Biomedical subjects

C G Barbeito

Publications and source records attributed to C G Barbeito.

14 recordsLinked to original sources

PubMed2006

DNA synthesis in tongue keratinocytes of hepatectomized and tumor-bearing mice.

Tongue keratinocytes have high S-phase and mitotic indices with evident circadian variation. Transplanted tumors modify the intensity and temporal structure of the S-phase index in cell populations in tumor-bearing animals; also, partial hepatectomy changes the concentrations of substances involved in cellular proliferation, leading to compensatory liver hyperplasia. The aim of our study was to analyze the interaction between tumor growth and the liver regeneration that follows partial hepatectomy, and the effects of both these processes on lingual keratinocytes. We used 380 adult male mice divided into six groups: tumor-free and tumor-bearing mice without surgery, with sham hepatectomy, and with partial hepatectomy. Each group was divided into six subgroups, which were killed at 4-h intervals until a circadian cycle was completed (from 26 until 50h post-surgery in the operated animals). Each animal was injected with 5-bromodeoxyuridine (50mg/kg) 1h before it was killed, and tongue samples were obtained and processed for histology. The sections were placed on silanized slides and incubated with the primary antibody Bu 20a (1/100 dilution). The reaction was developed using diaminobenzidine and staining was detected visually. SIs were measured as the number of labeled nuclei per thousand cells. The mean+/-S.E. of each group was calculated. Differences among experimental groups were analyzed by ANOVA and the Student-Newman-Keuls Multiple Comparisons Test. The results show that the presence of a tumor alters the normal circadian curve of SI in lingual keratinocytes, irrespective of whether the mice underwent surgery. This finding has to be considered in drug treatments for neoplasms and in experiments related to growth.

Animals↗

PubMed2006

Characterization of interstitial cells of Cajal in bowel of cattle (Bos taurus).

Interstitial cells of Cajal (ICC) have been described in the gastrointestinal tract of different mammals including humans, horses, pigs, rats, dogs, mice and guinea-pigs. In the present study, ICC were identified in the jejunum of Bos taurus using polyclonal anti-c-Kit antibodies in immunohistochemical assays. Vimentin and desmin intermediate filaments were also determined using monoclonal antibodies. ICC were found in the tunica muscularis either in a palisade distribution pattern between the outer longitudinal and the inner circular layers (ICC-MP) or freely distributed in clusters in the longitudinal layer (ICC-LM). Morphometric studies determined that ICC have a fusiform shape presenting cytoplasmic prolongations. ICC were positive to c-Kit and vimentin antigens but negative to desmin. We have observed and described for the first time the presence of ICC in a ruminant. As observed in the aforementioned mammals, bovine ICC were associated with the myenteric plexus. Nevertheless, the presence of widespread ICC in the longitudinal muscular layer of the jejunum differs from previously described studies of other mammals.

Animals↗

PubMed2006

An Argentine equine herpesvirus strain with special restriction patterns protect mice challenged with a pathogenic strain.

Equine herpesvirus 1 (EHV-1) was first isolated in Argentina in 1979. This strain SPv has special restriction patterns, but a previous study demonstrated that SPv did not modify its growth in cell culture. In addition, it showed low virulence in the mouse respiratory model consistently with results found in female BALB/C at different state of gestation. This study evaluates in a mouse respiratory model, if primary infection with SPv strain protects animals from subsequent challenge with a pathogenic strain. Body weight loss was not observed in mice intranasally inoculated with SPv strain and challenged with HH1 Japanese strain. The SPv primary infection does not completely prevent clinical presentation by HH1 infection but the SPv inoculated animals recovered more quickly, with less intense and less persistent histological lesions. The challenge infection caused a rapid and prolonged increase in anti-EHV-1 antibodies in the mice previously infected with SPv, along with a more rapid reduction of viral titres in lungs. In this work it was demonstrated that this EHV-1 strain constitute a good immunogen. These results show that this SPv strain could be considered to produce an EHV-1 vaccine.

Animals↗

PubMed2005

Cyclic related and pathological changes in the lectin-binding sites on the swine oviduct.

