TROP-2 expression in papillary thyroid cancer: a preliminary cyto-histological study.

OBJECTIVE: TROP-2 (human trophoblast cell surface marker) is a gene-related protein expressed in trophoblastic cells, which is also present in a variety of epithelial cancers and whose overexpression has been found to correlate with a poor prognosis. We analysed the possibility of using the expression of TROP-2 to detect papillary thyroid carcinoma (PTC) on cytological and histological samples, and compared it with Hector Battifora mesothelial antigen-1 (HBME-1). METHODS: From 127 patients, 127 fine needle aspirates (FNAs), in which HBME-1 was detected by immunocytochemistry (ICC), were re-classified according to the Bethesda system for reporting thyroid cytopathology (TBSRTC): 20% were benign, 56% were atypical cells/follicular lesion of undetermined significance (AUS/FLUS), 4% were follicular neoplasm/suspicious for a follicular neoplasm, 5% were suspicious for malignancy and 16% were malignant. Sufficient material to test for TROP-2 was available in 64 FNAs, 22 of which had a histological control. Including six additional cases in which the FNAs were not available, immunohistochemistry (IHC) was carried out with both markers on 94 cases. RESULTS: Among 88 FNAs with histological control, the sensitivity of HBME-1 to predict PTC was 87.5% (28/32) and the specificity was 86% (48/56), whereas, in 22 FNAs, TROP-2 sensitivity was 100% (13/13) and specificity was 89% (8/9). In 94 histological specimens in which IHC was carried out with both markers, the sensitivity and specificity were 82% and 86%, respectively, for HBME-1 and 87% and 89%, respectively, for TROP-2. The difference between the markers was not significant. Concordance between IHC and ICC was 76% for HBME-1 and 91% for TROP-2. CONCLUSION: TROP-2 can be used as well as HBME-1 in thyroid cytology to detect PTC. Positivity for either or both markers could help to stratify the risk of malignancy in indeterminate FNAs. Larger studies are need to analyse its role in the behaviour of PTC and its variants.

Optical coherence tomography and focal electroretinogram evaluation of cystoid macular edema secondary to retinitis pigmentosa treated with intravitreal triamcinolone: case report.

PURPOSE: To assess macular structure and function by optical coherence tomography (OCT) and focal electroretinogram (FERG) before and after intravitreal triamcinolone acetonide (IVTA) administration for cystoid macular edema (CME) in a patient with retinitis pigmentosa (RP). METHODS: A 33-year-old man with RP and refractory bilateral macular edema was treated with IVTA in his left eye and evaluated with visual acuity, OCT, and FERG for 6 months. RESULTS. Compared to the fellow eye, after IVTA mean retinal thickness significantly decreased, while FERG amplitude and phase did not show significant changes at the end of follow-up. Visual acuity showed a significant tendency to improve. CONCLUSIONS: In this case report, IVTA improved macular anatomy and visual acuity; this result, however, was not associated with a similar electrophysiologic response.

BRCA1 mutations and polymorphisms in a hospital-based consecutive series of breast cancer patients from Apulia, Italy.

INTRODUCTION: Hereditary breast cancer has been partly attributed to germline mutations in the BRCA1 gene that are deleterious for BRCA1 protein activity. This paper analyzes the incidence and characteristics of detectable BRCA1 mutations and polymorphisms in a hospital-based consecutive series of breast cancer patients from southern Italy to investigate the incidence and the association of these molecular alterations with breast cancer biology and family history. METHODS: One hundred cases with familial characteristics were selected from a consecutive series of 511 patients with a first diagnosis of breast cancer. DNA from peripheral blood was screened for whole BRCA1 gene mutations utilizing dHPLC as a pre-screening analysis and automatic DNA sequencing for the identification of specific alterations. RESULTS: In the overall series of 511 patients, 100 had a family history of breast cancer and were investigated for BRCA1 mutations. Two types of BRCA1 mutations were identified, 5382insC in six cases and 4566delA in one case. The 5382insC mutation was present in two out of six cases with ovarian cancer while 4566delA in one case of male cancer. The most frequent missense polymorphisms were E1038G, P871L, K1183R in exon 11, S1613G, M1652I in exon 16 and D1778G in exon 22. Confirming what found in previous studies, patients in whom pathological BRCA1 mutations were detected had early-onset breast cancer (p=0.05), positive nodal status (p=0.05), lower ER (p=0.02) and PgR (p=0.01) content. Interestingly, the K1183R polymorphism and, less strongly, S1613G polymorphism were associated to mutational risk (K1183R: OR 0.1 p=0.03; S1613G: OR 2.7 p=0.08). CONCLUSION: Mutations in the BRCA1 gene are frequent also in our consecutive series of patients from southern Italy. An association between two detected single nucleotide polymorphisms (SNPs) and BRCA1 mutational risk was ascertained. Finally, we confirm the fact that peculiar clinical-pathological features seem to characterize patients with a family history of breast cancer and BRCA1 alterations.

