Increased liver mast cell recruitment in patients with chronic C virus-related hepatitis and histologically documented steatosis.

Hepatitis C virus (HCV) is still one of the major causes of chronic viral infection worldwide, and hepatic steatosis is a frequent pathological finding in patients with chronic HCV-related diseases. It is unclear whether the steatosis is associated with host factors or the virus itself, although a consistent relationship has been found between steatosis and a necro-inflammatory reaction with the increased secretion of immuno-regulators. A primary sources of inflammatory mediators are mast cells (MCs) bone marrow-derived cells that are detected in both normal and diseased livers. We determined MC density and correlated it with the fibrosis, inflammatory reaction and steatosis observed in the liver biopsies of patients affected by HCV with or without steatosis. All the histological features were assessed using a computer-aided image analysis system. There was a statistically significant difference in MC density between the HCV-infected patients with and without steatosis, with the lower mean value being detected in those without (P < 0.02). Furthermore, a nonstatistically significant difference in fibrosis and inflammation between the two patient groups was found. In conclusion, this is the first study showing a significant increase in MC density in the tissues of patients with chronic HCV infection and histologically documented steatosis.

Computer-aided morphometry of liver inflammation in needle biopsies.

AIM: To introduce a computer-aided morphometric method for quantifying the necro-inflammatory phase in liver biopsy specimens using fractal geometry and Delaunay's triangulation. METHODS: Two-micrometer thick biopsy sections taken from 78 chronic hepatitis C virus-infected patients were immunohistochemically treated to identify the inflammatory cells. An automatic computer-aided image analysis system was used to define the inflammatory cell network defined on the basis of Delaunay's triangulation, and the inflammatory cells were geometrically classified as forming a cluster (an aggregation of a minimum of three cells) or as being irregularly distributed within the tissue. The phase of inflammatory activity was estimated using Hurst's exponent. RESULTS: The proposed automatic method was rapid and objective. It could not only provide rigorous results expressed by scalar numbers, but also allow the state of the whole organ to be represented by Hurst's exponent with an error of no more than 12%. CONCLUSION: The availability of rigorous metrical measures and the reasonable representativeness of the status of the organ as a whole raise the question as to whether the indication for hepatic biopsy should be revised by establishing clear rules concerning the contraindications suggested by its invasiveness and subjective interpretation.

[Fractal analysis of liver fibrosis]. / Analisi frattale della fibrosi epatica.

This study realized by two different study groups use of Fractal geometry to quantify the complex collagen deposition during chronic liver disease. Thirty standard needle liver biopsy specimens were obtained from patients with chronic HCV-related disease. Three mu-thick sections were cut and stained by means of Picrosirius stain, in order to visualise collagen matrix. The degree of fibrosis was measured using a quantitative scoring system based on the computer-assisted evaluation of the fractal dimension of the deposited collagen surface. The obtained results by both study groups, show that the proposed method is reproducible, rapid and inexpensive. The complex distribution of its collagenous components can be quantified using a single numerical score. This study demonstrated that it is possible to quantify the collagen's irregularity in an objective manner, and that the study of the fractal properties of the collagen shapes is likely to reveal more about its structure and the complex behaviour of its development.

Mast cell density, hepatic stellate cell activation and TGF-beta1 transcripts in the aging Sprague-Dawley rat during early acute liver injury.

