Don Kristt

Capsular collagen staining of follicular thyroid neoplasms by picrosirius red: role in differential diagnosis.

A key criterion in the diagnosis of thyroid follicular carcinoma is capsular invasion, but invasion cannot always be demonstrated histologically. Since invasion is likely to evoke reactions in the capsular collagen, we examined the effects of invasion on capsular collagen with the picrosirius orange-red (PSR) staining technique for collagen. Under polarized light, the color of PSR-stained collagen varies as a function of the structural and biochemical properties of the collagen fibers. Capsules of widely invasive carcinomas (n = 10), minimally invasive carcinomas (n = 10), and adenomas (n = 28) were stained with the PSR method. Carcinomas were assessed along the thickened capsule for sites of definite invasion, minimal invasion, and no evidence of invasion. In adenomas, sites of thickened capsules (similar to carcinomas) were compared to sites of thin capsules. All foci were evaluated for the color and color intensity of collagen fibers. We found a significantly higher frequency of yellow-green collagen fibers than of orange-red fibers at sites of invasion, whereas orange-red fibers significantly predominated at non-invaded sites. In a minority of cases both colors occurred but the non-dominant color was of lesser intensity in all but 1 case. There were no significant differences in staining between minimally and widely invasive carcinomas. Thick capsules of adenomas consistently stained with an intense orange-red color, although weakly stained yellow-green fibers were also observed in some of these cases. We conclude that PSR staining can provide diagnostically useful information in capsular samples of carcinomas, when both color and color intensity of PSR staining are evaluated at the same site. Specifically, intense yellow-green birefringence of collagen in a thickened capsule is additional evidence for capsular invasion.

Overexpression of cyclin D1 mRNA in colorectal carcinomas and relationship to clinicopathological features : An in situ hybridization analysis

Increased expression of a key cell cycle regulator, cyclin Dl, may have relevance to carcinogenesis and clinicopathological characteristics of some cancers. This study represents the first application ofin situ hybridization, ISH, to detect cyclin Dl mRNA in tissue sections from colorectal carcinomas. This approach was selected because of its unique potential to clarify whether increased expression of cyclin Dl mRNA correlates with clinical and pathological parameters. The ISH of a non-radioactive oligonucleotide probe (Biogenex) was immunocytochemically detected in paraffin embedded sections from biopsy or resection specimens. Tumors ranged from well to poorly differentiated, and from stages A, B, C, and D. Ten year survival data were available on the majority of patients. Intensity of tumor and background (smooth muscle) signals were independently scored from 0 to 3. Overexpressed cyclin Dl mRNA was seen in 86% of cases compared to back-ground. This frequency is similar to that reported for pancreatic carcinoma. The average signal intensity score in tumor foci was 1.9 with a background score of 0.05 (p < 001). All cases showed specific staining judged by the cytoplasmic localization and a tumor signal:background ratio > 1. Expression did not differentiate cancers based on grade, stage or survival (p>l), but did differentiate carcinoma and severe dysplasia from mild dysplasia. We conclude that ISH of cyclin Dl mRNA is an effective and relatively specific means of detecting activity of this gene in colonic neoplasms. The high frequency of overexpression implies that gene activity by itself is not likely to predict a tumor’s biological or clinical behavior. On the other hand, these data suggest that increased cyclin Dl gene activity may be an early event in colorectal carcinogenesis. They also are consistent with findings showing cyclin Dl is inducible by a variety of oncogene products.

Rectal Prolapse: A Possibly Underrecognized Complication of Anorexia Nervosa Amenable to Surgical Correction

Objective: Rectal prolapse is a complication of AN that may be more common than previously recorded experience would suggest. Method: In this report we document, for the first time, the association of anoxia nervosa (AN) and rectal prolapse in a series of three patients seen in the past three years. An extensive review of the literature using Medline over the period from 1966 to Jan 2000 failed to reveal any previous example of this association. Results and Conclusion: The finding could have significant health care implications if confirmed. It would suggest that patients with either the psychiatric or surgical problem may not be receiving the appropriate complementary referrals: psychiatrist to surgeon and vice versa. The importance of recognition of this association in anorectic patients is the availability of effective surgical therapy.

The spectrum of laryngeal neoplasia: the pathologist's view.

Several types of neoplastic change with different prognostic implications typically involve the laryngeal squamous epithelium. The purpose of this review is to examine the spectrum of these changes, as well as their relationship to benign squamous epithelial proliferative states. Since these pathological changes are apt to occur in regions where the epithelial lining is typically squamous, it is important to recognize that the epithelium of the larynx varies from stratified squamous to respiratory-type, depending on the location. The lingual (anterior) surface of the epiglottis is lined by a stratified squamous type, while the laryngeal (posterior) surface is stratified squamous merging into respiratory-type. In the larynx, the supraglottic and infraglottic portions are a respiratory-type, which contrasts with the stratified squamous epithelium of the glottis. This typical distribution does show some degree of variability in those patches of squamous epithelium and is frequently seen within the respiratory-type epithelial regions. The junction between the two epithelial types may be abrupt or separated by a transitional zone.

