is because of intracellular glycogen accumulation in type-2 diabetics. It can convey the glycaemic status of an individual in the recent past, thus a beneficial role in screening and therapeutic monitoring.

Research on purification and activity characterization of AmPR-10 ribonuclease from Astragalus membranaceus / 中草药

Objective To establish a stable and rapid separation and purification method for Astragalus membranaceus (Am) pathogenesis-related protein-10 (AmPR-10) using an automatic intelligent protein purification system AKTA Avant 25, and analyze its physiochemical and biological activity. Methods Am was extracted by Tris-HCl buffer. The crude extract was captured by anion exchange chromatography, and finely separated by hydrophobic chromatography and gel filtration chromatography. The relative molecular weight of AmPR-10 was measured by MALDI-TOF/TOF mass spectrometry, the protein identification was determined by mass spectrometry and MS/MS Ion Search, the glycoprotein identification was estimated by periodic acid-Schiff method, and the ribonuclease activity and effect factors were analyzed by agarose gel electrophoresis. Results The electrophoretically pure AmPR-10 was obtained by three-step purification of Q Sepharose Fast Flow, Butyl Sepharose High Performance and SuperdexTM 75 10/300 GL from the crude extraction. The relative molecular weight of AmPR-10 was 16 801. AmPR-10 was highly homologous to PR-10 and has no carbohydrate chains. Incubated at 56 ℃ for 30 min, AmPR-10 exhibited significant ribonuclease activity to total RNA of mammalian cells. The activity was insensitive to NaCl, pH value and mental ions, and weekly inhibited by 0.5 mol/L NaCl, pH 9.0, Mg2+ and Co2+. The activity was the same at EDTA as high as 20 mmol/L. Conclusion The three-step method of exchange chromatography-hydrophobic chromatography-gel filtration chromatography, a stable and rapid separation and purification method of AmPR-10, can be applied for other Chinese herbs. AmPR-10 might play an important role in resistance against virus.

Epithelial ovarian tumors in a premenarchal girl: a rare case report

Ovarian tumors are relatively uncommon in paediatric age group and mostly occur in the period after menarche and are extremely rare prior to menarche. However, in children and adolescent, the epithelial ovarian tumors account approximately 10% to 28% of all ovarian tumors. In the present case, the patient was presented with abdominal pain for 1 day and no mass was felt on physical examination. Contrast-enhance computed tomography abdomen and pelvis showed a left ovarian cystic mass measuring 7.3×5.1 cm in size and unilateral oophorectomy was done. Tumor shows multiloculated cysts of varying sizes filled with mucinous fluid. Cysts were lined by tall columnar epithelial cells and show predominantly periodic acid-Schiff positive neutral mucin favouring benign nature of lesion. We present this case due to its uncommon age of presentation and the patient was premenarchal girl.

Fluorescence of Candida in diagnosis of oral candidiasis.

Background: Many pathogenic fungi fluoresce in hematoxylin and eosin stained sections, and Papanicolaou (PAP)‑stained smears under ultraviolet illumination. In theory, this phenomenon could aid in the diagnosis of common fungal infections without the delay which is usually associated with special stains. Objective: To evaluate the role of fluorescence as a rapid screening technique for oral infections caused by Candida organisms in exfoliative smears of oral candidiasis. Materials and Methods: Two smears and one swab were collected from each of 62 clinically diagnosed cases of oral candidiasis. Smears were stained with (PAP) and periodic acid–Schiff stain (PAS). Both smears were evaluated under light microscopy (LM). Later, PAP smears were observed under fluorescent microscopy (PAP‑FM). The swab was inoculated on Sabouraud’s agar plate. Each technique was evaluated for sensitivity and specificity. Results: It was found that the PAS‑stained smears were more reliable for detection of Candida species than other methods (sensitivity = 100%; specificity = 66.7%). The PAP‑LM and PAP‑FM showed less sensitivity (67.9% and 85.7%) and specificity (66.7% and 33.3%), respectively. Combined results of both light and fluorescent microscopy of PAP (LM + FM) showed increased sensitivity (89.3%) but reduced specificity (16.7%). Conclusion: PAP autofluorescence is less sensitive than PAS, still it accentuates the distinct morphological features of Candida.

