Biomedical subjects
Publications and source records attributed to C A Rubio.
A baboon died of chronic, intractable diarrhoea. At autopsy gross examination revealed oedema, and signs of chronic inflammation in the entire colon. At histology, marked lymphocytic infiltration within the colonic epithelium covering the crypts was found. The term cryptal lymphocytic colitis is proposed for this previously unreported disease in baboons.
The histologic phenotype of the dysplastic lesion juxtaposing colorectal carcinomas was assessed in 100 consecutive colectomy specimens: in 50 patients with inflammatory bowel disease (IBD) and in 50 controls (non-IBD patients). Adenomatous growths (AG) were regarded both Dysplasia Associated Lesion or Mass (DALM) and sporadic adenomas. AGs juxtaposing carcinomas were found in 76% (n = 38) of the IBD cases: 52.3% (20 out of 38) were villous, 28.9% (11 out of 38) serrated, 5.3% (2 out of 38) tubular and the remaining 13.2% (5 out of 38) were mixed AGs. Juxtaposing AGs (sporadic adenomas) were also found in 58% (n = 29) of the control cases: 51.7% (15 out of 29) were villous, 6.9% (2 out of 29) tubular, 3.4% (1 out of 9) serrated and the remaining 37.9% (11 out of 29) were mixed. The majority or 81.2% (31 out of 38) of the dysplastic lesions juxtaposing IBD carcinomas were villous or serrated AGs, but only 55.1% (16 out of 29) in control cases. Serrated AGs in particular accounted for nearly 29% of the non-invasive dysplastic lesions abutting IBD carcinomas but only for 3% in control specimens. It would appear that villous and serrated AGs are the most common non-invasive neoplastic lesions from which IBD carcinomas originate.
In recent years, a substantial number of baboons died at the Southwest Foundation for Biomedical Research following protracted intractable diarrheas. The histopathologic diagnosis at autopsy was chronic colitis. The diarrhea could last for more than one year and occurred in infants, juveniles or young adult baboons, i.e. < or = 8 years of age (a healthy baboon may live up to 27 years). The aim was to assess the histopathologic subtype of chronic colitis in a relatively large number of colonic specimens obtained at autopsy (n = 132). In 88 out of 132 baboons with chronic diarrhea, the colonic mucosa was well preserved for histological examination. At review, various histological phenotypes of chronic colitis were disclosed in 86 of the 88 baboons. Chronic lymphocytic-plasmacytic colitis was found in 54.6% (47 out of 86), chronic ulcerative colitis in 15.1% (13 out of 86), Crohn's colitis in 12.8% (11 out of 86), superficial lymphocytic colitis in 10.5% (9 out of 86), cryptal lymphocytic colitis in 5.8% (5 out of 86) and collagenous colitis in 1.2% (1 out of 86). Chronic ulcerative colitis, Crohn's colitis, superficial lymphocytic colitis and collagenous colitis in baboons closely mimic corresponding histological phenotypes of chronic colitis in humans. Recently, cryptal lymphocytic colitis has also been found in humans. The awareness that chronic colitis in baboons is not one disease but a series oi chronic inflammatory changes, having common clinical symptoms and similar gross appearance, may lead to the correct diagnosis of the subtype of the disease. Only then would it be feasible to systematically initiate the search for the corresponding etiologic agent(s), aiming to tailor specific therapeutic strategies in those animals.
A Macaca fascicularis died following chronic, intractable diarrheas. At autopsy, the entire colon was macroscopically red and swollen. At histology, the most conspicuous histologic finding was a marked accumulation of intraepithelial apoptotic granules in the lower one third of the epithelium of the colonic crypts. The cells in those crypts showed various degrees of swelling or necrobiosis. Since no other explanation was found at autopsy, it was concluded that the marked apoptosis with marked epithelial cell destruction was the cause of the colonic disease which apparently resulted in untreatable protracted diarrheas in this animal. The term apoptotic colonic disease is proposed for this previously unreported condition.
