Intrauterine inflammation at term: a histopathologic study.

This study was undertaken to examine the presence of inflammation of the uterine and gestational tissues as defined by histopathology in clinically noninfected women with term gestation and intact fetal membranes and to evaluate its correlation with measured clinical variables and neonatal and maternal clinical outcome. Two hundred sixteen clinically noninfected term parturients who underwent cesarean section with intact membranes were analyzed for the presence of inflammatory lesions of the gestational tissues and uterus. Nine hundred eighty-one histologic samples were studied, including 212 samples from both chorion membranes and umbilical cords, and 209 placental, 192 myometrial, and 156 decidual samples. In 208 (96%) cases, either amniotic fluid (AF) or endometrial swab samples were cultivated for bacteria. In 148 (69%) cases, the AF leukocyte count was analyzed by the Gram stain method, and in 77 (36%), AF leukocyte esterase activity (LEA) was evaluated. Leukocytic infiltrations were present in samples from 41 (19%) women, varying from 2% to 10% in the different anatomic sites examined. However, after onset of labor, low-grade decidual inflammation was observed in 29% of cases. Cervical dilation (odds ratio, 4.7; 95% confidence interval, 2.4 to 9.3; P < .00003) and meconium-stained AF at the operation (odds ratio, 5.3; 95% confidence interval, 2.2 to 12.5; P < .00015) were associated with the histologic inflammatory lesions observed in decidual samples, independently of AF or endometrial microbial detection, AF leukocytes, or LEA.

Alterations in mast cells showing tryptase and chymase activity in epithelializating and chronic wounds.

Mast cells can be found in contact with epidermis in certain circumstances; especially in chronic inflammatory skin diseases and chronic ulcers, but the significance of this association is obscure. In this study, the association of mast cells with wound healing was studied by counting mast cells in the wound edges at different stages after wounding the donor site skin for pinch-grafting. Chronic venous leg ulcers were biopsed for comparison. Tryptase- and chymase-positive mast cells were stained enzyme-histochemically for active proteinases. Both the number of tryptase-positive, i.e. total mast cells, and chymase-positive mast cells decreased during wound healing, but only the change in chymase-positive mast cells was statistically significant (P< or =0.03) the maximal decrease being 63% on day 7. No mast cells could be found in the vicinity of epithelialization margin. In venous leg ulcers, significantly more mast cells were present in the perilesional skin near the epithelium margin than in the wound bed (P=0.03), and mast cells were also seen in close contact with the basement membrane. Immunoreactivity for IL-4 and TNF-alpha in mast cells was studied to see if either of these molecules was associated with wound healing. In normally healing wounds, only a minority of mast cells were immunoreactive for these cytokines and no change in positive mast cell numbers could be seen during wound healing. In chronic wounds, IL-4 was absent in mast cells, and TNF-alpha positive mast cells were present only in perilesional skin and in small numbers. These results show that mast cells especially chymase-positive - decrease in number and can not be found in the epithelialization zone in normal wound healing, whereas tryptase-positive mast cells are associated with delayed wound healing and epithelialization in chronic wounds. Thus it seems, that mast cells attempt to control hyperproliferation of epidermis in chronic wounds.

Quantitative analysis of tryptase- and chymase-containing mast cells in atopic dermatitis and nummular eczema.

The distribution of mast cells (MCs) containing tryptase (T) and chymase (C) was studied in the non-lesional and lesional skin of 26 patients with atopic dermatitis (AD) and 23 patients with non-atopic nummular eczema (NE), and in the skin of eight healthy controls. T and C activities were demonstrated enzymehistochemically using Z-Gly-Pro-Arg-MNA and Suc-Val-Pro-Phe-MNA as substrates, respectively. The T- and C-containing MCs were counted separately in the epidermis, in contact with the basement membrane, in the papillary dermis and in different dermal levels (0.2 mm each). Also, the C protein was determined immunohistochemically. T-positive MCs were similarly distributed in non-lesional and lesional skin of both AD and NE. The MC number was relatively high in the upper dermis (papillary dermis and levels I and II) of non-lesional and lesional skin of AD. In the upper dermis of non-lesional AD and NE skin and in normal skin, about 50% of T-positive MCs displayed C activity, whereas the percentage in lesional AD and NE skin was only about 30%. In this respect, the non-lesional and lesional samples differed significantly from each other in both dermatoses (in AD p = 0.003; in NE p = 0.002, Students' t-test). In all samples the MC number decreased in the deeper dermal levels, although numerous T-containing MCs were still counted in the deeper dermis (dermal levels IV-VII) of lesional AD and NE skin, differing significantly from the MC number in normal skin (In AD p = 0.005, In NE p = 0.041). In the deeper dermis, the percentage of MCs containing active C was about 70% in non-lesional and lesional AD and NE, and about 90% in normal healthy skin. However, in the upper dermis of non-lesional and lesional skin of both AD and NE, about 80% of all MCs contained the C protein, which differed significantly from the value of 100% in normal skin (p < 0.05). In conclusion, the increased number of T-positive MCs in the upper dermis of non-lesional and lesional AD contributes to promoting inflammation. C apparently loses its activity in the upper dermis of lesional AD and especially in NE. Thus, the enzyme partially lacks its capability to suppress inflammation, such as degradation of neuropeptides and proteins. The dysregulation of these proteinases exists already in non-lesional skin of AD and NE.

