Nuclear morphometry and chromatin textural characteristics of basal cell carcinoma

Abstract: Histological subtypes of basal cell carcinoma have biological, evolutionary and distinct prognostic behavior. The analysis of characteristics of the nucleus can provide data on their cellular physiology and behavior. The authors of this study evaluated nuclear morphological parameters and textural patterns of chromatin from different subtypes of basal cell carcinoma: nodular (n=37), superficial (n=28) and sclerodermiform (n=28). The parameters were compared between neoplasms' subtypes and with unaffected adjacent basal epithelium. Nuclear area and diameter of sclerodermiform neoplasms were superior to the other subtypes. Chromatin's color intensity and fractal dimension were less intense in superficial subtypes. Nuclear roundness and chromatin's entropy presented lower values in tumors than in normal epithelium. There was significant correlation between morphological and textural variables of normal skin and tumors. Morphometric elements and textural chromatin's homogeneity of basal cell carcinomas may be related to evolutionary, biological and behavior particularities related to each histotype.

Estresse pós-privação de sono: análises morfométricas do fígado / Stress caused by sleep deprival: liver morphometric analysis

OBJETIVO: Avaliar o efeito da exposição ao estresse pós-privação de sono em características celulares e nucleares do fígado de ratos. MÉTODOS: 16 ratos (Wistar) machos adultos (200-260 g) foram mantidos em ciclo de luz controlado recebendo dieta com quantidades usuais de sal e livre acesso à água e alimento. Os animais foram divididos em dois grupos de oito animais (grupo experimental e grupo controle). Os animais permaneceram na mesma gaiola (dois de cada vez) durante sete dias e, após esse período, foram pesados e separados. O animal do grupo controle continuou na mesma gaiola e o animal do grupo experimental foi colocado em aparato de privação de sono. Após as 96 horas, os dois ratos foram pesados,anestesiados, sacrificados com dose excessiva de anestésico e os fígados foram retirados. RESULTADOS: Não há diferença significante entre o grupo experimental e grupo controle em relação ao peso, embora haja diminuição de peso no grupo experimental. Nas análises cariométricas, houve diferença significante em relação ao diâmetro menor (p=0,03) e volume (p=0,04) do núcleo dos hepatócitos do grupo experimental. Nas análises esteriológicas, houve diferença significante do grupo experimental no núcleo (maior, p=0,036), do citoplasma (menor p=0,009) e outras estruturas (maior p=0,008). CONCLUSÃO: O estresse parece contribuir para alteração na estrutura celular hepática.

OBJECTIVE: To evaluate the effect of stress induced by sleep deprivation on nuclear and cellular features of rat liver. METHODS: 16 adult male Wistar rats (200-260 g) in controlled light cycle received diet with customary salt quantity and free access to water and food. The animals were divided into two groups with 8 animals each (experimental group and control group). Animals stayed in the same box (two at a time) during seven days andafter that period they were weighted e separated. The control group animal continued in the same box and the experimental group animal were transferred to a sleep deprivation apparatus. After the 96 hours the animals were sacrificed by an excessive anesthetic dose; animals were weighted and their liver extracted. RESULTS: There was no significant difference between experimental and control group regarding weight, although therewas a decrease of weight on the experiment group. Karyotipical analysis showed significant smaller diameter (p=0.03) and volume (p=0.044) of hepatocyte nuclei in experimental group. Stereological analyses showed significant differences in experimental group for nucleus (larger, p=0.036), cytoplasm (smaller, p=0.009) and other structures (larger, p=0.008). CONCLUSION: Stress seems to contribute to alteration of hepatic cell structure.

Effect of sodium cyclamate on the rat fetal exocrine pancreas: a karyometric and stereological study / Efectos del ciclamato de sodio en el pancreas exocrino fetal de ratas: estudio cariométrico y estereológico