Population studies in the Pacific Basin showed that gastric carcinomas of intestinal type often concur with distant mucosal changes (DMCs). In the present work, the presence of DMCs was investigated in populations dwelling in the Atlantic Basin. A total of 1737 gastrectomy specimens were reviewed: 627 in New York, 435 in Reykjavik, 198 in Buenos Aires, 186 in Florence, 174 in London and the remaining 117 in Stockholm. A total of 17,282 sections were carefully scrutinized. The following DMCs were investigated: intramucosal glandular cysts, gastric cells with ciliated metaplasia, with large or small mucus negative vacuoles, and extensive intestinal metaplasia (IM). The highest frequencies of DMCs were found in Florence for specimens with intestinal type carcinoma: 41.3% had intramucosal cysts, 22.4% had cells with ciliated metaplasia, 12.9% cells with large vacuoles, and 50.9% had high IM. The highest frequency of gastric cells with small vacuoles was recorded in New York (9.1%), also in specimens with intestinal type carcinoma. Significantly lower DMCs percentages were found in specimens with carcinomas of diffuse type, and miscellaneous gastric diseases. The occurrence of DMCs was not influenced to a significant degree by the number of sections available per gastrectomy. Since environmental factors trigger the evolution of intestinal type carcinomas and as DMCs also occurred in specimens without carcinoma-although at a significantly lower rate--it is conceivable that DMCs are also evoked by environmental factors (before a gastric carcinoma ensues). DMCs were found in specimens having intestinal carcinomas either in the cardia, the corpus or the antrum. Thus, DMCs seem to provide the adequate "soil" for the development of gastric carcinomas of intestinal type, independently of the future localization of that tumor in the stomach.
Clinical reports indicate that colonic neoplasias may be overlooked at colonoscopy. We investigated, in an experimental model, whether both gross and high power microscopical examinations could detect all induced colonic neoplasias. Forty-nine Sprague-Dawley rats received 1,2-dimethylhydrazine for 27 weeks. The colons were scrutinized grossly and at high-power microscopy complemented with chromoscopy. Biopsies were obtained from all mucosal alterations seen at gross and at high power microscopy. The gross examination of the colonic specimens in the 49 animals revealed 196 polyps or suspected polyps. Histology showed that 85.2% (167 out of 196) were adenomas or adenocarcinomas. The remnant polyp-free colons were subsequently examined by high-power stereo microscopy (46 magnifications). Fifty-five new lesions were detected and removed. Histology showed 48.9% (n = 24) additional neoplastic lesions in the 49 colon specimens. The remnant polyp-free colons were subsequently processed for histology. The sections showed 57 additional neoplastic lesions in the 49 colon specimens: 93.0% (n = 53) were adenomas (n = 35) and crypt dysplasias (n = 18), while the remaining 7.0% (n = 4) were invasive carcinomas. Experimentally-induced colonic neoplasias may be overlooked even after a careful scrutiny of the colonic mucosa using high-power optical devices.
BACKGROUND: Previous studies showed that the DNA synthesis (DS) in the esophagus of rats significantly increased at 4 weeks of stress, but decreased to normal values at 8 weeks. The possibility that this DS adaptation (DSA) could be maintained beyond 8 weeks was explored. MATERIALS AND METHODS: Sixty-five rats were investigated: 60 were transported to the stress laboratory (30 were water-plunged and 30 were controls) for 8 weeks. After 8 weeks, groups of stressed and control animals (n = 5) were allowed to rest one, two, three and four weeks, ending with single water-plunging or sham-handling. All animals (including 5 untreated) received i.p. injection of 3H-thymidine before sacrifice. The ratio radioactive DNA/total DNA reflected the DS of the esophagus. The isotope mainly labeled mucosal cells. RESULTS: DS significantly increased at 10 and 11 weeks (p < 0.05), but decreased below 8 week values at 12 weeks. CONCLUSION: DSA values at 8 stress weeks did not prevail since DS substantially increased in stressed rats at 10 and 11 weeks. At 12 weeks, DS re-adaptation (DSRA) occurred. Importantly, it took 8 weeks to achieve DSA, but only 4 weeks to accomplish DSRA. Control animals also showed DS fluctuations, but to a lower degree. The model described may prove of value in studies aimed to abate the disparate fluctuations of DNA synthesis in the esophageal mucosa during the various phases of protracted stress.
