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Background: Excessive tidal volume, respiratory rate, and positive end-expiratory pressure (PEEP) are all potential causes of ventilator-induced lung injury, and all contribute to a single variable: the mechanical power. The authors aimed to determine whether high tidal volume or high respiratory rate or high PEEP at iso-mechanical power produce similar or different ventilator-induced lung injury., Methods: Three ventilatory strategies-high tidal volume (twice baseline functional residual capacity), high respiratory rate (40 bpm), and high PEEP (25 cm H2O)-were each applied at two levels of mechanical power (15 and 30 J/min) for 48 h in six groups of seven healthy female piglets (weight: 24.2 ± 2.0 kg, mean ± SD)., Results: At iso-mechanical power, the high tidal volume groups immediately and sharply increased plateau, driving pressure, stress, and strain, which all further deteriorated with time. In high respiratory rate groups, they changed minimally at the beginning, but steadily increased during the 48 h. In contrast, after a sudden huge increase, they decreased with time in the high PEEP groups. End-experiment specific lung elastance was 6.5 ± 1.7 cm H2O in high tidal volume groups, 10.1 ± 3.9 cm H2O in high respiratory rate groups, and 4.5 ± 0.9 cm H2O in high PEEP groups. Functional residual capacity decreased and extravascular lung water increased similarly in these three categories. Lung weight, wet-to-dry ratio, and histologic scores were similar, regardless of ventilatory strategies and power levels. However, the alveolar edema score was higher in the low power groups. High PEEP had the greatest impact on hemodynamics, leading to increased need for fluids. Adverse events (early mortality and pneumothorax) also occurred more frequently in the high PEEP groups., Conclusions: Different ventilatory strategies, delivered at iso-power, led to similar anatomical lung injury. The different systemic consequences of high PEEP underline that ventilator-induced lung injury must be evaluated in the context of the whole body.

Objective: Tidal volume (VT) and volume of gas caused by positive end-expiratory pressure (VPEEP) generate dynamic and static lung strains, respectively. Our aim was to clarify whether different combinations of dynamic and static strains, resulting in the same large global strain, constantly produce lung edema., Design: Laboratory investigation., Setting: Animal unit., Subjects: Twenty-eight healthy pigs., Interventions: After lung computed tomography, 20 animals were ventilated for 54 hours at a global strain of 2.5, either entirely dynamic (VT 100% and VPEEP 0%), partly dynamic and partly static (VT 75-50% and VPEEP 25-50%), or mainly static (VT 25% and VPEEP 75%) and then killed. In eight other pigs (VT 25% and VPEEP 75%), VPEEP was abruptly zeroed after 36-54 hours and ventilation continued for 3 hours., Measurements and Main Results: Edema was diagnosed when final lung weight (balance) exceeded the initial weight (computed tomography). Mortality, lung mechanics, gas exchange, pulmonary histology, and inflammation were evaluated. All animals ventilated with entirely dynamic strain (VT 825±424 mL) developed pulmonary edema (lung weight from 334±38 to 658±99 g, p<0.01), whereas none of those ventilated with mainly static strain (VT 237±21 mL and VPEEP 906±114 mL, corresponding to 19±1 cm H2O of positive end-expiratory pressure) did (from 314±55 to 277±46 g, p=0.65). Animals ventilated with intermediate combinations finally had normal or largely increased lung weight. Smaller dynamic and larger static strains lowered mortality (p<0.01), derangement of lung mechanics (p<0.01), and arterial oxygenation (p<0.01), histological injury score (p=0.03), and bronchoalveolar interleukin-6 concentration (p<0.01). Removal of positive end-expiratory pressure did not result in abrupt increase in lung weight (from 336±36 to 351±77 g, p=0.51)., Conclusions: Lung edema forms (possibly as an all-or-none response) depending not only on global strain but also on its components. Large static are less harmful than large dynamic strains, but not because the former merely counteracts fluid extravasation.

