Lipoteichoic acid stimulates the proliferation, migration and cytokine production of adult dental pulp stem cells without affecting osteogenic differentiation.

AIM: To model in vitro the contact between adult dental pulp stem cells (DPSCs) and lipoteichoic acid (LTA), a cell wall component expressed at the surface of most Gram-positive bacteria. METHODOLOGY: Human DPSCs obtained from impacted third molars were cultured and exposed to various concentrations of S. aureus LTA (0.1, 1.0 and 10 µg mL-1 ). The effects of LTA on DPSCs proliferation and apoptosis were investigated by MTT assay and flow cytometry. Mineralization of DPSCs was evaluated by alizarin red staining assay. Migration was investigated by microphotographs of wound-healing and Transwell migration assays. Reverse transcription polymerase chain reaction was used to examine the effects of LTA on p65 NF-κB translocation and TLR1, TLR2 or TLR6 regulation. Enzyme-linked immunosorbent assay was used to investigate LTA-stimulated DPSCs cytokine production. One-way or two-way ANOVA and Tukey post hoc multiple comparison were used for statistical analysis. RESULTS: DPSCs expressed TLR1, TLR2 and TLR6 involved in the recognition of various forms of LTA or lipoproteins. Exposure to LTA did not up- or down-regulate the mRNAs of TLR1, TLR2 or TLR6 whilst LPS acted as a potent inducer of them [TLR1 (P ≤ 0.05), TLR2 (P ≤ 0.001) and TLR6 (P ≤ 0.001)]. Translocation of p65 NF-κB to the nucleus was detected in LTA-stimulated cells, but to a lesser extent than LPS-stimulated DPSCs (P ≤ 0.001). The viability of cells exposed to LTA was greater than unstimulated cells, which was attributed to an increased proliferation and not to less cell death [LTA 1 µg mL-1 (P ≤ 0.001) and 10 µg mL-1 (P ≤ 0.01)]. For specific doses of LTA (1.0 µg mL-1 ), adhesion of DPSCs to collagen matrix was disturbed (P ≤ 0.05) and cells enhanced their horizontal mobility (P ≤ 0.001). LTA-stimulated DPSCs released IL-6 and IL-8 in a dose-dependent manner (P ≤ 0.0001). At all concentrations investigated, LTA did not influence osteogenic/odontoblastic differentiation. CONCLUSIONS: Human DPSCs were able to sense the wall components of Gram-positive bacteria likely through TLR2 signalling. Consequently, cells modestly proliferated, increased their migratory behaviour and contributed significantly to the local inflammatory response through cytokine release.

Between history and gossip: the state of women relative to syphilis, a traditionally male disease.

Despite the new World Health Organization data remind us how syphilis is a disease which affects both sexes equally, this pathology has always been mainly considered a male disease. While several famous men are known to be affected by syphilis, there are very few women affected by this pathology of which we have historical records. Through the lives of Mary Todd Lincoln, Catherine of Aragon, Karen Blixen and Florence Foster Jenkins, this article would like to grant dignity to all the women who contracted syphilis in the wrong historical era where, for a woman, admitting to contracting it was shameful. Through the important women who lived their lives fighting against this disease and its complications, we would like to pay tribute to all those women who still today, in an era of antibiotics and where syphilis is a treatable pathology, they cannot be cured because they are both geographically and socially disadvantaged.

A short novel about the spread of two important diseases in history: syphilis and SARS.

Throughout the centuries, the undefended nature of man has very often manifested in cases of epidemics, which have determined scenarios of suffering and death. Through "historical" observation lens, it is possible to understand the relationship between the spread of viruses, bacteria and parasites responsible for these epidemics and the displacement of ancient or contemporary travellers. We have analysed two infective diseases, syphilis and SARS, and their role in history and in medicine.

Multifocal phaeohyphomycosis caused by Exophiala xenobiotica in a kidney transplant recipient.