This study was carried out to compare the lectin-binding pattern in the normal and pathological oviduct of sows during the ovarian cycle. Lectin-binding patterns showed differences between segments, phases of ovarian cycle and presence of morphologic lesions. In the infundibulum, it was observed that the cysts, in the follicular phase, reduced Ricinus communis-I (RCA-I) and Ulex europaeus-I (UEA-I) binding. Furthermore, in the pathological oviducts of the luteal-phase group, there is a reduction of Concanavalia ensiformis (Con-A) reactivity in this segment of the tube having wall cysts, adenomyosis and diverticulus. The Arachis hypogaea (PNA) binding in the infundibulum, during the luteal phase, decreased in the tube having adenomyosis. In animals with wall cysts, the Con-A, Triticum vulgaris (WGA) and RCA-I reactivity was minor in the glycocalyx of the isthmus epithelium during follicular phase. Con-A and Dolichos biflorus (DBA) binding pattern was minor in the luteal-phase isthmus of the tube having wall cysts, adenomyosis and diverticulus. In the ampulla, the wall cysts impaired the Con-A reaction only in the basal region of the epithelium, in the follicular phase. Binding with Con-A was decreased in the ampulla of animals in the luteal phase in the tube lesions with cysts and diverticulus. In addition, the diverticulus observed in the ampulla, during the luteal phase, reduced the PNA tubaric binding. The results of this study showed that the morphologic alterations modify the sugar pattern in the oviduct of sows. These modifications in glycoconjugates may be one of the reasons for the failure of fertility in sows.

Animals↗

PubMed2004

Argentine strain of equine herpesvirus 1 isolated from an aborted foetus shows low virulence in mouse respiratory and abortion models.

The equine herpesvirus 1 (EHV-1) was isolated in Argentina from an aborted equine foetus in 1979. This virus (SPv) has special restriction patterns (RP) in comparison with other Argentine isolates. In addition, SPv could be distinguished on the basis of its pathogenicity in baby mice inoculated intracerebrally. We studied the growth properties of the SPv in cell culture and its effects in a mouse respiratory and abortion model. We observed that SPv did not modify its capacity to grow in cell culture with respect to reference HH1 strain. Nevertheless, we found significant differences between the titres of the two strains at 8-14 h post-infection (PI). In this work we demonstrated that SPv showed low virulence in female at different stages of gestation, consistently, with results found in the mouse respiratory model. We considered that this low virulence of SPv could be related to its RP because the RP of HH1 strain are similar to those of the HVS25A strain and both showed effect on pregnant mice. More specific studies about genomic alterations to the SPv are necessary for identifying, more clearly, if the intra-strain variations have relation with the low virulence in the mouse respiratory and abortion model.

Animals↗

PubMed2004

Lectin binding patterns in hyperplastic and metaplastic bullock prostate tissues after diethylstilbestrol administration.

Hyperplasia and squamous metaplasia of the prostatic epithelium are conditions induced by oestrogens. Diethylstilbestrol (DES) has been banned from cattle used for beef production because of the health risks. The potential use of molecular markers for the detection of illegal oestrogen administration was evaluated by taking samples of prostatic tissue from control bullocks, bullocks which had been treated with oestrogens, and bullocks sacrificed 21 and 90 days after a single dose of DES. The expression of the glycoconjugates was examined by lectinhistochemistry and the lectin binding pattern was characterised in epithelium and connective tissue. In the animals sacrificed after 21 days there was an increase in the binding of one lectin (JAC) and there was an increase in the binding of one of the other lectins (DBA) in the animals sacrificed after 90 days. An increase in SWGA lectin staining was observed in the bullocks that had probably been treated with oestrogen and in the animals sacrificed 90 days after the inoculation with DES. There were also differences between the binding of SWGA in the control bullocks and the other groups.

Animals↗

PubMed2002

Development of compensatory hepatic hyperplasia in mice carrying the hepatocellular carcinoma ES12a.

In a previous paper we reported that the presence of the hepatocellular carcinoma SS1K in host mice resulted in an earlier appearance of the hepatocyte mitotic peak during liver regeneration after a partial hepatectomy as well as in an increase in the amplitude of that mitotic wave. In the present work we analyse the effect of another hepatocellular carcinoma, the ES12a (HCES12a). Adult male mice of the C3H/S strain standardised for circadian-periodicity analysis, were used. One group received a subcutaneous graft of the HCES12a tumor, while another group served as control. Fifteen days later, all animals were submitted to a partial (70%) hepatectomy at 10:00 h and beginning at 16:00 h lots of between 5 and 9 host and control animals each were sacrificed at 4 h intervals until 16:00 h on the third day thereafter. All mice were injected with 2 microg/g colchicine 4 hrs before killing, and samples of livers were processed for hematoxylin-eosin staining. We determined the hepatocyte mitotic index for each animal and the mean value +/- the standard error of the mean for each lot. The peak of mitotic activity in the tumor-bearing animals took place four hours earlier than in control mice but the average values of hepatocytic mitotic activity were similar in both groups

Animals↗

PubMed2001

Circadian rhythm of DNA synthesis and mitotic activity in tongue keratinocytes.

