Incontinentia pigmenti diagnostic criteria update.

In 1993 diagnostic criteria for incontinentia pigmenti (IP), a genodermatosis in which skin changes are usually combined with anomalies of other organs, were established. Approximately a decade ago, IKBKG gene mutation was discovered as a cause for IP. This finding has not been included in IP diagnosis so far. In addition, literature data pointed out a few other clinical findings as possible IP diagnostic criteria. Literature facts concerning IP diagnosis were analyzed. Different organ anomalies, their frequency and severity, were analyzed in the context of applicability as IP diagnostic criteria. Taking into account analyzed data from the literature, the proposal of updated IP diagnostic criteria was presented. We propose as major criteria one of the stages of IP skin lesions. As updated IP minor criteria in our proposal we included: dental, ocular; central nervous system (CNS), hair, nail, palate, breast and nipple anomalies; multiple male miscarriages, and IP pathohistological findings. In the diagnosis of IP, the presence of IKBKG mutation typical for IP, and existence of family relatives with diagnosed IP are taken into account.

Incontinentia pigmenti with ultrastructurally disordered leucocytes.

Incontinentia pigmenti (IP) is a rare X-linked genodermatosis in which skin changes are combined with anomalies of other tissues, mainly of ectodermal origin. Mutations of the IKBKG gene are responsible for IP. Haematological disorders among IP patients are rare. Four female patients from a single family, with typical clinical characteristics of IP, are reported. In addition, all affected family members show a distinct haematological phenotype: hypogranular granulocytes, leucocytes with pseudoplatelets, and different anomalies of nuclei. Pseudoplatelets are a typical finding in patients with leukaemia. As there is dysfunction of the IKBKG gene in leukaemia, it is hypothesised that mis-regulation of the NEMO pathway may cause the appearance of pseudoplatelets in acute leukaemias as well as in IP. These observations suggest that IP may not be only linked to skin and organs of the ectodermal origin.

Mesothelial paracrystalline inclusions in continuous ambulatory peritoneal dialysis patients.

OBJECTIVE: Uremia is known to be followed by changes in the serous membranes of pleura, pericardium, and peritoneum. During continuous ambulatory peritoneal dialysis (CAPD), the peritoneum is exposed to altered body conditions as well as to the influence of dialysate. The aim of the present study was to examine the ultrastructure of the mesothelial cells in CAPD patients, and to compare the findings with those from studies of the peritoneum in uremic controls. Paracrystalline intracytoplasmic inclusions in mesothelial cells were objects of special interest. METHODS: Biopsies of human parietal peritoneum were studied. These were taken from 12 uremic patients during catheter implantation before the start of CAPD, and from 7 CAPD patients during catheter removal for infection or malfunction. The samples were prepared in the standard way to be studied by transmission electron microscopy (TEM). RESULTS: Paracrystalline intracytoplasmic inclusions were seen in mesothelial cells only by TEM. They appear as filamentous structures at the outer part of the inclusions, and as pearl-like structures at the core of the inclusions. Sacculate dilatations of rough endoplasmic reticulum cisternae with partly destroyed membranes and only few ribosomes were also seen, with and without densely osmiophilic filaments within the cisternae. We have found paracrystalline intracytoplasmic inclusions in mesothelial cells from uremic and CAPD patients both. According to the literature, these changes are present in one third of biopsies from uremic patients. Until now, however, they have not been mentioned in CAPD patients.

[Changes in peritoneal mesothelial cells in patients on peritoneal dialysis]. / Promene celija mezotela peritoneuma kod bolesnika na peritonealnoj dijalizi.

