Placental histological patterns and uterine artery Doppler velocimetry in pregnancies complicated by early or late pre-eclampsia.

OBJECTIVES: To study placental patterns in pregnancies complicated by pre-eclampsia (PE) and to verify whether the findings are related to gestational age (GA) at PE onset and second-trimester uterine artery (UtA) Doppler. METHODS: For all pre-eclamptic women who delivered between January 2010 and December 2013, we collected retrospectively data related to placental findings and UtA Doppler velocimetry performed at PE onset. The study cohort was divided into groups according to early-onset (EO) or late-onset (LO) PE. Regression analysis was performed to test the ability of UtA Doppler velocimetry to predict subsequent development of placental lesions, after correcting for possible confounders. RESULTS: Placental abnormalities occurred with a significantly lower incidence in the LO-PE group (n = 72) than in the EO-PE group (n = 105) (P = 0.02). Irrespective of GA at PE onset, UtA pulsatility index (PI) was able to predict macroscopic infarctions (P = 0.001), distal villous hypoplasia (P = 0.03), decidual arteriolopathy (P = 0.03), villous infarcts (P < 0.001), syncytiotrophoblast 'knots' (P = 0.02), microcalcifications (P = 0.02), perivillous fibrin deposition (P = 0.02) and placental hemorrhage (P = 0.01). CONCLUSIONS: Similar placental abnormalities were present in both EO-PE and LO-PE groups, although with quantitative differences according to GA and UtA Doppler velocimetry at PE onset. Histological patterns were predicted by UtA-PI, independently of GA, supporting the use of UtA Doppler velocimetry as the key criterion in PE classification. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.

Mammaglobin B (SCGB2A1) is a novel tumour antigen highly differentially expressed in all major histological types of ovarian cancer: implications for ovarian cancer immunotherapy.

BACKGROUND: We studied the genetic fingerprints of ovarian cancer and validated the potential of Mammaglobin b (SCGB2A1), one of the top differentially expressed genes found in our analysis, as a novel ovarian tumour rejection antigen. METHODS: We profiled 70 ovarian carcinomas including 24 serous (OSPC), 15 clear-cell (CC), 24 endometrioid (EAC) and 7 poorly differentiated tumours, and 14 normal human ovarian surface epithelial (HOSE) control cell lines using the Human HG-U133 Plus 2.0 chip (Affymetrix). Quantitative real-time PCR and immunohistochemistry staining techniques were used to validate microarray data at RNA and protein levels for SCGB2A1. Full-length human-recombinant SCGB2A1 was used to pulse monocyte-derived dendritic cells (DCs) to stimulate autologous SCGB2A1-specific cytotoxic T-lymphocyte (CTL) responses against chemo-naive and chemo-resistant autologous ovarian tumours. RESULTS: Gene expression profiling identified SCGB2A1 as a top differentially expressed gene in all histological ovarian cancer types tested. The CD8+ CTL populations generated against SCGB2A1 were able to consistently induce lysis of autologous primary (chemo-naive) and metastatic/recurrent (chemo-resistant) target tumour cells expressing SCGB2A1, whereas autologous HLA-identical noncancerous cells were not lysed. Cytotoxicity against autologous tumour cells was significantly inhibited by anti-HLA-class I (W6/32) monoclonal antibody. Intracellular cytokine expression measured by flow cytometry showed a striking type 1 cytokine profile (i.e., high IFN-γ secretion) in SCGB2A1-specific CTLs. CONCLUSION: SCGB2A1 is a top differentially expressed gene in all major histological types of ovarian cancers and may represent a novel and attractive target for the immunotherapy of patients harbouring recurrent disease resistant to chemotherapy.

HER-2/neu overexpression and amplification in uterine serous papillary carcinoma: comparative analysis of immunohistochemistry, real-time reverse transcription-polymerase chain reaction, and fluorescence in situ hybridization.