Análise histoquímica das glândulas anexas do aparelho reprodutor do macho de paca (Cuniculus paca) / Histochemical analysis of the accessory glands of male paca (Cuniculus paca)

A paca (Cuniculus paca) é um roedor típico de regiões tropicais. Com a finalidade de estudar esta espécie selvagem para manejo adequado e sua preservação, objetivou-se neste trabalho caracterizar a histoquímica das glândulas anexas do trato reprodutor do macho da paca. Para este fim, cortes histológicos dessas glândulas foram submetidas às reações histoquímicas com Ácido Periódico de Schiff (PAS), Alcian blue (AB), PAS. + AB. e PAS + Amilase. Na glândula bulbouretral foi constatado que o epitélio produz secreção rica em glicoproteínas neutras e ácidas, glicosaminoglicanas, e em algumas regiões produz mais de um tipo de secreção. Não foi observada a presença de glicogênio no epitélio. Na glândula vesicular, seu epitélio em borda em escova corou-se por glicoproteínas neutras e também por substância de composição desconhecida, não contendo glicoproteínas ácidas, glicogênio ou glicosaminoglicanas. Verificou-se presença de pequena quantidade de glicoproteínas ácidas e neutras na próstata, em especial na mucosa, além de glicoproteínas ácidas carboxiladas e sulfatadas em pequena quantidade no tecido conjuntivo da lâmina própria dessa glândula. Por fim, a glândula coaguladora apresentou pequena quantidade de glicoproteínas neutras na borda em escova de seu epitélio e substância de composição desconhecida, sendo ausente o glicogênio. Conclui-se que as glândulas anexas do trato reprodutor da paca apresentam características histoquímicas que compartilham certa similaridade com outras espécies da ordem Rodentia, com a presença de glicoproteínas neutras e ácidas em algumas glândulas, principalmente no epitélio da glândula bulbouretal e na borda em escova do epitélio das demais glândulas.(AU)

Paca is a wild rodent typical of the tropical region. In order to study this species with focus on the proper management and preservation, this study aims to characterize the histochemistry of the accessory glands of the reproductive tract of the male paca. For this purpose, histological sections were stained with Periodic Acid-Schiff (PAS), Alcian Blue (AB), PAS + AB e PAS + Amilase. In the histochemical reaction of the bulbouretral gland was found that the epithelium produces secretion rich in neutral and acid glycoproteins, glycosaminoglycans, and in some regions produces more than one type of secretion. The epithelium did not contain any glycogen. In the vesicular gland the epithelium had bush border stained by neutral glycoproteins, but no acid glycoproteins, glycogen or glycosaminoglycan. The coagulating gland showed small amount of neutral glycoproteins in the brush border of the epithelium, without glycogen. It is concludes that the accessory glands of paca have histochemical characteristics that share some similarity with other species of the Order Rodentia, with some glands with neutral and acid glycoproteins, mainly in the epithelium of the bulbouretral gland and in the brush border of the other glands.(AU)

Análise histoquímica das glândulas anexas do aparelho reprodutor do macho de paca (Cuniculus paca) / Histochemical analysis of the accessory glands of male paca (Cuniculus paca)

Paca is a wild rodent typical of the tropical region. In order to study this species with focus on the proper management and preservation, this study aims to characterize the histochemistry of the accessory glands of the reproductive tract of the male paca. For this purpose, histological sections were stained with Periodic Acid-Schiff (PAS), Alcian Blue (AB), PAS + AB e PAS + Amilase. In the histochemical reaction of the bulbouretral gland was found that the epithelium produces secretion rich in neutral and acid glycoproteins, glycosaminoglycans, and in some regions produces more than one type of secretion. The epithelium did not contain any glycogen. In the vesicular gland the epithelium had bush border stained by neutral glycoproteins, but no acid glycoproteins, glycogen or glycosaminoglycan. The coagulating gland showed small amount of neutral glycoproteins in the brush border of the epithelium, without glycogen. It is concludes that the accessory glands of paca have histochemical characteristics that share some similarity with other species of the Order Rodentia, with some glands with neutral and acid glycoproteins, mainly in the epithelium of the bulbouretral gland and in the brush border of the other glands.