In recent years, black fungi have been increasingly reported as causing opportunistic infections after solid organ transplantation. Here, we report a case of insidious, relentless, and multifocal Exophiala xenobiotica infection in a kidney transplant recipient that eventually required multiple surgical excisions along with oral and intravenous antifungal combination therapy using liposomal amphotericin B and posaconazole. We compare the present case with all previously reported cases of Exophiala infection after kidney transplantation.

Severe obstructive sleep apnoea syndrome and erectile dysfunction: a prospective randomised study to compare sildenafil vs. nasal continuous positive airway pressure.

BACKGROUND: A high incidence of erectile dysfunction (ED) among patients with obstructive sleep apnoea syndrome (OSAS) has been reported, with a strong correlation between obstructive sleep apnoea, ED, and quality of life (QOL), and it has been estimated that 10-60% of patients with OSAS suffer from ED. In this prospective randomised controlled trial, we investigated 82 men with ED consecutively who were referred to the outpatient clinic for sleep disorders and had severe OSAS (AHI> 30 events/h) without any other comorbidities as a possible cause of ED. The aim of this study was to evaluate and compare the efficacy of sildenafil vs. continuous positive airway pressure (CPAP) in men with ED and severe OSAS. METHODS: Eighty-two patients were randomised to two main treatment groups: group 1 patients (n = 41) were treated with 100-mg sildenafil 1 h before sexual intercourse without CPAP, and group 2 patients (n = 41 men) were treated with only nasal CPAP during night time sleep. Both groups were evaluated with the same questionnaires (International Index of Erectile Function-EF domain; Sex Encounter Profile; Erectile Dysfunction Inventory Treatment Satisfaction) 12 weeks after treatment. RESULTS: In patients receiving sildenafil treatment, 58.2% of those who attempted sexual intercourses were successful compared to 30.4% in the CPAP group. The mean number of successful attempts per week was significantly higher in the sildenafil group compared with the CPAP group (2.9 vs. 1.7, respectively; p < 0.0001). The mean IIEF-EF domain scores were significantly higher in the sildenafil group compared with the CPAP group (p < 0.0001). The overall satisfaction rate was 68% with sildenafil treatment and 29% with CPAP treatment. CONCLUSIONS: This study confirms that severe OSAS is strongly associated with erectile dysfunction. CPAP and sildenafil (100 mg) are safe and effective therapies for OSAS-related ED patients. In the present study sildenafil was more effective than CPAP in treating ED associated with OSAS, as indicated by a significantly higher rate of successful attempts at intercourse and higher IIEF-EF domain scores. Our study, to date, is the only that has investigated sildenafil in patients with severe OSAS.

Clinical variables associated with late-onset thrombotic and cardiovascular events, after SARS-CoV-2 infection, in a cohort of patients from the first epidemic wave: an 18-month analysis on the "Surviving-COVID" cohort from Bergamo, Italy.

Importance: Population studies have recorded an increased, unexplained risk of post-acute cardiovascular and thrombotic events, up to 1 year after acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Objectives: To search for clinical variables and biomarkers associated with late post-acute thrombotic and cardiovascular events after SARS-CoV-2 infection. Design: Retrospective cohort study. Setting: Third-level referral hospital in Bergamo (Italy). Participants: Analysis of an existing database of adult patients, who received care for SARS-CoV-2 infection at our institution between 20 February and 30 September 2020, followed up on a single date ("entry date") at 3-6 months. Exposure: Initial infection by SARS-CoV-2. Main outcomes and measures: Primary outcome: occurrence, in the 18 months after entry date, of a composite endpoint, defined by the International Classification of Diseases-9th edition (ICD-9) codes for at least one of: cerebral/cardiac ischemia, venous/arterial thrombosis (any site), pulmonary embolism, cardiac arrhythmia, heart failure. Measures (as recorded on entry date): history of initial infection, symptoms, current medications, pulmonary function test, blood tests results, and semi-quantitative radiographic lung damage (BRIXIA score). Individual clinical data were matched to hospitalizations, voluntary vaccination against SARS-CoV-2 (according to regulations and product availability), and documented reinfections in the following 18 months, as recorded in the provincial Health Authority database. A multivariable Cox proportional hazard model (including vaccine doses as a time-dependent variable) was fitted, adjusting for potential confounders. We report associations as hazard ratios (HR) and 95% confidence intervals (CI). Results: Among 1,515 patients (948 men, 62.6%, median age 59; interquartile range: 50-69), we identified 84 endpoint events, occurring to 75 patients (5%): 30 arterial thromboses, 11 venous thromboses, 28 arrhythmic and 24 heart failure events. From a multivariable Cox model, we found the following significant associations with the outcome: previous occurrence of any outcome event, in the 18 months before infection (HR: 2.38; 95% CI: 1.23-4.62); BRIXIA score ≥ 3 (HR: 2.43; 95% CI: 1.30-4.55); neutrophils-to-lymphocytes ratio ≥ 3.3 (HR: 2.60; 95% CI: 1.43-4.72), and estimated glomerular filtration rate < 45 ml/min/1.73 m2 (HR: 3.84; 95% CI: 1.49-9.91). Conclusions and relevance: We identified four clinical variables, associated with the occurrence of post-acute thrombotic and cardiovascular events, after SARS-CoV-2 infection. Further research is needed, to confirm these results.