Uterine serous papillary carcinoma (USPC) is a rare and highly malignant form of endometrial cancer (EC) characterized by early metastasis, chemoresistance, and high mortality rate. Little is known about USPC tumorigenesis even if recently a HER-2/neu role has been suggested in its development and progression. The aim of the present study was to evaluate HER-2 expression by immunohistochemistry (IHC) in 12 USPC formalin-fixed, paraffin-embedded (FFPE) samples. Moreover, we looked at the correlation between HER-2 protein expression and HER-2/neu gene amplification by fluorescence in situ hybridization (FISH), other than HER-2/neu messenger RNA expression by quantitative real-time reverse transcription (RT)-polymerase chain reaction (PCR). Finally, these results have been compared with commonly evaluated clinical features in EC patients, in order to define the potential prognostic value of HER-2/neu overexpression in USPCs. A high expression of HER-2 protein by IHC was noted in 2 of 12 patients (16.6%), and the same cases showed specific HER-2/neu gene amplification by FISH. All the samples investigated displayed a perfect concordance between IHC and FISH data. Five (41.6%) of 12 tumors demonstrated polysomy of chromosome 17 and, focusing on the 2 USPCs that showed HER-2/neu overexpression, one of them (50%) was polysomic for chromosome 17. All the other USPC cases (58.4%) showed to be disomic for chromosome 17. Quantitative RT real-time PCR performed on complementary DNA obtained from all FFPE USPC samples showed a complete correlation with FISH and IHC data. Moreover, HER-2/neu overexpression was associated with a poorer overall survival and a very low relapse-free survival time, thus being considered a candidate marker of worse overall prognosis in USPC. The use of trastuzumab (Herceptin), a monoclonal antibody directed against HER-2/neu, for the therapy of patients with HER-2/neu-positive USPCs should be further investigated in clinical trials.

Correlation between serological immune response analyzed by a new ELISA for HPV-16/18 E7 oncoprotein and clinical characteristics of cervical cancer patients.

Human papillomaviruses (HPVs), particularly HPV-16/18, are linked to cervical cancer development. Full-length, recombinant HPV-16/18 E7 oncoproteins were used in a new streptavidin-biotin capture ELISA method to investigate anti-HPV E7 antibody prevalence in serum. Sera from 99 healthy women, 70 cervical cancer patients, and 30 patients with cervical pre-invasive neoplasia were analyzed. Anti-HPV-16/18 E7 positivity was found in 53% of cervical cancer patients, in 40% with cervical pre-invasive neoplasia, and in 8% of healthy women. Serum samples from 12 cervical cancer patients were obtained at different time intervals during the treatment. Eleven out of 12 showed a correspondence between HPV-E7 antibody levels (decreasing versus increasing) and the type of response (clinically complete or partial response versus progression or stable disease) at each serological evaluation. Five patients with recurrent HPV-16/18-positive cervical carcinoma were analyzed before and after vaccination with HPV-16/18 E7-pulsed autologous dendritic cells; anti-HPV-16/18 E7 positivity was found in 3 out of 5 women. In conclusion, this assay could potentially be used as an adjunctive tool to monitor the type of response to treatment and possibly to detect antibody induction in cervical cancer patients after vaccination, as a potential marker to evaluate its efficacy.

Disseminated histoplasmosis with atypical cutaneous lesions in an Italian HIV-infected patient: another autochtonous case.

Disseminated histoplasmosis is recognized as a common AIDS-defining opportunistic disease in endemic areas (Americas, Africa, East Asia), while it is rarely described in Europe, usually in individuals returning from endemic regions, or following endogenous reactivation of a latent infection imported long before from overseas countries. However, reports of autochtonous cases in Europe suggest the possible, endemic presence of Histoplasma capsulatum in some European regions, such as the South of France or the Po valley in Italy. A case of disseminated histoplasmosis with atypical, papular and ulcerate skin lesions in an Italian HIV-infected patient, without history of travels outside his native region, is described. Our patient represents the fifth autochtonous case of AIDS-associated histoplasmosis described in Italy.

A comparison of immunocomplex-dissociated serum HIV-1 p24 antigenemia and plasma HIV-1 viral load: assessment of 3,129 paired assays.

In order to compare HIV-1 p24 antigenemia and plasma HIV-1 RNA levels as markers of viral replication, 3,129 paired determinations of alkaline immunocomplex-dissociated serum HIV-1 p24 antigenemia (performed with an immunoenzymatic assay), and plasma HIV-1 RNA levels (carried out with a branched DNA method, a reverse transcriptase-coupled polymerase chain reaction, and a nucleic acid sequence-based assay) were assessed over a two-year-period. When excluding samples with undetectable plasma HIV-1 RNA levels (which tested negative or borderline positive at serum p24 antigen assay in 97.9% of cases), immunocomplex-dissociated p24 antigenemia proved significantly less sensitive than viral load at all considered HIV-1 RNA reference levels, although the profile of positive serum p24 antigen assays (values above 10 pg/ml) paralleled the trend of plasma HIV-1 viral load, especially at higher levels. However, serum HIV-1 p24 antigenemia (even after immunocomplex dissociation) can be longer suggested as a the sole virological tool in the laboratory management of HIV-1 infection, due to its significantly lower sensitivity levels compared with viral load assessment.

[Helicobacter heilmannii: anatomo-clinical study of 14 new cases]. / Helicobacter heilmannii: studio anatomo-clinico di 14 nuovi casi.