Rationale: Unphysiologic strain (the ratio between tidal volume and functional residual capacity) and stress (the transpulmonary pressure) can cause ventilator-induced lung damage., Objectives: To identify a strain-stress threshold (if any) above which ventilator-induced lung damage can occur., Methods: Twenty-nine healthy pigs were mechanically ventilated for 54 hours with a tidal volume producing a strain between 0.45 and 3.30. Ventilator-induced lung damage was defined as net increase in lung weight., Measurements and Main Results: Initial lung weight and functional residual capacity were measured with computed tomography. Final lung weight was measured using a balance. After setting tidal volume, data collection included respiratory system mechanics, gas exchange and hemodynamics (every 6 h); cytokine levels in serum (every 12 h) and bronchoalveolar lavage fluid (end of the experiment); and blood laboratory examination (start and end of the experiment). Two clusters of animals could be clearly identified: animals that increased their lung weight (n = 14) and those that did not (n = 15). Tidal volume was 38 ± 9 ml/kg in the former and 22 ± 8 ml/kg in the latter group, corresponding to a strain of 2.16 ± 0.58 and 1.29 ± 0.57 and a stress of 13 ± 5 and 8 ± 3 cm H(2)O, respectively. Lung weight gain was associated with deterioration in respiratory system mechanics, gas exchange, and hemodynamics, pulmonary and systemic inflammation and multiple organ dysfunction., Conclusions: In healthy pigs, ventilator-induced lung damage develops only when a strain greater than 1.5-2 is reached or overcome. Because of differences in intrinsic lung properties, caution is warranted in translating these findings to humans.

Penicillium marneffei is an intracellular opportunistic fungus causing invasive mycosis in AIDS patients. T cells and macrophages are important for protection in vivo. However, the role of T-cell cytokines in the immune response against P. marneffei is still unknown. We studied by semiquantitative reverse transcription-PCR and biological assays the patterns of expression of Th1 and Th2 cytokines in the organs of wild-type (wt) and gamma interferon (IFN-gamma) knockout (GKO) mice infected intravenously with P. marneffei conidia. At 3 x 10(5) conidia/mouse, a self-limiting infection developed in wt BALB/c mice, whereas all GKO mice died at day 18 postinoculation. Splenic and hepatic granulomas were present in wt mice, whereas disorganized masses of macrophages and yeast cells were detected in GKO mice. The infection resolved faster in the spleens than in the livers of wt mice and was associated with the local expression of type 1 cytokines (high levels of interleukin-12 [IL-12] and IFN-gamma) but not type 2 cytokines (low levels of IL-4 and IL-10). Conversely, both type 1 and type 2 cytokines were detected in the livers of wt animals. Disregulation of the cytokine profile was seen in the spleens but not in the livers of GKO mice. The inducible nitric oxide synthase mRNA level was low and the TNF-alpha level was high in both spleens and livers of GKO mice compared to wt mice. These data suggest that the polarization of a protective type 1 immune response against P. marneffei is regulated at the level of individual organs and that the absence of IFN-gamma is crucial for the activation of fungicidal macrophages and the development of granulomas.

Aims: We evaluated by immunohistochemistry HSP70 expression in reactive lymph nodes since its morphological expression and location have not been previously described and correlated with lymphocyte kinetics., Methods and Results: Ninety-six cases of non-specific lymphadenitis were immunostained for HSP70, CD20, CD3, Ki67, Bcl-2, CD21. The type and the location of HSP70-positive cells were determined. Their number out of 2000 cells in each germinal centre and in each paracortical area was counted at 60x magnification with the help of a quantitative grid. Seventeen percent of germinal centre cells and 7.6% of the paracortex cells were positive. This difference was highly significant. The positively reacting cells were B-cells and had a blast (centroblast or immunoblast) morphology, with negative mantle and marginal lymphocytes and T-cells. Lymphoplasmacytoid cells and plasma cells reacted only weakly or were negative. Germinal centre antigen-presenting cells and interdigitating dendritic cells reacted from lightly to moderately., Conclusions: HSP70 was selectively over-expressed by B-blasts mainly located within germinal centres with a lower number in the paracortex. The difference in the mean number between the two sites was statistically highly significant. No correlation was found with bcl-2 and Ki67 expression. Mantle, marginal and T-lymphocytes were always negative. The biological meaning and role of this over-expression in centroblasts and immunoblasts remain to be elucidated.