The first-generation phosphodiesterase 5 inhibitors and their pharmacokinetic issue.

BACKGROUND: Erectile dysfunction (ED) is a relatively frequent disease that negatively impacts the overall quality of life, well-being, and relationships. Although the use of phosphodiesterase 5 inhibitors (PDE5is) has revolutionized the treatment of ED, a high percentage of ED patients discontinue PDE5i treatment. OBJECTIVES: (i) To analyze the reasons for patient dissatisfaction leading to PDE5i discontinuation; (ii) analyze the pharmacokinetics of new formulations focusing on the time needed to reach an effective plasma concentration of PDE5is (Tonset ) following drug intake; and (iii) summarize the physicochemical properties of sildenafil to understand which excipients may increase the absorption rate. MATERIAL AND METHODS: An online PubMed literature search was conducted to identify English language publications from inception to January 2019. RESULTS: The main reasons for patient dissatisfaction when using PDE5is on demand are the relatively long Tonset after taking vardenafil and sildenafil, including formulations such as film-coated tablets, fine granules, orally disintegrating tablets (ODTs), and oral thin films (ODFs). The relatively long Tonset , further worsened when accompanied by eating, highlights the following: (i) the need for planning intercourse, determining partner-related issues; (ii) issues when having sex before the maximum effect of the drug; and (iii) lower drug-related placebo effects. Some data suggest that sildenafil is a 'difficult' molecule, but Tonset can be improved following absorption by buccal mucosa using appropriate excipients. CONCLUSIONS: We conclude that several ODT and ODF formulations can improve the 'discretion' issue because they are taken without water, but they have similar pharmacokinetics to corresponding film-coated tablet formulations. One ODF formulation of sildenafil was characterized by a shorter Tonset and could potentially increase patient satisfaction following treatment. However, more clinical studies are needed to confirm the findings. Surfactants and ascorbic acid appear to be crucial excipients for achieving a high absorption rate, but more studies are needed.

Evaluation of intralesional injection of hyaluronic acid compared with verapamil in Peyronie's disease: preliminary results from a prospective, double-blinded, randomized study.

