[Why are diabetic patients treated at rheumatological hospitals?].

Retrospective analysis included 316 case histories of diabetic patients treated at the Silesian Rheumatology Hospital in 1987-1988. An analysis included causes of disorders, calcium-phosphorus metabolism disturbances, lipid and purine disorders. Statistical parameters were compared with the type of diabetes mellitus, duration of the disease, sex, age and obesity. There were 10% of inflammatory rheumatic disorders (6.4% rheumatic arthritis, 1.7% of rheumatoid spondylosis and 2% of other disorders) in the analysed case histories, and 32% of degenerative disorders (19% of vertebral column joints and 12.7% of other joints). Degenerative disorders were noted more frequently in patients with diabetes mellitus type 2, treated with insulin, while spondylopathies were particularly frequent in female patients of this group. Biochemical disorders in the form of hypocalcemia and hypophosphatemia, hypertriglyceridemia, hyperuricemia, signs of lesions to the liver and kidneys were more increasing with the duration of the disease and the degree of insulin-dependence. Locomotive system disorders are not related only to primary articular lesions. They depend also on diabetic neuro-vascular complications and osteopenia.


[Barrett esophagus diagnosed after appendectomy in a patient with diabetes mellitus].

A pathogenetic outline of oesophageal inflammatory lesions with special emphasis on Barrett's oesophagitis has been presented. As recorded in a history of a 65-year old patient at first the stenocardia-mimicking pains occurred a few days after appendectomy. Gastrobulboscopy performed (after coronary insufficiency had been excluded) revealed extensive inflammatory and erosive lesions of the oesophagus and multiple ulcerations of the duodenum. According to the several authors the rapid manifestation of the inflammatory lesions of the oesophagus may be related to the intubation necessary for laparotomy. It may refer particularly to diabetic patients. Control endoscopies showed that a six-week treatment resulted in a total disappearance of mucosal changes. It might be thoughtful to take into consideration a possibility of oesophageal inflammation in clinical differentiation of stenocardia mimicking pains in diabetics who underwent the intubation.


[Adaptation reaction to the health resort treatment in patients with diabetes mellitus].

Physical, clinical, biochemical, and functional examinations have been carried out in 70% of patients with diabetes mellitus type 2 treated in health resorts. Patients were treated with diet, drugs, and physiotherapy during a 3-5-week period. It was found, that the whole period of the treatment may be divided into 3 subperiods: period I is characterized by the subjective symptoms produced by the process of adaptation to different environment. This period lasts for 4 days, on average. Periods II and III are characterized by the set of symptoms reflecting a sum of therapeutical physical stimuli. Such symptoms are known in the literature as bath- or spa-reaction. Symptoms of spa-reaction were noted in about 50% of the examined patients. In 21% of them late reaction, i.e. after about 3 weeks, was seen. No worsening of diabetes mellitus was observed during spa-reaction. No correlation between the onset of this reaction and duration of the disease as well as the results of therapy was found. However, spa-reaction was more frequent in the elderly. Period III was followed by the decrease of the symptoms of spa-reaction with marked improvement in glycaemia. Authors' observations lead to some practical conclusions concerning the programs of physiotherapy in dependence on the course of patients' adaptation to the spa. Regulations concerning the routine 24-day treatment period of all patients in health resorts should be revised.


[Relative effectiveness of tolbutamide, chlorpropamide and gliclazide].

In clinical practice of hypoglycemic therapy of diabetes mellitus the problem of the optimal selection of oral hypoglycemic agent, corresponding to the individual patterns of regulatory and metabolic disturbances is of primary importance. The individual, pathophysiological basis should be met as much as possible by the pharmacodynamic properties of the selected, hypoglycemic drug. For this reason group of 23 diabetics type II underwent a prolonged, open trial of controlled pharmacotherapy with 4 sulphonylurea derivatives. Pertinent clinical and metabolic parameters were assessed before, during and after planned periods of therapy with Tolbutamide or Chlorpropamide to Gliclazide and Gliclazide to Glibenclamide. In the same time the levels of serum insulin fasting and after breakfast were determined. Also the comparative efficacy of the exchange of the drug in a subgroup of diabetics with fasting glycemia below and above 160 mg% was assessed. It was shown, that the change of Tolbutamide or Chlorpropamide to Gliclazide or Glibenclamide improved the therapeutical effectiveness in general. The individual responses to such a change were however individually differentiated. The change of Tolbutamide or Chlorpropamide to Gliclazide increased the therapeutical efficacy only in these patients, in whom such a change was associated with an increase of prandial, reactive serum insulin level (IRI). In practice they were patients with the fasting glycemia lower than 160 mg%. In patients with fasting glycemia higher than 160 mg% the change of oral compounds under study was not connected with an increase of prandial serum insulin. The metabolic parameters have not improved either. Perhaps they were patients with diabetes mellitus type II, who should be primarily qualified to insulin.


[Serum N-acetyl-beta-D-glucosaminidase (NAG) activity in patients with diabetes mellitus with reference to diabetic microangiopathy].

In 110 diabetics type I and type II and in 30 control subjects the total serum activity of NAG as well as of its isoenzymatic forms A and B was determined. In cases with diabetic nephropathy the GFR and serum creatinine was also analysed. It was found that NAG activity is correlated to the degree of clinical symptoms of diabetic vascular changes. Therefore it could be suggested that NAG plays a role in development of microangiopathy. NAG determination may also serve as an index differentiating the diabetic microangiopathy from other forms of microvessels.


[Evaluation of surgical results in lower limb atherosclerotic ischemia comparing men and women].

Multicenter studies involved 1449 male and 131 female patients operated for the atherosclerotic ischemia of the lower limbs. About 50% of all limbs were in the III or IV stage of ischemia (according to Fontaine's classification) before surgery. Atherosclerotic lesions involved mainly aortoiliac segment in the majority of patients. Early results of surgery were similar in both men and women. An improvement during a 5-year follow up period was noted more frequently in women than in men--86% and 68.5% of limbs respectively. Late result of lumbar sympathectomy was similar in both groups. Limb amputation and mortality rates were similar in both groups in the early postoperative period and during a 5-year follow up as well.


[Evaluation of the results of the treatment of children with insulin-dependent diabetes mellitus fully or partially trained in self care].

The study was aimed at assessing the results of diabetes mellitus therapy in 162 children, who underwent 1) the full programme of diabetes education (114 children) and 2) only the part of it (48 children). The first group was under care of Cracow Medical School Pediatrics Institute since the disease and had 2 weeks of formal diabetes mellitus teaching. The second group was treated in a less unified way in different hospitals and did not have the chance of regular teaching programme. The degree of metabolic control was assessed by series of Hb A1c determinations in whole 1987 year. The levels of Hb A1c were also compared with the children age, diabetes mellitus duration and the period elapsing from the time of last teaching. Children who were fully educated in the problems of their disease achieved significantly better results. In all group a deterioration of diabetes control during puberty was however observed. In children not fully educated such a deterioration was also noted in correlation with diabetes mellitus duration.


[Comparison of early and delayed results of unilateral patency restoration and transplantation in aortoiliac obstruction].

Early and delayed results of the treatment of aortoiliac unilateral incompetence in 144 patients were compared. Eighty seven (60%) patients were treated with aortoiliac prosthesis whereas 57 (40%) with arterial patency restoration. Fifty seven patients of group I reported to the control examination. Favourable effect was found in 47 (82.5%) patients, i.e. in 54% of all operated patients. Thirty six patients of group II reported to the control examination. Favourable effect was noted in 27 (75%) patients, i.e. in 47.4% of all treated patients. Twenty two (25.3%) of patients of group I and 9 (15.8%) of group II died. It was noted also that in 20% of operated patients with unilateral aortoiliac incompetence the disorders in the opposite limb develop despite the type of surgery. Therefore, indications to the implantation of bifurcated prostheses should be extended in some cases.


[Delayed results after implantation of bifurcated prosthesis in aorto-iliac incompetence].

Early results and those seen after a 5-year follow up are discussed. Three hundred fifty one patients with aortoiliac incompetence were treated with the implantation of bifurcated aortoiliac prosthesis. Very favourable early result was achieved in 284 (81%) patients. Very favourable delayed effect was noted in 133 patients, i.e. in 75% of patients who reported for the control examination. In the follow up period, amputation of the limb was performed in 30 (8.5%) patients because of progressing ischemia. Hundred thirty seven (39%) followed up patients died during 5 years. Causes of so high mortality rate are discussed in view of risk factors and postoperative complications.


[Assessment of late results in surgical treatment of lower limb ischemia in obliterative atheromatosis of the femoral-popliteal region].

The results of treatment of arterial incompetence localized in femoro-popliteal region in the selected 14 centres have been assessed after 24 and 60 months. Eight hundred fifty five patients were operated in the selected centres during 12 months i.e. 50% of all patients operated for obliterative atheromatosis of the lower limbs. Perioperative mortality rate was 4%. During a 5-year follow up, 30% of patients died, and 15% did not return supplied questionnaire, and their fate is unknown. The limb was preserved in 70% of patients operated with any technique except the primary amputation. Markedly worse results were noted in patients operated for necrosis or resting pain and in patients who underwent surgery which did not directly improve blood flow in the affected limb. The authors conclude that the most favourable results are produced by the implantation of vascular prosthesis during the period of intermittent claudication distance shortening before the development of resting pain or necrosis. Such operation are loaded with the lowest rate of failures and complications.


[Anticipated results of arterial surgery in chronic atherosclerotic ischemia of the lower limb].

In 1983-1988, the results of conservative and surgical treatment of patients with atherosclerotic ischemia of the lower limbs were assessed with the aid of a questionnaire. A possibility of prognosis was assessed with the use of mathematically processed data obtained with such approach. An effect of clinical symptoms (intermittent claudication distance, resting pain, necrosis) and stage of the disease (duration, K/R index) and risk factors (blood cholesterol, triglycerides, diabetes mellitus, ischemic heart disease, arterial hypertension) on the result of surgical treatment was analysed. The obtained results suggest that clinical symptoms and risk factors may predict the results of surgical treatment in the atherosclerotic ischemia of the lower limbs.


[Suppurating lesions following surgical treatment of lower limb ischemia due to atherosclerosis].

