Brain stem lesions in the sudden infant death syndrome: variability in the hypoplasia of the arcuate nucleus.

In the present study we investigated quantitatively the incidence of hypoplasia of the arcuate nucleus (ARCn) of the medulla oblongata, reported earlier [Gozal D, Hathout GM, Kirlew KAT (1994) J Appl Physiol 76:207], as well as its distribution in 62 cases of sudden infant death syndrome (SIDS; mean age 14 postnatal weeks, 39 male and 23 female) and 25 controls (mean age 16 postnatal weeks, 14 male and 11 female), using detailed histopathological and morphometric analyses performed on serial sections of medulla oblongata. The SIDS cases were divided into four subtypes: SIDS A (27 cases, 43%) with histologically well-developed ARCn; SIDS B (16 cases, 26%) with severe bilateral hypoplasia along the whole length; SIDS C (11 cases, 18%) with partial bilateral hypoplasia, located mainly in the lateral portions of the caudal two thirds of the nucleus, and SIDS D (8 cases, 13%) with right monolateral hypoplasia of the ARCn. ARCn hypoplasia was detected in 56% of cases (35 cases). Three-dimensional volume reconstruction showed that in the SIDS A victims the mean volume was analogous to controls, whereas in the SIDS group with ARCn hypoplasia, severe or partial, the mean volume was significantly different from controls on both sides of the medulla oblongata (SIDS B group: P=0.003, P=0.002; SIDS C group: P=0.007, P=0.008). The mean ARCn volume in the SIDS D group was statistically significant only on the right side ( P=0.005). We also observed reduced neuron density of the ARCn, associated with a decrease in the total number of neurons over the whole length of the nucleus itself. On the basis of the morphometric results of neuronal population in the different portions of the ventrolateral medulla in SIDS cases, we hypothesized that infants without the full complement of neurons and neuropil (ARCn hypoplasia) are at risk for SIDS because they are unable to develop appropriate cardioventilatory control during this crucial developmental period.

The role of different biomarkers (DNA, PCNA, apoptosis and karyotype) in prognostic evaluation of superficial transitional cell bladder carcinoma.

BACKGROUND: In order to clarify the variable behaviour of transitional cell bladder carcinomas (TCBC) with same clinico-pathologic pattern, we investigated the prognostic significance of various biomarkers (PCNA, DNA, apoptosis, karyotype). MATERIALS AND METHODS: We studied 177 superficial TCBC (stage T1) undergoing transurethral resection (TUR). Analysis of biological indicators was performed on serial paraffin sections: DNA by static cytometry, karyotype by fluorescence in situ hybridisation (FISH), PCNA and apoptosis by immunohistochemistry. RESULTS: The most salient results are represented by prevalence of diploidy (56%), a mean PCNA labeling index (PCNA-LI) of 8.2%, nonrandom trisomies and tetrasomies of chromosome 7, and a high presence of apoptosis (in 72% of cases). These data were not related to histological grading. Multivariate analysis showed that only PCNA-LI is a an independent prognostic marker for patient survival (p = 0.01). Besides, we observed a worse prognosis in the presence of both very high PCNA indices and low or absent apoptosis. CONCLUSIONS: The results of this study suggest that tumor prognostic potential in TCBC should be evaluated on the basis of the association between PCNA cell kinetics and cell apoptosis information.

Prolonged QT interval and sudden infant death--report of two cases.

In the two cases where infants died suddenly and unexpectedly the electrocardiogram (ECG) of a younger sibling (case 1) and of a living twin (case 2) led to the suspicion that the two infants could have died from long QT syndrome (LQTS). In case 1, a His bundle (HB) dispersion and a pronounced hypoplasia of the right external nucleus arcuatus were detected. In case 2, a severe interstitial pneumonia and an accompanying mild myocarditis were found by histology. Molecular genetic investigations of the coding regions of the genes, HERG, KVLQT1 and SCN5A gave no indication for the mutations, thus, affecting related myocardial ion channels as possible sources of inhomogeneity of repolarisation. Since a molecular genetic deviation could not yet be elaborated the possible role of related disturbance remains unknown.

Severe hypoplasia of medullary arcuate nucleus: quantitative analysis in sudden infant death syndrome.