The cyclamate, a sweetner substance derived from N-cyclo-hexyl-sulfamic acid, is largely utilized as a non-caloric artificial edulcorant in foods and beverages as well as in the pharmaceutical industry. The objective of this study was to evaluate karyometric and stereological alterations in the rat fetal pancreas resulting from the intraperitoneal administration of sodium cyclamate. The exocrine pancreas of ten fetuses of rats were evaluated, five treated and five controls chosen at random, in which five rats that received from the 10th to 14th days of pregnancy an intraperitoneal daily injection of sodium cyclamate at 60 mg/Kg of body weight during 5 days. At the 20th day of gestation, the animals were removed and weighed, as were their placentas; the length of the umbilical cords also were measured. After the laboratory processing, semi-seriated 6mm cuts stained with haematoxyline and eosine were performed. In seven karyometric parameters (major, minor, and medium diameters, volume, area, perimeter, and volume-area ratio), the increase was statistically significant in the treated group when compared with control group. Stereological parameters showed in the treated group a significant increase in the cellular volume and a significant reduction in the numerical cellular density. These results showed that the sodium cyclamate in pregnant rats led to retardation of fetal development and hypertrophy in the exocrine pancreas of the rat fetuses.

El ciclamato, es una substancia derivada del ácido N-ciclo-hexil-sulfámico, bastante usada como edulcorante no calórico en los alimentos y bebidas, así como en la industria farmacéutica. El objetivo de este estudio fue evaluar las alteraciones cariométricas y estereológicos en páncreas fetal de rata tras la administración intraperitoneal de ciclamato de sodio. El páncreas exocrino de diez de los fetos de rata fueron evaluados, cinco tratados y cinco controles seleccionados al azar, en el que cinco ratas recibieron del día 10 al día 14 de preñez una inyección intraperitoneal diaria de ciclamato de sodio a 60 mg/Kg de peso corporal durante 5 días. En el día 20 de gestación, los animales fueron retirados y pesados, al igual que sus placentas. Asimismo, se midió la longitud de los cordones umbilicales. Después del procesamiento de laboratorio, cortes semi-seriados de 6µm, se tiñeron con hematoxilina-eosina. En siete parámetros cariométricos (diámetros mayor, menor y medio, volumen, área, perímetro y relación área/volumen). El aumento fue estadísticamente significativo en el grupo tratado comparado con el grupo control. Los parámetros estereológicos mostraron en el grupo tratado un aumento significativo del volumen celular y una reducción significativa en la densidad numérica celular. Estos resultados mostraron que el uso del ciclamato de sodio en las ratas preñadas causa retardo en el desarrollo fetal e hipertrofia en el páncreas exocrino de los fetos de rata.

Estudio cariométrico de placentas de ratones con infección aguda por diferentes cepas de Trypanosoma cruzi / Karyometric study of placentas from mice with acute infection by different strains of Trypanosoma cruzi

El objetivo de este trabajo fue evaluar cariometricamente las alteraciones causadas por diferentes cepas de T. cruzi en la placenta del ratón. Ratones hembras de 60 días, grávidas, fueron inoculadas, intraperitonealmente, con 2 x 10(5) tripomastigotes sanguíneos de las cepas colombiana, Y, Solivia o RC del T. cruzi. Fueron observadas claras diferencias en las alteraciones cariométricas de las células trofoblásticas gigantes y de las células trofoblásticas de la zona esponjosa. Los resultados demostraron que las cepas colombiana y RC causan alteraciones tanto en las células trofoblásticas gigantes como en las células del trofoblasto esponjoso, mientras que las cepas Y y Bolivia provocan alteraciones solamente en las células trofoblásticas gigantes. Es posible concluir que cada cepa posee características propias y que, a pesar del tipo similar de transmisión, presenta matices diferenciales en el proceso de la patogénesis placentaria.

The objective of this work was to evaluate karyometrically the alterations caused by different strains of Trypanosoma cruzi in the mouse placenta. Pregnant mice, 60-day old, were intraperitoneally inoculated with 2 x 10(5) bloodstream trypomastigotes of the Colombian, Y, Bolivia or RC strain of T cruzi. There were observed clear differences in the karyometric alterations of the trophoblast giant cells and in the spongiotrophoblast cells. The results demonstrate that the Colombian and RC strains cause alterations both in the trophoblast giant cells and in the spongiotrophoblast cells, whereas the Y and Bolivia strains provoke alterations only in the trophoblast giant cells. It is possible concluding that each strain has its own characteristics and that, in spite of the similar type of transmission, it show differential nuances in the placental pathogenic process.