The antimicrobial activities of chloroquine (CQ) and several 4-aminoquinoline drugs were tested against Penicillium marneffei, an opportunistic fungus that invades and grows inside macrophages and causes disseminated infection in AIDS patients. Human THP1 and mouse J774 macrophages were infected in vitro with P. marneffei conidia and treated with different doses of drugs for 24 to 48 h followed by cell lysis and the counting of P. marneffei CFU. CQ and amodiaquine exerted a dose-dependent inhibition of fungal growth, whereas quinine and artemisinin were fungistatic and not fungicidal. The antifungal activity of CQ was not due to an impairment of fungal iron acquisition in that it was not reversed by the addition of iron nitrilotriacetate, FeCl3, or iron ammonium citrate. Perl's staining indicated that CQ did not alter the ability of J774 cells to acquire iron from the medium. Most likely, CQ's antifungal activity is due to an increase in the intravacuolar pH and a disruption of pH-dependent metabolic processes. Indeed, we demonstrate that (i) bafilomycin A1 and ammonium chloride, two agents known to alkalinize intracellular vesicles by different mechanisms, were inhibitory as well and (ii) a newly synthesized 4-amino-7-chloroquinoline molecule (compound 9), lacking the terminal amino side chain of CQ that assists in drug accumulation, did not inhibit P. marneffei growth. These results suggest that CQ has a potential for use in prophylaxis of P. marneffei infections in human immunodeficiency virus-infected patients in countries where P. marneffei is endemic.

Objective: In order to investigate the possible association between mast cells (MCs) and the fibrous plaque of La Peyronie's disease, the number of MCs in normal penile tissue and in the fibrous plaque was determined., Methods: The control group consisted of 5 total and 3 partial penectomies with no fibrotic lesions, while the study group consisted of 23 excisional biopsies from cases of La Peyronie's disease dating back to at least 2 years earlier and with no signs of activity. The biopsies included tissues from the tunica albuginea (TA), the areolar tissue (Br) between the tunica and the erectile tissue (CC) and from the latter. The number of MCs was counted with the aid of an image analysis program following staining the antibody antitryptase., Results: In the cases of La Peyronie's disease the number of MCs/mm(2) was significantly higher in the TA and Br but lower in the CC. The MCs were related to fibroblasts and vasculonervous channels in the TA, and were concentrated around the fibrous plaques and granulation tissue between the TA and BR and between the latter and the CC., Conclusion: Our data indicate that MCs have a role in the genesis of the fibrous plaque in the TA and in the persistent inflammation in the Br. Medical treatment aimed at repressing MC activation and proliferation locally might be useful in La Peyronie's disease.

Aims: Apoptosis in prostate cancer was evaluated after three months of combined endocrine therapy to investigate the association with tumour grade, tumour stage, and the immunohistochemical detection of p53 and bcl-2 in tumour cells before and after therapy., Methods: Twenty six formalin fixed, paraffin wax embedded core biopsies and corresponding prostatectomy specimens, excised after three months of combined endocrine therapy, were analysed for the presence of apoptotic cells by the terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labelling (TUNEL) method, and for p53 and bcl-2 overexpression by immunohistochemistry., Results: All 26 adenocarcinomas were clinically localised at diagnosis. In biopsies performed before combined endocrine therapy, the apoptotic indices varied between 0.09% and 1.73%, while the tumour grade fell between Gleason score 1 and 8. The mean (SD) apoptotic count pretherapy was 0.71% (0.50). There was a significant association between elevated apoptotic counts and higher Gleason scores in the biopsies (p = 0.005). After three months of therapy, the percentage of apoptotic tumour cells increased independently of tumour stage, while a significant association with Gleason grade was found (p = 0.0018) and all the tumours had Gleason scores of < 7. In eight cases the apoptotic index was more than twice its pretherapy value. The remaining tumours showed less of an increase in the apoptotic index (five cases) or a reduction in the percentage of apoptotic cells. The overall moderate increase in apoptotic index after combined endocrine therapy was not statistically significant (p = 0.8). Immunoreactivity to p53 was absent in all cases, before and after therapy, while a slight increase in the number of cells overexpressing bcl-2 was observed in five of the 13 tumours (38.1%) with reduced apoptotic indices after therapy., Conclusions: After three months of combined endocrine treatment a minority of clinically localised prostate neoplasms showed regressive epithelial alterations, associated with an increase in apoptotic tumour cells; an increase in cells overexpressing bcl-2 was observed in five of the 13 tumours with reduced apoptotic indices.