The human arcuate nucleus (ARCn) is postulated to be homologous to ventral medullary cells involved in chemoreception, and respiratory and blood pressure responses. Abnormalities in central respiratory control may result from dysfunction of this anatomic ventral area. We evaluated the changes of the neuronal population of the medullary ARCn in infants victims of the sudden infant death syndrome (SIDS). In this study we tested the hypothesis that anatomical deficiency of the ARCn is associated with SIDS. The volume and neuronal density of the ARCn were morphometrically quantified with an image analyzer in 36 cases of SIDS and 12 age-matched controls. We found a marked hypoplasia in the SIDS ARCn compared to controls and, particularly, in 11 SIDS cases (30%) in which the ARCn exhibited a severe hypoplasia, being almost totally absent. Three-dimensional reconstructions and morphometric measurements of ARCn confirmed this marked hypoplasia in all the serial sections examined (P = 0.0001) and the reduced neuronal density (P = 0.0025) in relation to control cases. In conclusion these abnormalities observed in the ARCn are consistent with the idea that ARCn dysfunction plays an important role among the causative factors of sudden infant death. The hypoplasia of the ARCn represents the most frequent congenital abnormality in our experience, and can be a plausible morphological substrate for a subset of SIDS.

Prognostic significance of different biomarkers in non-small cell lung cancer.

The purpose of our study was to examine the prognostic significance of different biomarkers [DNA content, proliferating cell nuclear antigen labeling index (PCNA-LI), p53 mutation and apoptosis], in 152 surgically resected non-small cell lung cancer (NSCLC). The ploidy was carried out by densitometry; PCNA-LI, p53 and apoptosis were determined with immunohistochemistry. The results were correlated to histology, stage and patient survival. A considerable variability of the PCNA indices, ranging from 0 to 33.5% with a mean value of 7.0%, was found. DNA evaluation showed a prevalence of aneuploid tumors (62%) with a DNA index >1. Overexpression of p53 protein and apoptotic positivity were observed in low percentages of cases (16% and 32% respectively). Only stage and PCNA-LI were found to be significant prognostic factors on multivariate analysis. PCNA was superior to stage in predicting shortened survival of patients with NSCLC. PCNA immunostaining can be applied on a routine basis in formalin-fixed, paraffin-embedded samples of NSCLC to predict patient prognosis and thus to identify patients in need of additional postoperative therapies.

Hyperplasia of the aorticopulmonary paraganglia: a new insight into the pathogenesis of sudden infant death syndrome?

In this study, the Authors search for morphologic alterations in the aorticopulmonary paraganglia (APP) of sudden infant death syndrome (SIDS) victims when compared with age-matched controls. Morphometric studies on serial sections of the APP were performed with an image analyzer in combination with a standard microscope attached to a video camera. APP hyperplasia, characterized by an increase in some parameters such as number, mean lobule diameter and total glomic tissue volume when compared with age-matched controls, was observed in 23.8% of SIDS victims. Similar alterations have been reported in peripheral chemoreceptors of animals and human beings who are chronically hypoxemic. In SIDS, it could reflect an abnormal chemoreceptor function, contributing to an altered respiration control.

Prognostic significance of different biological markers (DNA index, PCNA index, apoptosis, p53, karyotype) in 126 adenocarcinoma gastric biopsies.

BACKGROUND: The different clinical evolution of gastric adenocarcinomas with the same clinico-pathologic characteristics prompted the authors to investigate the prognostic significance of different biological markers. METHODS: One hundred twenty-six preoperative cancer gastric biopsies, selected according the stage evaluated after following gastrectomy, were examined for DNA content by means the static cytometry, proliferating cell nuclear antigen (PCNA), p53 mutation, apoptosis by immunohistochemistry and karyotype using fluorescence in situ hybridization (FISH) techniques. RESULTS: The gastric cancers, ail belonging at stage III, were adenocarcinomas histologically typed as: 26 well differentiated (G1), 45 moderately differentiated (G2), 43 poorly differentiated (G3) and 12 of undifferentiated type with signet-ring cells. The tumours showed a prevalence of diploidy (68%), a mean PCNA-LI of 4.8%, trisomy of chromosome 7 in 40% of the cases, low presence of apoptosis (30%) and p53 mutation (17%). Only apoptosis was significantly correlated to histological diagnosis (p = 0.009). A multivariate analysis showed that the DNA content and PCNA-LI were the only independent prognostic markers for survival (p = 0.005 and p = 0.0002 respectively). CONCLUSIONS: The evaluation of these two biological variables, especially of the PCNA index, on gastric cancer biopsies may be useful in predicting the aggressiveness of each tumor and in identifying patients in need of additional perioperative therapies.

Detection of trisomy 7 with fluorescence in situ hybridization and its correlation with DNA content and proliferating cell nuclear antigen-positivity in prostate cancer.