Evaluation of epithelial cell proliferating activity and fibroblast nuclear kariometry in recurrent pterygium treated with mitomycin C / Avaliação da atividade proliferativa das células epiteliais e da cariometria do núcleo dos fibroblastos em pterígios recidivados previamente tratados com a mitomicina C

PURPOSE: To evaluate the effectiveness of mitomycin C (MMC) in preventing recurrence of pterygium following conjunctival autograft transplantation (CAT). Ki-67 antigen to evaluate epithelial cell proliferation and fibroblast nuclear kariometry were used to assist treatment evaluation. METHODS: Twenty-nine patients with recurrent pterygium were divided into three groups: Group (G) 1 - CAT and placebo eyedrops (PED); G2 - CAT, 0.015 percent MMC subconjunctivally, and PED; G3 - CAT and 0.02 percent MMC eyedrops. Immunohistochemistry for the Ki-67 antigen and fibroblast nuclei kariometry were performed on the excised tissue, divided into nasal and temporal sides. Kariometry was evaluated in terms of volume (Vl) and area (Ar) using at least 50 cells/patient. RESULTS: The percentage of positive epithelial cells for the Ki-67 antigen on the nasal and temporal side after treatment of the three groups were: nasal (5.39 percent G1, 4.49 percent G2, and 3.88 percent G3); temporal (3.30 percent G1, 4.46 percent G2, 4.14 percent G3), did not show significant differences. Fibroblast nucleus kariometry was: nasal Vl (792.1 µ3 G1, 605.1 µ3 G2, and 549.9 µ3 G3) and Ar (100.58 µ2 G1, 83.13 µ2 G2, and 78.41 µ2 G3). The three groups showed significant differences: p=0.039 and p=0.035, for respectively Vl and Ar, on the nasal side. After a six month of treatment, the three groups presented the following recurrence rates: G1, 22.22 percent, G2, 18.18 percent and G3, 33.33 percent, respectively. CONCLUSION: MMC did not reduce the number of positive epithelial cells for the Ki-67 antigen in recurrent pterygium, but decreased fibroblast nucleus volume and area on the nasal side of the pterygia. The number of positive epithelial cells for the Ki-67 antigen seemed not to be related to pterygium recurrence observed over a six-month post-surgery period. The role of epithelial cell proliferation in pterygium recurrence should be evaluated by further studies.

OBJETIVO: Avaliar a eficácia da mitomicina C (MMC) na prevenção da recorrência quando previamente utilizada no transplante autólogo de conjuntiva (TAC). A avaliação da proliferação celular epitelial pelo antígeno Ki-67 e a cariometria do núcleo dos fibroblastos foram usados como auxiliares na avaliação do tratamento. MÉTODOS: Vinte e nove pacientes com pterígio recidivado foram divididos em três grupos: Grupo (G) 1-TAC e colírio placebo (PLA); G2-TAC, MMC 0,015 por cento subconjuntival e PLA; G3-TAC e colírio de MMC 0,02 por cento. A imuno-histoquímica foi realizada no tecido excisado para o antígeno Ki-67, como a cariometria dos núcleos dos fibroblastos (divididos em lado nasal e temporal). A cariometria dos núcleos dos fibroblastos foi avaliada de acordo com os seguintes parâmetros: volume (Vl) e área (Ar) em pelos menos 50 células por paciente. RESULTADOS: A porcentagem das células epiteliais positivas para o antígeno Ki-67 no lado nasal e temporal após o tratamento dos três grupos estudados foi: nasal (3,30 por cento G1, 4,49 por cento G2 e 3,38 por cento G3) e temporal (3,30 por cento G1, 4,46 por cento G2 e 4,14 por cento G3) não mostrando diferença significativa. A cariometria do núcleo dos fibroblastos foi: Vl nasal (792,1 µ3 G1, 605,1 µ3 G2, e 549,9 µ3 G3) e a Ar (100,58 µ2 G1, 83,13 µ2 G2, e 78,41 µ2 G3). Os três grupos mostraram uma diferença significativa p=0,039 e p=0,035, respectivamente do Vl e da Ar no lado nasal. Após seis meses de tratamento, os três grupos apresentaram a seguinte taxa de recidiva: 22,22 por cento G1, 18,18 por cento, G2 e 33,33 por cento G3 respectivamente. CONCLUSÃO: O uso da MMC não interferiu nas células epiteliais positivas para o antígeno Ki-67 no pterígio recidivado, mas acarretou diminuição do volume e área dos núcleos dos fibroblastos no lado nasal do pterígio. As células epiteliais positivas para o antígeno Ki-67 parecem não ter relação com a recidiva do pterígio após seis meses...