INTRODUCTION: Some thirty years ago peritoneal dialysis (PD) became a respectable modality of renal replacement therapy. That is why peritoneal membrane attracted interest of investigators. Certain changes, known as uremic serositis, appear in morphology of serous membranes in end stage kidney disease (ESKD). The aim of our investigation was to examine the morphology of peritoneal lining cells in control group of healthy persons and morphology of peritoneal lining cells in patients on PD. MATERIAL AND METHODS: Peritoneal biopsies were taken in 10 healthy volunteers during the kidney donation and in 15 patients on PD during clinically indicated extirpation. Biopsy samples were prepared for standard routine HE staining and for plastic embedded fine sections studying. Sections were mounted in an ultramicrotome, stained with Toluidine blue (TB) and studied by light microscope (SM), while fine sections were mounted in an ultramicrotome and studied by transmission electron microscope (TEM). RESULTS: One layer mesothelium of the cuboidal or flattened lining cells were present over the lamina propria connective tissue. Mesothelial cells were overlapped like tiles on the roof. These cells were interconnected with different types of cell junctions (unpermeable, adhesion and communication junctions) positioned on lateral parts of the interdigitated cell membranes. A great number of microvilli were often present on the appical surface, as well as a kinocilia and lamellar bodies. Nuclei were euchromatic with well developed nucleoli. Many ribosomes, mitochondria, cisternae of rough endoplasmic reticulum (RER) and Golgi apparatus, lamellar bodies and lipid inclusions were present in the cytoplasm. Using TEM in analyzing fine sections of biopsies of patients on PD, characteristic ultrastructural changes including epithelial defects with only remaining parts of destroyed cells were established, as well as significantly greater number of rough endoplasmic reticulum (RER) cisternae and immature mesothelial cells in lamina propria indicating intensive regeneration of this epithelium. The cytoplasm of new mesothelial cells were of less electron density on TEM photomicrographs, whereas the nuclei of mesothelial cells in these patients were euchromatic with prominent nucleoli and numerous perichromatic granules and fibrogranular nuclear bodies, indicating cells of great activity. Cytoplasmic protrusions of different shape and content were often recognized on the apical surface of cells. Lamellar bodies were also present in this group of patients within the mesothelial cells, as well as between two mesothelial cells or on their apical surface. Mitochondria were picnotic in many of the mesothelial cells of peritoneum in this patient group. In these mesothelial cells intracytoplasmic paracrystaline inclusions were established. TEM photomicrographs showed basal lamina multiplication in this epithelium. CONCLUSION: Our findings comply with reports of other authors. It should be stressed that TEM examination detects characteristic ultrastructural changes in mesothelial lining cells of peritoneum in patients on PD, which could compromise the function of peritoneum as a membrane for dialysis.

[Ultrastructure of peritoneal mesothelial cells]. / Ultrastruktura mezotelnih celija peritoneuma.

The introduction of peritoneal dialysis (PD) as a respectable modality of renal replacement therapy some three decades ago, suddenly drew attention of many authors to peritoneal membrane as insufficiently investigated structure. In order to explain the pathological changes in peritoneum due to renal diseases, it became necessary to explore the normal peritoneal structure. The aim of this study was to examine the morphology of peritoneal lining cells in healthy persons. Biopsies of the peritoneum were performed on 20 volunteer kidney donors. Tissue samples were taken during renal transplantation. Special care was taken in getting appropriate samples without artificial damage because of the extreme fragility of the peritoneal tissue. The preparing procedure was standard for routine HE staining and for plastic embedded semifine and fine sections studies. Semifine sections were made on ultramicrotome, stained with Toluidin blue and studied by light microscope, while fine sections were made by ultramicrotome and studied by transmission electron microscope. One layer of cuboidal or flattened lining cells present over the lamina propria connective tissue presented mesothelium. The cells were overlapped like tiles on the roof. Lateral parts of their interdigitated membranes were interconnected with different types of cell junctions: unpermeable, adhesion and communication junctions; inhibiting intercellular transport. Cell surface was often covered with great number of microvilli and lamellar bodies. A single kinocilia was also often present on apical cell surface. Nuclei were euchromatic with well developed nucleoli. Cytoplasm was filled with a great number of ribosomes, mitochondria, cisterns of rough endoplasmatic reticulum and Golgi apparatus, lamellar bodies and lipid inclusions. Numerous pinocytic vesicles on all parts of the membrane as well as in the cytoplasm indicating active endocytosis, egsocytosis and transcytosys in the process of secretion and reabsorption of serous liquid in peritoneal cavity, were visible. Euchromatic nuclei with prominent nucleoli and numerous mitochondria indicate cells of great metabolic activity.

Ultrastructural changes of peritoneal lining cells in uremia.