Association of psychic stress with clinical severity and symptoms of psoriatic patients.

The association of stress with psoriatic skin and joint symptoms was studied in 38 patients with psoriasis by dividing the patients into low-stress and high-stress groups based on their answers to four questionnaires (General health questionnaire, Somatization questionnaire, Depression questionnaire, and Life change questionnaire), measuring minor psychiatric disorder, psychosomatic reactivity, depression and life changes, respectively. Compared to the low-stress group (n = 21), the patients in the high-stress group (n = 17) had more severe skin and joint symptoms and a higher score, which in established psoriatic General severity score that was calculated by judging Psoriasis Area and Severity Index, activity of psoriasis and the presence of joint symptoms. All the four questionnaires showed higher morbidity frequency and higher score points. Actively spreading psoriasis was significantly associated with stressful life events for men but not for women. However, in the low-stress group, men had experienced significantly fewer stressful events than women. Psoriasis Area and Severity Index showed strong correlation with the Somatization score, but no differences between men and women. The General severity score correlated with the Somatization score, and the higher the General severity score (score > or = 6) the higher were all the four questionnaire scores. Also, patients having joint symptoms had higher scores in all of the four psychic questionnaires than patients without joint symptoms. By the chi-test, female patients with joint symptoms showed a significant correlation with the high General health questionnaire. The presence of joint symptoms showed a tendency for correlation in women with high depression level and in men with high somatization level. This study suggests that psychic stress is associated with exacerbation of psoriasis, and more attention to mental well-being should be paid at least in the case of those psoriatic patients seeking actively for medical care.

Mast cell tryptase and chymase are potential regulators of neurogenic inflammation in psoriatic skin.

BACKGROUND: Tryptase and chymase are proteinases present only in mast cells. In psoriatic lesions, mast cells are increased in number. Certain neuropeptides are also more abundant in lesional than nonlesional psoriatic skin. Based on some earlier results as well as the results of the present study, a hypothesis is presented concerning the regulatory action of mast cell tryptase and chymase on neuropeptides in psoriatic inflammation. METHODS: Forty patients were biopsied, 13 for a mature psoriatic plaque and 9 patients of 27 for a developing (1-3 weeks) psoriatic lesion induced by tape stripping (Koebner reaction). Each lesion had its nonlesional control from the same patient. Mast cell tryptase and chymase, and the neuropeptides Substance P (SP) vasoactive intestinal polypeptide (VIP), and calcitonin-gene-related peptide (CGRP) were stained by enzyme- and immunohistochemical methods. Morphological contacts between mast cells and neuropeptides were visualized using double stains and quantitated in the upper dermis. RESULTS: As the lesion aged, MCTC mast cells displaying tryptase activity increased in number, whereas chymase activity in these cells decreased. All neuropeptides showed some increase along with the development of the lesion, but SP was most abundant in mature lesions. Substance P-positive nerves had also more contacts with mast cells compared to VIP- or CGRP-containing fibers, the contact count being highest in mature lesions. CONCLUSIONS: Tryptase is known to degrade VIP and CGRP, but not SP. Chymase is capable of cleaving both SP and VIP, but is rendered partially inactive in psoriatic skin. These data together with the results of the present study strongly suggest that SP has potency to act as an important mediator in different stages of the psoriatic inflammation.

Screening of effects of selenomethionine-enriched yeast supplementation on various immunological and chemical parameters of skin and blood in psoriatic patients.