Differences in histochemical characteristics of gingival collagen after ER:YAG laser periodontal plastic surgery.

OBJECTIVE The aim of this study is to evaluate gingival collagen for the effect of treatment with the Erbium:YAG Kesler handpiece. The handpiece is designed for gingival resurfacing in cases of hypertrophic gingiva and gingival pigmentation. BACKGROUND DATA Lasers represent recent technological advances that afford new options for the treatment of periodontal diseases. However, lasers used for esthetic gingival soft tissue resurfacing require careful histopathological evaluation of the effects on tissue. In particular, it is important to determine the effect of laser irradiation on connective tissue, especially the collagen fibers. To date, no stage-wise clinical or histological studies have been performed addressing this issue. METHODS Ten patients underwent irradiation with the following parameters: energy per pulse, 500 mJ; repetition rate, 10 pps; spot size, 3 mm. Gingival biopsy specimens were derived from 6 patients with hypertrophic gingiva and 4 with gingival pigmentation. The patients were examined before laser treatment and at 7 and 14 days after laser treatment. The tissues were fixed in tymph node revealing solution (LNRS), embedded in paraffin, sectioned at 5 microm, and stained with hematoxylin & eosin. The status of collagen in the treatment site was examined under polarized light after picrosirius red (PSR) staining. PSR is a collagen stain that differentiates collagen fiber density or size by means of a spectrum of color changes under polarized light. The major colors are red, orange, yellow, and green. RESULTS We found a significant difference in the properties of collagen fibers at the first week and at 14 days post-treatment. In the normal gingiva, the predominant polarization colors were in the red-orange range, signifying tightly packed, mature collagen. During the first postoperative week, collagen fibers exhibited polarization colors in the green to green-yellow range, implying loosely packed collagen fibers. After 2 weeks, collagen fibers reacquired their preoperative PSR characteristics. CONCLUSIONS We conclude that sequential series of changes accompany photothermal treatment of the gingiva. The occurrence of this sequence in all successful outcome cases may suggest the importance of these temporally sequenced changes in collagen during gum healing. In any event, the status of PSR staining of gum collagen provides a useful adjunct in the assessment of gingival health.

Successful cell-mediated cytokine-activated immunotherapy for relapsed acute myeloid leukemia after hematopoietic stem cell transplantation

Acute myeloid leukemia (AML) is an extremely aggressive disease with a high relapse rate even after allogeneic hematopoietic stem cell transplantation (HSCT). We report the successful outcome of cell‐mediated cytokine‐activated immunotherapy in a high‐risk pediatric AML patient who relapsed shortly after allogeneic HSCT. Donor lymphocyte infusion along with interferon induced a graft‐versus‐leukemia effect, presenting as a reversible episode of graft‐versus‐host disease, which led to stable complete donor chimerism and total eradication of AML for over 24 months, at the time of this report. The curative potential of immunotherapy in hematological malignancies is discussed. Am. J. Hematol., 2009.

Colonic aberrant crypts may originate from impaired fissioning: relevance to increased risk of neoplasia.

Colonic aberrant crypt foci (ACF) can be identified on the unembedded mucosal surface as clusters of abnormal crypts with enlarged, surface opening. Because dysplasia is frequent, and may be a precursor of carcinoma, epithelial changes have been well studied. However, the basis for the distinctive changes in crypt architecture remain unclear. We hypothesized that some of the architectural alterations of aberrant crypts may reflect impaired fissioning during normal crypt duplication cycles. Fissioning begins at the crypt base. Using morphometric and immunocytochemical approaches, we examined 55 human ACF, both dysplastic and nondysplastic, for their architectural features. Non-ACF mucosa was compared. Microscopically, all lesions contained crypts that were attached, paired, dilated, and angulated. In 3 dimensions, these features related to multiple, individual complexes of connected crypts, referred to as connected crypt structures (CCSs). CCSs terminated in enlarged surface openings (2 to 5 x normal) which are morphometrically equivalent to the macroscopic aberrant crypts (P > .1). These openings trap marker dye. Support for an origin of CCSs in impaired basal fissioning is 3-fold. Crypt profiles in ACF are twice as frequent in basal mucosa as superficially (P < .001); in normal mucosa, the ratio is 1. In a CCS with vertically connected, co-planar crypts, the upper parent crypt diameter was the sum of diameters of inferiorly attached daughter crypts (P > .1). Proliferating cell marker, Ki-67, is not expressed at attachment points. In non-ACF mucosa, isolated CCSs consistently occur at foci of mechanical crypt distortion such as mucosal folds. We conclude that a CCS is a fundamental component of ACF of all histotypes. Impairment of normal crypt fissioning is probably a major factor in the histogenesis of CCSs, which often occurs in settings of mechanical distortion of the mucosa. The pathological significance of this process may be in the formation of enlarged crypt openings. The latter could trap dietary carcinogens as they trap dye, and thereby predispose the CCS to dysplasia.