Patients with suppurative lesions complicating surgical reconstruction of the arteries have been analysed. Such complications have been noted in 110 (102 men and 8 women) out of 311 operated patients. Considering the difference in the number of male and female patients, the risk of suppurative lesions complicating vascular surgery is proportional in both sexes. Thousand three hundred sixty six surgeries included: 361 recanalizations, 944 transplantations, and 61 arterial plasties. Percentage of suppurative complications ranged from 8.8% after transplantations to 9.8% after arterial plasties. More than one surgery has been performed in some patients. The risk of infectious complications has been higher in these patients. Despite antibiotic treatment suppurative infections have been noted in 108 (101 men and 7 women) out of 1244 operated patients, i.e. in 8.6%. Intravenous administration of antibiotics during surgery has proven the most effective prophylaxis. An infection of postoperative wound is the most severe local complication in vascular surgery. It has also been most frequent in the analysed group of patients, being 31.3% of all local complications.


[Results of surgical and conservative treatment for multi-level atheromatous arterial occlusion of the lower limb after five years].

After 5 years of observation, materials of 239 patients treated operatively and 96 patients treated conservatively because of multisegmental arterial occlusion of lower limbs were obtained within the framework of multi-central programme of investigation. The results of the treatment with these methods have been evaluated according to the site of the atheromatous changes and the degree of ischemia of lower limbs. It has been ascertained, that after operative therapy, the percentage of good results and the percentage of death in the case of occlusion comprising the aorto-ilio-femoral segment is higher than in the case of occlusion of femoro-popliteal and peripheral segments, whereas in the case of conservative therapy, the situation is reverse. The results of therapy (operative and conservative) of multilevel arterial occlusion, in all kinds of location, depend upon blood supply in the limbs. The results are best (the highest percentage of good results and without improvement, the least number of amputation and death) in the II period of disease and they become worse when the degree of ischemia increases. It appears that the operative therapy is less effective than the conservative therapy in the case of atheromatous multilevel changes manifesting clinically in the form of intermittent claudication, whereas in the case of the IV degree of ischemia, only the operation can save the limb or even the patient's life.


[Results of conservative treatment in patients with atheromatous obstruction of arteries in the leg].

Late results of the treatment of 107 patients with crural arteries occlusion (21 (20%) with isolated lesions and 86 (80%) with multisegmental lesions) were evaluated. Favourable result of the conservative treatment was achieved in 38% of patients with isolated lesions, no improvement in 38%, and worsening in one patient were noted during a 5-year follow up. Percentage in the group of patients with multisegmental lesions (including crural arteries) was 23%, 50%, and 27%, respectively. No patients with isolated lesions died during follow up period, whereas 11 (13%) patients with multisegmental lesions died.


[Late results of surgical and conservative treatment for atheromatous ischemia of the lower limb in young patients].

The study was carried out within a multicenter program. Hundred fifty four patients under 40 years were treated (73 conservatively, and 81 surgically) for the atheromatous ischemia of the lower limbs in 14 Departments of Vascular Surgery in Poland within 5 years. The results of the treatment were evaluated according to the site of atheromatous lesions, type of surgery, and mainly according to the progress in pathologic process expressed by the degree of limb ischemia. It was found that surgery performed in the II period of the disease produces the best results. Surgery produces positively better results than conservative treatment in patients with the II period of the disease whereas both surgical and conservative therapy are ineffective in the III and IV degrees of ischemia. Sympathectomy may be effective in the occlusion of crural arteries even of the IV degree ischemia. In case of the II degree ischemia better results are produced by the operation of aorto-iliaco-femoral segment (transplantation or thrombendarterectomy) than that involving distal segment (sympathectomy) whereas the results of conservative therapy are better in peripheral and iliaco-femoral occlusion than those in aortofemoral and multisegmental arterial occlusion. An opinion of several authors on the specificity of atheromatous ischemia of the lower limbs in young patients was not confirmed.


[Effect of extended anticoagulant therapy on long-term results of bypass for arterial occlusive disease].

Data selected from the questionnaire supplied by the 14 centres of vascular surgery have been analysed. All operated patients have been followed-up for 5 years. An analysis revealed that long-term oral anticoagulant therapy following vascular grafting statistically significantly improved the outcome of surgery in comparison with patients who have not been given oral anticoagulants. There has been no such a relationship, if antiaggregation agents had been used for the treatment.


Interactive cytometry, chance or evil of bias?

Interactive selection of a limited number of cells in imaging cytometry for determining the DNA histogram of breast cancer cells as the best known prognosticator at the moment, implies statistical and systematic sampling problems. Analysis of histograms of 361 breast cancer aspirate specimens measured in two laboratories demonstrate the expected high statistical variations in view of the only 100 cells measured per case but also slight systematic differences. Controlled systematic sampling without pathological bias results in a somewhat higher malignancy grading than selective biased sampling. For this finding we have no explanation. The main result is, however, that we did not find the expected contrary which makes the argument for at least this application invalid that expert pathologists are needed for reliable interactive sampling.


Construction of the knowledge file for an image understanding system.

To enable an image understanding system to provide an automated interpretation of diagnostic imagery it must have access to all of the concepts, procedures and methods used by human experts. The paper describes information elicitation from experts of different domains and the construction of a knowledge file. Uncertainty management is based on Bayesian belief network methods.


Time dimension in histopathology.

In the liver acinus hepatocytes and littoral cells stream from the portal tract toward the terminal hepatic vein. Their average displacement trajectory was denominated as tissue radius. Since cells advance on the tissue radius in one direction, the farther a cell is the older it is, and cell age may be estimated from its position. This property is regarded as time dimension of a tissue. Since distance may be estimated by image cytometry, age determination may be incorporated in the image cytometer software. Distance may be expressed in two types of units: 1. metric e.g. microns, and 2. cell location or the number of cells separating a given cell from tissue origin. Cell age may also be expressed by two units: chronological and biological. Chronological age may be derived from the cell's displacement velocity. A hepatocyte advances daily 2 microns so that a cell at 100 microns distance is 50 days old. Biological age is defined as the cell's location on the radius. Cell location may serve as biological age unit since cells ranked according to their location are also ranked by their ages. The average acinus radius is 22 location long so that in biological age units, maximal hepatocyte life span is 22 locations. One may even say that a cell at location 5 is 2 biological units older than a cell at location 1. In order to estimate chronological age the cell has to be followed for a period of time, while biological age may be read off once from the section. There is yet another advantage for utilizing cell location as unit.(ABSTRACT TRUNCATED AT 250 WORDS)


An electronic primer in dermatopathology.

We present an electronic primer in dermatopathology based on a full-text retrieval system which contains all the features of traditional handbooks of dermatopathology. The system including high-resolution black and white pictures with coloured overlays runs on every personal computer with VGA card and needs no additional software and hardware equipment. The representation of pictures is improved by using high-resolution VGA cards. The system is able to work like an electronic diagnostic consultant in dermatopathology.


[Long-term results of aorto-iliac occlusive disease treatment with implantation of the Polish vascular prosthesis].

The author analyses distant results of the Polish vascular prostheses implantation to 227 patients with aorto-iliac occlusive disease. Eighty two (29.6%) patients died within 5 years following the operation. Therefore, an analysis of the distant results of therapy included 145 patients. An excellent result was achieved in 20.7% of the treated patients, satisfactory result in 53.9%, no improvement or worsening in 20.7% of cases. Statistically significant relationship between the degree of pre-operative ischemia and outcome of surgery has been noted. Considering blood hypertension, diabetes mellitus, obesity, hypercholesterolemia, and tobacco smoking prior to and after surgery as risk factors, no statistically significant relationship between the distant result of the treatment and the number of risk factor in a single patient has been observed.


A computer-based decision support system for diagnostic histopathology of the breast.

Accurate histological diagnosis of breast lesions is essential for the appropriate management of the patient. However, the technique of histological typing is problematic due to the large number of histological patterns, often of a complex and variable nature, which occur in breast disease. The introduction of the Breast Screening Programme has increased the burden on pathologists, and emphasised the need for training. Problems arise because mammographic screening detects a greater proportion of special histological types, with their attendant difficulties of identification, when compared to clinically palpable lesions. A computer-based decision support tool has been developed to assist pathologists in the histological diagnosis of breast disease. Unlike conventional expert systems, which seek to recreate the problem-solving processes of an expert, this system has been designed to act as an intelligent assistant to the pathologist. The system represents knowledge in the form of 'disease profiles', and utilises a novel inference model based upon the mathematical technique of hypergraphs. Initial trials with this system have demonstrated that a high level of diagnostic accuracy can be achieved.


Development of an automatic program for the stereological estimation of the volume weighted mean volume of cell nuclei.

A stereological method was recently developed for the estimation of the volume weighted mean volume of particles from 2D sections. Several authors successfully applied this manual method to paraffin sections of bladder cancers and malignant melanomas and found a good correlation between the mean nuclear volume and the histological grading and prognosis of these tumors. The implementation of this measuring method on an automatic image analyser, devoted to routine work in tumor pathology, is presented. The accuracy of the automatic measurement has been evaluated together with the first results obtained from human esophageal cancers.


MIPSY: real-time morphometry to quantify the time course of rapid epithelial restitution.

The gastrointestinal mucosa has the ability to repair itself rapidly following superficial mucosal injury by rapid epithelial restitution. The mechanism consists of cell migration and does not involve mitosis. This study reports the mechanisms of rapid epithelial restitution in the rabbit colon in vivo and in the human colon in vitro, describes a new computerized real-time morphometry system to investigate the time course of restitution and presents a new method to calculate the migration speed of epithelia during the repair process. Superficial mucosal damage to the rabbit colon in vivo was produced by luminal exposure to 100 mM HCl for 5 min (80% of mucosal surface), a comparable injury in the human colon in vitro was obtained by luminal exposure to 10 mM HCl for 10 min (96% of mucosal surface). After detachment of the damaged tissue the intact epithelial cells in the vicinity of the necrosis extended pseudopodia and migrated over the denuded basal lamina. The morphological appearance of rapid epithelial restitution was the same in the rabbit and the human, only the time course was postponed in the human. The time course of rapid restitution was assessed by a newly developed computerized morphometry system (MIPSY). When tissues were examined after various time points following acid damage, 61% of the mucosa were damaged in the rabbit after 1 h, 10% after 2 hs and 20% after 5 hs. In the human colon 85% of the mucosal surface were damaged after 2 hs and 20% 5 hs after the end of acid exposure.(ABSTRACT TRUNCATED AT 250 WORDS)


Cell contour extraction on multi-threshold images.