INTRODUCTION: Helicobacter pylori is one of the most common causes of human gastritis. Recently, a new agent has been isolated, which also causes a gastritis. It has been initially named Gastrospirillum hominis and renamed Helicobacter heilmannii (Hh). Hh is extremely rare. In spite of the rarity it is important to recognize and diagnose it, as it requires a proper therapy, different from Hp therapy. Clinical presentation and serological results of Hh are superimposable to those of HP. Therefore differential diagnosis resides on histological grounds. PURPOSE of the present paper is to report 14 new cases of Hh gastritis, which constitutes the first italian series. RESULTS: Cases constituted 0.01% of all gastric biopsies seen in the period 1994-1998. Nine patients were male and five were female; age ranged from 32 to 76 years (50 years on average). All patients presented a mild to moderate gastritis. Hh is a spiral bacterium, being about 10 micra in length, localized in single or small groups in the glandular mucus. Two cases were associated with Hp. One case was associated with gastric adenocarcinoma. Two cases were diagnosed during the follow-up of duodenal ulcer. In CONCLUSION, the incidence of Hh gastritis in the present series seems consistent with that from other European countries. In all cases the presence of Hh was associated with features of gastritis. This confirms the pathogenetic role of Hh.

[The diagnosis of new cases of HIV infection: a reversal of the trend?]. / Diagnosi di nuovi casi di infezione da HIV: un'inversione di tendenza?

In order to assess the trend of new cases of HIV infection, spontaneous requests of HIV counseling and testing made at our outpatient Center since 1993 were retrospectively evaluated. During the considered 6-year period, 7,492 subjects had voluntary HIV testing, and 225 proved HIV-positive (3.0%). While the overall number of spontaneous requests of HIV serology did not show remarkable variations over time, a tendency towards a significant increase of newly diagnosed HIV disease was observed during the past two years (1997 and 1998). In particular, among the 57 subjects with HIV infection disclosed in 1998, a predominance of young adults and a relatively elevated frequency of females compared with males was demonstrated, while heterosexual (38 cases) and homo-bisexual contacts (18 cases) accounted for the great majority of newly identified retroviral infections, and 66.1% out of the 56 patients who acquired HIV infection by sexual route were stable partners of individuals with known HIV disease. Although epidemiological estimates foresaw a progressive reduction of cases of HIV infection since 1990, a surprising increase of newly diagnosed HIV disease among patients who spontaneously asked for HIV testing was observed in our experience. The apparent temporal coincidence of this phenomenon with the remarkable advances obtained in both diagnostic and therapeutic strategies of HIV disease just since 1996, could not be a fortuitous event. The non-decline of newly diagnosed cases of HIV infection should re-orient strategies of communication and prophylaxis dealing with HIV disease, and involving the general population and people with persisting high-risk behavior.

Evaluation of three different commercial procedures for quantifying human immunodeficiency virus type-1 RNA levels.

A branched DNA method for the quantification of human immunodeficiency virus type 1 (HIV-1) RNA levels (Quantiplex HIV RNA 2.0) was compared with a reverse transcriptase-coupled polymerase chain reaction method (Amplicor HIV-1 Monitor) and a nucleic acid sequence-based assay (Nuclisens HIV-1 QT) in plasma samples from a group of HIV-1 seropositive patients. We found a high correlation between Nuclisens and Quantiplex (r = 0.89; p < 0.001) and between Amplicor and Quantiplex (r = 0.94; p < 0.001), a shift of RNA viral load to higher Nuclisens and Amplicor values compared with the Quantiplex results and a significant positive correlation (rS = 0.60; p < 0.001) between the p24 antigen level and the RNA viral load determined with the Quantiplex assay. We also found higher sensitivities of the Nuclisens and the Amplicor procedures compared with the Quantiplex assay. The total sensivity of the Quantiplex assay in our study was 70% whereas that of the p24 antigen was only 29%.

Typhoid fever and HIV infection: a rare disease association in industrialized countries.