A total of 31 cases with Ulcerative Colitis (UC) and colorectal carcinoma were retrieved from the files of the Karolinska Hospital, Stockholm between 1951 and 1998. Sections from 16 colectomy specimens (operable cases) and 15 biopsies obtained at laparotomy (inoperable cases), were available for the study. Of the 31 patients reported here, 22 (71%) were 49 years of age or younger at the time of surgery for carcinoma. In comparison only 47 (5.5 %) of the 855 colorectal carcinomas without UC reported in the Stockholm area in 1990 were 49years of age oryounger. When this hospital was a referral Center (1951 through 1969) 18 cases of carcinoma in UC were operated between 1951 and 1960 (1.8 patients/year), but only 4 between 1961 and 1969 (0.44 patients/year). During the surveillance period of 29 years (1970 to March 1998) only 9 patients (0.31 cases/year) were found to have carcinoma complicating UC. Notably, 8 of the 9 patients were operated on between 1970 and December 1989 (0.42 patients/year), but only one case between January 1990 and March 1998 (0.11 patients/year). The data presented indicate that the frequency of carcinoma cases in pancolitics has decreased at this hospital, not only during the referral period, from 1.8 patients/year during the 50's to 0.40 patients/year during the 60's, but also during the surveillance period (from 0.44 patients/year/during the 70's and 80's to 0.11 patients/year between 1990 and March 1998). This, despite the incidence of UC in the Stockholm County remained stable for the past 40 years (4.2 to 5 patients/10(5) inhabitants) and that the population in the Stockholm County has steady increased since 1950. A review of the present literature indicated that the ris for colorectal carcinoma in pancolitics is presently decreasing, not only in Sweden but also in other Scandinavian countries.
Sixty-seven gastric adenomas found at gastrectomy in two ethnic groups (53 in Japanese and 14 in Icelandic patients) were investigated. Micrometric measurements were carried out on tissue sections. Adenomas being not higher than twice the measured height of the non-dysplastic adjacent gastric mucosa were regarded as non-protruding (n = 40) and those surpassing that limit as protruding (n=27). Measurements showed that 59.7% (n=40) of the 67 adenomas were non-protruding. The mean width in protruding adenomas (11.6 mm) surpassed the mean width of non- protruding adenomas (7.1 mm), suggesting that non-protruding adenomas may evolve into protruding forms. On the other hand, 13.6% (3 out of 22) of the protruding adenomas in Japanese patients were very small (< or = 5mm in width), implying that some small gastric adenomas grow from the outset in a protruding fashion. Conversely, 22.5% (n =9) of the 40 non-protruding adenomas measured 10 to 25 mm in width, suggesting that a proportion of the larger gastric adenomas may grow in a non-protruding, horizontal fashion without "becoming" protruding. Thus, similarly to colorectal adenomas, gastric adenomas may be classified into protruding and non-protruding. Micrometric measurements permit the correct assessment of the type of adenoma.
The nm23 gene is thought to play a role as an inhibitor of metastatic progression in several human cancers and its down-regulation has been associated with increased metastasis and reduced survival in some studies, though not in others. To better investigate the role of nm23 in gastric cancer (GC), the expression and prognostic impact of this gene was examined in 107 radically operated GC patients in a high risk area. The expression of nm23 was determined immunohistochemically by using the rabbit antibody anti-human nm23 protein. The expression of nm23 was detected in 40.2% (n = 43) of 107 gastric tumours and correlated with a poorer clinical outcome. In a survival analysis at 5 years, patients with nm23-positive tumours had significantly worse prognosis than patients (n = 64) with nm23-negative tumours (p < 0.05). The prognostic significance of nm23 expression was confirmed by multivariate analysis including terms for tumour stage and lymph node involvement. Our results suggest that the expression of the nm23 gene in gastric carcinoma is significantly related to tumour progression and poor prognosis at 5 years.