Mast cells (MCs) have been indicated as a source of various inflammatory cytokines, chemokines and growth factors. This study evaluates liver tissue MC density as a quantitative marker of acute liver inflammation in 2- and 19-month old rats treated with carbon tetrachloride (CCl4) toassess the relationships between MC density, hepatocellular damage, mRNA encoding TGF-beta1, hepatic stellate cell (HSC) activation and collagen levels. Consecutive histological sections from each age group were stained with toluidine blue to identify granulated MCs, Direct Red 80 to recognize collagen matrix, and by immunohistochemistry to identify activated hepatic stellate cells (HSCs), which were subsequently counted by means of a computer-aided image analysis. Histology showed hepatocellular necrosis with inflammatory cell infiltration and collagen matrix deposition. Two and 24 hours after intoxication, MC density had considerably increased in the younger rats, but less in those aged 19 months. Although the untreated older rats had a larger area occupied by activated HSCs than the untreated younger rats, the increase in the number of HSCs was greater in the younger rats both two and 24 hours after intoxication. The greater MC density in younger rats suggests that older rats have a reduced immune response or recruit fewer MCs. The activated HSCs and TGF-beta1 transcripts did not increase significantly during the study period, thus indicating that these are later events in chemically induced hepatic toxicity. In conclusion. MC density may be an index of acute liver inflammation after CCl4 intoxication.

Simultaneous staining of cytoplasmic iron and collagen matrix in human liver biopsy specimens.

One of the major liver fibrogenic activators is the cellular iron overload that can severely damage parenchymal and non-parenchymal cells. The aim of this study was to investigate a histochemical staining technique that allows the simultaneous detection of the irregular deposition of matrix collagen and both the amount and distribution of iron in liver cells on the same histological section. The method was evaluated using 3-microm histological sections obtained from ten standard liver biopsy specimens taken from patients with hepatitis C virus-related diseases and simultaneous liver cell iron overload. The results indicate that the double-staining technique is simple, sensitive and rapid, and can be routinely applied to liver biopsy specimens for diagnostic purposes. Furthermore, it may also facilitate the study of the complex relationship between hepatic fibrosis and iron overload during common genetic or secondary liver metabolic disorders.

Fractal geometry: a useful tool for quantifying irregular lesions in human liver biopsy specimens.

Irregularity and complexity are the main features of every biological system, including human tissues, cells and sub-cellular components. These two properties of the organized biological matter cannot be quantified by means of the classical Euclidean geometry, which is able to measure regular object, practically unknown in Nature. The aims of our paper were a) to underline the importance of the shape of a biological structure, b) to investigate the fractal geometry for quantifying the liver histo-pathological structures, and c) to explain the significance of several terms used in the fractal analysis of complex biological systems.

[Geometrical analysis of benign and malignant breast lesions]. / Analisi geometrica delle lesioni benigne e maligne della mammella.

PURPOSE: The aim of this study was to describe a computer-assisted method based on fractal dimension and the coefficient of roundness, two mathematical descriptors of irregularly shaped objects, in order to discriminate benign and malignant mammographic lesions. MATERIAL AND METHODS: Twenty-three digitized mammograms were classified as containing benign lesions (n=12) or as containing malignant lesions (n=11) on the basis of readings by two independent radiologists. Morphometrical and fractal analysis of the breast lesions were automatically performed by a computer-assisted image analysis system. RESULTS: The method shows, for the first time, two parameters that allow the classification of irregularly shaped objects, such as benign and malignant breast lesions. A significant increase was obtained when comparing the fractal dimension and the coefficient of roundness of benign versus malignant mammographic lesions. DISCUSSION AND CONCLUSIONS: Benign and malignant breast lesions are characterized by complex morphologies. Complexity is the main property of all biological systems, including natural and pathological structures. In this study, we showed that fractal geometry allows quantitative measurement of the complex morphology of benign and malignant mammographic lesions. Furthermore, this mathematical approach may be helpful for the study of the non-linear dynamic processes involved in breast carcinogenesis.

Fractal and spectral dimension analysis of liver fibrosis in needle biopsy specimens.

