Determination of the minimal representative number of microscopic fields to quantify apoptosis in canine lymph nodes

Moro, Luciana; Vasconcelos, A.C.; Santos, F.G.A.; Alves, Camille Marques; Nunes, J.; Sampaio, Ivan Barbosa Machado

doi:10.1590/s0102-09352004000300020

Cited by 18 publications

(15 citation statements)

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“…The total number of cells and the number of apoptotic cells in 35 fields (100X) per slide were counted and apoptotic indices (number of apoptotic cells/ total number of cells) were calculated. was reached through the analysis of the instability of the standard deviation of the mean (Sampaio, 1998;Moro et al, 2004). Firstly, a thymus of an infected dog was collected and processed routinely for histological examination and sections 5µm thick were stained by Shorr.…”

Section: Methodsmentioning

confidence: 99%

“…Apoptosis is an organized ATP-dependent form of cellular death which requires de novo protein synthesis and gene transcription, and is often called programmed cell death (Orlowski, 1999). Morphologically it is characterized by shrinkage of the cell, chromatin condensation or compaction against the nuclear periphery, active membrane blebbing (zeiosis), and ultimate fragmentation of the cell into membrane enclosed vesicles (apoptotic bodies) (Wyllie et al, 1980).…”

Section: Introductionmentioning

confidence: 99%

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Morphometric analysis of the thymus of puppies infected with the Snyder Hill Strain of canine distemper virus

Alves¹,

Vasconcelos²,

Martins³

et al. 2006

Arq. Bras. Med. Vet. Zootec.

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The thymic morphometry analysis was used for determining apoptosis and atrophy of the thymus of eight puppies inoculated with canine distemper virus (CDV). Three healthy dogs were used as negative controls. Sections, 5µm thick, were stained by HE and Shorr, and the latter were evaluated by morphometry. CDV nucleoprotein was detected by immunohistochemistry. Morphometric results confirmed lymphoid hypotrophy in CDV inoculated dog thymuses, more stroma, less parenchyma and higher apoptotic index/field than negative control (not inoculated) puppies. Apoptosis plays a role in the mechanism of thymus atrophy that develops in canine distemper.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Morphometric analysis of the thymus of puppies infected with the Snyder Hill Strain of canine distemper virus

Alves¹,

Vasconcelos²,

Martins³

et al. 2006

Arq. Bras. Med. Vet. Zootec.

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“…As imunomarcações foram quantificadas em um número mí-nimo representativo de campos definido pela avaliação da evolução dos erros padrões de acordo com o aumento da amostragem (Moro et al 2004). Brevemente, determinou-se o Índice de marcação a partir da contagem de células no número máximo obtido de campos tomados numa lamina.…”

Section: Methodsunclassified

Apoptose e maturação placentária bovina: um estudo imunohistoquímico e morfométrico

et al. 2016

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Placental release after birth involves loss of maternal-fetal adhesion and occurs only after complete maturation of the placentoma related to the decrease in cellularity of fetal and maternal tissues. Apoptosis is required for both the normal maturation and release of the placenta after birth. The aim of this study was to evaluate the occurrence of apoptosis in samples of the placenta of cows in different stages of gestation. Samples of 15 healthy cow placentomas at 4 (n=5), 6 (n=5) and 9 (n=5) months of gestation were harvested and processed for routine histology, immunohistochemistry and histochemistry. The slides were stained with HE, PicroSirius Red and subjected to immunohistochemical analysis of proteins Caspase 3, Caspase 8, Bax and Bid. Increase in number of vessels was not associated with increase in vascular area during progression of gestation. The results of histomorphometry revealed increased labeling for Bax and Caspases 3 and 8 in trophoblastic binucleated cells in late pregnancy, where the Bid remained without significant change. Histomorphometry analyzing the mononuclear trophoblast cells showed a high expression for Bax in early pregnancy, but decreased at 6 months of gestation. Immunolabeling revealed increased Caspases 3/8 and Bid with advancing of gestation. Further evaluation of type I and III collagen showed a decrease of both types of collagens at the end of gestation, what is very important for the reduction of maternal-fetal adhesion. These results confirm that Caspases 3 and 8 and Bax are involved in the mechanisms of activation of apoptosis through mitochondrial intrinsic and/or extrinsic pathway during pregnancy in trophoblastic binucleated cells. In mononuclear trophoblast cells Bax looses importance in the apoptosis process, awhile Bid and caspases 3 and 8 become the most significant.

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“…A cut-off value of the minimal representative number of microscopic fields was established by the standard deviation (SD) instability analysis described elsewhere (Moro et al, 2004). The AgNORs counting was carried out with a digital image analyzer.…”

Section: Methodsmentioning

confidence: 99%

“…Studies using experimental transplantation have demonstrated that integral viable cells can produce TVT in dogs of all ages (Rogers, 1997). The routes of inoculation include: subcutaneous, intraperitoneal and subaracnoideal (Nielsen and Kennedy, 1990;Das and Das, 2000). Following subcutaneous inoculation, the TVT cells induce Arq.…”

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confidence: 99%

Cell proliferation markers in the transplanted canine transmissible venereal tumor

Santos

Moro

Cassali

et al. 2011

Arq. Bras. Med. Vet. Zootec.

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Adult male mongrel dogs were subcutaneously transplanted with the canine transmissible venereal tumor (TVT) on the hypogastric region. Twelve specimens of tumors were collected, half during the proliferative phase and the other half during the regressive phase. Fragments of the tumor were fixed in 10% buffered formalin and routinely processed for light microscopy. Sections of 4µm were stained by Schorr or AgNOR or either immunostained for MIB1 (Ki67). Schorr stain, AgNOR and MIB1 showed an increased proliferative activity through mitotic index, nuclear argyrophilic protein stain and cycling tumoral cells in the growing tumors, respectively. All of the three cell proliferation markers were able to distinguish the TVT in both evolution phases. MIB1 monoclonal antibody was the best in the morphologic evaluation of growth and regression of TVT. This resulted in higher values than AgNORs counting and mitotic index. MIB1 immunostaining was the most effective parameter of the proliferative activity of TVT. However, a significant correlation has been detected only between mitosis counting and AgNORs. Keywords: dog, venereal tumors, MIB1, AgNOR RESUMO Cães machos, adultos, mestiços, foram transplantados com células do tumor venéreo transmissível canino (TVTC), na região hipogástrica. Foram coletados doze espécimes do TVTC

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Determination of the minimal representative number of microscopic fields to quantify apoptosis in canine lymph nodes

Cited by 18 publications

References 1 publication

Morphometric analysis of the thymus of puppies infected with the Snyder Hill Strain of canine distemper virus

Morphometric analysis of the thymus of puppies infected with the Snyder Hill Strain of canine distemper virus

Apoptose e maturação placentária bovina: um estudo imunohistoquímico e morfométrico

Cell proliferation markers in the transplanted canine transmissible venereal tumor

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