The authors applied fluorescence in situ hybridization (FISH), using specific DNA probes for chromosome 7, to routine paraffin-embedded tissue sections obtained from 35 radical-prostatectomy specimens. Proliferative activity was also evaluated using static cytometry to assess DNA content and immunohistochemistry for proliferating cell nuclear antigen (PCNA) positivity. These results were correlated with each other and with the morphologic parameters. The presence of three or more of chromosome 7 was observed in 71% of the cases, as was a high S phase, with a triploid prevalent DNA content and a PCNA index above mean value in 66% of the cases. No correlation was detected between these findings and histologic grade; conversely, there was a significant correlation with stage (chi-square = 5.33; p = 0.021). From these results, the authors maintain that the presence of an extra chromosome 7-correlated in most cases with an increase in cell kinetics and an advanced stage-may be an additional prognostic marker of aggressive behavior.

Discrete T-lymphocytic leptomeningitis of the ventral medullary surface in a case of Sudden Unexpected Infant Death.

The Authors report the case of a 4 month old female infant dying suddenly and unexpectedly. At post-mortem examination, nothing significant was found, except a focal T-lymphocytic brainstem leptomeningitis, involving the ventral medullary surface (VMS) particularly coincidental with the pyramids, with involvement of the outer layer of the nucleus arcuatus (NARC). Moreover, the NARC itself presented, on serial sections, interdigitated subdivisions. To the present authors' best knowledge, no case has been recordered in the literature, as yet, exhibiting VMS inflammation, evidently acquired (likely viral) in nature, associated to a NARC developmental defect, altogether bespeaking for the etio-pathogenic importance of such a combined pathology of the central chemosensitive field among the long debated mechanisms of SIDS.

Significance of the DNA synthesis in hypertrophic cardiomyopathies.

DNA content and proliferating cell nuclear antigen (PCNA) expression were investigated in normal hearts, in hypertrophic from hemodynamic overload hearts and in hypertrophic cardiomyopathy (HCM). The aim of this study was mainly to determine whether the hyperdiploid myocardial cells in all cases are in dynamic or static phase. The percentage of PCNA positive cells only in the HCM group was significantly higher (mean value = 25.4%) than the percentage of hyperdiploid cells (mean value = 9.3%). Therefore, the DNA replication occurs through a different process from that of normal cell cycle which lead to an increase in ploidy and eventually mitosis. These data should be interpreted not only as the result of a periodic amitotic DNA renewal and not even as the result of an increased apoptosis, but especially as a repair process of the DNA molecules affected by a various types of damages in HCMs.

DNA content and thymidine labeling index correlate with prognosis in squamous cell carcinoma of the larynx.

The prognostic value of proliferative activity was determined in 48 cases of squamous cell carcinoma of the larynx. Tumor samples were obtained during surgery: DNA content was determined by static cytometry with an image analysis program, and cell kinetics were analyzed by the in vitro thymidine labeling index (T-LI). The DNA content analysis visualized a diploid cellular population in 32 cases and an aneuploid cellular population in 16 cases. The DNA content was not significantly related to the sex and the age of the patients or the TN stage of the tumors, but to histological grade (p = 0.03). Similarly significant correlation was found between T-LI and age of the patients, indeed the median value was higher in patients older than 59 (p = 0.024). T-LI was not related to the sex of the patients, or to the histological grade or the TN stage of the tumors. In multivariate analysis, using the Cox model, T-LI was the most important prognostic parameter (p = 0.015), superior to DNA content (p = 0.021) and to TN stage (p = 0.047; p = 0.043). From the data obtained in this study it can be concluded that evaluation of DNA content and thymidine labeling index in squamous cell carcinoma of the larynx may be new independent parameters of prognosis and may contribute to optimal and individualized therapy.

Thymidine labeling index in colorectal carcinoma: relation to clinicopathological parameters and to mitotic index.

Two cell-kinetic parameters, the thymidine labeling index (T-LI) and the mitotic index (MI) were determined for a prospective series of 74 primary colorectal cancers. Dukes' stages were used to define the extent of the tumor: 10 tumors were classified as Dukes' A, 36 as Dukes' B, and 22 as Dukes' C. Of these tumors, 14 were well-differentiated, 44 moderately differentiated, and 15 poorly differentiated adenocarcinomas. The H3dT-LI values obtained varied widely, from 2.5 to 38.9% (median value: 16.8%). The MI shows a range of from 0 to 13.2% with a median value of 3.2%. The T-LI values were independent of the sex and age of the patients and of the histological grading and Dukes' stage. Conversely, the MI was related to histological grading. This study indicates that the T-LI, whose application has reached the gold standard, still represents an independent parameter and the most sensible, objective, and specific complementary method for the analysis of the S phase.