Selenium (Se) is known to affect the immune system, and decreased Se-levels in blood of patients with moderate or severe psoriasis have been reported. In this study, the effect of Se-supplementation (400 micrograms/day for 6 weeks as Se-yeast, containing about 70% selenomethionine, SeMet) on skin and blood Se-content, on skin glutathione peroxidase activity and on various chemical and immunological parameters of blood and skin was investigated in 7 psoriatic patients. Before the SeMet-supplementation, serum and blood Se-levels were at the normal range, but they increased 42-45% during the Se-dosage, while zinc levels remained unchanged. Se-dependent glutathione peroxidase activity in both normal and lesional psoriatic skin remained unchanged during the trial, although a small net Se-uptake was detected. At the same time, a slight but statistically significant increase in the number of CD4+ T-cells was observed in the reticular dermis of the psoriatic lesions whereas the numbers of CD8+, CD11c+, and CD1+ cells were not significantly altered. Also, a relatively high number of patients (3 out of 7) showed a strongly reduced number of gamma/delta T-lymphocytes or increased CD8+ T-cells (2 patients) in peripheral blood. However, SeMet-supplementation was not related to these abnormalities or to the number of other peripheral blood immunocytes or to serum immunoglobulin levels. In addition, no marked effect on the clinical condition of the patients was observed. This pilot study suggests that SeMet may be able to modulate the immunological mechanism of psoriatic lesions by increasing the number of CD4+ T-cells.

Immunohistochemical analysis of sensory nerves and neuropeptides, and their contacts with mast cells in developing and mature psoriatic lesions.

The distribution of the neuropeptides substance P (SP), vasoactive intestinal polypeptide (VIP) and calcitonin gene related peptide (CGRP) was studied immunohistochemically in psoriatic skin during the Koebner response (6 h, 2 days, 7 days, 14 days, 21 days), and in mature psoriatic plaques, of 37 psoriatic patients. The morphological association of sensory nerves, SP and VIP with papillary mast cells was also monitored. The nerves containing SP, VIP or CGRP were very scanty in control skin, and in non-lesional and Koebner-negative psoriatic skin. The first psoriatic lesions were seen 7 days after tape stripping the symptomless psoriatic skin. SP- and VIP-containing nerves were slightly increased in Koebner-positive specimens, but the increase was very prominent in dermal papillae of mature psoriatic plaques. In the plaques, nerve-mast cell contacts were significantly increased (p < 0.001) compared with non-lesional psoriatic skin. Only SP-positive fibres were detected in the epidermis and in contact with papillary mast cells. VIP was mainly located around capillaries where SP was also found. No change was noted in CGRP-positive fibres between lesional and non-lesional specimens. The appearance of SP and VIP in the capillary walls is morphological evidence for their function as vasodilators in psoriatic lesion. A slight increase in SP- and VIP-positive fibres in Koebner-positive specimens suggests that these neuropeptides may participate in the inflammatory reaction at an early stage. Their prominence in mature psoriatic plaques in turn indicates a role for them in the maintenance of psoriatic lesions.(ABSTRACT TRUNCATED AT 250 WORDS)

Mast cell tryptase and chymase in developing and mature psoriatic lesions.

The number and distribution of mast cells in non-lesional and lesional skin samples from 13 psoriatic patients were analyzed enzyme- and immunohistochemically. Mast cell tryptase was stained with the sensitive substrate Z-Gly-Pro-Arg-4-methoxy-2-naphthylamide, and chymase with Suc-Val-Pro-Phe-MNA and monoclonal B7 anti-chymase antibody. In addition, healthy-looking skin from 27 psoriatic patients was tape-stripped resulting in induction of the Köbner response in 9 patients. Sequential biopsies were taken before and after (7, 14 and 21 days) tape-stripping, and both tryptase and chymase were stained enzyme-histochemically. In non-lesional psoriatic skin, 70 +/- 24% (mean +/- SD) of the mast cells contained chymase enzyme activity, and 78 +/- 18% chymase immunoreactivity. About 10% of the chymase-immunoreactive cells lacked chymase activity. In lesional psoriatic skin, tryptase-positive cells were increased in number throughout the dermis but especially beneath the epidermis. Chymase immunoreactivity paralleled the tryptase activity, whereas chymase activity was strongly diminished both in terms of mast cell numbers and in staining intensity in the papillary dermis. The apparent inactivation of chymase may be due to the action of the chymase inhibitors, alpha 1-antitrypsin and alpha 1-antichymotrypsin, localized immunohistochemically in mast cells of lesional and non-lesional psoriatic skin. In the developing psoriatic lesion, mast cells displaying chymase activity were already 27-38% decreased in number in the upper dermis on day 7 after tape-stripping, along with the first clinical signs of psoriasis.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantitative analysis of contact sites between mast cells and sensory nerves in cutaneous psoriasis and lichen planus based on a histochemical double staining technique.