A histological and morphometric study was conducted on 372 placentae out of a total 440 delivered in Zanzibar. Fibrin (F), intervillous space (IVS) and Villi (V) relative volumes were determined by the point-counting system and the ratio of syncytium to blood capillaries by the linear intercept method. Parasitemia load and inflammatory reaction were graded semiquantitatively by the use of a 1 mm square grid. Parasitised red cells identified active malaria (AM), the presence of malarial pigment only identified past malaria (PM), and the absence of both characterized non-malarial placentae (NM). AM(17.87%), PM(21.61%) and NM(60.52%) placentae did not vary significantly in weight. Newborns from AM had a significantly lower weight than those from PM and NM. Peripheral and placental parasitemia were not coincident. Placental parasitemia load increased parallel with birthweight. The latter decreased with the increasing severity of the inflammation, particularly with the prevalence of lymphocytes in the IVS. Significantly increased volume of F was found in AM and PM placentae, while no significant variation was noticed in IVS and V volumes. The syncytium/capillaries ratio was significantly increased in AM. We conclude that low birthweight in malaria is linked to IVS inflammation but not to F deposits or parasitemia load. Non-leukotactic lymphokines might play some role. Morphologic aspects bespeak for a less mature placenta than expected and this might represent an adaptive change.

In urban Zanzibar, Tanzania, 389 women with full-term pregnancies were studied to see what effect their infection with malaria (at delivery) had on the birthweight (BW) of their infants. The overall prevalence of low birthweight (LBW) (i.e. < 2500 g) was 3.9% (15 out of 389). Overall, 21.3% (82/384) of the women only had peripheral parasitaemias at delivery, 17.6% (58/329) only had active placental infections and 47.9% (157/328) had both. The youngest women (< 20 years), the primiparae and those with Plasmodium infection gave birth to neonates with relatively low mean BW. The lowest mean BW (2967 g) was found among the offspring of women with active placental infection (N = 58). The women with past/chronic infection (N = 73) or no infection (N = 201) generally produced heavier infants, with mean BW of 3242 and 3338 g, respectively. The women with active placental infection were also far more likely to have babies of LBW (15.5%) than those with past/chronic infection (1.4%) or no infection (1.5%). Multivariate analysis indicated that the highest relative risk of LBW (10.1, with a 95% confidence interval of 2.9-35.4) was associated with active placental infection, with no significant difference between primiparae and multiparae. In the study population, therefore, with its low prevalence of LBW, malaria infection increased the risk of LBW in full-term neonates by about 10-fold, with a population-attributable proportion of 55.4%.