A paca (Cuniculus paca) é um roedor típico de regiões tropicais. Com a finalidade de estudar esta espécie selvagem para manejo adequado e sua preservação, objetivou-se neste trabalho caracterizar a histoquímica das glândulas anexas do trato reprodutor do macho da paca. Para este fim, cortes histológicos dessas glândulas foram submetidas às reações histoquímicas com Ácido Periódico de Schiff (PAS), Alcian blue (AB), PAS. + AB. e PAS + Amilase. Na glândula bulbouretral foi constatado que o epitélio produz secreção rica em glicoproteínas neutras e ácidas, glicosaminoglicanas, e em algumas regiões produz mais de um tipo de secreção. Não foi observada a presença de glicogênio no epitélio. Na glândula vesicular, seu epitélio em borda em escova corou-se por glicoproteínas neutras e também por substância de composição desconhecida, não contendo glicoproteínas ácidas, glicogênio ou glicosaminoglicanas. Verificou-se presença de pequena quantidade de glicoproteínas ácidas e neutras na próstata, em especial na mucosa, além de glicoproteínas ácidas carboxiladas e sulfatadas em pequena quantidade no tecido conjuntivo da lâmina própria dessa glândula. Por fim, a glândula coaguladora apresentou pequena quantidade de glicoproteínas neutras na borda em escova de seu epitélio e substância de composição desconhecida, sendo ausente o glicogênio. Conclui-se que as glândulas anexas do trato reprodutor da paca apresentam características histoquímicas que compartilham certa similaridade com outras espécies da ordem Rodentia, com a presença de glicoproteínas neutras e ácidas em algumas glândulas, principalmente no epitélio da glândula bulbouretal e na borda em escova do epitélio das demais glândulas.

Identification and glycobiological characterization of circulating immune complexes in patients with visceral leishmaniasis and post kala azar dermal leishmaniasis.

Here, we investigated the quantitative and qualitative differences in antibody classes and subclasses in serum immune complexes (ICs) of Visceral Leishmaniasis (VL), Post Kala-azar Dermal Leishmaniasis (PKDL) and different cross reactive diseases like Malaria, Leprosy, Vitiligo as compared to control subjects. IC levels were measured through a newly developed PEG ELISA, using L. donovani promastigote membrane antigen coated plate. Antibody classes and subclasses were identified using polyspecific sera and monoclonal antibodies, respectively. ICs were purified using polyethylene glycol (PEG) precipitation. Conditional logistic regression showed an association between IgG1-containing ICs and increased risk of PKDL (OR=75, P <0.05) and an association of IgG-containing ICs with VL (OR=621, P=0.001). PEG ELISA demonstrated almost 13-15 fold higher IgG containing ICs titers in VL as compared to control (P <0.001). The assay further established a significant (P <0.05) difference in the IgG containing ICs titers between VL and PKDL. The isolated ICs were further analyzed by subjecting them to one-dimensional PAGE and subsequently stained with combination of periodic acid schiff (PAS) with silver. A differential banding pattern between VL and PKDL was obtained. Four distinct bands with carbohydrate rich glycoconjugates were identified in PKDL ICs, which were absent in VL and control group. It suggests the scope for developing a novel differential diagnostic assay.

Application of special staining techniques in pathological diagnosis of fungi infections in HIV/AIDS patients / 中华临床感染病杂志

Objective To apply special staining techniques in pathological diagnosis of fungal infections in HIV/AIDS patients.Methods Pathological data of 20 HIV/AIDS patients complicated with fungi infections in Shanghai Public Health Clinical Center during February 2010 and November 2013 were retrospectively analyzed.Tissue specimens were stained with hematoxylin and eosin (HE),Periodic acidSchiff (PAS) and methenamine silver nitrate (MSN),and the sections were observed under optical microscope.Results Among 20 HIV/AIDS patients complicated with fungi infections,2 were infected with pulmonary cryptococcosis; 3 were penicillium marneffei infections in skin,lung and abdominal mesenteric lymph nodes; 5 were histoplasma capsulatum infections in epiglottis,neck lymph nodes,oral cavity,abdominal cavity and skin; 4 were aspergillus infections in maxillary sinus,lung and vocal cords,and 3 of them were combined with tuberculous lesions; 6 were candida albicans infections in liver,pharynx,esophagus and stomach.In tissues stained with HE the infiltration of inflammatory cells,granuloma formation and coagulative necrosis were observed,and the shape of fungi needed careful observation to avoid missed diagnosis and misdiagnosis.In tissues stained with PAS,fungal spores and pseudohypha were presented in bright amaranth,and cell nucleus was in purple-blue.In tissues stained with MSN,fungal spores and pseudohypha were identified clearly in brown-black.Conclusion HE plus PAS and MSN staining will help pathological diagnosis of fungi infection.