Hemodiafiltration with post-dilution reinfusion of the regenerated ultrafiltrate: a new on-line technique.

AIMS: All convective hemodiafiltration techniques require a replacement fluid, which must have an adequate electrolytic composition and must be sterile and pyrogen-free. Using an integrated adsorption cartridge, the ultrafiltrate can be "regenerated" and used as a replacement fluid (hemo-filtrate reinfusion; HFR). The aim of this study was to evaluate whether the HFR technique as suggested in its original configuration could be improved by inverting the purification sequence (post-dilution HFR; PDHFR) in order to increase the purification efficiency of the whole system. METHODS: We performed standard HFR in 6 uremic patients during 6 months and, subsequently, during further 6 months, PDHFR. The dialytic efficacy of the two techniques and the filter blood loss were evaluated. Moreover, we studied how both techniques affected cytokine levels. RESULTS: We observed a significant increase of urea extraction and of Kt/V values in PDHFR. An equally significant improvement was observed in regard to the extraction of beta2-m and the blood loss. Furthermore, IL6 and TNFalpha decreased significantly after PDHFR treatment. CONCLUSIONS: HFR has proven to be an easy-to-perform hemodiafiltration technique, capable of resolving the typical problem of the other hemodiafiltration technique, the availability and production of a sterile and ultrapure reinfusion solution. The inversion of its configuration has allowed us to improve three aspects that have characterized, in our experience, the treatments performed in the original geometry: the removal of both urea and beta2-m, and the filter. Finally, it's notable that the decrease in cytokines levels achieved with PDHFR might attenuate the uremic micro-inflammatory state.

The association between retinal microvascular changes, metabolic risk factors, and liver histology in pediatric patients with non-alcoholic fatty liver disease (NAFLD).

BACKGROUND: The prevalence of childhood obesity is increasing worldwide. Studies in adult populations show that retinal microvascular changes are associated with obesity and components of the metabolic syndrome. In our study we have assessed the effect of body mass index (BMI), metabolic parameters, and adiposity on the retinal microvasculature in children. METHODS: Fifty-four consecutive children with biopsy-proven NAFLD were enrolled in this study. Anthropometric and laboratory parameters were obtained using standardized protocols. Retinal caliber was quantified from digital retinal images using well-known computer-based programs. Twenty-four-hour ambulatory blood pressure monitoring was measured using a standard protocol. RESULTS: In our population, the prevalence of retinopathy was of 53 % (13 males). The 29 patients with retinopathy (mean age 10.91 ± 3.10) showed significantly higher values of triglycerides (mg/day) (105.57 vs. 90.20, p = 0.04), basal insulin (mUI/ml) (17.20 vs. 12.97, p = 0.02), and HOMA-IR (3.37 vs. 2.76, p = 0.04). The patients with a HOMA-IR >2.5 (OR = 3.34, p = 0.02; 95 % IC, 1.07-10.39), and systolic non-dipping (OR 4.16, p = 0.028, 95 % IC, 1.11-13.67), have an increased risk of retinopathy. Moreover, the study of correlation between all stages of liver biopsy (CRN criteria) and the grade of retinopathy showed a positive correlation with fibrosis (r = 0.31) and an NAS score (r = 0.28). CONCLUSIONS: We found an association between metabolic parameters and nocturnal blood pressure on the retinal microvasculature among the obese children with NAFLD. Furthermore, for the first time, we report the positive relationship between hepatic fibrosis in pediatric NAFLD patients and the degree of retinopathy signs.

nm23 expression in malignant ascitic effusions of serous ovarian adenocarcinoma.