A procedure for the extraction of cell and nuclear contours from digital images is presented. The procedure is simple and suitable for efficient implementation in an interactive environment where the user selects a region of interest containing the cell to be segmented. Segmentation is facilitated by multi-thresholding where the threshold values are determined by an iterative algorithm using the mean square error criteria to cluster the image pixel values. Edges are detected on the threshold images by employing a binary morphological filter and the desired contours are selected by considering closed contours in the edge image. The cell region is first extracted by performing segmentation, edge detection and contour extraction on the region of interest. The nuclear region is subsequently extracted by reapplication of the algorithms within the extracted cell region. Segmentation examples are presented.


Evolutionary somatic cell changes in cervical tumour progression quantitatively evaluated with morphological, histochemical and kinetic parameters.

The somatic cell changes which characterise malignancy evolution in human cervical preneoplastic and neoplastic lesions have been assessed on histological sections by means of a computerised image analyser. Many features have been simultaneously measured on each cell of the lesions studied, and the following results have been obtained: Some features, mainly kinetic, show continuously increasing values which express changes correlated to the increasing malignancy; other features, especially related to nuclear atypia, cellular heterogeneity and the degree of aneuploidy, have values dropping at the level of early stromal infiltration, which can be morphometrically characterised as composed of relatively homogeneous phenotypes; these features seem to express the degree of genetic instability and relate to the evolutionary somatic cell changes; tumour progression evolves through sequential discontinuous steps, each of them characterised by specific phenotypical features of the neoplastic cell population; the neoplastic cells in the foci of early stromal infiltration and vascular invasion, phenotypically more homogeneous than the parent cell populations of carcinoma in situ and infiltrating carcinoma, seem to possess a greater genetic stability.


A simplified histoscore for the estrogen receptor assay in breast cancer.

Different histoscores combining the number of positive cells and the intensity of staining have been used to evaluate the estrogen receptor immunocytochemical assay (ER-ICA). Our aim was to investigate if the simple estimation of the amount of positive cells could be sufficient for the semiquantitative analysis of ER-ICA. Tissue from 51 women with ductal breast carcinoma was used. Half of each sample was processed with the quantitative assay (ER-EIA) and the other half with ER-ICA. Microscopical analysis was performed by two independent observers and classified on a simple scale from 0 to 4+. With EIA 31 cases (60.78%) were positive and 20 (39.21%) negative. With ER-ICA 29 (56.86%) had immunostaining, whereas 22 (43.13%) did not. 95.83% of the ER-ICA positive cases and 77.7% of ER-ICA negative had a good correlation with EIA values. Statistical analysis showed a high degree of correlation (r = 0.88 p 0.001). Hence, simple semiquantitative estimation in ER-ICA is sufficient to provide useful information for clinical use about ER content in tissue sections.


An image cytometric DNA-analysis in breast neoplasms. Parameters of DNA-aneuploidy and their relationship with conventional prognostic factors.

In this prospective study, an image cytometric DNA-analysis was performed in 86 women with breast neoplasms (72 primary invasive carcinomas and 14 benign lesions). Four DNA ploidy parameters were analysed: histogram type (according to AUER classification), DNA-index, tumor cells with DNA content above the 5n limit and DNA malignancy grade (DNA-MG, calculation according to Böcking). Their correlations with well established prognostic factors in breast carcinomas (tumor size, lymph node status, histologic grade, hormone receptor content) were studied. All but one benign lesions were diploid (13/14 cases), whereas the majority of the primary invasive breast carcinomas were aneuploid (58/72 cases). A predominance of carcinomas with a percentage of cells superior or equal to 1% with DNA content above the 5n limit was observed (54 cases out of 58). Most of the aneuploid tumors had a histogram type III or IV (53 cases) or a high DNA-index (50 cases). Of these 58 aneuploid cases, only 26 tumors had a DNA-MG superior to 1. Interestingly, 26 tumors had the 4 criteria of aneuploidy, 19 had 3 and 9 had 2 and only 4 tumors had one parameter. The DNA-MG was significantly related to hormonal receptors (p less than 0.001) and tumor size (p less than 0.01). The histogram types (Auer classification) and the DNA content above the 5n limit were correlated with histologic grade (SBR or SBRM) (p less than 0.02). Concerning the DNA-index no correlation was observed with well established prognostic factors. On the other hand no significant correlation was found between these new biologic variables and lymph node status.(ABSTRACT TRUNCATED AT 250 WORDS)


Two types of endometrial papillary neoplasm. A morphometric study.

Fourteen cases of Papillary Endometrial Carcinoma (EC) were analyzed by Interactive Computerized Morphometry. Seven cases were diagnosed as well differentiated adenocarcinomas with papillary features (PF) and belonged to a group of EC with associated adenomatous hyperplasia (AH). Seven cases were diagnosed as uterine papillary serous carcinomas (PA) and belonged to a group of EC without associated AH. Two morphometric procedures were used. DRAW for the characterization of individual nuclei (area, perimeter, chord) and NU-MEAS for tissue architectural features (crowding and stratification). Using a stepwise discriminant multifactorial analysis, both methods proved to be accurate for the two diagnostic categories, as shown by the 100% posterior probabilities and by the two diagnostic categories, as shown by the 100% posterior probabilities and by the distances between group means. A doubtful case was analyzed and classified using a K-nearest neighbor procedure, compared to the individual case in the database. The distinction between the two types of papillary EC is important for the differential diagnosis of the two lesions. Well differentiated adenocarcinoma with papillary features is seen usually in the context of a well-differentiated adenocarcinoma, in a group of patients known to have estrogen-related less aggressive tumors. Uterine papillary serous carcinoma was described to have a biological behavior similar to that of papillary ovarian carcinoma and is encountered in a group of patients with more invasive and less differentiated EC2. Computerized interactive morphometry is a valuable method to use for the accuracy of this differential diagnosis in doubtful cases.


Densitometric and morphometric study of immunocytochemical estrogen receptors detection in breast carcinomas.

Immunohistochemical quantitative evaluation of estrogen receptors (ER) detected in tissue sections from 30 breast tumors by monoclonal antibody was performed using a densitometric method. In particular, ER concentration was calculated by nuclear mean optical density (nMOD), while heterogeneity in ER content was calculated by the coefficient of variation (CV) of the nuclear optical density histogram. Tumors which showed more than 60% of positive cells had a mean value of ER-nMOD of 0.116 +/- 0.002 a.u. and of ER-CV of 33.74 +/- 0.68. Tumors which showed 30% to 60% of positive cells had a mean value of ER-nMOD of 0.082 +/- 0.006 a.u. (arbitrary units) and of ER-CV of 36.25 +/- 3.44. Tumors showing less than 30% of positive cells had ER-nMOD of 0.052 +/- 0.009 a.u. and ER-CV of 48.49 +/- 5.61. These results indicate that the greater the concentration the lower the ER heterogeneity within the tumor sample. No significant differences between ER-ICA results, nuclear size and form factors were found.


Immunohistological quantitative analysis of S100 protein-positive cells in T-cell malignant lymphomas, especially in adult T-cell leukemia/lymphomas.

S100 protein-positive cells (S100+ cells) in 36 cases of T-cell lymphoma (T-ML) in the lymph node and 15 cases of T-ML in the skin were analyzed immunohistologically in order to study their quantitative features in adult T-cell leukemia/lymphoma (ATLL). The T-MLs were categorized according to the updated Kiel classification, and the T-cell pleomorphic type (Pleo) was subcategorized into 3 subtypes: Pleo-ATLL, Pleo-clear and Pleo-others. The population of S100+ cells and the first to fifth minimal distances of every S100+ cell were measured on micrographs of paraffin sections that had reacted to anti-S100 protein antibody according to the ABC method. Lymphoblastic and chronic lymphocytic leukemia types showed low populations of S100+ cells and long values of the first minimal distance. T-zone lymphoma without follicles and angioimmunoblastic lymphadenopathy with dysproteinemia-type T-ML had high populations and low values of the first minimal distance. Among the three subtypes of Pleo in the lymph node, Pleo-ATLL gave the highest population and the shortest value of the first minimal distance of S100+ cells, but this trend was not found in the skin. Clusters of more than five S100+ cells were more common in the Pleo-ATLL subtype than in the other two subtypes. The increase and clustering of S100+ cells in Pleo-ATLL suggests that the lymphoma cells act on S100+ cells as a helper.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparative quantitative study of Ki-67 antibody staining in 78 B and T cell malignant lymphoma (ML) using two image analyser systems.

Total Ki-67 stained area percentage was studied in 32 B and 46 T malignant lymphomas (ML) using two different image analyser systems (TAS, Leitz; SAMBA TM 2005, TITN) respectively. The total Ki-67 area percentage was highly correlated to the number of Ki-67 positive cellular profiles (B-ML, r = 0.93; T-ML, r = 0.88), indicating that area percentage is a reliable alternative method to the manual cell counting. Image analysis allows quicker measurements, appropriate to large and strictly lymphomatous regions. The cell image processor (SAMBA TM 2005, TITN) linked to a color video camera was more suitable for immunohistochemical sections and allowed more automated and faster measurements than the texture analyser (TAS, Leitz) linked with a black and white camera. Alkaline phosphatase technique with fast red as chromogen was more suitable for the detection of Ki-67 stained area by thresholding than peroxidase technique with aminoethylcarbazol or with diaminobenzidine as chromogens. Significant differences were found between low and high grade in B and T ML according to the Kiel classification (mean values +/- SD of 7.7 +/- 3.8% and 16.6 +/- 6.2% in B-ML and of 10.2 +/- 7.9% and 25.6 +/- 16.3% in T-ML respectively). In follicular B-ML, considering follicular areas only, values were comparable to high grade ML; angioimmunoblastic-lymphadenopathy-like (AILD-type) T-ML belonging to low grade ML showed similar values to pleomorphic T-ML with medium and/or large cells belonging to high grade ML.(ABSTRACT TRUNCATED AT 250 WORDS)


Nuclear DNA density of parathyroid lesions.

Nuclear DNA density analysis was carried out on six normal control human parathyroid glands and forty-six pathological lesions of human parathyroid tissue. The pathological glands had originally been classified as twenty hyperplasias, twenty-one adenomas, and five carcinomas in accordance with clinical-pathological information. Specimens had been obtained through the Surgical Pathology Department of the University Hospital, London, in the form of either imprints from the cut surface of a lesion, or as cell separation samples from a paraffin block. Nuclear DNA ploidy analysis of the normal glands showed, as expected, a diploid cell population. Sixty-one percent of nuclei of the carcinomas were above 2C, extending to 7C. There was a significant difference in nuclear DNA content between normal and benign tissue versus tissue classified as carcinoma. No significant difference in nuclear DNA ploidy could be established between the benign adenoma and hyperplasia tissue; however, marked significant DNA ploidy differences were found between tissue from the latter two and carcinoma. DNA ploidy from cell image analysis using Feulgen's reaction can be useful in establishing a differential diagnosis between carcinoma and benign lesions.