Avaliação histopatológica e histoquímica das células claras em lesões odontogênicas císticas / Análisis histopatológico y histoquímico de células claras en lesiones quísticas odontogénicas / Histopathological and histochemical analysis of clear cells in odontogenic cystic lesions

Alterações degenerativas e metaplásicas são usualmente observadas no revestimento epitelial dos cistos odontogênicos. Nos tumores odontogênicos estes processos são considerados mais raros. Objetivo: demonstrar a presença de células claras no revestimento epitelial de uma série de lesões odontogênicas císticas. Métodos: realizou-se um estudo descritivo, em uma amostra de 22 lâminas histológicas, do total de lesões odontogênicas císticas procedentes do Laboratório de Patologia Bucal da Universidade Federal da Paraíba. As lâminas foram examinadas por dois avaliadores previamente calibrados e os blocos parafinados correspondentes às lesões em que se observaram células claras, foram novamente processados, e as lâminas foram coradas pela técnica do Ácido Periódico de Schiff após digestão pela diástase e avaliadas quanto à presença de células mucosas. Resultados: células contendo mucina estavam presentes em 12 (54,5 porcento) das lesões coradas pelo Ácido Periódico de Schiff com diástase. Células mucosas foram observadas em 5 (41,66 porcento) dos 12 casos de cistos radiculares, 1 (50 porcento) dos 2 casos de tumores odontogênicos ceratocísticos, nos 4 (100 porcento) casos de cistos dentígeros e em 2 (100 porcento) casos de ameloblastomas unicísticos. Células claras foram muitas vezes observadas em áreas de inflamação. Conclusão: na amostra estudada, células mucosas puderam ser evidenciadas ocasionalmente no epitélio de lesões odontogênicas císticas(AU)

Cambios metaplásicos y degenerativas se observan generalmente en el revestimiento epitelial de los quistes odontogénicos. En tumores odontogénicos estos procesos son considerados raros. Objetivo: demostrar la presencia de células claras en el epitelio de revestimiento de una serie de lesiones quísticas odontogénicas. Métodos: se realizó un estudio descriptivo, en una muestra de 22 cortes histológicos, del total de lesiones quísticas odontogénicas procedentes del Laboratorio de Patología Oral de la Universidad Federal de La Paraiba. Los cortes fueron examinados por dos examinadores calibrados previamente y los bloques de parafina correspondientes a las lesiones que se observaron con células claras se volvieran a procesar y los cortes fueron teñidos por el Ácido Periódico de Schiff después de la digestión por la diastasa y evaluados para la presencia de células mucosas. Resultados: células contiendo mucina estaban presentes en 12(54,5 por ciento) de las lesiones teñidas por el Ácido Periódico de Schiff con diastasa. Células mucosas fueran observadas en 5(41,66 por ciento) de los 12 casos de quistes radiculares, 1 (50 por ciento) de los 2 casos de tumores odontogénicos queratoquísticos, en los 4 (100 %) casos de quistes dentígeros y en los 2(100 por ciento) casos de ameloblastomas uniquísticos. Células claras fueron muchas veces observadas en zonas de inflamación. Conclusión: En la muestra estudiada, las células mucosas pudieron ser encontradas ocasionalmente en el epitelio de lesiones quísticas odontogénicas(AU)

Metaplastic and degenerative changes are usually observed in the epithelial lining of odontogenic cysts. These processes are considered rare in Odontogenic tumors. Objective: to demonstrate the presence of clear cells in the epithelial lining of a series of odontogenic cystic lesions. Methods: a descriptive study was done, in a sample of 22 histological slides of the total odontogenic cystic lesions originating from the Oral Pathology Laboratory at the Federal University of Paraiba. The slides were examined by two calibrated examiners and the paraffin blocks, corresponding to the lesions, where clear cells were observed, were re-processed and the slides were stained by the Periodic Acid-Schiff technique after digestion by diastase and evaluated for the presence of mucous cells. Results: mucin-containing cells were present in 12 (54,5 percent) of the stained lesions by Periodic Acid-Schiff technique with diastase. Mucous cells were observed in 5 (41.66 percent) of 12 root cysts cases, 1 (50 percent) of 2 keratocystic odontogenic tumors, in 4 (100 percent) cases of dentigerous cysts and in 2 (100 percent) cases of unicystic ameloblastomas. Clear cells were often observed in inflammation areas. Conclusion: In the study sample mucous cells could be evidenced occasionally in the epithelium of cystic odontogenic lesions(AU)