OBJECTIVE: To evaluate the usefulness of a reliable and reproducible mathematical scoring system based on fractal geometry for quantifying the irregular pattern in fibrosis commonly seen in liver biopsy specimens from chronic liver diseases. STUDY DESIGN: The study used 26 standard liver biopsy specimens obtained from patients with chronic hepatitis C virus-related liver disease. The degree of fibrosis in each specimen was estimated using a quantitative scoring system based on the computer-assisted evaluation of both the fractal and spectral dimensions of deposited collagen. The fractal dimension was then compared with the percent area of collagen measured using an image analysis system. RESULTS: The fractional dimension of its irregular shape defines fibrosis as a natural fractal structure. The complex distribution of its collagenous components (unmeasurable by means of the usual morphometric parameters) can be optimally quantified using a single numerical score that seems to be a better alternative to the semiquantitative methods adopted so far. The proposed method is reproducible, rapid and inexpensive; furthermore, supported by specific software, its mathematical approach excludes subjectivity and eliminates the external factors capable of influencing staging and classification. CONCLUSION: This study demonstrated that it is possible to quantify the irregularity of the structures of the liver in an objective manner and that the box-counting fractal dimension does not depend on the amount of collagen deposited on the slide. Furthermore, as has been found in other fields of investigation, study of the fractal properties of the liver is likely to reveal more about its structure and the pathogenesis of liver diseases.

Geometry of liver system functioning: from a helix to a trajectory on curved space.

Autoisodiasostasis of the liver, i.e., its self-maintenance without change over the course of time, is characterized by a bistable equilibrium between two extreme states called homopoiesis (during which the liver system repairs its worn structures and replicates its cells) and homoerhesis (during which it satisfies the needs of the body). Retinol binding protein traffic within the hepatocyte, made visible by means of immunohistology, has been used as a prototype for studying the competition processes and dynamics of autoisodiasostasis. An account is given of the oscillation of autoisodiasostasis between homopoiesis and homeorhesis through intermediate phases. The phase cycle of autoisodiasostasis is illustrated in the form of bi-dimensional and tri-dimensional diagrams. The temporal behaviour of the systems is represented as a helix trajectory obtained by projecting homopoiesis-homeorhesis phase cycles onto a time series. A unified topological representation of the uncoupled but combined processes of competition and autoisodiasostasis is proposed.

Hepatocellular carcinoma in Italian patients with cirrhosis.

BACKGROUND AND METHODS: Patients with cirrhosis of the liver are recognized as being at risk for hepatocellular carcinoma. The magnitude of the risk, the natural history of this disease, and the possibilities for detecting potentially curable tumors in patients in the Western world are unknown. To address these questions, we examined 447 Italian patients with well-compensated cirrhosis (which was of viral origin in 62 percent of them) from 1985 through 1990, performing serum alpha-fetoprotein assays and real-time ultrasonography every 3 to 12 months. RESULTS: Hepatocellular carcinoma was found in 30 patients (7 percent) at base line and in another 29 patients (7 percent of 417 patients free of tumor at base line) during follow-up periods averaging 33 months (range, 1 to 48). The cumulative hazard of the development of hepatocellular carcinoma during follow-up was higher among patients with persistently elevated serum alpha-fetoprotein levels (12 with tumors among 42 with such levels) than among those with fluctuating levels (11 among 82) or those with consistently normal levels (6 among 255). Only 17 patients had potentially operable tumors. The proportion of potentially operable tumors among those detected during follow-up was significantly lower than the proportion at enrollment (4 of 29 vs. 13 of 30, P = 0.027). The survival at one year of the 12 patients who underwent surgery was 67 percent, and the tumor-recurrence rate was 60 percent. Outcome was not appreciably different for the five patients who refused surgery. CONCLUSIONS: In the West, as in Asia, patients with cirrhosis of the liver are at substantial risk for hepatocellular carcinoma, with a yearly incidence rate of 3 percent. Our screening program did not appreciably increase the rate of detection of potentially curable tumors.

[Neurotic and depressive status related to organic pathology in patients in thermal therapy]. / Stati nevrotici e depressivi in rapporto con la patologia organica in utenti termali.