Study of the Nuclear Volume of Testicular Cells of Rats Receiving Ofloxacin Through Lactation / Estudio del Volumen Nuclear de Células Testiculares de Ratas que Recibieron Ofloxacin Durante la Lactancia

Ofloxacin presents an ample spectrum of antimicrobial action, including combating Mycobacterium leprae, and is currently employed as a substitute when the use of rifampicin is impossible. The objective of this work was to study alterations in testicular cell nuclei of suckling rats, whose dams were submitted to oral application of ofloxacin, and respective control groups. The method utilized was morphometry by the karyometric technique. The main structures observed in histological preparations of the testicles were interstitial cells, spermatogonias, and sustentacular cells. 10 Wistar rats were utilized, four treated and five controls, in the period of the first 25 days of life, whose dams received ofloxacin 12 mg/ Kg of body weight / day orally, before being killed on the 25th day after birth. The karyometric study of interstitial cells and spermatogonias revealed that there were no changes in the form of their nuclei (p > 0.05). Since sustentacular cell nuclei presented increased major diameter, minor diameter, mean geometric diameter, volume, area, volume/area ratio and perimeter, as well as an augmented and statistically different eccentricity (p < 0.05) in suckling pups whose dams were administered ofloxacin, the nuclei presented larger size and more elongated form. It was concluded that the sustentacular cells were more sensitive to the ofloxacin effect at the administered dose.

El ofloxacin presenta un amplio espectro de acción antimicrobiana, incluyendo el combate a.Mycobacterium leprae, y es frecuentemente empleado como un sustituto cuando el uso de la rifampicina es imposible. El objetivo del trabajo fue estudiar las alteraciones en el núcleo de las células testiculares en ratas que se encontraban amamantando y que fueron sometidas a la aplicación oral de ofloxacin. El método utilizado fue la técnica morfométrica de la cariometría. Las principales estructuras observadas en las preparaciones histológicas fueron las células intersticiales, espermatogenias y células sustentaculares. Se utilizaron 10 ratas Wistar, cinco fueron el grupo control y cinco sometidas al tratamiento, cuyas madres recibieron ofloxacin en dosis oral diaria de 12 mg/Kg de peso corporal los primeros 25 días de vida, para luego ser sacrificadas al día 25 después del nacimiento. El estudio cariométrico de las células intersticiales y de la espermatogénesis revelaron que no hubo cambios en la forma de sus núcleos (p > 0,05). Las células sustentaculares presentaron un incremento en su diámetro mayor, diámetro menor, diámetro geométrico promedio, volumen, área, razón volumen/área y perímetro, también hubo aumento con una diferencia de la excentricidad estadísticamente significativa (p < 0,05) en las crías amamantadas, a las cuales se les administró ofloxacin. Los núcleos presentaron un gran tamaño y una forma más alargada. Esto concluye que las células sustentaculares son más sensibles al efecto de la administración de ofloxacin.

Effect of Sodium Cyclamate on the Rat Fetal Liver: A Karyometric and Stereological Study / Efectos del Ciclamato de Sodio en el Hígado Fetal de Ratas: Estudios Cariométrico y Estereológico

The cyclamate, a sweetner substance derived from N-cyclo-hexyl-sulfamic acid, is largely utilized as a non-caloric artificial edulcorant in foods and beverages as well as in the pharmaceutical industry. The objective of this study was to evaluate fetal hepatic karyometric and stereological alterations in the rat fetal liver resulting from the intraperitoneal administration of sodium cyclamate. The livers of ten rats were evaluated, five treated and five controls chosen at random, in which five rats that received from the 10th to 14th days of pregnancy an intraperitoneal daily injection of sodium cyclamate at 60 mg/Kg of body weight during 5 days. At the 20th day of gestation, the animals were removed and weighed, as were their placentas, on a precision balance; the length of the umbilical cords also were measured. After the laboratory processing, semi-seriated 6mm cuts stained with haematoxyline and eosine were performed. In seven karyometric parameters (major, minor, and medium diameters, volume, area, perimeter, and volume-area ratio), the increase was statistically significant in the treated group when compared with control group. Stereological parameters showed in the treated group a significant increase in the cytoplasmatic and cellular volume, and a significant reduction in the nucleus-cytoplasm ratio as well as in the numerical cellular density. These results showed that the sodium cyclamate in pregnant rats led to retardation of fetal development and hepatic-cellular hypertrophy in the offspring, suggesting toxicity in liver of rat fetuses.