Tongue keratinocytes have a high mitotic index (MI) with an evident circadian variation. Our study set out to compare and contrast two phases of the cell cycle: DNA synthesis (S-phase), with inmunocytochemical detection by bromodeoxyuridine (BrdU), and mitosis (M-phase), by the colchicine-arrest of metaphase method, exploring both the dorsal and ventral surfaces of the mouse tongue throughout a circadian period. Adult male mice standardized for light periodicity used for MI experiment were injected intraperitoneally with colchicine. Other animals were injected intraperitoneally with 5-BrdU for S-phase determination. Animals given both treatments were divided into six groups and killed at 4 h intervals until 20:00 h. Tongue samples were processed for histology and immuno-histochemistry. S and M indices were expressed as labelled nuclei or colchicine metaphases, respectively, per 1000 nuclei. Peak MI occurred at 12:00, with the minimum value at 20:00 on dorsal and ventral tongue surfaces. Peak S-phase was at 04:00, whereas the minimum value was at 16:00 for both surfaces. These results show that the proliferative activity of the tongue epithelium is of similar intensity and temporal distribution on both surfaces.

Animals↗

PubMed2001

Effect of partial and sham hepatectomy on the growth of a hepatocellular carcinoma.

We examined the effect of partial hepatectomy on the proliferation of hepatoma ES12a grafted into C3H/S mice compared to tumor growth in sham-hepatectomized controls. The animals were sacrificed every 4 hrs during three days from the 6th to the 78th h following each type of surgery. Unoperated tumor-bearing mice were likewise killed as controls, but only during one complete circadian period. All animals received 2 microg of colchicine per g of body weight intraperitoneally 4 hrs before decapitation. Measurement of mitotic indices in hematoxylineosin-stained tumor samples revealed a decrease in proliferation and a modification of the diurnal mitotic-activity profile in the hepatectomized and sham-operated animals from the first day after surgery. These differences persisted by the third postoperative day only in the hepatectomized animals. Thus, although surgical stress may initially affect tumor growth, the latter results must be the effect of the influence of the hepatic regeneration.

Animals↗

PubMed2000

Mitotic activity of the pars intermedia in the female mouse: age-associated variations in proliferation rate and circadian periodicity.

We previously reported daily variations in the mitotic activity of the endocrine cells in the pars intermedia of 21- and 28-day-old male mice. Since cellular proliferation might be affected by factors such as sex and age, we undertook the present experiments to study the mitotic activity of the pars intermedia from 14-, 28-, and 150-day-old female mice. Inbred C3H/S mice, grouped according to age, were housed under standard conditions (12h each of light and dark [LD 12:12]) for periodicity analysis and were killed in lots of 5-11 animals every 4h over a single 24h cycle, with each mouse receiving 2 microg/g of colchicine 4h before decapitation. Pituitaries were excised, extracted, fixed in buffered formaldehyde, embedded in celloidin-paraffin, sectioned at 5 microm, and stained with hematoxylin and eosin. We counted the total number of nuclei to estimate the total number of cells monitored and then calculated the mitotic index (metaphases/1000 nuclei). Differences were analyzed for statistical significance by the Student t test. While the 14-day-old animals manifested no significant changes in mitotic activity during the 24h cycle, the 28- and 150-day-old mice showed higher mitotic indices during the period of darkness. The average mitotic activity over the entire cycle, however, was higher in the two groups of younger animals than in the 150-day-old mice. Moreover, the averages for the 28-day-old females were higher than the corresponding values previously reported by us for male mice of the same age.

Aging↗

PubMed2000

Intermediate filament protein expression and sugar moieties in normal canine placenta.