Females and males with no psychiatric history, 25-75 years old, attending thermal facilities in Boario Terme (N = 449) and in Sangemini Terme (N = 40) were administered, on the 2nd-3rd day of their admission, a questionnaire expressly conceive in order to declare conscious motivations for thermal treatments and, in addiction, the Middlesex Hospital Questionnaire (MHQ), the Zung Self Rating Scale for Depression (ZUNG-D) and the State and Trait Anxiety Inventory (STAI), in order to detect neurotic symptoms and/or anxious or depressive complaints in this kind of population. A re-test was performed after 10 days by 73 subjects, in order to verify the impact of thermal treatments on psychoneurotic profiles. Baseline data were compared with those from normal controls and psychiatric outpatients. All statistical comparisons (probands versus matched controls; test versus re-test) were analysed by means of two-tailed Student "t" test (lever of significance: p less than 0.05). In the light of the data so obtained, subjects attending thermal facilities show a typical MHQ psychoneurotic profile, more marked in females and in subjects with gastrointestinal referred somatic disease and partially responding to thermal treatments, which supports the hypothesis that a particular form of neurosis may play a significant role in motivating request of thermal therapies. The results are largely discussed.

Modeling of biological and medical systems: a systemic strategy.

A strategy in the frame of General System Theory is proposed for the study of biological systems for medical purposes. Its definition and use requires in each experiment the collaboration between physician and system scientists and hence the definition of a common language, by which the real system under study is described. The strategy is based on three intermingled steps: first an ingenuous model is proposed, gathering all the medical knowledge about the studied system, organized within an informal frame derived from the state space approach. Next, a functional model is derived, enlightening the organization of the relations in the medical model. Finally, this organization is formalized by the most suitable algebraic tools, which are thereafter translated into APL programs. This last version is used for simulation, which is exploited not only as a tool to describe and make provisions on the dynamics of the models, but also to deepen and improve the knowledge about the observed system.

Liver system. IV. More on the liver micro-unit: physics and geometry of its action.

The geometry of liver functional micro-units is described in the light of the immunohistologic features of human hepatocytes stained with a retinol-binding protein antibody. The stained cells were distributed into two ordered sets which provided a provisional representation of the law regulating both the motion and the alternation of the rest-work phases (homopoiesis and homorhesis configurations, respectively) which reflect bistable liver micro-unit equilibrium (auto-isodiasostasis). Oscillations between the HP and HR states generate a relaxation cycle that can be assumed to be shared by all micro-units and to be regulated by feedback mechanisms. A model illustrating the compactness of the liver's global action as a result of the activity of each micro-unit is also proposed.

Liver system. V. Activation-extinction line of cyclic hepatocyte activities.

The excitation-extintion line of hepatocytes from an inert state towards the stabilization of a given activity is described. Within the cell, the switching on of any given activity is a competitive process among different activities. The process is driven by the influence field created in the environment of the Rappaport acinus by sinusoidal blood which changes its characteristics during its passage from the portal zone to the central vein. Every step of the excitation-extintion pathway follows the so-called law of autoisodiasostasis (AIS), i.e. it is characterized by an oscillatory motion between restoring (homopoiesis or HP) and working (homeorhesis or HR) states. Since the cyclical bistable equilibrium of AIS characterizes all conditions of hepatocyte activities, the AIS cycle can be defined a limit cycle.

A computer-assisted study of macrophage behavior in HBV and nAnB related infectious chronic active hepatitis.

Activated intralobular macrophages were quantified as their lysozyme contents in liver biopsies from patients with chronic HBV and nAnB active hepatitis, and subjects with no evident liver histological damage. There were more macrophages (0.62 +/- 0.06/2,500 mu 2) with a larger area (20.36 +/- 0.87 mu 2) in HBV-related chronic active hepatitis than in controls (0.21 +/- 0.04/2,500 mu 2 and 16.01 +/- 1.48 mu 2), whereas in nAnB chronic active hepatitis their numbers were similar to that in the controls (0.22 +/- 0.03/2,500 mu 2) and their area was smaller (11.68 +/- 0.79 mu 2).