The aim of the present study was to test further our previous hypothesis that the inflammatory reaction in psoriasis is neurogenic. For this purpose, contact sites between mast cells and sensory nerves were morphometrically analysed in the basement membrane zone, papillary dermis and three dermal zones of lesional/non-lesional psoriatic and lichen planus skin as well as in healthy control skin. The analyses were made on sections stained with a histochemical double stain developed for this study. With the double stain, active mast cell tryptase was stained blue enzyme histochemically, and the sensory nerves black using specific monoclonal anti-neurofilament antibodies with immunogold. In psoriatic lesions, both mast cells and mast cell--nerve contacts were markedly more frequent in the basement membrane zone and in the papillary dermis when compared with the corresponding areas in the other groups. Mast cell numbers were increased in both lesional and symptom-free skin in lichen planus, but no increase was found in the mast cell--nerve contacts. Increased contacts between mast cells and sensory nerves indicate that the elements exist for neurogenic inflammation in psoriatic lesions. These increased contacts are not due to the extensive inflammatory reaction only, because they were not observed in lichen planus lesions.

Quantitation of tryptase- and chymase-containing mast cells in cutaneous lichen planus.

The distribution and density of tryptase- and chymase-positive mast cells in lesional and non-lesional cutaneous lichen planus (LP) was analysed. For this, enzyme-histochemical staining techniques and morphometrical measurements were applied. In non-lesional LP skin, chymase-positive cells (TC mast cells) showed a distribution similar to that found in both non-lesional psoriatic skin and in normal skin. Tryptase-positive cells (reflecting both T and TC mast cells), however, were increased in number in the upper dermis of non-lesional LP skin. In lesional LP skin, there were fewer chymase-positive cells in the upper dermis, whereas there were more tryptase-positive cells. In the upper dermis, no differences in the number of tryptase containing cells were detected between lesional and nonlesional LP skin. In lesions of LP and psoriasis, tryptase-positive mast cells are increased but differ in their distribution in the papillary dermis. In psoriatic lesions, tryptase-positive cells are frequently observed in epidermal contact, a feature very rarely seen in LP lesions. The present results suggest that the increased numbers of T mast cells in the upper dermis of nonlesional LP skin may be involved in initiating the LP lesion. It seems unlikely that mast cells could be responsible for the epidermal basal cell damage, though T mast cells do participate in the general inflammatory reaction.

Quantitative enzyme-histochemical analysis of tryptase- and chymase-containing mast cells in psoriatic skin.

Tryptase-containing mast cells have recently been found to be increased in the upper dermis of psoriatic lesions. In the present study, the distribution of chymase- and tryptase-containing mast cells was morphometrically analysed at different dermal levels of lesional and non-lesional psoriatic skin (12 patients) as well as normal human skin. Mast cell tryptase was identified enzyme-histochemically, using Z-Gly-Pro-Arg-MNA as the substrate. For demonstrating mast cell chymase, a simple and specific enzyme-histochemical staining method was developed, using Suc-Val-Pro-Phe-MNA as the substrate. All mast cells positive for chymase were also positive for tryptase and Giemsa stain. Although the number of tryptase-positive mast cells was slightly increased throughout the dermis of lesional psoriatic skin, this increase was most pronounced in the upper dermis immediately beneath, and in close contact with, the epidermis. In contrast, the number of chymase-positive mast cells was clearly decreased in the upper dermis of psoriatic lesions, but not in the deeper dermis, as compared with non-lesional psoriatic skin. In addition, all chymase-positive mast cells observed in the upper dermis were very weakly stained when compared with those in the deeper dermis. No differences were found between non-lesional psoriatic skin and normal skin in which the number of mast cells containing chymase was 72-73% of the number containing tryptase. The present results suggest that T mast cells particularly, containing tryptase but no chymase, proliferate in psoriatic lesions, and that the increase in tryptase activity and the decrease in chymase activity in the upper dermis may lead to an imbalance in the biochemical regulatory systems.

Cellularity of the ovarian stroma in patients with endometrial carcinoma.

Cellularity of the ovarian stroma was determined morphometrically in 124 patients with endometrial carcinoma and was correlated with histologic and clinical features of the neoplasm. High cellularity of the ovarian stroma was found to be significantly associated with poor histologic differentiation of the tumor. There was a correlation between patient age and the degree of differentiation and depth of myometrial invasion of the tumor, whereas age did not correlate with clinical stage of the carcinoma or with cellularity of the ovary. Since the ovarian stroma produces androgens, which after peripheral conversion may exhibit estrogenic activity, hyperplasia of the ovarian stroma may contribute not only to the development of well-differentiated endometrial carcinomas, but also to that of poorly differentiated carcinomas. So far, the latter have been considered mostly unrelated to estrogenic influence.