Background: The PTH-calcium sigmoidal curve is shifted to the right, the slope of the curve is steeper, and the set point of calcium is increased in dialysis patients with secondary hyperparathyroidism, compared to patients with low-turnover bone disease. These findings could be related to increased parathyroid cell mass and increased sensitivity of parathyroid cells to serum calcium variations in these patients. Calcitriol therapy has been documented to reduce PTH levels by shifting the curve to the left and downward. The effect of a surgical reduction of parathyroid gland mass on the PTH-calcium curve has not yet been investigated. In this study we compared the effects of calcitriol and subtotal parathyroidectomy (PTH) on the dynamics of PTH secretion in response to acute changes of serum calcium in two groups of dialysis patients with severe hyperparathyroidism., Methods: Fourteen dialysis patients treated for 6 months with high-dose i.v. calcitriol (1-2 micrograms thrice weekly, and 10 dialysis patients who underwent subtotal PTx were studied. The PTH-calcium relationship obtained by inducing hypo- and hypercalcaemia means of low and high calcium dialysis was evaluated before and 2-6 months after treatment., Results: Both calcitriol and subtotal PTx significantly decreased PTH (respectively from 797 +/- 595 to 380 +/- 244 and from 1036 +/- 250 to 70 +/- 34 pg/ml), as well as maximal PTH response to hypocalcaemia (PTHmax), and maximal PTH suppression during hypercalcaemia ( PTHmin). When the PTH-calcium curves were constructed using PTHmax as 100% to factor for differences in absolute PTH levels and to provide an assessment of individual parathyroid cell function, a shift of the sigmoidal curve to the left and downward, and a significant decrease in the set point of ionized calcium (from 1.31 +/- 0.05 to 1.26 +/- 0.05 and from 1.36 +/- 0.09 to 1.22 +/- 0.07 mmol/l) was documented with both treatments. However, the slope of the PTH-calcium curve increased after subtotal PTx indicating that the sensitivity of the parathyroid cell to serum calcium changes increased with PTx, while on the contrary it decreased with calcitriol., Conclusions: PTH secretion decreases proportionally more with calcitriol than with surgery for a given decrease in the functional mass of parathyroid cells. The change in the PTH-ICa sigmoidal curve induced by subtotal PTx is due to the removal of a large mass of parathyroid tissue with advanced hyperplasia.

The expression of the p53 gene product was investigated immunocytochemically in a series of 51 fine-needle aspiration (FNA) samples of breast carcinomas. Results were compared with those obtained by immunocytochemically on paraffin embedded tissue sections of the corresponding surgical specimens. Cytological samples showed a variable degree of p53 immunoreactivity in 14 tumors (27.6%), all of ductal type, while p53 immunoreactive tumor cells were present in tissue sections from 15 carcinomas (29.4%). The only discordant case was a signet-ring cell carcinoma. Abnormal p53 expression was significantly associated with high nuclear grade in ductal carcinomas. No association was seen with tumor size, lymph node status, and age of the patient. Detection of p53 altered expression in FNA samples of breast carcinoma may play a role in the assessment of tumoral grading and is predictive of p53 immunoreactivity in histological specimens.

Severe anaemia in pregnancy is a major obstetric problem in malaria-endemic areas. This study reports a cross-sectional investigation of malaria infection and haematological values at delivery in 440 women from urban Zanzibar. Severe anaemia [with haemoglobin (Hb) < or = 7 g/dl] was identified in 36 women (9.3%) and mild anaemia (7 < Hb < 10 g/dl) in 269 (69.7%) while 81 women (21.0%) had normal Hb values. Malaria infection was diagnosed in 187/385 women (48.6%) on the basis of either peripheral blood examination or placental histology. Univariate analysis indicated that the proportion of women with anaemia was similar in those with (85%) and without (80%) malaria infection. However, when primigravidae were considered alone, malaria infection was significantly more frequent among anaemic women (65.2%) than in those with normal Hb values (40.0%). In the logistic regression analysis, including age, parity, education level, malaria and free serum iron as independent variables and anaemia (Hb < or = 10 g/dl) as response variable, the odds ratio (OR) for malaria infection was 1.2 (P > 0.1). However, a similar analysis indicated that malaria was significantly associated with anaemia in the primigravidae, with an OR of 3.2 (95% confidence interval: 1.1-9.6; P < 0.05). In conclusion, this cross-sectional investigation indicated that malaria plays a significant role in the determination of anaemia in primigravidae, but not in multiparae, in the urban study area.