The aim of this study was to evaluate nm23 expression as detected in malignant cells of neoplastic ascites and to verify its relationship with the presence of tissue nm23 and p53 in primitive neoplasia. Using an immunocytochemical assay with a specific anti-nm23 polyclonal antibody, nm23 expression was evaluated in ascitic effusions of 45 patients with ovarian serous adenocarcinoma and ascites in normal and/or hyperplastic mesothelial cells from 37 women with various neoplasms (12 ovarian neoplasms) free of malignant cells as controls. nm23 and p53 tissue expression was also detected in 21 corresponding tumor samples, including 11 bilateral lesions. nm23 was expressed in 57% of malignant effusions compared to 43% in controls: the two groups were not correlated. nm23 in effusions agreed with tissue expression (p=0.02) but a direct correlation was not demonstrated. The incidence of nm23 was more frequent in stage III than in stage IV disease (p=0.08) and was associated with mutated p53 expression (p=0.01). Using the Wilcoxon test for unpaired data, a higher incidence (p=0.05) of p53-positive tumors in bilateral cancers was found while a higher expression of nm23 in effusions of patients with monolateral lesions was observed (p=0.08). The presence of p53 was correlated with that of nm23 in both cytologic (p=0.005) and histological samples (p=0.01). Our findings, together with the diversity in biological behaviour present in various tumors, suggest that nm23 is a family of genes with differing biological functions which act as tumor-specific inhibiting factors within a complex process also involving other genes. Due to the analogies and correlations between nm23 and p53, the role of nm23 as a potential predictive factor of response to chemotherapy and in DNA repair is emphasized.

nm23 expression in human breast cancer: correlation with cell proliferation (S-phase) as observed with a double-labeling immunocytochemical-autoradiographic technique.

In 51 unselected breast cancer samples, a double-labeling immunocytochemical technique was utilized to observe the phenotypic expression of the nm23 gene during S-phase. The feasibility of the method was confirmed by comparison with routine evaluations for both thymidine-labeling index and nm23 (p<0.001). No correlation was found between the two parameters in the overall series or when subgroups regarding menopausal status, nodal involvement, tumor size, hormone receptor content were considered. Tumors with a higher nuclear grade showed a significant correlation only with TLI (p=0. 02). A trend for an inverse relationship between the two parameters was noted but was not statistically significant. A significant association between TLI and nm23 expression was found by Chi-square test using median values as cut-off. Our data based on morphological and in situ observations do not confirm a correlation between nm23 and cell proliferation, even if this correlation cannot be completely excluded. Multicentric trials of high power with these same techniques are still necessary to definitely establish the prognostic role of nm23 in breast cancer.

Timing of breast cancer surgery within the menstrual cycle: tumor proliferative activity, receptor status and short-term clinical outcome.

We verified the variations of primary tumour steroid receptor status and proliferative activity at different times and phases (follicular vs luteal) of the menstrual cycle and their relationship with short clinical outcome in a cohort of 248 N- breast cancer patients. Steroid receptor content (ER and PgR) was evaluated by DCC assay and proliferative activity by 3H-Thymidine autoradiographic assay (TLI). Median age was 44 years, 60% of tumors were T1, and cytohistological grade was G1-2 in 54% of cases. At surgery, 57% were in the luteal phase while 43% were in the follicular phase. No significant variations were found in mean TLI or ER and PgR characteristics of the primary tumors surgically treated in different periods of the menstrual cycle; however, the ER level resulted significantly higher in 4th with respect to the 3rd week of menstrual cycle, while PgR level was higher in PgR+ cases treated during the 3rd week. The number of relapses and disease-free survival curves after 36 months median follow-up did not differ significantly for patients treated in different periods of the menstrual cycle (12% and 9% of disease relapses in luteal and follicular phases; p=n.s.). We can conclude, therefore, that TLI, ER and PgR expressions could vary significantly during menstrual cycle only in certain specific tumor subgroups.

Epidemiological surveillance of staphylococcal resistance.

The aim of our investigation was to characterize better the possible changes in staphylococcal susceptibility to some antimicrobial agents. We considered the staphylococcal strains isolated between June 1986 and June 1989 and held responsible for proved bacterial infections. In this relatively short time we observed a moderate increase in methicillin resistance. Compared with methicillin-sensitive strains, methicillin-resistant isolates showed minor susceptibility to most antimicrobial agents; nevertheless, vancomycin, teicoplanin, cefamandole, imipenem, fluoroquinolones and netilmicin, according to our results, appear at present the most reliable antistaphylococcal antibiotics.

[Secretory IgA of the cervical mucus in neoplastic and nonneoplastic conditions of the uterine cervix]. / IgA secretorie del muco cervicale in condizioni neoplastiche e non neoplastiche della cervice uterina.