Morfología y citoquímica de cultivos celulares de Aedes aegypti (Diptera: Culicidae) y susceptibilidad a Leishmania panamensis (Kinetoplastida: Trypanosomatidae)

Morphology and cytochemistry of Aedes aegypti’s cell cultures (Diptera: Culicidae) and susceptibility to Leishmania panamensis (Kinetoplastida: Trypanosomatidae). The first cellular line of Aedes aegypti was developed by Grace in 1966; afterwards, other cellular lines of this species have been generated. These have been used for the study of pathogenic organisms like viruses, bacteria and parasites, which demonstrates their importance in biomedical applications. This research describes, for the first time, some cytochemical characteristics of A. aegypti cell cultures, that were infected with (MHOM/CO/87CL412) strain of Leishmania panamensis. A morphological study of the cell culture was also carried out. Maintenance of the cell culture, parasites and infection in vitro were carried out in the Laboratory of Entomology, Cell Biology and Genetics of the Universidad de La Salle. The cell cultures infected with the parasite were maintained in a mixture of mediums Grace/L15, supplemented with 10 % fetal bovine serum (FBS) at pH 6.8 and a temperature of 26 ºC, during 3, 6 and 9 post-infection days. After this, these cell cultures were processed through High Resolution Light Microscopy (HRLM) and Transmission Electron Microscopy (TEM) based on standard protocols defined by the Group of Microscopy and Image Analyses of the Instituto Nacional de Salud. Semi-fine slices of 1 µm colored with toluidine blue were used for the morphological analysis of the culture, and ultra fine cuts of 60 to 90 nm stained with uranyl acetate and lead citrate where used for the ultrastructural study. In addition, PAS and peroxidase staining was carried out in cells fixed with methanol. The morphometric study was analyzed with software ImageJ (NIH). In the semi-fine slices, small cells were observed showing fibroblastic appearance 10.84±2.54 µm in length and 5.31±1.26 µm wide; other cells had epithelial appearance with a great peripheral nucleus, voluminous and vacuolated cytoplasm, 23.04±4,00 µm in length and 13.96±3.70 µm wide. These last ones predominated over the ones with fibroblastic appearance. Regarding the PAS coloration, 7.08 % of the cells presented abundant PAS positive cytoplasmatic granules which indicated polysaccharides presence. The peroxidase test gave a negative result. The greatest percentage of infection (18.90 %) of one total of 101 cells, turned up by day 6. Some cells analyzed by TEM presented a vacuolated aspect cytoplasm; some contained parasites, other fibrillar material and others were empty. The results indicate that A. aegypti cell culture can support the internalization and transformation of the parasite, which demonstrates the capacity that these cell cultures have to be infected with L. panamensis and to maintain the infection for approximately one week. Rev. Biol. Trop. 56 (2): 447-458. Epub 2008 June 30.

La primera línea celular de Aedes aegypti fue establecida por Grace en 1966 y desde entonces se han utilizado para el estudio de virus, bacterias y parásitos. En el presente trabajo se describen, por primera vez, algunas características citoquímicas de los cultivos celulares de A. aegypti, infectados con la cepa (MHOM/CO/87CL412) de Leishmania panamensis. También se realizó un estudio morfológico de las células del cultivo. Se observaron 30 células pequeñas con apariencia fibrolastoide de 10.84±2.54 µm de largo y 5.31±1.26 µm de ancho; otras 30 presentaron apariencia epitelioide con 23.04±4.00 µm de largo y 13.96±3.70 µm de ancho; éstas últimas predominaron sobre las de apariencia fibroblastoide. De 113 células, un 7.08%, presentaron abundantes gránulos citoplasmáticos positivos con la coloración de PAS, indicando presencia de polisacáridos. La prueba de peroxidasa dio un resultado negativo. El mayor porcentaje de infección (18.90%), de un total de 101 células, se presentó el día 6. Ultraestructuralmente, las células presentaron un citoplasma con aspecto vacuolado; algunas contenían parásitos, otras material fibrilar y otras estaban vacías. Los resultados indican que los cultivos celulares de A. aegypti pueden ser infectados por L. panamensis y mantener dicho proceso por aproximadamente una semana.