Three study groups of pregnant CD1 albino mice were inoculated intraperitoneally with Plasmodium berghei on the 6th (Group I), 13th (Group II) and 18th (Group III) day of gestation. Two control Groups were included, one of non pregnant mice (Group IV) and the other of pregnant non inoculated mice. Group IV was inoculated in the same day of group I. All mice of these two Groups died. Of the 20 mice in Group II 8 died, 7 delivered prematurely and 5 gave birth of low birth weight offspring. The 15 mice in Group III delivered normally with only 4 low birth weight offspring and no deaths. Histological examination revealed total placental necrosis, resorption of the products of conception and widespread foci of hepatic necrosis, malarial pigment accumulation in Group I. Such changes were also present in 8 mice of Group II, but the remainder of the Group retained the product of conception, had marked morphological changes in the placenta and moderate focal hepatic necrosis and malarial pigment accumulation. Group III had mild placental changes and moderate pigment accumulation. The products of conception displayed marked hepatic hematopoiesis in Group II while it was moderate in Group III. No parasitized red cells were observed in the fetal circulation. The stage of gestation at which the malarial infection was contracted was decisive. Malarial infection does not cross the placental barrier so its pathogenetic effects are felt in the maternal circulation with variable effects on the products of conception.

Nucleolar organizer regions (NOR) have DNA loops containing genes that encode for ribosomal RNA; some proteins located near the NORs are argyrophilic and by Ploton's one step method we could quantify the NORs in sections of paraffin embedded tissues. We have evaluated the number of intranuclear dots in 46 parathyroid glands from 18 parathyroidectomies. The glands were weighed and the stromal fat was evaluated by a point-counting method. AgNORs were counted in 100 cells from each gland. The mean count was 3.03 in six adenomas (A), 2.39 in six cases of primary hyperparathyroidism (PH) and 2.63 in six cases of secondary hyperparathyroidism (SH). Statistical analysis performed by Mann-Whitney U test showed that differences between A and PH groups were significant. The difference between A and SH was also significant. The residual parathyroid tissue surrounding the adenoma had a significantly lower AgNOR mean count than adenomatous cells. The AgNOR mean number can be useful in assessing normal or suppressed parathyroid tissue.

A total of 722 nuclei from 22 fine needle aspirates (FNA) of breast carcinomas have been subjected to quantitative nuclear analysis. Form Factor, Nuclear Area and Nuclear Major Diameter were determined for each case. An equal number of nuclei from as many aspirates from benign breast nodules was used as control. The aim of the study was to evaluate the predictive value of these parameters on the result of the immunohistochemical determination of the c-erb B-2 antigen in the tumour mass, since 6 of the 22 carcinomas were positive to this antigen. None of these parameters singly or in combination proved of any value in predicting the positive or negative result of the c-erb B-2 antigen determination. Nuclei both positive and negative for the antigen were located within the large cell carcinomas. Nucleolar Organizer Regions were also determined on the histologic sections of both groups to assess any statistically significant difference not only between controls and carcinomas, but also between c-erb B-2 positive and negative cases. Both NOR mean numbers and NOR cytograms were of no use, since no difference was found between c-erb B-2 positive and negative cases.

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.

Nucleolar organizer regions (NORs) were counted on ten cases each of normal ectocervix, CIN 1, 2 and 3 to verify the possibility of a differentiation between the various grades of CIN and between them and condylomata. Counts were performed on the full thickness of the tissue, layer by layer (stratified counts). A significant difference (p < 0.05) was found between the means of normal tissue toward condylomata and CIN 2 and 3 and between CIN 1 and CIN 2 and 3. There was no significance (p < 0.05) between normal tissue and CIN 1, between CIN 2 and 3 and between condylomata and CIN 2 and 3. The range of variations on the counts was associated with overlapping between the various cases. Our data showed also a progressive rise in mean NOR values from normal tissue to CIN 3. The stratified counts showed in all the groups a rise from basal to parabasal cells. Counts on parabasal and intermediate layers distinguished two groups of cases. In one there was either the same number of dots or a further rise from one layer to the next, while in the other a definite decrease was seen. The former pattern may be related to a potential for malignant evolution of the lesion. NORs should be counted in all cases of CIN and condylomata to treat more aggressively those lesions which present the pattern of a progressive rise of NORs from basal to intermediate cells.