Cell kinetics of pleomorphic adenomas of the parotid gland.

The aim of the present study is to characterise the cell kinetics of pleomorphic adenoma of the parotid gland by assessing DNA content and proliferating cell nuclear antigen (PCNA) positivity. In 22 parotid adenomas, DNA content was measured by densitometry in histological serial sections stained with Feulgen's method and PCNA positivity was determined by immunohistochemistry with the monoclonal antibody PC10. To assess the proliferative activity, DNA index and PCNA index were evaluated. It was possible to distinguish two types of adenoma. In Group I there was a prevalence of diploid cells with a low PCNA index. Group II is represented by adenomas with a large percentage of triploid cells and a PCNA index significantly higher than that of Group I. Our findings suggest that the possibility of recurrence or malignant transformation depends on intrinsic biological properties of each adenoma.

PCNA--a cell proliferation marker in vocal cord cancer. Part II: Recurrence in malignant laryngeal lesions.

Laryngeal squamous cell carcinoma constitutes the most frequent carcinoma found in the head and neck region. A precise prediction for recurrence potential cannot be done on site, treatment and histologic grading. Since Proliferating Cell Nuclear Antigen (PCNA) and DNA-cytometry have shown a good correlation between premalignant lesions and their progressive potential towards full-fledged carcinoma in the larynx as described in part I of this work, we have analyzed the PCNA index and DNA cytometry in specimen taken from vocal chord carcinomas with a 5-year follow-up, in order to assess its relationship with the presence or absence of tumour progression. 42 cases with (21) and without (2) recurrence have been examined. The DNA-index ranged from 1.01 to 1.43 (mean 1.10) in the group without and from 1.02 to 1.59 (mean 1.38) in the group with recurrent carcinoma (p = 0.002). The PCNA-index ranged from 0.00% to 18.90% (mean 6.97%) in the nonrecurrent group and from 0.00 to 3g.50% (mean 16.35%) in the patients with recurrence (p = 0.001). Both indices also correlated in a highly significant way. From these data emerges a highly significant correlation between the cytometric indices of cell proliferation and PCNA immunostaining. Furthermore the high correction between PCNA and DNA-index is of special interest for single case assessment. High DNA aberration and PCNA-index in vocal chord carcinoma may indicate a higher cellular aggressiveness of the tumour, resulting in a greater overall risk of metastases and local recurrences. Our results support the thesis that the indices of cellular proliferation within some cancers can define subsets of patients of high risk and help in isolating a population in which a more aggressive clinical protocol may be proposed.

Densitometry of Pautrier's microabscess cells in cutaneous T cell lymphoma.

BACKGROUND: Epidermotropic lymphoid T cell infiltrates are part of a continuous spectrum of lesions ranging from "benign" parapsoriasis to frank cutaneous T cell lymphoma (CTCL, mycosis fungoides). Either the clinical or histologic differentiation between these entities prove often difficult and the prognosis may be difficult to assess. PATIENTS AND METHODS: We studied 15 patients, men aged 50 to 81 years, mean +/- SD 68 +/- 12 years, with patch-plague CTCL. Sections from punch biopsies from active lesions were stained with Feulgen reagent, coupled with a "twin" adjacent hematoxylin and eosin (H&E)-stained section and analyzed with a VIDAS Zeiss-Kontron Image Analyzer. At least 50 dermal infiltrating cells and 50 epidermotropic Pautrierian cells per specimen were counted and the biopsies were repeated periodically. RESULTS: Nine patients with aneuploid Pautrierian cell DNA patterns dis well after conventional phototherapy (dermal cell ploidy was irrelevant), whereas six patients with euploid Pautrierian cell DNA patterns had to be treated aggressively (IFN + retinoids, COP). Clinical and histopathologic aspects of the first group were comparable to those of the second group. CONCLUSIONS: The classical cytophotometric aphorism seems to be reversed in this sample: "The more abnormal the ploidy of epidermotropic Pautrierian cells, the better the prognosis." Euploid epidermotropic cell prove more efficient in invading the skin and other areas and this efficiency may be reflected in more aggressive trends in the evolution of the lymphoma. Thus, DSM analysis of epidermotropic cells could prove very useful as an inexpensive tool for routine CTCL grading.

PCNA--a cell proliferation marker in vocal chord cancer. Part I: Premalignant laryngeal lesions.