El ciclamato, es una substancia derivada del ácido N-ciclo-hexil-sulfámico, bastante usada como edulcorante no calórica en alimentos y bebidas, así como en la industria farmacéutica. El objetivo del trabajo fue evaluar los efectos del ciclamato de sodio en hígados de fetos de ratas, considerándose las alteraciones cariométricas y estereológicas. Fueron utilizadas 10 ratas adultas (Rattus norvegicus) variedad Wistar, con peso medio de 240 g, siendo 5 el grupo control y 5 tratadas con ciclamato de sodio. Entre el 10 y 14 día de la preñez, 5 ratas recibieron una inyección diaria intraperitoneal de 60mg/Kg/día de ciclamato de sodio durante 5 días. En el 20 día, los animales fueron sacrificados y los fetos fijados en solución de Alfac, incluidos en parafina, cortados a 6 µm y teñidos com H-E. Hubo aumento estadísticamente significativo en siete parámetros cariométricos (diámetros mayor, menor y medio, volumen, área, perímetro y relación área/volumen) en el grupo tratado con ciclamato de sodio comparado con el grupo control. Parámetros estereológicos mostraron aumento significativo en los volúmenes citoplasmático y celular y disminución significativa en la relación núcleo/citoplasma y densidad numérica celular. Los resultados mostraron que el uso del ciclamato de sodio en las ratas preñadas causó retardo en el desarrollo fetal e hipertrofia celular hepática en los fetos, sugerente de toxicidad en el hígado fetal de las ratas.

Acción del Etanol sobre el Epitelio Nasal y Glándulas Septales de Ratas, Durante la Lactancia / Action of Ethanol on Rat Pups Nasal Epithelium and Septal Glands, During Lactation

El consumo materno de etanol durante la lactancia altera la composición de la leche, provoca la aparición de etanol y acetaldehído en la leche y agrava los efectos del etanol en las crías de ratas. De este modo, fueron estudiados los efectos del etanol, administrado a las ratas madres durante la lactancia, sobre el epitelio respiratorio y en las glándulas septales anterior y posterior de las crías lactantes con 21 días de vida postnatal. Las ratas recibieron etanol al 20% en el bebedero, ad libitum durante los 21 días que amamantaron. Los controles recibieron un volumen similar de agua sin alcohol. Las crías fueron sacrificadas en el 21 día. Las cabezas fueran cortadas frontalmente. Los cortes seriados de 6 µm de grosor fueron teñidos con hematoxilina y eosina. Fueron determinados los parámetros nucleares de los epitelios respiratorio y glandular, así como los volúmenes citoplasmático y celular, relación núcleo-citoplasma, densidades numérica y superficial y espesor del epitelio. El peso corporal medio de las crías control fue 34,9 g y en las tratadas 20,2 g. Histologicamente, el epitelio respiratorio se mostró adelgazado en el grupo tratado, constituido por células abundantes y menores, con núcleos menores. La glándula septal posterior presentó ácinos menores. En este trabajo, el etanol indujo un cuadro de hipotrofia epitelial (respiratorio y glandular), indicando una acción directa sobre las células de la mucosa nasal, además de retardar el desarrollo de las crías intoxicadas.

Maternal ingestion of ethanol during lactation alters the chemical composition of milk, results in the appearance of ethanol and acetaldehyde in milk, and exacerbates the effects of ethanol on the rat pups. So that, the effect on respiratory epithelium and anterior and posterior septal glands in 21-day-old suckling pups of ethanol treated mothers was studied. Female rats received dinking water ad libitum containing ethanol (20%) throughout the whole lactation. Control animals received a similar volume of water without ethanol. Lactent rats (21 day-old) were killed by lethal dose of anesthetic. The heads were serially sectioned (6µm thick) in a frontal plain, and stained with haematoxylin and eosin. Cell nuclear parameters were estimated, as well as cytoplasm and cell volume, nucleus-cytoplasm ratio, number and surface densities, and epithelial thickness. Mean body weight of the pups was 34.9 g for the controls and 20.2 g for the treated group. Histologically, the respiratory epithelium was thinner, with more numerous and smaller cells and small nuclei. The posterior septal glands showed smaller acini. In this paper, ethanol induced epithelial (respiratory and glandular) hypotrophy, indicating a direct action in nasal mucous cells, besides retarded development in pups.