Some thirty years ago, peritoneal dialysis (PD) became a respectable modality of renal replacement therapy. That is why peritoneal membrane attracted the interest of investigators. Uremia is followed by changes in the morphology of serous membranes (uremic serositis). Uremic effects on pleura and pericardia have been studied for a long time, but the peritoneum is affected as well. The aim of our study was to examine the morphology of the peritoneum in uremic patients before the start of PD and to compare the findings with those from examinations of peritoneum in healthy controls. We examined 12 uremic patients and 10 healthy controls (kidney donors). Biopsies were taken from parietal peritoneum. The samples were prepared in the standard way for study by transmission electron microscopy (TEM). Certain pathological changes--deformation of mesothelial cells, their detachment from the basement membrane, and unusual bulging of apical surface--were identified at the light microscopy level on semi-fine sections. Paracrystalline intracytoplasmic inclusions were seen in mesothelial cells only by TEM. We hypothesize that the inclusions were causing deformation of the mesothelial cells and detachment of those cells from the basement membrane. Sacculate dilatations of rough endoplasmic reticulum (RER) cisternae with partly destroyed membranes and few ribosomes were also seen, with and without densely osmiophilic filaments within cisternae. Although these changes are mentioned in the literature, the exact reason for their appearance remains unknown.

The acidophilic nature of neuronal Golgi impregnation.

The mechanisms of Golgi impregnation of neurons has remained enigmatic for decades. Recently, it was suggested that divalent (di)chromate anions play a role in the Golgi impregnation process. Therefore, we incubated slices of (para)formaldehyde-fixed rat brain tissue in solutions of potassium (di)chromate, phosphate, chloride or nitrate at pH 6 or 7. Slices were then immersed in solutions of silver nitrate and processed for light microscopical analysis. At pH 6, dichromate probes resulted in dense and homogeneous impregnation of neuronal cytoplasm (typical impregnation). At pH 7, chromate probes showed solely partial cytoplasmic and heavy nuclear-region neuron impregnation (atypical impregnation). Phosphate probes at pH 6 resulted in typical impregnation, whereas at pH 7 phosphate probes gave atypical impregnation. Both at pH 6 and 7, chloride and nitrate probes did not yield any Golgi impregnation. These findings confirmed the pH-dependence of silver-chromate Golgi impregnation as well as the correctness of corresponding acidic silver-phosphate impregnation. Our study revealed a previously unknown, strong anion-dependence of Golgi impregnation, suggesting that hydrogenated monovalent anions are carriers of the neuron impregnation.

Correlations between mitotic and apoptotic indices, number of interphase NORs, and histological grading in squamous cell lung cancer.

Proliferative activity of tumors is strongly associated with prognosis and response to therapy. The reason for faster and uncontrolled growth rate of tumors compared with normal tissue may be caused by the greater proliferation of cells, the smaller rate of cell death, or both. Cell production vs. cell loss rates, and their correlation with a grade of tumor cell differentiation (G) was estimated in 45 cases of squamous cell lung cancers (SCLC) by the use of mitotic indices (MI), number of interphase NORs, and apoptotic indices (AI) as parameters. The mitotic figures as well as apoptotic cells were observed on paraffin sections (4-microm thick) stained with haematoxylin and eosin, and with Feulgen reaction with Schiff-type reagent containing 0.5% Toluidine Blue. According to our results, all three parameters distinguish significantly (P < 0.05) between well and moderately or poorly differentiated groups, but not between the first two groups, and clearly discriminate between low- and high-grade malignancy. These results suggest classification of squamous cell lung cancers into two groups, a group of low and a group of high proliferative activity, despite their morphological appearance. Regression analysis revealed a significant (P < 0.0005) correlation between MI and AgNOR counts per cell nucleus as proliferative markers and AI as a marker of cell loss. The number of mitoses and apoptoses, especially when they are expressed as a percentage of the total number of tumor cells, are markers of tumor proliferation rate. They both can be used in biofunctional staging, based on cell kinetics, to provide more prognostic information about lung cancers than clinicopathological staging.

Incidence and role of apoptosis in myelodysplastic syndrome: morphological and ultrastructural assessment.