Nucleolar organiser regions (AgNOR) are chromosomal segments in which ribosomal RNA is codified. These structures can be identified in the nuclei of cells in routine histological sections using a silver staining technique which reveals the proteins associated with transcription RNA. The number of AgNOR identified depends on the state of cell proliferation. The purpose of the present study was to evaluate the distribution of AgNOR in normal laryngeal mucosa, dysplastic and neoplastic, in order to establish a possible correlation with the degree of dysplasia and/or the entity of the malignant lesions. Ten slides each of specimens of laryngeal carcinoma, serious dysplasia and middle dysplasia were prepared with the silver staining technique. A count of the NOR in the basal, parabasal, intermediate and superficial levels was made. Findings show an increase in the average value of NOR proceeding from normal tissue to carcinoma in situ and malignant carcinomas. The number of AgNOR was lower always than that observed in the parabasal level. In the intermediate cell section the number of AgNOR was inferior to that in the parabasal level in normal tissue as well as in middle dysplasia, while it remained high in serious dysplasia and in carcinomas, thus indicating a great deal of biological activity in these cases. This method can be employed in order to better individualize those dysplastic lesions which, because of their tendency towards malignancy, must be kept under observation by means of an extremely accurate follow-up.

Nucleolar organizer regions (NORs) were counted on normal tissue, condylomata, CIN 1, 2 and 3, to verify the possibility of a differentiation between the various grades of CIN and between them and condylomata. Counts were performed on the full thickness of the tissue, layer by layer (stratified counts). A significant difference (p < 0.05) was found between the mean of normal tissue in relation to condylomata and CIN 2 and 3 and between CIN 1, and CIN 2 and 3. There was no significance (p > 0.05) between normal tissue and CIN 1, between CIN 2 and 3 and between condylomata and CIN 2 and 3. The range of variations in the counts was associated with overlapping between the various cases. Our data showed also a progressive rise in mean NOR values from normal tissue to CIN 3. The stratified counts showed in all the groups a rise from basal to parabasal cells. Counts on parabasal and intermediate layers distinguished two groups of cases. In one there was either the same number of dots or a further rise while in the other a definite decrease was seen. The former pattern may be related to a potential for malignant evolution of the lesion. NORs should be counted in all cases of CIN and condylomata to treat more aggressively those lesions which present the patterns of a progressive rise of NORs from basal to intermediate cells.

The authors have evaluated in eight reactive and eight malignant pleural effusions the number of intranucleolar dots representing the nucleolar associated proteins stained with silver colloid technique (interphase nucleolar organizer regions-AgNOR). The mean number per nucleus in benign reactive mesothelium was significantly lower than in mesothelioma effusions. Other morphometric measurements of nuclear diameters and areas were effected. The authors evaluate the variability of the AgNOR counts and morphology in reactive and mesothelioma cells of pleural fluids.

Fifty bioptic endomyocardial samples from 15 patients with idiopathic dilatative cardiomyopathy have been stained with PicroSirius, Masson-Goldner, Van Gieson Verhoeff, AFOG. The extension of stained areas has been evaluated by morphometry using a videoscan. As control sections samples have been taken from lower third of the right ventricle at autopsies of healthy subjects dying in road accidents. Morphometric analysis of subendocardial interstitial and perivascular fibrosis has been performed. For the authors the most useful stain in the evidentiation of fibrosis was PicroSirius both in the bioptic samples and the autoptic ones.

Indirect immunoperoxidase with antichlamydia serum has been applied to 34 Pap-smears where cytologic diagnosis of Chlamydia infection was suspected. Only 4 cases were confirmed by this technique. The limited success could be due to the need for a more active antiserum or to an overestimation of Pap-smear images. Cytologic diagnostic features are critically outlined.