Several intralesional therapeutic protocols have been proposed for the treatment of Peyronie's disease. Among all, hyaluronic acid (HA) and verapamil have been differently tested. We aimed to evaluate the efficacy of intralesional verapamil (ILVI) compared with intralesional HA in patients with early onset of Peyronie's disease (PD). This is a multi-centre prospective double-arm, randomized, double-blinded study comparing ILVI vs. intralesional HA after 12-weeks. Sexually active men, older than 18 years and affected by the acute phase of PD were eligible for this study. Patients have been double-blinded randomly divided into two groups (1 : 1 ratio): Group A received intralesional treatment with Verapamil (10 mg in 5 mL of normal saline water) weekly for 12 weeks, while group B received intralesional treatment with HA (0.8% highly purified sodium salt HA 16 mg/2 mL) weekly for 12 weeks. The primary efficacy outcome was the change from the baseline to the endpoint (12 weeks after therapy) for the penile curvature (degree). The secondary outcome was the change in the plaque size and in the International Index of erectile Function (IIEF-5) score. The difference between post- and pre-treatment plaque size was -1.36 mm (SD ± 1.27) for Group A and -1.80 mm (SD ± 2.47) for Group B (p-value = NS). IIEF-5 increased of 1.46 points (SD ± 2.18) in Group A and 1.78 (SD ± 2.48) in Group B (p-value ± NS). No difference in penile curvature was observed in Group A, while in Group B the penile curvature decreased of 4.60° (SD ± 5.63) from the baseline (p < 0.001) and vs. Group A. According to PGI-I results, we found significant difference as concerning patient global impression of improvement (PGI-I) (4.0 vs. 2.0; p < 0.05). This prospective, double-arm, randomized, double-blinded study comparing ILVI vs. HA as intralesional therapy showed greater efficacy of HA in terms of penile curvature and PGI-I.

Recovery of sexual function after nerve-sparing radical retropubic prostatectomy: is cavernous nitric oxide level a prognostic index?

The preservation of NANC nerve fibers (producing nitric oxide, NO) is necessary for erection recovery after retropubic radical prostatectomy (RRP). Yet, it is impossible to establish when and if a patient will recover erections; therefore, we investigate the prognostic value of cavernous blood NO levels on this parameter. Nerve-sparing RRP was performed on 14 patients for localized prostate cancer. We evaluated all patients 3 months after surgery by IIEF score: no patients had erections. A cavernous blood sample was also taken to determine NO levels (as nitrite). Patients were evaluated again 18 months after surgery. In six cases, erectile function was compromised, whereas in seven cases, potency was restored. Statistical analysis showed a relationship between nitrite levels in cavernous blood 3 months after surgery and the recovery or erectile function at 18 months. We propose that cavernous NO blood levels are a prognostic index of erection recovery.

[Efficient prevention of overweight and obesity in the experience of family practitioners and "nutrition units" of the public health system in Lombardy]. / La prevenzione efficace del sovrappeso e dell'obesità sul territorio, esperienza in Lombardia fra u.o. nutrizione del SIAN e medici di famiglia.

General practitioners (GP's) presence is homogeneously distributed in the Lombardy territory. GP's are easy accessible by people so they could play a key role in the prevention of overweight and obesity. In this study GP's included in everyday medical activity a primary prevention strategy regarding nutrition and lifestyle. The goal was to maintain a constant BMI for one and a half year for at least 50% of the patients (stable means without progression within BMI's range). During the same time another endpoint was to decrease from 1 to 3 units the BMI in overweight patients in half of the observed population. Subjects were randomly recruited during routinely ambulatory activity, without a specific BMI based selection. Diet quality was assessed with a food frequency questionnaire regarding "protective foods" as vegetables, fruits and legumes. Some "basic" information about nutrition and lifestyle where then provided through explanation of issues printed on A4 page brochure. Percentage BMI's range variation before and after the intervention show a stability in the observed population, with an increase 0,3% for normal weight males and 0,9% for females. BMI was constant in 72,2% of the subjects considering a 1 unit variation and in 92,2% considering 3 units variation. Within subjects who varied 1 unit (27,7%) more subjects decreased rather than gained weight (16% vs 11,7%); within subjects who varied 3 units (7.8%), 4,5% decreased and 3,3 increased their BMI. GP's demonstrate to have an efficient role in weight gain control with a simple and regular prevention strategy towards healthy lifestyles and simple nutritional tips. It is important for patients to comprehend the importance the doctor give to the overweight and obesity problem, apart from the medical issue considered during the visit. GP's have a key role for healthy life-style change programs among their patients. These results provide new arguments about the opportunity to invest public resources towards population (managed in collaboration with GP's coordinated by a public health department such as NU), rather than towards a single patient. The project management was supervised by the Nutrition Unit (NU) of the public heath system SIAN (ASL di Brescia).