In 39 women affected by cervical intraepithelial neoplasia (CIN) concentration of the cervical mucus secretory IgA (sIgA) was significantly higher than in 414 controls matched for age, reproductive status and smoking habit (196 healthy women; 109 with "specific" and "aspecific" cervicitis or cervico-vaginitis; 51 with benign epithelial disorders of the cervix; 40 with adenomatous cervical polyp and 18 with early invasive exocervical squamous carcinoma). CIN carried the highest proportion of cases with sIgA detectable or beyond the cut-off value and sIgA concentration was inversely related to CIN grade. Present data confirm our preliminary investigation on the cervical total IgA and explain that any increased levels of these IgA are induced by the secretory and not serum fraction of this immunoglobulin. We still don't know the exact meaning and value of the present results. It is only possible to suppose that they are the expression of a vigorous local immune activation adverse to CIN, which is elevated for as long as the architecture of cervical glandular epithelia is preserved.

Ultrastructural alterations of choroid plexuses of lateral ventricles of rats (Rattus norvegicus) submitted to experimental chronic alcoholism.

Adult male rats (Wistar lineage) were alcoholized with sugar cane liquor diluted at 30(0) GL during 300 days and sacrificed every 60 days in 5 stages. Samples of choroid plexuses of lateral ventricles were collected and examined at transmission electronic microscope to detect possible ultrastructural alterations and to raise possible pathological correlations. Gradual changes were observed in these animals during all the experiment: dilatation and enlargement of cisternae of Golgi complex, dilatation of RER, presence of digestive vacuoles and a large amount of pinocytic vesicles as well as vesicles with electronlucent content throughout cytoplasm, as well as an enlargement of intercellular space between basolateral interdigitation of the cells and of the connective tissue. The changes observed in the epithelium and connective tissue of choroid plexuses specially in 240 and 300 days of treatment are presumably due to a disturbance in hydroelectrolitic homeostasis, contributing to several morpho-functional disturbs of central nervous system. No changes were observed in the control group animals.

Scanning electron microscopy study of the choroid plexus in the monkey (Cebus apella apella).

The cells of the choroid plexus of the lateral ventricles of the monkey Cebus apella apella were examined through scanning electron microscopy at contributing to the description of such structures in primates. The animals were anesthetized previously with 3% hypnol intraperitoneally and after perfusion with 2.5% glutaraldehyde, samples of the choroid plexus were collected after exhibition of the central portion and inferior horn of the lateral ventricles. The ventricular surface of those cells presents globose form as well as fine interlaced protrusions named microvilli. Among those, it is observed the presence of some cilia. Resting on the choroid epithelial cells there is a variable number of free cells, with fine prolongations which extend from them. They are probably macrophages and have been compared to Kolmer cells or epiplexus cells, located on choroid epithelium. The choroid plexus of the encephalic lateral ventricles of the monkey Cebus apella apella at scanning electron microscopy is similar to that of other primates, as well as to that of other species of mammals mainly cats and rats, and also humans.

[Comparison between two different on-line dialytic techniques: Standard HFR vs post-dilutional HFR]. / Emodiafiltrazione con reinfusione di ultrafiltrato rigenerato in pre- e post-diluizione: due tecniche on-line a confronto.

PURPOSE: Among hemodiafiltration (HFD) techniques, hemodiafiltration reinfusion (HFR) seems unable to achieve an optimal depurative efficacy. This study aimed to evaluate whether the HFR technique as suggested in its original configuration could be improved by devising a new technique (post-dilution HFR (PDHFR)) inverting the purification sequence to increase the purification efficiency of the entire system. METHODS: We performed standard HFR in six uremic patients during 6 months and, subsequently, during a further 6 months, PDHFR was performed. The dialytic efficacy of the two techniques and the filter blood rest were evaluated. In addition, we studied the behavior of cytokines during the inverted HFR sessions. RESULTS: We observed a significant increase in urea extraction and in Kt/V. An equally significant improvement was observed with regard to beta2-microglobulin (beta2-m) extraction and blood rest. Furthermore, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) significantly decreased after inverted HFR treatment. CONCLUSIONS: The inversion of the original configuration allowed us to improve the depurative efficacy of standard HFR, increasing the removal of both urea and beta2-m, and reducing the blood rest. Finally, it was notable that the reduction in cytokine levels could attenuate the uremic microinflammatory state.

BRCA1 expression and molecular alterations in familial breast cancer.