Nuclear morphometry of primary B cell thyroid lymphoma.

Thyroid lymphoma is usually distinguished from anaplastic thyroid carcinoma and from Hashimoto's thyroiditis by morphological and immunohistochemical assessment of tissue sections. Our objective was to assess the value of nuclear morphometry in the differential diagnosis of these conditions. Nuclear area measurements were performed on 10 cases of thyroid lymphoma using an IBAS 2000 Image Analyser and compared with similar measurements performed on 10 cases of Hashimoto's thyroiditis and 2 of anaplastic thyroid carcinoma. It was found that karyometry demonstrated differences between all three conditions, the cases of thyroiditis being distinguishable from lymphoma on the basis of mean nuclear area alone. Mean nuclear area for lymphomas was greater than for Hashimoto's thyroiditis and lower than for anaplastic carcinomas. The mean nuclear area also reflected the grade of lymphoma, with the exception of one case which had a large reactive T cell population. It is concluded that nuclear morphometry provides valuable information in the diagnosis and assessment of thyroid lymphomas.


Cytologic diagnosis of non-oxyphil follicular neoplasias of the thyroid. Multivariate classification procedure based on quantitative nucleolar features.

The study was done on cytologic material of 58 non-oxyphil follicular neoplasias of the thyroid, 32 of which were adenomas and 26 carcinomas. Three groups of nucleolar features were quantified using a routine microscope with an ocular micrometer: frequency-, size-, and margination-related features. Since value overlap was present between two categories for all the variables, stepwise discriminant analysis was applied. The following three features were selected by the computer for calculation of one canonical discriminant function: percentage of marginated nucleoli, percentage of nuclei with one nucleolus, mean major nucleolar diameter. The percentage of agreement between morphologic and computer classifications was 95%. Two follicular adenomas were allocated to the carcinoma category, whereas one follicular carcinoma was allocated to the adenoma category. Out of 58, 52 were diagnosed by the computer into one of the two diagnostic categories with a very high probability, i.e. P greater than 0.75, the remaining 6 being considered intermediate.


Prognostic factors in squamous cell carcinoma of the oral cavity. A retrospective study of 80 cases.

A multiparameter analysis of 80 squamous cell carcinomas of the oral cavity was performed to determine the relative prognostic weight of the location of the tumor, tumor size (T), microscopic grade (G) and DNA content measured by means of flow cytometry. Tumors of the lip have an invariably favourable outcome, while at the other locations they have high rates of mortality (total 5 years of survival less than 35%). A simple statistical analysis was performed using the concepts of death sensitivity (DS), death specificity (DSp), life sensitivity (LS) and life specificity (LSp): the best measure of unfavourable outcome was represented by G3 (DS = 0.69; DSp = 1) and by DNA aneuploidy (DS = 0.76; DSp = 0.45), while the best measure of favourable outcome was represented by G1 (LS = 0.53; LSp = 0.96), T1 (LS = 0.81; LSp = 0.78) and by DNA diploidy of the tumor cells (LS = 0.45; LSp = 0.86). A survival analysis using a step by step regression model according to Cox was carried out in order to evaluate more precisely the relative importance of prognostic factors: traditional prognostic factors (histological grade = G and tumor size = T) showed a strong statistical significance, while DNA content added some additional prognostic information only if associated to the macroscopic features of the tumor.


Laryngeal intraepithelial neoplasia (LIN). An analytical morphometric approach.

Laryngeal Intraepithelial Neoplasia (LIN) is graded in 3 levels (LIN Grade I-II-III), corresponding to the classic aspects of mild, moderate and severe dysplasia-in situ carcinoma, on the basis of the number and position of mitoses and of the undifferentiated or atypical cells limited to the basal or extended to the intermediate or the superficial layers of epithelium. In order to reduce the subjective imprecision of these parameters we have applied not only traditional dimensional evaluators but also procedures of analytical morphometry to the nuclear shape. By using the software system S.A.M. (Shape Analytical Morphometry) we have examined fifty nuclei of the basal layer in LIN grade I, II and III, fifty nuclei in normal laryngeal mucosa and fifty nuclei in invasive carcinoma of the larynx (twenty-five cases in all). Normal and dysplastic nuclei did not show any dimensional differences, while the carcinomatous nuclei were significantly larger. An asymmetric distortion of the nuclear contour was noted in the moderate and severe dysplasia, but not in carcinomatous cells. Also the Fourier parameters, increased in severe dysplasia, decreased dramatically in carcinomatous cells which showed nuclei with minor contour irregularities than the normal cells. These findings outline the discriminative power of the analytical morphometry and suggest a possible correlation between nuclear shape and cell biology.


Quantitative histopathological evaluation of vocal cord dysplasia with particular emphasis on nuclear orientation.

We have applied morphometry on formaldehyde-fixed, H & E-stained diagnostic laryngeal biopsies from 7 patients with mild dysplasia and 7 with severe dysplasia/carcinoma in situ, in search of objective parameters required for reproducible histopathological grading of epithelial dysplasias. Special emphasis has been put upon the individual nuclear polarity as a spatial variable. Also included were 4 specimens with normal epithelium. By means of a semiautomatic digitizing tablet, the nuclear and epithelial area, formfactor and the polarity variation between the longitudinal axes of adjacent nuclei were measured in the basal, parabasal, middle and luminal layers of the epithelium. N:C-ratio, mean values of nuclear area, formfactor or their coefficient of variation could not distinguish between mild and severe dysplasia. The variations in neighboring nuclear polarity, however, revealed a highly significant distinction between mild and severe dysplasia (p less than 0.001). This parameter may therefore have diagnostic potential.


Quantitative analysis of nucleoli and nucleolar organizer regions in cultured primary human normal, reactive and malignant mesothelial cells.

The number and the size of silver-stained intranuclear granules, which correspond to the nucleolus and nucleolar organizer regions, have been determined by means of quantitative methods in cultured primary human mesothelial cells obtained from normal, reactive and malignant mesothelium. The mean values per nucleus of the number, the total area, the average area, and the relative area of the silver-stained granules and the mean nuclear area were determined for each of the three conditions. Normal, reactive and malignant mesothelial cells differed significantly in all the features. These findings at the optical level reflect the differing rate of the nucleolar biosynthetic activity related to the different biological properties of the three cell types, and the features can be useful morphometric descriptors in the diagnostic pathology of the mesothelium.


Evaluation of AgNOR count in distinguishing benign from malignant mesothelial cells in pleural fluids.

The authors have evaluated in eight reactive and eight malignant pleural effusions the number of intranuclear dots representing the nucleolar associated proteins stained with silver colloid technique (interphase nucleolar organizer regions-AgNORs). The mean number per nucleus in benign reactive effusions was 1.56 (SD 0.77) while in mesotheliomatous effusions it was 2.81 (SD 1.44). The statistical analysis of values, by Mann-Whitney U Wilcoxon Rank Sum W Test, revealed a significant difference of AgNOR counts in the two cytological samples. The variability of AgNOR areas and morphologies in reactive and mesotheliomatous nuclei in pleural fluids is evaluated applying automatic image analysis.


Intra-tumor DNA ploidy distribution pattern and its relation to histologic type in gastric carcinoma.

The pattern of intra-tumor DNA ploidy distribution was analyzed in stepwise sections in 64 cases of surgically resected gastric carcinoma. Five varying patterns were identified: Type A comprised only diploidy in all stepwise sections, Type B comprised only aneuploidy with the same DNA Index(DI), Type C comprised diploidy in the great majority of sections with aneuploidy in some parts, Type D comprised aneuploidy in the great majority of sections with diploidy in some parts and Type E comprised only aneuploidy, but with varying DI. These 5 patterns could be grouped into 2 categories; predominantly diploid (Types A and C) and predominantly aneuploid (Types B, D and E). The former category included 34 cases (12 differentiated carcinomas) while the latter included 30 cases (22 differentiated carcinomas). Thus, a statistically significant correlation was detected between the histologic type and the intra-tumor DNA ploidy distribution pattern; in the majority of cases, differentiated carcinoma exhibited predominantly aneuploidy, while undifferentiated carcinoma exhibited predominantly diploidy (p less than 0.01). This tendency was the same for all depths of invasion, except for submucosal carcinomas which exhibited predominantly diploidy.


Epithelial dysplasia of the gastric mucosa. A morphometric and ploidy pattern study.

60 cases of gastric dysplasia (20 mild, 20 moderate and 20 severe) were quantitatively studied and compared with cases of hyperplastic polyp (No 10), intestinal metaplasia (No 20), peptic ulcer (No 10) and invasive adenocarcinoma (No 20). In each case the area, the perimeter and the integrated optical density of 100 nuclei were measured in Feulgen stained tissue sections by means of a Leitz TAS. The stepwise discriminant analysis demonstrates that severe dysplasia and adenocarcinoma can be well distinguished from mild and moderate dysplasia. The ploidy pattern study demonstrates that the presence of severe dysplastic changes corresponds to a sharp increase in the degree of aneuploidy. In peptic ulcer, intermediate values between "benign" and "malignant" lesions were found.


Cytometry in histological sections of colon carcinoma.

A short description of a project of cytometry in histological sections of colon carcinoma is given with emphasis on the methodical aspects. Possible strategies of cytometric measurement and problems related to it (focus, overlap, segmentation of objects) are described. The main effort concerns interactive selection of tumor cells and the segmentation in cases of densely distributed and overlapping nuclei. All other succeeding processing steps are performed fully automatically. The resulting quantitative features are stored together with the original images on an optical disk for further examinations and reexaminations, allowing the direct relation of feature values to visual image content. The evaluation of the features as well as their interpretation is only at the beginning. Especially the problem of relating section information with true 3-dimensional information is not described here and necessitates further research. In a first investigation only a few tumors without and with metastases were analyzed. The preliminary results correspond with findings of Kunze et al.


Quantitative study of the precancerous and malignant lesions in colorectal adenomas.

150 endoscopically removed colorectal adenomas are quantitatively studied. In 90 cases dysplastic glands constituted the entire lesion (30 mild, 30 moderate, 30 severe); in 60 cases foci of adenocarcinoma were present (30 intramucosal, 30 submucosal). In each case the area, the perimeter and the integrated optical density of 100 nuclei were measured from Feulgen-stained paraffin sections. The results show a continuous neoplastic progression in the trends of the morphometric parameters as well as in the ploidy pattern. The stepwise discriminant analysis defined the morphometric features of the atypical nuclei, which were present in small numbers also in mild dysplasia; they progressively increased in number, reaching the maximum in submucosal carcinoma.