Muciphages are mucin-rich phagocytes believed to evolve as a result of the disruption of colorectal crypts. In a previous work we found, in rectal biopsies from patients with chronic ulcerative colitis, muciphages having not only mucin but also lysozyme, an enzyme with a potent antimicrobial activity. Recently we detected lysozyme-rich muciphages in the normal mucosa of the stalk of colonic adenomas. Filed hematoxylin and eosin (H&E)-stained sections from 30 consecutive colorectal adenomas with a stalk (lined by normal colorectal mucosa) were stained with PAS (for mucopolysaccharides), with CD68 (to label macrophages) and with lysozyme (Muramidase). Of the 30 adenomas, 16 (40%) showed muciphages in the mucosa of the stalk. Those muciphages were PAS- CD68- and lysozyme-positive. Although the significance of these findings remains elusive, it is conceivable that lysozyme-rich muciphages mirror increased cell destruction in colorectal adenomas with a high cell turnover. The possibility that lysozyme-rich muciphages surrounding adenomas are instrumental in a novel molecular mechanism of host defense, effective at the early stages of colorectal carcinogenesis, was also entertained. Such a mechanism would prevent the lateral expansion of the dysplastic epithelium of the adenoma into the surrounding normal mucosa of the stalk.
The normal basement membrane (BM) of the colon and the collagenous colitis band (CCB) seen in H&E-stained sections were recently found to be autofluorescent. Measurements of the thickness of the BM and of the CCB were taken using transmitted-light (TL) and incident-light fluorescence (IFL). The mean thickness of the BM in control cases was 3.2 microns with TL and 3.4 microns with IFL. The mean thickness of the CCB was 50.3 microns with TL and 55.8 microns with IFL. In 10 consecutive cases of CC, the thickness of the CCB was repeatedly measured in the same area, on different occasions. Substantial variations in the vertical size of the CCB were recorded in the same area when using TL but not with IFL. Good demarcation of the CCB with ILF and poor demarcation of the deep border of the hyaline band, because of overlapping inflammatory cells, were the apparent causes. Autofluorescence was induced by the eosin stain. The results suggest that the simple observation of H&E--stained biopsies with ILF may be sufficient to diagnose CC. Re-sectioning for special staining can be avoided, thus reducing final diagnosis-time and laboratory costs.
In recent years, a substantial number of macaques have died at the Southwest Foundation for Biomedical Research, USA, following protracted intractable diarrhea. The diarrhea could last for up to two years and occurred in infant, juvenile or young adult animals. The histopathological diagnosis at autopsy was chronic colitis. The histopathological subtype of chronic colitis is here assessed in a relatively large number of colonic specimens obtained at autopsy (n = 90). The colonic mucosa was well preserved for histological examination in 83 of the 90 macaques. At review, various histological phenotypes of chronic colitis were found. Chronic lymphocytic-plasmacytic colitis occurred in 78.3% (65 out of 83), chronic ulcerative colitis in 20.5% (17 out of 83) and Crohn's colitis in 1.2% (1 out of 83). In macaques, some histological phenotypes of chronic colitis (ulcerative and Crohn's colitis) closely mimic chronic colitis in humans. The awareness that chronic colitis in macaques is not one disease but a series of chronic inflammatory changes, with common clinical symptoms and similar gross appearances, may lead to the correct histological diagnosis of the subtype of the disease.
A patient with intractable diarrhea showed, in 6 out of 7 biopsies taken at various colonic levels, a rich number of apoptotic granules at the base of the colonic crypts. Those apoptotic granules stained positive with Feulgen, a reaction that stains nuclear DNA. The nuclei of the cells at the base of the crypts stained positive with TUNEL, a method that labels nuclei undergoing apoptosis. No associated inflammatory reaction was present in the biopsies. As no other explanation was found for the phenomenon, the possibility that marked cryptal apoptosis was the cause of the untreatable protracted diarrhea was entertained. A similar histological finding had previously been recorded in a non-human primate dying of intractable diarrhea. That phenomenon was designated apoptotic colonic disease. We propose a similar denomination for this previously unreported finding in a human subject.