Laryngeal hyperkeratotic lesions can progress to fully developed malignant carcinoma in some cases. These premalignant lesions are proliferative disorders whose potential for further tumour progression is perhaps difficult to assess by mere histology. Immunostaining with PCNA, a protein correlated with cell proliferation, has been used to study tissue behavior in 30 cases of premalignant laryngeal vocal chord lesions treated by epithelial stripping in microlaryngoscopy, 15 of whom had no progression and 15 had recurrence and final development of full malignancy. The results showed a statistically significantly higher PCNA-index in the cases which underwent further tumour progression towards malignancy. PCNA testing may thus be suggested as a marker for tumour progression potential and help in determining clinical treatment choices.

Morphometric and densitometric approach in hypertrophic cardiomyopathy (HCM).

The aim of this study was to apply an easy method for the quantitative evaluation of changes in myocardial fibrocell size (area, diameter, circular shape factor, nuclear area, DNA content), and in the fibrous-interstitial area in hypertrophic cardiomyopathy (HCM) by mean of a computerized image analysis. A Vidas image analyser was employed for the morphometric study. The following parameters were assessed: area, maximum and minimum diameters, circular shape factor of myocytes; percentage of fibrosis; DNA content (integrated optical density) nuclear area of myocytes. The morphometric results in HCM compared to the ones in normal hearts, indicate an increase in the myocyte area and the transverse diameters, especially in the septum and left ventricle, altogether illustrating the hypertrophic condition of the myocytes, and increased fibrotic area was found in the left ventricular wall and septum (22.6 +/- 2.1% and 15.9 +/- 2.9% respectively). Densitometric analysis showed a significant increase in all test samples then compared with controls. The increase in the two nuclear parameters (area and DNA) also suggests hyperplasia. It is concluded that the morphometric determination of the morphologic abnormalities occurring in HCM by a rapid and less laborious approach, is an extremely useful method to describe the characteristics of this pathologic condition.

Glomerular morphometry of twenty-three biopsied patients with IgA nephropathy.

The aim of the present study was the process of an easy method for quantitative evaluation of the alterations of the mesangial matrix, cellularity and urinary space in IgA glomerulonephritis (IgA-GN) by the image analysis technique. A Vidas (Kontron-Zeiss) image analyzer was employed for the morphometric study of renal biopsies from 23 patients with IgA-GN. The following parameters were assessed: mesangial matrix index, expressed as the ratio between mesangial area and total glomerular area x 100; mesangial cellularity index, considered as the ratio between the total number of nuclei contained in the glomerulus and the overall glomerular area in mm2 x 10(4); urinary space/glomerular area ratio. The morphometric results compared to the respective values observed in normal glomeruli indicate an increase in the overall area of glomeruli with IgA-GN and, conversely, a decrease in the total area occupied by the urinary space (p = 0.0001). It can therefore be concluded that the morphometric determination of the morphologic abnormalities occurring in the renal glomerulus with IgA-GN is an extremely useful method to describe the characteristics of this pathologic condition with a rapid and less laborious approach.

[Exercise rhinomanometric test in the diagnosis of vasomotor rhinopathy]. / Il test da sforzo nella diagnosi di rinopatia vasomotoria.

The Authors examined 55 patients divided into two groups, one of 30 vasomotor rhinopatic patients and the other of 25 normal subjects. The aim of the study was to set up an objective diagnostic rhinomanometer test through observation and evaluation of reaction brought about by physical exertion following an exertion test of 5 minutes which employed a computerized climb simulator. Nasal resistance was measure before and immediately after exertion and then at intervals of 10, 20 and 50 minutes. Our data reveal a noteworthy decrease in nasal resistance brought about by the sympathetic nervous system in normal and rhinopathic subjects. Return to pre-exertion values, however occurred difference in the two groups: in the control group nasal resistance returned to pre-exertion values slowly, while in the rhinopathic group, it decreased rapidly and a rebound effect was observed. In vasomotor rhinopathy the orthosympathetic system is overcome by the para sympathetic system with regard to nasal vascularization control and, therefore, in rhinopatic patients exertion stimulates efficacions activation of the orthosympathetic system a phenomenon which does not occur in healthy subject. On the basis of our results an intensive stimulus such as physical exertion seems to have a different effect on the two groups suggesting that the vessels of nasal mucosa do not always respond in the same way, and that in vasomotor patients there is an alternate reactivity of neural control of nasal mucosa. The rhinomanometric exercise test seems to be useful in revealing this phenomenon and to be diagnostically reliable. The method is easily reproducible, non-invasive and can be usefully applied in clinical practice associated with the normal rhinomanometric challenge test.