Texture analysis of histological images of giant cell tumor of bone.

To gain an objective evaluation of histological sections of giant cell tumors of bone (GCT) and osteosarcomas, microscopic pictures were taken and their grey-tone image measured, using a flying spot scanner and computer. Various values of eight parameters expressing certain characteristical brightness distribution patterns were computed, and comparatively examined among the three groups of benign GCT, malignant GCT and osteosarcoma. As a result, some parameters could facilitate differentiation between the histological images of these bone tumors. Especially, "angular second moment (ASM)", "Contrast" and "Coefficient of variation (COV)" were useful even for discrimination between malignant and benign GCT. After factor analysis of the values of these parameters, scores of each factor for a number of histological scene images were plotted on a 2-dimensional factor plane. On this plane, which was considered to be a histological feature plane, cases of benign GCT were separated from those of osteosarcoma. Cases of malignant GCT were distributed between the two groups. These results suggest that this method could be valuable for computer evaluation of histological images of benign GCT and osteosarcomas.


Morphometric analysis of cartilaginous tumors.

144 chondromas and chondrosarcomas have been investigated. Morphometry has been performed: the relative volume density of tumor cell nuclei (Vv%n) and the nuclear area have been measured. The results show that the relative volume density of nuclei is a useful parameter, regarding the localisation of the tumors, to discriminate chondromas and chondrosarcomas, while the nuclear area is no safe criterion to separate these entities.


Morphometrical investigation of medulloblastoma nuclei by S.A.M. (Shape Analytical Morphometry) software system.

In order to characterize medulloblastomas and to get over the difficulties sometimes encountered in differential diagnosis, a double morphometric procedure has been applied to its nuclei. The first consisted of size measurements (maximum diameter, area and perimeter), the latter is represented by S.A.M. (Shape Analytical Morphometry) software-system specifically implemented to describe shape of biological structure by analytical parameters. Analytical and dimensional parameters submitted to Hotelling's multivariate discriminant analysis gave the best results when used together in convenient discriminant subsets, thereby allowing a good distinction between medulloblastoma in comparison with neuroblastoma, Ewing's tumor, lymphoblastic and lymphocytic lymphoma. These results underline the usefulness of morphometric characterization also for practical diagnostic purposes.


Significance of variation in DNA flow cytometric analyses from paraffin-embedded breast cancers. Evaluation of the grading efficiency of ploidy determination, DNA index, and S-phase fraction.

116 paraffin-embedded breast cancer samples were analyzed by flow cytometry. From each sample 3 consecutive 50 microns sections were cut for the study. The presence of neoplastic tissue was verified from light microscope sections cut before and after the adjacent sections. One laboratory started with one section from each block and was allowed extra sections when needed for analysis. At the end the laboratory obtained results from all 116 cases. The rest of the samples were studied by 2 other laboratories. Samples with results from 2 or 3 laboratories then allowed variability analysis and the estimation of the grading efficiency in a 2-grade system. Inconsistency in diploidy/aneuploidy distinction was present in 36 of 111 (32.4%) cases studied by two or three laboratories. This inconsistency was less obvious in samples graded as multiploid by at least one of the laboratories. The grading efficiency as analyzed from the results of 3 laboratories was 0.89, and of 2 laboratories, 0.84. The DNA index showed a slightly higher grading efficiency. At the cutoff point of 1.3, 91% of cases could be expected to be correctly classified into low ploidy and high ploidy groups (grading efficiency 0.91). The S-phase fraction had a mean grading efficiency of 0.89, a performance comparable to that of diploidy/aneuploidy distinction. In the light of the available data the flow cytometric analysis can add to the consistency of grading, especially when compared with subjective histological grading. However, the data do not suggest that flow cytometric analysis of paraffin sections as a grading method would be more consistent than quantitative histopathology from sections.


Biological image analysis for microscopical diagnosis: the work-station S.A.M. (Shape Analytical Morphometry).

The S.A.M. (Shape Analytical Morphometry) software system and its related work-station give a generalized and easy-to-handle tool to face a classic and intriguing problem in biomedical morphological diagnosis. What is the shape of an object in a microscopic image? How can we understand the relationship between size and shape? According to Holloway: "Measurements such as length, width, height, whether in chords or arcs only describe space, ... and further run into the abyss of allometric correction ... if additional information (shape?) to size is expected, some method of allometric correction must be used." The S.A.M. software system assumes a logic architecture able to separate and to parametrize independently shape characteristics in terms of allometry and local pertubation by analytical procedures (polynomials, parabolic fitting, Fourier analysis) in addition to the classic evaluations of size and density carried out by image analyzers for microscopical diagnosis.


Silver-stained nucleolar organizer proteins in urothelial bladder lesions. A morphometric study.

The diagnostic value of image analysis of silver-stained nucleolar organizer regions (AgNORs) has been investigated in 170 urothelial bladder lesions obtained by transurethral biopsies in the same number of patients. According to the WHO classification 25 specimens showed a flat non-invasive growth pattern with simple hyperplasia or mild dysplasia (H/D1 = 10), as well as moderate (D2 = 10) to severe atypia (Cis = 5). 135 samples were of papillary and/or infiltrating type including polypoid cystitis (pC = 5), true papillomas (G0 = 7), and carcinomas of different malignancy grade (G1 = 30, G2 = 55, G3 = 38). 10 biopsies served as normal controls. Benign non-neoplastic urothelium (controls, H/D1, pC) exhibited few but large AgNORs (mean number [MNN] = 3.3 +/- 0.5, mean area [MNA] = 0.29 +/- 0.08 micron 2), whereas carcinomatous lesions (Cis, G1-G3) showed numerous small silver-stained dots within their nuclei (MNN = 6.9 +/- 1.1; MNA = 0.12 +/- 0.04 micron 2). Both parameters, MNN and MNA, were inversely correlated (r = -0.69, p less than 0.001); their quotient (NQ = MNN/MNA) revealed a clear cut difference in the AgNOR content of benign (control, H/D1:NQ = 12.8 +/- 2.5) and moderate to severe atypical flat urothelial lesions (D2, Cis: NQ = 44.2 +/- 12.8, p less than 0.001). This parameter also discriminated between G1 (NQ = 40.7 +/- 17.3), G2 (NQ = 57.5 +/- 18.8), and G3 carcinomas (NQ = 79.0 +/- 21.8, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Quantitative immunohistologic study of lip biopsies. Evaluation of diagnostic and prognostic value in Sjögren's syndrome.

In a group of 45 patients with Sjögren's syndrome (SS) and 80 controls the high specificity (95%) and sensitivity (100%) of a recently proposed bivariate quantitative immunohistologic (QIH) criterion for SS, based on percentages of IgA and IgG-containing plasma cells in labial salivary gland (LSG) tissue, was confirmed. The best univariate QIH criterion for discrimination between LSG biopsies of SS patients and controls appeared to be based on the percentage of IgA containing plasma cells, and had a specificity of 99% and a sensitivity of 96%. A criterion based only on the percentages of IgM-containing plasma cells, proposed in another recent study, resulted in a high number (31%) of false negatives. Interobserver reproducibility of QIH diagnoses was excellent. Moreover it was demonstrated that accuracy, precision and the interobserver reproducibility of plasma cell counting depends on the choice of tissue fixation and immunohistologic staining procedure. The combination of formol sublimate fixation and peroxidase anti-peroxidase procedure appeared to be the best combination for QIH examination. Furthermore, in 2 SS patients systemic monoclonal IgM/kappa gammopathy was preceded by high predominance of IgM and kappa containing plasma cells in the plasma cellular infiltrate of the LSG tissue.


Quantitative analysis of quadriceps muscle biopsy in systemic sclerosis.

The lesser diameter of the fibers, separately of type 1 and type 2 fibers, and the capillary density evaluated as number of capillaries/area and number of capillaries/fiber were quantitatively estimated in muscle biopsies from 12 patients with systemic sclerosis. Nine patients with polymyositis or rheumatoid arthritis and six patients with muscle dystrophy and neurogenic atrophy served as controls. The results showed that patients with systemic sclerosis had low values of the lesser diameter of the fibers. This reduction was most obvious in type 2 fibers. The capillaries were also significantly reduced when compared with the control groups. When considering the capillaries/fiber ratio, seven out of eight patients with values lower than 0.7 belonged to the systemic sclerosis group. In systemic sclerosis the capillary density values were not significantly correlated with those of the lesser diameters of the fibers.


Enlargement of synaptic size as a compensative reaction in aging and dementia.

A quantitative investigation has been carried out on synaptic contact zones of dentate gyrus supragranular layer and cerebellar glomeruli in autoptic samples from adult, old and demented patients. During physiological aging and senile dementia, the synaptic average area was significantly increased as compared to adult values in both the CNS areas investigated. Conversely, the number of contacts and their total surface contact area per unit volume of tissue were decreased. Current literature reports that, in animal models, enlarged synapses undergo perforations and splitting to modify synaptic connectivity. As against these assumptions, the increased synaptic size observed in our study appears to represent a compensative reaction of old and demented CNS to counteract the reduction in number and in total contact area of the synaptic junctions.


The value of simple morphometric techniques in the diagnosis of osteoporosis.

In order to assess the value of simple morphometric methods in the diagnosis of osteoporosis we measured trabecular bone volume (TBV), trabecular number per unit area and relative osteoid volume in two separate iliac crest biopsies in 20 patients with conclusive autopsy evidence of osteoporosis and 20 controls. In osteoporotic patients trabecular bone volume ranged between 3.0-19.2% (median 10.7%) in the anterior and 3.0-14.3% (median 9.7%) in the posterior biopsy and in controls between 2.5-24.8% (median 13.5%) in the anterior and 5.8-25% (median 12.4%) in the posterior. Trabecular numbers in osteoporotic patients ranged from 1.7-8.3/10 mm2 (median 3.5) and in controls 1.6-17/10 mm2 (median 5.63). Relative osteoid volume showed a similarly wide range. In adjacent biopsies trabecular bone volume and trabecular number could differ by as much as 100% and this casts doubt on the value of sequential bone biopsies in the management of individual patients. We conclude that simple morphometric techniques cannot accurately diagnose osteoporosis. However, if the TBV is greater than 18% osteoporosis is unlikely to be present (upper 95% confidence limit = 17.8%). However, a low value for TBV is not diagnostic for osteoporosis because of a large overlap between control and osteoporotic groups.