Peak systolic velocity thresholds of cavernosal penile arteries in patients with and without risk factors for arterial erectile deficiency.

To simultaneously assess the peak systolic velocity (PSV) and the end-diastolic velocity (EDV) at dynamic duplex examination of the cavernosal penile arteries, and penile rigidity in subjects with satisfactory erectile function with and without risk factors for arterial erectile deficiency (ED). This multicenter prospective study examined two populations having satisfactory sexual function with dynamic duplex examination of the cavernosal arteries; one population had risk factors for arterial ED (65 patients, Group 1) and the other (60 patients, Group 2) had no risk factors. Penile rigidity was assessed using the Schramek grading system score (SGSS). The PSV, the EDV, and the SGSS values of Group 1 and of Group 2 were measured and compared using analysis of variance. The EDV and PSV data presented in this abstract are the arithmetical means of the data of the left and right cavernosal arteries. The Group 1 patients showed a mean ± standard deviation PSV of 26.4 ± 13.2 cm/sec and the Group 2 patients showed a PSV of 44.7 ± 9.6 cm/sec (p = 0.002). The EDV of Group 1 was -15.6 ± 16.1 cm/sec and the EDV of Group 2 was -14.9 ± 13.7 cm/sec (p = 0.329). The SGSS in Group 1 was 3.2 ± 0.3 and the SGSS in Group 2 was 4.8 ± 0.2 (p = 0.008). Intra- and inter-operator variability were not statistically significant. The PSVs and the SGSSs of patients with risk factors for ED and satisfactory erectile function were subnormal and significantly lower than the PSVs and the SGSSs of patients without risk factors. It has been hypothesized that compensatory mechanisms, probably of psychological origin, might allow satisfactory erectile response, even in the presence of a subnormal PSV.

Plasma and skin concentration of 5-methoxypsoralen in psoriatic patients after oral administration.

The aim of this work was to investigate the distribution of 5-methoxypsoralen in the skin after oral administration of the drug and to examine the correlation between skin and plasma concentrations. 5-Methoxypsoralen skin concentration was measured in both healthy and psoriatic sites of 10 psoriatic patients after single and multiple oral doses. The results obtained show that 5-methoxypsoralen accumulates at higher levels in the more external layers of the skin after oral administration. The high affinity of drug for the stratum corneum was confirmed by in vitro skin affinity measurements. The concentration of 5-methoxypsoralen in the skin was similar in both psoriatic and healthy sites, indicating that the pathology does not influence drug distribution in the skin. After single dose administration, a linear correlation was found between skin and plasma drug concentration. After multiple dose administration, drug concentration in the skin was fairly constant despite the variable plasma concentrations in different subjects.

Differing expression of enzymes of the glyoxalase system in superficial and invasive bladder carcinomas.

This work aimed to study the activities of the glyoxalase system enzymes (glyoxalase I (GI) and glyoxalase II (GII) and their gene expression in human bladder carcinomas compared with the corresponding normal mucosa. Samples of these tissues were collected from 26 patients with superficial (SBC) or invasive bladder cancer (IBC) and used to evaluate enzyme activity and gene expression by northern blot analysis. In keeping with the electrophoretic pattern and the expression level of the respective genes, GI activity significantly increased in SBC samples, while it remained unchanged in IBC samples compared with the normal mucosa. In contrast, GII showed a higher activity in the tumour (either SBC or IBC samples) versus normal tissues. These results confirm the role of the glyoxalases in detoxifying cytotoxic methylglyoxal (MG) in bladder cancer. The differing levels of GI activity level and gene expression of GI between the SBC and IBC samples could help in their differential diagnosis.

Detection of apoptosis by the TUNEL technique in clinically localised prostatic cancer before and after combined endocrine therapy.

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.

No-mortality liver resection for hepatocellular carcinoma in cirrhotic and noncirrhotic patients: is there a way? A prospective analysis of our approach.