By application of morphological and ultrastructural methods for identification of apoptosis, we analyzed the incidence of morphologically evident apoptosis in the bone marrow of 30 patients with myelodysplastic syndrome (MDS), and in the bone marrow of 12 healthy individuals. According to FAB classification, out of 30 patients, eight (26.6%) had refractory anemia, three (10%) had refractory anemia with ringed sideroblasts, 14 (46.6%) had refractory anemia with excess of blasts and two (6.8%) had refractory anemia with excess of blasts in transformation. Three patients (10%) had chronic myelomonocytic leukemia. Cells in apoptosis were examined on semithin slides and expressed as the apoptotic index (AI) (percent counted on at least 1000 cells). An overall increase in apoptosis in patients with MDS was found (median AI in patients vs controls, 3.13% vs 1.05%, P < 0.01 by Mann-Whitney U test). Also, negative correlation between AI and white blood cell count was found (linear r= -0.53, or Spearman rank R= -0.52, both P < 0.01). In patients with evident karyotype changes AI was not higher than in patients with normal karyotype. This suggests that discrete alterations in apoptosis are present even in karyotypically 'normal' clones. Our results strongly support the hypothesis that apoptosis has a role in ineffective hematopoiesis and may be a mechanism responsible for the paradox of hypercellular bone marrow and peripheral blood pancytopenia in MDS.

Ultrastructural and cytochemical characteristics of monocytes in various types of acute leukemias.

We have examined ultrastructural and cytochemical characteristics of monocytes in lymphoblastic leukemia (ALL) patients and compared them to monocytes in acute monoblastic leukemia (AMoL) patients and acute myeloid nonmonoblastic leukemias (AML), respectively. Blood samples were prepared in the standard way for ultrastructural and cytochemical (alpha-naphthyl butyrate esterase and myeloperoxidase) analyses. Our results indicate that monocytes in ALL and acute phase AML have the same characteristics as the malignant ones in AMoL.

[Pseudothrombocytes in acute leukemias]. / Pseudotrombociti u akutnim leukemijama.

Pseudoplatelets are the cytoplasmic fragments in the size of platelets originated not from megakaryocytes but from other blood cells which were observed in some patients with leukemia and infection. The clinical importance of pseudoplatelets is manifested in the falsely increased number of platelets in these patients. Transmission electron microscopy was used in the cytologic examination of 23 patients with different types of acute leukemia. The presence of pseudoplatelets in the form of budding of the cytoplasm of the granulocytic, monocytic and lymphocytic strains of various maturity has been shown.

Myocardial bridges over coronary arteries in Cercopithecus.

The presence of myocardial bridges over the coronary arteries has been studied in 29 monkey (Cercopithecus aethiops) hearts. The great resemblance between the Cercopithecus subepicardial arterial net with the corresponding one in humans has been revealed. There is a high incidence (83%) of myocardial bridges only over the ventricular branches of both coronary arteries. Myocardial bridges are usually (90%) located over the left coronary artery branches, and the left anterior interventricular branch is the most frequently (69%) overbridged vessel. The bridges are always single over the vessel examined and their length varies from 0.5 mm to 31.6 mm. No statistically significant sexual difference in myocardial bridges distribution is reported.

[Origin of mature granulocytes in the active phase and in remission in various types of acute leukemias]. / Poreklo zrelih granulocita u aktivnoj fazi i remisiji raznih oblika akutnih leukemija.

The problem of provenance of the mature granulocytes in acute myeloblastic leukemia (AML) and in acute lymphoblastic leukemia in their active phase and in remission is still completely unresolved. Mature granulocytes could possibly arise in acute phase of AML from the remaining normal möelopoietic or granulopoietic stem cell or from the very leukemic clone of stem cell which still display, at least partially, the ability to sustain maturation to normal granulocyte level. In ALL, mature granulocyte probably arise from normal granulocyte precursors. In remitted AML, according to opinions of a majority of authors, mature neutrophils descend from normal stem cells. Following the study of 19 patients with AML and 19 patients with ALL, by utilizing the morphological, cytochemical and cytoenzymic properties of granulocytes et the light and electronomicroscopic level the authors have concluded: (1) in comparison with normal granulocytes, granulocytes show in the active AML, hypogranularity as well as a decrease in the activity of all studied enzymes (myeloperoxidase, alkaline and acidic phosphatase and the esterases). (2) In ALL, mature granulocytes do not feature any particular differences in the above pattern as compared with normal granulocytes. (3) In AML remission, the difference between the mature granulocytes and the granulocytes in the active disease is rather slight as compared with the morphological, cytochemical an cytonzymic characteristics. The authors conclude that in a number of patients with AML and in the remitted patients mature granulocytes could possibly originate from the leukemic stem cells which acquired the ability to respond to normal CSF by manner of the gross destruction of the leukemic tumour mass.(ABSTRACT TRUNCATED AT 250 WORDS)