Osteologic standardization of human coxarthrosis using histomorphometry and its relevance for HIP alloarthroplasty.

Using bone histomorphometry, in this study the osteologic status of 107 patients with coxarthrosis and femoral neck fracture [FNF] was assessed and compared with bone parameters from patients revised for aseptic loosening. Bone biopsies of the acetabulum and the proximal femur from patients with primary coxarthrosis [pCoxA] (69), dysplastic coxarthrosis [CDH] (19), rheumatoid arthritis [RA] (9), femoral head necrosis [FHN] (8), femoral neck fracture [FNF] (4) and aseptic loosening (12) were taken during hip alloarthroplastic surgery, prepared undecalcified and analysed using histomorphometry (according to MERZ). In pCoxA the following average figures of bone parameters of the acetabular biopsy were determined: trabecular bone volume (TBV) 39.6%, osteoid volume (OV) 3.9%, active osteoblastic surface (AOS) 6.5%, osteoclastic resorption surface (ORS) 2.4%, osteoid surface (OS) 17.4% and resorption surface (RS) 7.0%. As average figures of the femoral biopsy in pCoxA were assessed: TBV = 17.2%, OV = 1.3%, AOS = 0.9%, ORS = 0.4%, OS = 6.5% and RS = 2.5%. These data were compared with bone parameters of secondary coxarthrosis, osteoporosis (FNF) and aseptic loosening. Based on the study of Oettmeier et al. on femoral heads (Skel Radiol 18: 165-174, 1989), the investigated groups were subdivided into three osteologic types of the hip. The osteopenic type was found in 10% of pCoxA, 43% of RA and 28% of CDH. The hyperostotic type, predominantly occurring in males, was mostly demarcated in FHN (38%) and pCoxA (12%). The results demonstrate the individual osteologic status of patients before hip alloarthroplasty. This could influence planning of the operation and the bone-implant interface as well as long-term prognosis of artificial joints.


Macrophage and epithelioid cell nuclear morphology in leprosy.

The spectrum of disease in leprosy is characterised by the presence of macrophages in lepromatous lesions and epithelioid cells in tuberculoid granulomas. Since changes in nuclear shape occur during macrophage activation, we have measured nuclear morphology by planimetry in biopsies across the leprosy spectrum. The results show no significant correlation of any of the parameters of nuclear morphology measured between different lesions or between biopsies from the centre and edge of the same lesion. There were no differences between the Ridley-Jopling groups. However, several parameters which measure the degree of ellipticity of the nuclei showed strong correlation with granuloma size in untreated leprosy patients. This suggests that local pressure effects may influence epithelioid cell and macrophage nuclear morphology in leprosy lesions.


Age-related modification of average volume and anisotropy of vascular smooth muscle cells.

The aim of this study was to determine what changes in the arterial wall are related to age. In two groups of rabbits, one young and one adult, the aorta and carotid were studied using a morphometric approach based on stereological axioms and planimetric morphometry. The problem of anisotropy of smooth muscle cells is discussed in detail. Two forms of anisotropy must be distinguished, that of single cell and that due to the histological pattern of the smooth muscle cells in the arterial wall. Our results show in adult animals, as compared to the young ones, statistically significant decrease in anisotropy of the cell pattern which tends to become more regular. Moreover, in aorta and carotid of young and adult animals there is an increment of 95.39% and 80% of the absolute cell volume, respectively. We suggest that there may be a direct relationship between aging and phenotypical modulation of the smooth muscle cells and that the modification of the architectural cell pattern with age may represent an adaptive event related to the change in forces acting upon the arterial wall.


Age-dependent increase of rabbit aortic atherosclerosis. A morphometric approach.

Aging has been indicated as one of the major risk factors for development of atherosclerotic lesions, although the role aging plays, lacks accurate evaluation. Our study was aimed at quantitatively defining such a role by using morphometric analysis. Aged (median age 3 years and 8 months) and young (4 months) white New Zealand rabbits received a hyperlipemic diet enriched with a low dose of cholesterol for 16 months. At regular intervals, levels of serum lipemic parameters were checked. A Quantimet 920 image analyzer was used on paraffin-embedded sections of the entire aortas to measure the volume density of the tunica intima, the volume density of atheroma, the ratio intima/media and the surface area of the tunica intima. Our results indicated for aged hyperlipemic rabbits a statistically significant increase in all morphometric parameters examined as compared to young hyperlipemic animals, and no statistically significant differences in serum cholesterol, triglycerides and phospholipids.


Morphological, immunohistochemical, stereological and nuclear shape characteristics of proliferative Leydig cell alterations in rats.

Proliferative Leydig cell (LC) alterations (hyperplasia, adenoma) of laboratory rats often pose diagnostic problems because the progression from normal to hyperplasia to neoplasia is continuous. The LC compartments of 130 Wistar rats (kfm: WIST strain) of approximately 2 years of age were examined. Ten typical cases conventionally classified as being normal or as showing diffuse or focal hyperplasia or small or large adenomata were investigated in more detail. In large adenomata, areas with large and small LC nuclei were identified. Immunohistochemical characterization, EM examination, as well as stereologic and planimetric investigations were performed. Hyperplastic and neoplastic LC essentially retained their normal appearance and immunohistochemical characteristics, but were found to contain more lipid droplets, fibroblast-like cells and patches of collagen than normal LC at the EM level. LC proliferation was accompanied by significant LC hypertrophy. LC nuclei of hyperplastic LC compartments were slightly larger while those of LC adenoma were markedly larger than nuclei of normal LC. The values for circle-related and ellipticity factors indicated that the nuclei of normal and hyperplastic LC were more markedly oval than nuclei of neoplastic LC. Concavity factor and bending energy measurements revealed that the small and oval nuclei of normal and hyperplastic LC had significantly more and deeper indentations than the larger and somewhat rounder nuclei of neoplastic LC. It is concluded that LC proliferations conventionally diagnosed as hyperplasia or adenoma on the basis of their size were composed of cytologically different LC populations.


Changes in rat epiphyseal cartilage after treatment with dexamethasone and glycosaminoglycan-peptide complex.

In 3-month-old rats, systemic administration of glucocorticosteroids induced regressive changes in the epiphyseal plate cartilage. Dexamethasone treatment resulted in inhibition of both chondrocyte proliferation and cartilage matrix production. The inhibition of chondrocyte proliferation was determined histologically from the thickness of the epiphyseal plate, the number of cell columns and the ratio of proliferating to hypertrophied chondrocytes. The decrease in cartilage matrix production was measured autoradiographically by incorporation of radioactive 35S-sulphate. Concomitant treatment with glycosaminoglycan-peptide complex (GP-C = RUMALON) overcame the negative effect of dexamethasone. The values in the dexamethasone + GP-C group approached those of the control group and differed significantly from the group given dexamethasone alone.


Morphometry of parenchymal and vascular alterations in ARDS after extracorporal carbon dioxide removal therapy (ECCO2-R-therapy).

The lungs of 12 patients suffering from ARDS treated by ECCO2-R-therapy were examined. The thickness of the interlobular septa, area portions of honeycombing, bleeding and lung parenchyma with diffuse alveolar damage were assessed by semi-automatic image analysis. Medial thickness of the pulmonary arteries with an external radius ranging between 40 microns and 150 microns was assessed planimetrically. The area portion of honeycombing ranged from 0% to 66% (average: 28.8%, standard deviation: 17.0%). The thickness of interlobular septa was increased with values between 74.0 microns and 195.5 microns (average: 140.4 microns, standard deviation: 41.7 microns; normal controls: 72.0 microns +/- 19.5 microns). Parenchymal and vascular changes were closely related: Average medial thickness rose from nearly normal values (4.9%) in cases with low area portions of honeycombing and bleeding to the double (11.1%) of normal values in cases with area portions of honeycombing and bleeding greater than 40%. We conclude that parenchymal changes due to shock and high inspiratory O2 concentration lead to changes of the pulmonary arterial hemodynamics which result in a quickly developing medial hypertrophy.


Metastatic capacity and differentiation in murine melanoma cell lines. A morphometric study.

A morphometric analysis was carried out on electron micrographs of cells of the F1 (low metastatic capacity) and F10 (high metastatic capacity) variant sublines of the murine B16 melanoma, both in in-vitro cultures and in lung-metastatic nodules developed after the intravenous injection of neoplastic cells in syngeneic C57 black male mice. A group of 28 morphometric parameters was derived to describe quantitatively each neoplastic cell profile. No qualitative difference was observed between the two cell lines. The quantitative expression of subcellular organelles was dissimilar in the two sublines, being consistently characterized, both in in-vitro cultured cells and in lung-metastatic colonies, by a significant decrease in the mean values of parameters related to melanosomes in the high metastatic capacity cell line (B16-F10). Moreover, in in-vitro cultured cells, indices describing heterochromatin masses and cytoplasmic membranous compartments displayed statistically significant differences between the two sublines. In this experimental system, an inverse relationship between metastatic capacity and differentiation is detected, since cells with a more aggressive metastatic behavior exhibit a decreased degree of differentiation.


In situ analysis of adenohypophysis proliferative activity in beagle dogs. Preliminary results.

In order to establish control data on adenohypophysis cell cycle in beagle dogs, two different approaches were used. In the first approach, the adenohypophysis mitotic index (percentage of cells in mitosis: MI) was determined using pituitary glands from 50 control beagle dogs, 10 to 25 months old, from retrospective regulatory safety assessment studies. We found that the MI in males was 0.024 +/- 0.007% and in females 0.068 +/- 0.019%. The second approach involved the determination of the adenohypophysis cell fraction in S phase by means of bromodeoxyuridine (BrdU) anti-BrdU method in 16 control beagle dogs, 8 to 20 months old, from ongoing safety assessment studies. We found that the BrdU labeling index (percentage of BrdU-positive cells per total number of cells: BrdU LI) was 0.053 +/- 0.009% in males and 0.059 +/- 0.015% in females. Analysis of our data indicated that the results obtained from the BrdU LI were more reliable (coefficient of variation [CV] less than 0.07) compared to those obtained from the MI (CV greater than 1.4). No clear difference was found between male and female beagle dogs, regarding adenohypophysis cell proliferation. In both studies, adenohypophysis cell proliferation was found to be age-dependent, with the highest value in immature dogs. No correlation between estrus cycle in females and adenohypophysis cell proliferation was noted.