The aim of the study was to evaluate the performance of immunohistochemical MS110 expression in a series of familial and sporadic breast cancer patients. An immunohistochemical study was performed on TMA samples from 93 sporadic and 94 familial breast cancer patients with (7/94) and without BRCA1 germline mutations. BRCA1 protein expression level was evaluated using the monoclonal MS110 antibody. Immunohistochemistry, performed on TMA samples, showed positive nuclear staining for BRCA1 in 34 sporadic and 37 familial breast tumours, respectively. All the tumours from patients carrying BRCA1 mutations showed complete loss of both BRCA1 and ERalpha expression, regardless of the type of mutation. The percentage of MS110 positive cases was significantly lower in mutated versus wild type BRCA1 familial cases (p=0.02) while the percentage of patients with higher ERalpha expression was significantly lower in BRCA1-mutated versus BRCA1-wild type familial patients (p=0.05). Interestingly, the presence of the E1038G polymorphism in BRCA1 exon 11 was significantly associated with protein expression (p=0.029). The frequency of MS110 negative cases also detected in BRCA1-wild type tumours, points to the inability of the BRCA1 IHC expression in discriminating between familial and sporadic breast cancer.

Study of the trabecular projections (Chordae Willisii) of the superior sagittal sinus.

The trabecular projections or Chordae Willisii were studied in the internal surface of the human superior sagittal sinus, being defined and quantified as to having relation with afferent veins to the sinus, as to their lateral or medial location and also in the anteroposterior direction. The results indicate a predominance of trabecular projections with no relation to afferent vessels to the sinus in its medial portion and central region, as well as predominance of these projections on the left side. The number of trabecular projections related to afferent vessels was nearly the same when left and right sides of the superior sagittal sinus were compared.

[Expression of steroid receptors and DNA synthesis in male breast cancer]. / Espressione dei recettori steroidei e sintesi di DNA nel carcinoma della mammella maschile.

Using a double-labeling immunocytochemical-autoradiographic assay we studied 18 male breast carcinomas to evaluate the cell kinetic and receptor status in neoplastic cells during S-Phase and to detect possible differences with respect to 21 female breast cancers, from a previously, published study. In male breast cancer, the tumor receptor content and ER/PgR expression in neoplastic cells during S-Phase was higher (p = 0.01) than that in corresponding female tumor while tumoral cell proliferation was lower, but not significantly. In the previous reported study on female breast cancer we demonstrated that proliferative activity was higher in receptor negative cell population both for ER and PgR. Conversely, in male tumor, that difference was only present in relation to the expression of PgR: in fact, the proliferative activity was higher in PgR negative than in PgR positive cells (Anova Test: p = 0.04) while no difference was evidenced between ER negative versus ER positive cells. Moreover, the arrest of DNA synthesis, expressed as percentage of cells without 3H-Tdr labeling, was not related to either the ER or PgR expression, while in female breast cancer it was higher in PgR positive than ER positive cell population. Our data confirmed differences between males and females regarding the receptor status and cell cycle S-Phase in breast cancer. The poorer prognosis of the male breast carcinoma might be related to ineffective therapies which do not consider these differences in the biological profile of the male tumor. The Authors indicate that prognostic and predictive tissutal markers, detected by immunocytochemical methods and useful for therapeutic programming in the female breast cancer have a different significance in the male breast cancer and stress the need for different therapeutic strategies specific for male breast cancer.

Imipenem and meropenem induced resistance to beta-lactam antibiotics in Pseudomonas aeruginosa.

The ability of imipenem and meropenem in subinhibitory concentrations to influence the results of disk diffusion susceptibility tests was assessed. Selection of stably derepressed mutants resistant to beta-lactam antibiotics other than carbapenems was also investigated. Beta-lactams were shown to be subject to carbapenem-mediated antagonism in the disk diffusion test. On the other hand in vitro selection of stably derepressed mutants resistant to other beta-lactams could not be demonstrated.

[Dna synthesis and expression of steroid receptors in cells of human breast carcinoma]. / Sintesi di DNA ed espressione dei recettori steroidei in cellule di carcinoma mammario umano.

In order to observe receptor expression (ER and PgR) in human breast cancer cells during the S-phase of the cellular cycle, we evaluated 21 tumoral samples using a double labeling immunocytochemical-autoradiographic assay. In 16 of these we have verified that expression of ER and PgR is not influenced by incubation with 3H-Thymidine for 60 minutes. The results also demonstrated that receptor expression during the S-phase is more common in postmenopausal tumors and is principally dependent on percentages of ER+ (p < 0.01) and PgR+ (p < 0.01) cells; moreover, proliferative activity occurs almost exclusively in ER- (p = 0.003) and Pgr- (p = 0.001) cellular sub populations and the presence of PgR, but not of ER, is related to arrest of DNA synthesis (p = 0.02).