In the female dog, the placenta is considered zonal, endotheliochorial and labyrinthic. The distribution of the intermediate filaments as well as the surface glycoproteins in the canine placenta are still unknown. The aim of the present study was to provide this information for further understanding of pathological conditions in the bitch. Samples were obtained from normal uterine horns at the end of gestation. Tissues were routinely fixed and stained. Monoclonal antibodies against cytokeratins, vimentin and desmin were used for immunohistochemical staining. UEA-1, PNA, RCA-1, SBA, DBA, WGA and ConA were used for the lectin histochemical staining. A computer morphometrical analysis was made. Statistical analysis was then accomplished. The results showed the maximum immunohistochemical percentage for vimentin in the supraglandular connective tissue, for pancytokeratin in the spongy zone and for desmin in miometrium. SBA showed the highest staining percentage in the gland cells of the spongy zone, while ConA was the highest in the syncytiotrophoblastic cells and gland cells of the deep glandular zone. The results obtained indicate that the lectin binding pattern is partially different from other animal species. On the contrary, the intermediate filament data coincide with similar observations from other mammals.

Animals↗

PubMed1998

Characterisation of cytotrophoblastic-like cells present in subinvolutioned placental sites of the bitch.

This paper describes an approach to study the cells present in the subinvolution of placental sites (SIPS), a pathological post partum condition of the bitch that causes persistent hemorrhage of the genital tract. The expression of intermediate filament proteins was examined to determine the fetal or maternal origin of the cytotrophoblastic-like cells found in this entity. Lectin binding on tissue sections were also studied to characterise cellular glycoconjugates. Image processing and morphometrical analysis of the histological images were done. The results revealed that the cells observed in bitches with SIPS expressed pancytokeratins but neither vimentin nor desmin, in coincidence with normal cytotrophoblasts. The lectin binding pattern of both types of cells was similar, with the only exception of Arachis hypogaea agglutinin (PNA) and Triticum vulgaris agglutinin (WGA). These observations, in addition to the non statistically significant differences between morphometrical characteristics of cytotrophoblastic and cytotrophoblastic-like cells in SIPS, might suggest the fetal origin of the latter cells which could play a role in the pathogenesis of this entity.

Animals↗

PubMed1997

[Mitotic activity of duodenal-crypt enterocytes in mice with hepatocarcinoma].

Tumors grafted into mice may modify the proliferation of normal cell populations. In this paper, we have studied the evolution of mitotic activity (MA) in duodenal-crypt enterocytes of ES12a hepatocarcinoma-bearing mice; a total of 87 28-day-old female animals of the C3H/S strain were used after standardization for circadian-periodicity analysis. The mice were distributed into two groups: those remaining intact and those receiving tumor grafts. Each group was then divided into six batches (n = 6-10), one of which was sacrificed every 4 h over a period of one day. A dose of colchicine (2 micrograms/g) was administered to each animal 4 h before killing. Samples of duodenum were fixed in 10% (v/v) buffered formalin and processed for assessment of mitotic activity. The number and topographic localization of the colchicine-arrested metaphases were recorded among the entero-cytes within 20 longitudinal sections of the duodenal crypts in each animal. From these data the mitotic indices over the total crypt-cell population as well as within each previously-established zone were determined along with mean +/- SEM for each experimental group. The statistical significance of the differences among the data were analyzed by Student t test. The results show that the presence of ES12a tumor inhibits the mitotic activity of the duodenal-crypt enterocytes and produces an apparent temporal shift in the peak and trough within the circadian curve for this growth parameter.

Animals↗

PubMedDate not supplied

Lectin binding patterns and immunohistochemical antigen detection in the genitalia of Tritrichomonas foetus-infected heifers.

Heifers inoculated intra-vaginally with Tritrichomonas foetus were examined after long-term infection (70 days) and short-term infection (20 days) by lectin-histochemical, immunohistochemical and cultural techniques. The organism was recovered from the genital tract and T. foetus antigens were detected immunohistochemically in the lumina of uterine glands and cytoplasm of vaginal subepithelial macrophages. An increase of galactosylated residues (galactose and N-acetyl galactose), binding to PNA, was observed in the genital epithelium (vagina, uterus and oviduct) from infected animals. In the oviductal epithelium of short- but not long-term infected heifers, mannose (binding to Con A) was detected, suggesting that the persistent presence of T. foetus and its virulence factors or inflammatory processes result in a change in the glycoproteins of the epithelial surface. The findings have implications for the adhesion of T. foetus to cells and for the pathogenesis of bovine trichomonosis.

Animals↗