Morphometric analysis on myocardial injury related to the use of high volume potassium cardioplegic solution during ischemic arrest.

We correlated the effects of high volumes of K+ cardioplegic solution on myocardial structure and function in 16 dogs following open-heart surgery. Eight animals received high volume potassium cardioplegic solution (25 cc/kg body weight, every 30 min) during 90 min of ischemic arrest (HVK-C group). The others received sufficient cardioplegic solution to maintain complete electrical arrest as defined by voltage monitoring criteria (VM group). Cardiac index (CI), left ventricular stroke work index (LVSWI), and myocardial contractility (dp/dt) were determined before arrest and after 90 min of ischemia and 45 min of reperfusion. Biopsies were taken for EM ultrastructure and ATP estimation. Morphometric analysis of EM micrographs found increased volume of damaged mitochondria (DMR) (p less than 0.025), damaged myofibrils (DMF) (p less than 0.001), intermyofibrilar edema (p less than 0.005), T-tubule and sarcoplasmic reticulum (p less than 0.05) in the HVK-C group. Left ventricular (LV) function was more depressed in animals receiving HVK-C. CI decreased by 1.8 +/- 0.4 l/min/square meter (p less than 0.01), LVSWS fell by 3.3 +/- 0.8 gm-m/beat/Kg (p less than 0.01), dp/dt decreased by 684 +/- 135 (p less than 0.0025). ATP decreased by 26% in HVK-C and by 12% in VM group (0.1 less than p less than 0.05). Structural damage (scores of injured volume of mitochondria and myofibrils) correlated with post-ischemic depression of LV function (Cardiac output and myocardial contractility), r = -0.72 and -0.66 (p less than 0.001 and 0.004).


Comparison of the ECL-cell frequency in the stomachs of 3 different rat strains.

Three different rat strains, Sprague-Dawley, Wistar and Fischer 344, were treated for 3 months with 2 doses (0.8; 4 mg/kg) of the gastric acid suppressing ATPase inhibitor pantoprazole. The gastrin levels were determined, the height of the mucosa measured and the number of enterochromaffin-like (ECL) cells counted. Because these cells were stained according to the method of Grimelius they were designated as GPC (Grimelius positive cells). Under 4 mg/kg, the gastrin levels were increased 8 hours after administration, but fell again after 24 h. The Fischer rats showed the highest value. Also the height of the mucosa was increased under 4 mg/kg. A trend towards an increased mucosal height was noticeable even at 0.8 mg/kg. The number of GPC was determined in 2 ways: 1) without taking the mucosal height into account, 2) taking the height into account. An increase in GPC was observed at 4 mg/kg with both methods.


Stimulation of bone marrow by administration of excessive doses of recombinant human erythropoietin.

Recombinant human erythropoietin (rhEPO) was administered intravenously to Beagle dogs in daily doses of 100, 500 and 3000 units/kg/day for 3 months. The high dose was more than 200-fold the therapeutic human maintenance dose. Such excessive rhEPO doses elicited extreme erythropoiesis. There was a dose-dependent stimulation of bone marrow fibroblasts, leading to bone marrow fibrosis in some of the high dose animals. The extent of myelofibrosis was intra- and interindividually different in various bones. Sternebrae proved to be practical for morphometric studies. The point-counting method was used for measurement. The portion of fatty tissue, sinusoids and fibrous tissue in the medullary space as well as the number of blood vessels and megakaryocytes were calculated. Such experimental conditions are not of relevance in human patients whose rhEPO therapy is interrupted as soon as their PCV reaches 35 vol%. Experimentally induced myelofibrosis should therefore not be considered as a risk in patients receiving therapeutic doses of rhEPO.


Fractal dimension of pericellular membranes in human lymphocytes and lymphoblastic leukemia cells.

When blood cells are projected into an image plane their contours appear as borderlines of irregular shape with the property of the statistical self-similarity. The dimension D of these cell perimeters may be fractal rather than an integer as in the Euclidean space. The fractal dimension D for different q'order moments was evaluated by means of a probabilistic method after recording EM image profiles of normal peripheral blood mononuclear cells (PBMN) and mature lymphocytes and also of lymphoblasts isolated from humans with acute leukemia or obtained by lectin stimulation in vitro. PBMN cells, CD 4 and CD 8 T-lymphocytes showed a fractal dimension D quite similar among five moments, ranging from 1.23-1.17. Cells of hairy-cell leukemia with highly convoluted morphology displayed D comprised between 1.32-1.36, whereas blasts of T or B acute lymphoblastic leukemia were characterized by a smaller D of 1.11 and 1.13-1.16 respectively. When normal T-lymphocytes were transformed into blasts by lectin PHA stimulation in vitro, the fractal dimension D was significantly reduced and found close to the value recorded on pericellular contour of blasts from acute T-lymphoblastic leukemia.


Pharmacokinetics of inulin in blood and central lymph in the rat.

Inulin was used as a model drug for the study of factors influencing the transport of drugs across the blood-lymph barrier. The model drug was administered iv (0.75, 7.5 and 37.5 mg/kg) as a bolus and by infusion. Kinetic parameters in blood and central lymph including lymphatic bioavailability (FL) were determined. It has been found that FL is influenced (increased) by larger doses of inulin (FL extent 0.934 +/- 0.250 - 1.914 +/- 0.250), infusion concentration ratio lymph/blood in steady state was 0.941 +/- 0.013. Identical kinetic parameters were found in rats with differentiated content of total lipids in lymph (in fed and fasted group, FL were 0.861 +/- 0.167 and 0.934 +/- 0.250 respectively).


Stimulatory effect of low doses of mechlorethamine on humoral response of ovalbumin-immunized rabbits--comparison with levamisole.

The effect of low doses of mechlorethamine (1-10 micrograms/kg) and levamisole (2.5 mg/kg) on humoral response of rabbits immunized twice with ovalbumin (0.1 mg/kg) was compared. It was shown that mechlorethamine given in a dose of 5 or 10 micrograms/kg potentiates the increase in the level of serum anti-ovalbumin hemagglutinins; this increase is induced both after the first and the second (after 14 days) antigen administration. Levamisole acted similarly but with much lower efficiency.


Interaction of epidermal growth factor and transforming growth factor beta in human prostatic epithelial cells in culture.

The present study was conducted to study the interaction between epidermal growth factor (EGF) and transforming growth factor-beta (TGF-beta) in benign human prostatic epithelial cells in culture. Primary cultures of human prostatic epithelial cells were grown in complete WAJC, which consisted of WAJC-404 medium and, in addition to other defined additives, EGF and bovine pituitary extract (BPE). Incomplete WAJC contained the same composition except EGF and BPE were deleted. TGF-beta was added into media at concentrations of 0, 0.1, and 1.0 ng/ml. When cells were grown in complete WAJC, they proliferated rapidly. Cell proliferation was greatly suppressed when incomplete WAJC was used. Addition of TGF-beta to these cultures caused a significant reduction in the final cell number when either complete WAJC or incomplete WAJC was used. In additional experiments, cells were prelabeled with 3H-thymidine for 72 hr prior to treatment with TGF-beta. The percentage of radioactivity released into the medium at the end of a 6-day culture was used as an indication of the extent of cell death. Trypan blue exclusion test was also used to assess the extent of cell death. Addition of TGF-beta into complete WAJC did not significantly affect the extent of cell death beyond what was considered as the result of normal cellular turnover. Addition of TGF-beta into incomplete WAJC, however, caused a significant increase in the percent of cell death in the culture. These results demonstrated an interaction between EGF and TGF-beta in proliferation and cell death in human prostatic epithelia in culture. In the presence of EGF alone in the culture medium, prostatic epithelial cells were stimulated to proliferate. The rate of proliferation was greatly diminished when EGF was deleted from the medium or when TGF-beta was added in the presence of EGF. Finally, cell death was induced when TGF-beta was added into the medium in the absence of EGF.


Interaction of adenosine analogs with morphine in analgesic tests.

The effect of selective adenosine receptor agonists on nociceptive responses of mice and rats and on morphine analgesia was investigated. All compounds used: phenylisopropyladenosine (R-PIA), adenosine ethylcarboxamide (NECA), cyclohexyladenosine (CHA) and 2-chloroadenosine (2-CADO) exhibited antinociceptive action in mice and rats in the hot-plate (56 degrees C) and tail-immersion (52 degrees C) tests. R-PIA, CHA and NECA potentiated the antinociceptive action of morphine in mice, and R-PIA and NECA--in rats. 2-CADO did not affect the morphine action in the tests.


High-performance liquid chromatographic method for the simultaneous determination of propafenone and 5-hydroxypropafenone in human serum.

A simple, rapid and sensitive high-performance liquid chromatographic method for the simultaneous determination of propafenone and 5-hydroxypropafenone in human serum is described. Method involves a single-step extraction of the drug and its metabolite with dichloromethane:2-propanol (4:1 v/v) mixture from 0.2 ml of serum. Separation of the investigated compounds on deactivated Supelcosil LC18-DB column is accomplished by ultraviolet detection at 210 nm. The limit of detection is 10 ng/ml for propafenone and 4 ng/ml for 5-hydroxypropafenone. The method is useful for the routine monitoring of propafenone and its main metabolite in serum as well as for the pharmacokinetic studies.


Correlation between the prostatic vein and vertebral venous system under various conditions.

In dogs, the venous blood from the prostate gland was observed under X-ray fluoroscopy to drain into the vertebral venous system under conditions of abdominal compression, the addition of various intraabdominal pressures, and occlusion of the inferior vena cava by a balloon catheter. Pressure in the inferior vena cava and abdominal cavity were measured simultaneously. The venous blood draining from the prostate gland started to flow from the inferior vena cava into the vertebral veins at more than 25 mmHg of intraabdominal pressure with the animal in the supine position. The average pressure of the inferior vena cava draining into the vertebral veins was 12.8 +/- 1.3 mmHg in the supine position and 21.1 +/- 2.7 mmHg in the standing position. The average intraabdominal pressures were 35.5 +/- 3.9 mmHg and 30.1 +/- 2.8 mmHg, respectively. Under conditions of abdominal compression and balloon occlusion of the inferior vena cava, the materials flowed into the vertebral venous system from various routes, such as the internal iliac vein, common iliac vein, and inferior vena cava. It was suggested that the inferior vena caval blood easily enters the vertebral venous system in the standing position by adding high intraabdominal pressure, and that the vertebral venous system may be useful for experimental study of drug administration in bone metastasis of prostate cancer.