Patterns of proliferative changes in crypts bordering colonic tumors: zonal histology and cell cycle marker expression.

Proliferative crypt changes have been noted in mucosa bordering colonic carcinomas, but their biological significance is disputed. We anticipated that zonal patterning of histological changes and cell cycle marker expression would provide clues to the pathogenesis of these border changes. 81 specimens were examined including carcinomas, adenomatours polyps, adenomas with early carcinoma, flat adenomas and aberrant crypt foci. The spatial distribution and frequency of micro-architectural features, and mucosal thickness were determined in a border domain of 150-300 sequential crypts/specimen. Immunocytochemical expression of Ki67 and p53 antigens in crypts also was semi-quantitatively examined. We found that in 100% of carcinomas two histologically abnormal zones (Proximate and Middle) separated tumor from normal mucosa. Differences in the feature frequency between zones were statistically significant (p<0.05). Both zones showed mild increases in crypt cell expression of Ki67, with a statistically significant relationship to zonal patterning (p<0.005). Weak expression of p53 only appeared in rare cells. Crypt elongation with mucosal thickening (1.9x normal, p<0.001) in the Proximate and Middle zones distinguished carcinomas from border changes in all benign lesions, except flat adenomas. Since this change occurs in all cases of carcinoma, there is no correlation with tumor stage or grade. Also in carcinomas, elaborate complexes of attached crypts (connected crypt structures) were characteristic of the Middle zone, so that proximate zone was always architecturally simpler. We conclude, that despite continuous carcinoma growth, the invaded border mucosa maintains a prototypical zonal organization of molecular and histological crypt changes. This spatially organized reaction pattern is likely to reflect an interplay between regulated growth and destructive processes in response to advancing carcinoma. Compared to the edges of benign colonic tumors, the edges of carcinomas are distinctive and consistent enough to be diagnostically useful.

Novel Histochemical Stain for Tinctorial Detection of Cancer and Neoplastic Cells

Abstract Zetiq has introduced a histochemical stain that claims to tinctorially identify cancer and neoplastic cells. Because of the potential importance of such a capability, we undertook to investigate Zetiqs CellDetect® staining technology as applied to cultured cell lines as well as an initial sample of clinical cases. This goal was pursued by investigating four types of comparisons: 1) cancer cell lines before and after differentiation; 2) cervical squamous-cell carcinoma (SCC) biopsies to non-neoplastic squamous epithelium; 3) SCCs to neoplastic, nonmalignant squamous epithelium; and 4) neo-plastic squamous cells to non-neoplastic squamous cells in cytological preparations. The clinical material was also stained with hematoxylin and eosin (biopsies) or Pap (cytologies) for diagnostic purposes. We found that all CellDetect®-stained cells exhibited one of the two tinctorial outcomes. Cell lines with malignant phenotype uniformly had red/purple cytoplasm, whereas the differentiated phenotype changed the color to blue/green. Moreover, these two color values are sufficiently distinct that they can be readily distinguished quantitatively with an ELISA reader when applied to cells in tissue culture. Biopsies of SCC and non-neoplastic tissues exhibit the same two color values: cancers had red/purple cytoplasm, whereas non-neoplastic tissues were green/blue stained. In contrast, neoplastic, nonmalignant tissues (dysplasias) stained red/purple, similar to SCCs. Cytological preparations gave similar results: neoplastic cells stained red/purple, whereas non-neoplastic cells exhibited green/blue cytoplasm. The study demonstrated that the staining was rapid and reproducible. Conclusion: The CellDetect® stain allows detecting cancer and neoplastic cells tinctorially based on a rapid, reproducible histochemical process.

Genetic considerations in human sex-mate selection: partners share human leukocyte antigen but not short-tandem-repeat identity markers.

Previous studies support a role for MHC on mating preference, yet it remains unsettled as to whether mating occurs preferentially between individuals sharing human leukocyte antigen (HLA) determinants or not. Investigating sex‐mate preferences in the contemporary Israeli population is of further curiosity being a population with distinct genetic characteristics, where multifaceted cultural considerations influence mate selection.