The Evaluation of Usefulness of PAS Stain and GMS Stain as Clinical Diagnostic Methods of Tinea unguium / 대한의진균학회지

BACKGROUND: Tinea unguium is a common problem seen in clinical practice. Considering the many differential diagnoses of dystrophic nails, it is important to make a definitive diagnosis of dermatophyte infection before the initiation of antifungal therapy. Potassium hydroxide (KOH) preparation and fungal culture, which are commonly used in the diagnosis of these infections, often yield false-negative results. Recent reports have suggested that nail plate biopsy (Bx) using periodic acid-Schiff (PAS) (Bx/PAS) stain may be a very sensitive technique for the diagnosis of tinea unguium. OBJECTIVE: The purpose of this study was to evaluate the usefulness of PAS and Grocott's methenamine silver (GMS) staining of nail specimen in the diagnosis of tinea unguium as a standard method. METHODS: We evaluated 75 nail specimens from suspected tinea unguium using KOH preparation, biopsy using periodic acid-Schiff stain, and Grocott's methenamine silver stain. RESULTS: Of the 75 nails which were negative on potassium hydroxide mounting, 43 and 39 cases were tested positive respectively on periodic acid-Schiff stain and Grocott's methenamine silver stain. CONCLUSION: Bx/PAS and Bx/GMS are the sensitive methods for the diagnosis of tinea unguium. They are indicated if clinical suspicion of onychomycosis is high and KOH preparation shows no fungal elements.

The Effect of Botulinum Toxin Injection on Compound Action Potentials and Repetitive Nerve Stimulation in Rat Gastrocnemius Muscle

Botulinum toxin develops muscular paralysis through the inhibition of acetylcholine release from presynaptic membrane in neuromuscular junction. It has been used clinically to treat strabismus, blepharospasm and spasmodic dysphonia. Recently it was introduced for the treatment of limb spasticity as well. Serial compound muscle action potential(CMAP) amplitudes were measured and repetitive nerve stimulation test(RNST) was performed with 2Hz and 30Hz on the rat gastrocnemius muscle to observe the effect of muscle paralysis. Also, Periodic acid Schiff (PAS) staining sections of the muscle for glycogen was studied to quantify the degree of muscular paralysis. Thirty Sprague-Dawley rats, 10 for control and 20 for experimental group were studied for 12 weeks. Normal saline 0.025 ml and 0.125 ml was injected into gastrocnemius muscle in cotrol group 1 and 2, respectively. Botulinum toxin type A(Botox) was injected 5.0U/0.025 ml in experimental group 1, 2.5U/0.025 ml in group 2, 2.5U/0.125 ml in group 3, and 0.5U/0.025 ml in group 4. The amplitudes of CMAP declined markedly by 81.1% to 96.5% of basal amplitudes on the first week after Botox injection, but slightly recovered on 12th week by 20.8% to 42.2% with greater recovery in lower dose group. RNST with 2Hz produced no remarkable 1 : 5 amplitude change in experimental group. RNST with 30Hz produced marked increment in 1 : 5 amplitude up to 24.4%. PAS staining for muscle sections showed residual glycogen after tetanic stimulation due to neuromuscular block by Botox.

Malignant Mixed Mesodermal Tumors of Ovary: 3 cases report

Malignant mixed mesodermal tumor (MMMT) of the ovary is endometrioid tumor containing epithelial and mesenchymal components. The mesenchymal component may be homologous, in which case the term carcinosarcoma is often used, or heterologous, containing cartilage, striated muscle, osteoid, bone, or fat, or combination of these elements. MMMT usually afflicts the postmenopausal woman and occurs more often in the nulliparous female. The disease is usually advanced (stage III) when diagnosed. Cliniopathologic data were presented for 3 cases of MMMT of the ovary with review of literatures. Clinical history substantiated previous reports in postmenopausal women wit this tumor. Symptoms were the same as for ovarian malignancy in general. All the patients presented wit stage II disease, according to the International Federation fo Gynecology and Obstetrics (FIGO) classification. Two patients died of tumor 4 and 10 months after operation, respectively. One patient had been alive at 2 months after operation. None of 3 cases demonstrate evidence of endometriosis in the remaining ovaries. Microscopically, periodic acid-Schiff (PAS)-positive, diastase-resistant hyaline droplets were found frequently in undifferentiated mesenchymal stroma and epithelial structures. The clinical significance and origin of the hyaline droplets in MMMT should be further explored.