Presence of calcitonin-like immunoreactivity (iCT) in human prostate gland: evidence for iCT secretion by cultured prostate cells.

Immunoreactive calcitonin (iCT) has been detected in human prostate tissue extracts as well as seminal plasma. The present studies were undertaken to examine whether iSCT (immunoreactive salmon CT-like human peptide) co-exists with iHCT (thyroid CT-like substance) in human prostate tissue extracts, and whether these substances are secreted by primary prostate cells in culture. Since the local secretion of these substances seems to increase in some neoplasms, a second objective of the study was to examine whether basal secretion of iCTs from primary prostate cells is increased in carcinoma. The present results have shown that both iHCT and iSCT were present in prostate tissue extracts. The mean iHCT levels in extracts of benign hyperplastic prostates (BPH) were 0.59 ng/g prostate, and these were significantly lower than iHCT concentrations in prostatic carcinoma (PC) (2.53 ng/g). No significant differences in their iSCT contents were observed. However, the results from culture of over 90 individual prostate tissue specimens from BPH or PC indicate that primary prostate cells secreted detectable quantities of iSCT and the basal release of this material from PC prostate cultures was almost four-fold higher than that from BPH prostate cultures. These results suggest that a CT-like immunoreactive material is secreted by primary prostate cells in culture, and the basal secretion of this material is significantly higher in PC cells as compared to BPH cells. Endogenous secretion of prostatic CT, and the elevation of its expression in PC suggest that it may serve as a regulatory factor in the pathophysiology of the prostate gland.


Synthesis and some pharmacological properties of new V1/V2 antagonists of arginine-vasopressin with structural changes at their N-terminals.

In an effort to develop more effective and selective V2-antagonists of arginine-vasopressin (AVP) we designed and synthesized four new analogs of this hormone. The peptides were designed in order to explore how the combination of modification of thioacids occupying position 1 and substitutions of positions 2 and 4 by D-Phe and Ile respectively, will influence their antagonistic properties. Three of the reported analogs are moderately potent V1/V2 antagonists.


Synthesis and some pharmacological properties of 1,2-amino ethers with natural monoterpene structures.

Synthesis and physicochemical and pharmacological properties of 10 analogs of 2-[2-(6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)ethoxy]N, N-diethylamino ethane (3) have been described. The compounds possess toxicity close to or lower than the parent compound--myrtecaine, have no antiarrhythmic activity but some of them (15, 16, 22, 24), similarly as compound 3, show quite strong local anesthetic action.


Vascular action of natural vasopressin-like peptides in isolated rat tail artery.

The literature data regarding the vasoconstriction potency of natural vasopressin-like peptides are contradictory. The cumulative concentration-response curve for arginine-vasopressin (AVP), lysine-vasopressin (LVP), arginine-vasotocin (AVT), lysine-vasotocin (LVT) and phenypressin (PHP) on the isolated rat tail artery was determined. The potency rank of these peptides on the vascular smooth muscle of the rat tail artery was the following: AVP greater than LVP greater than AVT = LVT greater than PHP. The results are discussed in comparison to the data in the literature.


Current status of allografting for bone tumors.

Over the past 20 years the Orthopaedic Oncology Unit of the Massachusetts General Hospital has performed over 660 massive cadaveric allograft transplantations, mostly for patients with bone tumors. The overall success rate for such procedures is around 80%, with most of the failures occurring in the first 3 years as a result of fracture (19%), nonunion (14%), and infection (10%). With longer follow up, late occurring osteoarthritis has been noted in approximately 17% of those patients at risk, suggesting that the articular surface begins to "wear" at 5 years. Some problems remain with the procedure, and more study of such aspects as the immune mechanism, cryopreservation of cartilage, maintaining the status and safety of the bone banking system, and improving the techniques of surgery ultimately may lead to a more uniform and predictable success rate.


Current procedures for banking allograft human bone.

The demand for bone and soft tissues for surgical usage has increased rapidly as the efficacy and safety of these materials has been demonstrated. Advances in technology and procedures used to prepare the tissues have also grown in sophistication and remain a dynamic area. How tissue banks screen donors and prepare allografts impacts on the risks and benefits of these materials. Surgeons should understand how tissue banks supplying their graft materials work and have confidence in this treatment option. They will then be prepared to educate their patients to help them make informed decisions concerning allograft usage.


Immune responses to osteochondral allografts: nature and significance.

Osteochondral allografts evoke immune responses. The nature of these immune responses and their biologic significance are still only partially understood. It is clear, however, that cell surface antigens of the major histocompatibility complex represented on the cellular elements of bone grafts cause T-cell activation, specifically those of the suppressor/cytotoxic phenotype. In numerous animal models, the most immunogenic bone allografts (mismatched, fresh) have demonstrated the poorest clinical and biologic outcomes, while more closely matched and/or grafts treated to reduce immunogenicity (frozen, freeze-dried) have more successfully incorporated. These observations support the hypothesis that immune responses against bone-graft related antigens have biologic significance and that reducing these responses may improve clinical results.


Fresh and frozen articular cartilage allografts.

Fresh and frozen cartilage allografts both function well to relieve joint pain due to cartilage injury. Fresh cartilage allografts appear to remain alive for many months and are most applicable to small injuries such as osteochondritis dissecans, traumatic defects, and osteonecrosis. Frozen cartilage allografts have been used mostly for treatment of bone tumors which most often involve the subchondral and the metaphyseal areas of the long bones, particularly around the knee and hip. Frozen cartilage allografts appear to survive well and maintain joint space for several years. Deterioration of either type of graft does not appear to result in joint pain, making these grafts useful for long periods of time.


The long-term success of fresh, small fragment osteochondral allografts used for intraarticular post-traumatic defects in the knee joint.

Ninety-two fresh osteochondral allografts were implanted in 91 patients with posttraumatic osteoarticular defects of the knee joint. These patients have been prospectively followed since 1972. An analysis of long-term survival of these grafts has been performed to determine their success rates: 75% at 5 years, 64% at 10 years, and 63% at 14 years. An evaluation was made between unipolar grafts, which involve only one surface of the compartment, and bipolar grafts, which involve both surfaces. The unipolar grafts had a lower failure rate at all time periods compared to bipolar grafts, with 76% survival at 5 years, 69% at 10 years, and 67% at 14 years. While investigating other factors that might affect the survival of the fresh osteochondral allografts, we calculated the influence of the anatomical location of the graft, patient sex, and patient age. There was no meaningful impact on allograft survival by either the location of the allograft or the sex of the patient. However, there was a significant effect on allograft longevity in terms of patient age, with patients under age 60 doing better. The best indication for the use of unipolar fresh osteochondral allografts is for posttraumatic defects in relatively young, active patients. Joint malalignment should be corrected to achieve best results.


Clinical use of fresh, frozen soft tissue allografts.

Fresh frozen allograft tissue provides a good alternative to autogenous tissue for the treatment of patients with symptomatic ligamentous deficiency. While transmission of AIDS and other infectious diseases remains a small possibility, allografts provide the advantages of shorter operative time, small incisions, no damage to a patient's own tissues, and superior usefulness in multiple reconstructions, as well as revision of failed autogenous reconstructions. An independent, critical evaluation of the 4-year results of patients with allograft ACL reconstructions is presented. Issues important in allograft tissue recovery, banking, and intraoperative preparation are reviewed.


Autogenous bone grafts: nonvascular and vascular.

Nonvascularized autogenous bone grafts have been used for at least 150 years. Vascularized bone grafts were first done only 17 years ago. We now know that bone grafts act through osteoconductive and osteoinductive mechanisms to produce new bone. Autogenous bone grafts remain the gold standard against which allografts and artificial graft materials should be measured.


Immunofluorescence studies of lung tissue in cystic fibrosis.

Previous studies have suggested that immune mechanisms contribute to lung injury in cystic fibrosis (CF); however, there have been no comprehensive studies of immunofluorescent staining patterns in CF lung tissue. We performed immunofluorescence (IF) studies for immunoglobulins, C3, and fibrinogen on autopsy frozen lung tissue from 21 CF patients. Results were compared with lung findings in patients without CF. In CF-derived lung tissue fibrinogen was ubiquitous along the alveolar wall, alveolar space, and interstitium. Free immunoglobulin G (IgG) and IgA coated the alveolar surface segmentally in 14 and 6 cases, respectively. Unequivocal interstitial deposits were infrequent and IgM was present in blood vessels in one patient only. Intra-alveolar and interstitial inflammatory cells demonstrated cytoplasmic IgG, IgA, and IgM, respectively, in 18, 14, and 6 patients. C3 was seen only segmentally along the alveolar wall in two patients and in blood vessels in one. Antinuclear antibody (ANA) staining of interstitial cells for C3 and immunoglobulins was seen in five patients, four of whom had interstitial pneumonitis. Insignificant amounts of alveolar or interstitial fibrinogen and immunoglobulins in inflammatory cells were seen in controls in the absence of lung inflammation. The IF patterns were similar in the inflammatory lesions of CF and control specimens. The IF patterns observed in CF lung tissue are consistent with nonspecific vascular leakage and chronic inflammation with little evidence of immune complex deposition in the interstitium or blood vessels. This study confirms previous reports of ANA activity in CF patients, although the significance of this finding is unknown.


Fatty change of the pediatric myocardium.

The myocardium was studied for the presence of fat in 980 pediatric deaths over a wide age range during a 10-year period. Of these, only 67 cases (7%) were found to have fatty change. The cause of death varied widely and no common pathogenetic factor was identified. Infections and congenital disorders constituted the most frequent causes of death associated with fatty change. The degree of fatty change was variable. Severe fatty change correlated with severe panacinar fatty change of the liver. The pathogenesis of the lesion is discussed.


Tracheal cartilaginous sleeve.

Tracheal cartilaginous sleeve (TCS) is a rare congenital malformation in which discrete cartilaginous rings are replaced by a grossly uninterrupted cartilaginous sleeve. Seven previous patients with TCS have been reported in the world literature; in each instance, TCS was associated with craniosynostosis (CS). We report details of five additional patients with TCS, of whom four had a dominantly inherited CS. Gross examination of the available tracheas and bronchi demonstrates a cartilaginous sleeve with posterior interruption but lacking a normal pars membranacea. The stained and cleared tracheas all demonstrate variable ring formation, usually limited to the posterolateral aspect. The functional significance of TCS, if any, is unknown. No data are available on the prevalence of TCS in CS syndromes. The formation of TCS implies a common mesenchymal defect in which normally discrete structures fuse and is probably analogous to other mesenchymal abnormalities seen in these patients.