BACKGROUND: Low resectability rates and significant morbidity and mortality rates often make surgery for hepatocellular carcinomas (HCCs) unfeasible. HYPOTHESIS: Our policy for surgical treatment of cirrhotic and noncirrhotic patients with HCC is adequate and safe. DESIGN: Prospective validation cohort study. SETTING: University hospital. PATIENTS: One hundred seven consecutive patients with HCCs. Associated cirrhosis was present in 64 (59.8%), and only 7 (6.5%) had normal livers. INTERVENTIONS: The presence of ascites, serum bilirubin level, and indocyanine green retention rate at 15 minutes were considered when selecting patients for surgery. Preoperative recovery of liver function was achieved with portal venous branch embolization, liver volumetry, bed rest, and control of serum aminotransferase levels. The surgical techniques mainly involved bloodless dissection using intraoperative ultrasonography and intermittent warm ischemia. The main perioperative care regimen was fresh frozen plasma infusion and strict limitation of blood transfusion. MAIN OUTCOME MEASURES: The 30-day postoperative mortality and morbidity rates. RESULTS: All the patients underwent surgery (37 major resections, 45 segmentectomies, and 25 limited resections), with no 30-day postoperative mortality, overall morbidity of 26.2%, and no major complications. Multiple logistic regression analysis revealed that only the type of operation was associated with a significantly higher morbidity risk (P = .05). CONCLUSION: With high resectability, low morbidity, and no mortality, our policy represents a solution to the drawbacks of surgical resection for treatment of HCCs, especially in patients with associated liver cirrhosis.

Malaric placentas. A quantitative study and clinico-pathological correlations.

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.

Skin secretion of Siphonops paulensis (Gymnophiona, Amphibia) forms voltage-dependent ionic channels in lipid membranes.

The effect of the skin secretion of the amphibian Siphonops paulensis was investigated by monitoring the changes in conductance of an artificial planar lipid bilayer. Skin secretion was obtained by exposure of the animals to ether-saturated air, and then rinsing the animals with distilled water. Artificial lipid bilayers were obtained by spreading a solution of azolectin over an aperture of a Delrin cup inserted into a cut-away polyvinyl chloride block. In 9 of 12 experiments, the addition of the skin secretion to lipid bilayers displayed voltage-dependent channels with average unitary conductance of 258 +/- 41.67 pS, rather than nonspecific changes in bilayer conductance. These channels were not sensitive to 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid or tetraethylammonium ion, but the experimental protocol used does not permit us to specify their characteristics.

Further characterization of the "incipient lesion of chronic stationary type psoriasis vulgaris in exacerbation". The CD4-positive lymphocytes are the prominent cell population infiltrating the dermis.

In previous studies, others and we have demonstrated by immunohistological methods in light microscopy that the prominent cell population infiltrating the psoriatic lesional dermis is represented by CD4-positive cells, thus suggesting an implication of T4 lymphocytes in the development of psoriatic lesion. However, recent findings have indicated that not only T4 lymphocytes, but also macrophage-like cells, are CD4-positive. In the present study, therefore, a highly specific and sensitive 'in situ' gold-immunoelectronmicroscopy procedure developed by us was performed to identify ultrastructurally with precision the nature of the CD4-positive cells infiltrating the dermis of the incipient psoriatic lesion. Although a consistent proportion (28%) of slightly CD4-positive macrophage-like cells was detected, the strongly CD4-positive lymphocytes were the prominent cell subset (51%). The important role presumably played by such CD4-positive lymphocytes in the pathogenesis of the psoriatic lesion is discussed. Our gold-immunoelectronmicroscopy technique improved the study of psoriatic cell subpopulations in situ, since (a) in comparison with immunocytochemistry in light microscopy, it allowed us not only to recognize the ultrastructure of the labelled cells, but also to detect even small amounts of antigen on the cell surface; and (b) in comparison with the commonly used peroxidase immunoelectronmicroscopy techniques, it allowed us to quantitatively assess the expression of antigens (e.g. CD4) on different cell subpopulations (e.g. the low CD4 expression by macrophage-like cells versus the high CD4 expression by T4 lymphocytes).

p53 protein expression in fine-needle aspirates of breast cancer: an immunocytochemical assay for identifying high-grade ductal carcinomas.

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.