Using hematoxylin and eosin (H&E)-stained sections, Paneth cells are identified by the presence of eosinophilic granules. Several other methods have been proposed to stain Paneth cells. Recently, we noticed that Paneth cells were autofluorescent in H&E sections. To assess the number of Paneth cells by this method, 30 surgical specimens from the small bowel and 10 from the colon with IBD were investigated. Consecutive sections were stained with H&E and immunostained with lysozyme (Lz). Paneth cell counting was done in 20 well-oriented, vertically cut crypts per-case using alternatively transmitted light (TL) and incident-light fluorescence (ILF). When H&E and Lz sections were observed with TL, the number of Paneth cells was easily assessed in some crypts, but in crypts having numerous Paneth cells, their exact number was ascertainable. On the other hand, using IFL, the number of H&E-stained Paneth cells-crypt was easily assessed, even in crypts with Paneth cell clustering. Using H&E-ILF, a mean of 3.9 (range 3-8) Paneth cells-pe-crypt was found in the normal small bowel and a mean of 3.3 (range 1-14) in the colon with IBD. ILF provided the adequate visual conditions--i.e. a dark, non-autofluorescent background--for assessing the actual number of normal and metaplastic Paneth cells, even when arranged in tight cell groups. The present method will improve the enumeration of the actual number of normal and metaplastic Paneth cells both in experimental research and in clinical trials.
Sections from 130 DMH-induced carcinomas in rats were reviewed. In 78% (74 out of 95) of the gland-forming adenocarcinomas, neoplastic glands displayed at the growing edge either a thinner layer or a partial loss of tumor cells ("pore" formation). The thinner epithelium appears to be a stage preceding the focal "destruction" of the tumor cells ("pore" formation). Through those glandular "pores" mucins, inflammatory cells or necrotic elements were being released free into the surrounding host tissues. It is theorized that those non-neoplastic "pseudopodial" forerunners (rich in proteolytic enzymes) encourage the degradation of the surrounding host tissues. The malignant cells surrounding glandular "pores" eventually grow around the secreted products to restore the continuity of the gland. The remodelling of the glands will guarantee a stepwise but everlasting tumor progression deeper into the host.
Much research has centred around the histological characteristics of invasion in colorectal tumors, but the possible mechanisms involved in local tumor progression in individual cases have not been unveiled. We studied large histological sections from 10 consecutive patients operated for a colonic (n = 5) or a rectal (n = 5) carcinoma. Particular attention was paid to the outmost growing tumor-host edge (GT-HE). Results showed that the avant-garde tumor glands (at the GT-HE) had either thinner tumoral epithelium (n = 2), or "pores" (i.e. lacking tumor cells in the deeper portion of the gland), containing either mucin (n = 5), neutrophils or necrotic material (n = 3). The thinner epithelium appears to be a stage preceding the "destruction" of the tumor cells (i.e. glands with "pores"). The progression of colorectal adenocarcinomas at the GT-HE interphase would proceed by the release (free into the host tissues) of mucins, inflammatory cells or necrotic elements through those "pores". The non-neoplastic elements may be "pseudopodial" forerunners (rich in proteolytic enzymes) which permit the local degradation of the tissues of the host. The malignant cells surrounding those glandular "pores" would eventually grow around the secreted products, thus guaranteeing a stepwise but everlasting tumor progression in untreated cases.
Colorectal adenomas are usually classified into two categories: tubular and villous adenomas. A mixed (tubulovillous) phenotype is also recognized. Two other histological phenotypes, namely serrated and microtubular adenomas, have been reported in the literature but neither have been included in large series. In this survey, all colorectal adenomas seen at this Department between 1993 and 2000 (n = 3135) were classified into the above 5 histological categories. The results showed that 66% (n = 2074) were tubular, 9% (n = 281) villous, 18% (n = 551) mixed, 6% (n = 202) serrated and the remaining 1% (n = 27) microtubular. More than 80% of the tubular, villous, serrated and microtubular adenomas were located in the colon. None of the microtubular adenomas were found in the rectum. The molecular signals whereby the colorectal mucosa in some patients "etch" serrated or microtubular neoplastic structures whereas in others tubular or villous neoplastic structures, have not been explored. One possible explanation might be that the pathway followed by the cascade of committed molecular events is less "accessible" for the "etching" of serrated and microtubular structures or that the process is less complex for the molecular design of tubular and villous configurations.