The prognostic value of quantitative analysis in the endometrial carcinoma.

The epithelial and stromal components in 146 adenocarcinomas, adenocanthomas and adenosquamous carcinomas of the endometrium were quantified morphometrically on Feulgen-stained tissue sections. The classification of the tumors was based on the criteria of World Health Organization and International Federation of Gynecology and Obstetrics. As expected, the survival percentage of the patients fell with the advancing clinical stage and histological grade. The ratio of the epithelial cells to the stromal ones proved to be a good prognostic indicator and may be useful in identification of cases with the most favorable prognosis among grade 1 tumors. A high mitotic activity predicted poor prognosis to the patient although statistical significance was not achieved.

Mucosubstances in classification of serous and mucinous ovarian tumors: a morphometrical study.

108 serous and 106 mucinous ovarian cystadenomas of varying malignancy were studied with the saliva-periodic acid-Schiff (SPAS) reaction and with the Alcian blue (AB) pH 2.7 method, which stain neutral and acid mucosubstances, respectively. Positively stained mucin was quantitated morphometrically and the measured parameters were tested by discriminant analysis. As expected, the mucinous tumors were more positive for mucin than the serous ones. Also the borderline tumors contained more mucin than their benign or malignant counterparts. The best discriminating capacity for mucin was obtained in serous tumors with the AB stain and in mucinous tumors with the SPAS reaction. The ratio of neutral to acid mucosubstances decreased in mucinous cystadenomas with increasing malignancy.

Periodic acid-Schiff stain as a prognostic indicator in serous and mucinous ovarian tumors.

The prognostic significance of periodic acid-Schiff (PAS) stain in 112 serous: 43 benign, 25 borderline and 44 malignant cystadenomas: and in 106 mucinous: 60 benign, 32 borderline and 14 malignant cystadenomas of the ovary were investigated. The amount of positively stained mucin was estimated morphometrically. The outcome of most patients with benign or borderline lesion was good. One patient with benign mucinous cystadenoma died, however, of pseudomyxoma peritonei and another patient with borderline mucinous cystadenoma died of peritoneal carcinosis. Other patients were alive and free of the disease after a follow-up of 1-14 years, or had died of causes unrelated to the ovarian disease. Abundant PAS positive mucin predicted a longer survival both in serous and in mucinous malignant tumors. The 5-year survivals for the serous cystadenocarcinomas with and without PAS positive mucin were 21% and 13%, respectively (not statistically significant). For mucinous cystadenocarcinomas with mucin value over and below the median, the 5-year survival rates were 57% and 14%, respectively (P less than 0.10). High PAS positivity in both serous and mucinous cystadenocarcinomas clearly indicated better prognosis, although statistical significance was not achieved. Thus, further studies are needed for final evaluation of the prognostic significance of the PAS stain in these ovarian tumors.

Evaluation of carcinoembryonic antigen and periodic acid-Schiff stains in the diagnosis of serous and mucinous ovarian tumors: morphometrical study.

108 serous and 104 mucinous neoplasms of the ovary, classified according to the criteria of the World Health Organization, were studied with periodic acid-Schiff (PAS) method for mucin and glycogen, and with immunoperoxidase method for carcinoembryonic antigen (CEA). The amount of epithelium, the number of mitoses, and the staining positivities were quantitated morphometrically and the parameters were evaluated using the discriminant analysis. In general, CEA scores were higher in mucinous than in serous tumors and higher in malignant tumors than in their benign or borderline counterparts. High PAS positivities were found both in serous and mucinous borderline lesions. Mitotic activity correlated with the malignancy of the tumors. The best discriminator, however, was the amount of epithelium. The results show that simple morphometry with standard stains is an efficient method in the first-line classification of these tumors. The quantitation of CEA and PAS stains may be helpful in cases in which histological criteria do not allow definite classification.

Histopathological and morphometrical analysis applied to skin changes in NMRI mice induced by dithranol (anthranil) and its acyl analogs.

Histology and morphometric changes in skin lesions of NMRI mice caused by dithranol and its acyl analogs, with and without pretreatment with dimethylbenzanthracene (DMBA) are described. The course of neoplastic progression seems to have three distinct phases (normal skin, hyperplasia, papilloma), each characterized by typical architectural and cellular changes. The parameters reflecting the nuclear size increased in the above order and the differences between the groups were highly significant. Nuclear shape changes did not take place, as shown by the almost constant form factor. The study demonstrates the value of the morphometric methodology in characterizing quantitative nuclear changes in mouse skin models.