Typhoid fever is still a global health problem, mainly in tropical and subtropical areas of the world and in developing countries, where relatively elevated morbidity and mortality rates still are present, mostly because of persisting poor hygienic conditions. In the majority of Mediterranean regions, including Italy, the disease is constantly present, though with a low prevalence rate, as a result of an endemic persistence of Salmonella typhi infection.1-4 On the other hand, in industrialized countries, most cases of S. typhi infection are related to foreign travel or prior residence in endemic countries.4-6 In the United States, 2445 cases of typhoid fever have been reported in the decade 1985 to 1994, and the annual number of cases remained relatively stable over time: over 70% of episodes were acquired in endemic countries (mostly Mexico and India).6 The persisting morbidity of S. typhi also may be supported by the increasing resistance rate of this pathogen against a number of commonly used antimicrobial compounds. For instance, 6% of 331 evaluable S. typhi strains were resistant to ampicillin, chloramphenicol, and cotrimoxazole, and 22% of isolates were resistant to at least one of these three agents in a recent survey performed in the United States.6 The spread of antibiotic resistance among S. typhi isolates is emerging in many countries, and multidrug-resistant strains have been isolated, as well as isolates with poor susceptibility to fluoroquinolones,3-5,7-9 so that in vitro susceptibility should be determined for all cultured strains, and antimicrobial treatment should be adjusted accordingly. Nevertheless, fluoroquinolones (e.g., ciprofloxacin and pefloxacin) or third-generation cephalosporins, still represent the best choice for empirical treatment,2,4,6-8,10 and mortality remains rare in Western countries (less than 1% of episodes), although it is expected to be greater in developing areas of the world. The aim of this report is to describe two cases of typhoid fever that occurred in patients with human immunodeficiency virus (HIV) infection, a rarely reported disease association in industrialized countries.

[The finding of endocardial fibroelastosis in a 20-week-old fetus]. / Riscontro di fibroelastosi endocardica in un feto di 20 settimane.

Endocardial fibroelastosis is a rare disease that only sporadically has been diagnosed prenatally. The cases reported so far were found after the second trimester of pregnancy. We report a case of endocardial fibroelastosis found in a 20-week fetus, in whom the diagnosis was performed by echocardiography and, after voluntary interruption of pregnancy, was confirmed by necroscopy and histology. Early intrauterine detection of endocardial fibroelastosis allows to plan pregnancy, modality of delivery and a possible therapy.

Expression of cell adhesion molecules in jejunum biopsies of children with coeliac disease.

The composition of inflammatory cells as well as the expression of cell adhesion molecules (LFA-1/CD11a, ICAM-1/CD54, CD36) in jejunal biopsies from children with active, untreated coeliac disease (CD) has been analyzed and compared with control biopsies. In CD the number of intraepithelial and lamina propria T cells was greater than in controls. ICAM-1 was found on most cells in the lamina propria; many of them expressed LFA-1 as well. In contrast, no ICAM-1+ and a few LFA-1+ cells were noticed in the epithelium. The brush border from control biopsies reacted with HLADR and anti-CD36. In CD, de novo expression of HLADR was found in the cytoplasm of villous and cryptal enterocytes in contrast, CD36 disappeared from the brush border and no expression of ICAM-1 on the inflamed epithelium was noticed. The results indicate that adhesion molecules other than LFA-1, ICAM-1 and CD36 may be involved in the cellular interactions occurring in the intraepithelial compartment in CD; the lack of ICAM-1 and the reduced expression of LFA-1 on intraepithelial lymphocytes might reflect a defective activation of these cells. Several macrophages were found in the lamina propria in cases of CD; some of them were located beneath the surface epithelium, showed a spindle/dendritic morphology, and expressed the HLADR+, CD36+, CD11a- phenotype. These cells might be stimulated by luminal antigens and might play an important role in subsequent activation of T cells in the lamina propria.

Static cytometry of DNA in breast lump with reference to grading.

Fresh smears of 400 consecutive cases of breast lumps stained with Feulgen stain were measured with densitometric technique in static cytometry. A correlation was observed between DNA content expressed by the DNA index and histologic grading of malignant lesions; relatively high values were observed in the group of "hyperplasias", which were similar to G1. Values of DNA content in different categories: benign, hyperplastic (nodular and sclerosing adenosis, epitheliosis . . .) and all malignant types variously graded in G1, G2 and G3, show broad areas of overlapping, suggesting that the parameter is not always discriminating between benign, hyperplastic and malignant, but it can identify different amounts of DNA (high and low degrees) within a homogeneous category. Correlation between number of hypertetraploid nuclei (4c) and histologic grading was also evident. Therefore, fresh smears, in relation to DNA content and to the percentage of hypertetraploid elements allow a better definition of "high and low degree" lesions in a short time with possible prognostic involvement as regards tumoral progression.

[Epidermal nevus syndrome with multiple vascular hamartomas and malformations]. / Sindrome del nevo epidermico con malformazioni e amartomi vascolari multipli.

The authors described a 39 year old woman affected by epidermal nevus syndrome, with cutaneous (verrucous epidermal nevus), skeletal (thoracolumbar levoscoliosis and frontal bossing) and ocular (papillar coloboma and coroideal nevus) defects. Moreover the patient presented vascular malformations and hamartomas: lymphangioma circumscriptum of the mammary area, left peroneal Gorham's disease, artero-venous acral tumour of the left foot and multiple artero-venous shunts of the lower limbs. Since puberty, hemodynamic modifications have caused pseudo-Kaposi of Bluefarb-Stewart of legs and feet and malleolar painful ulcers. Solomon's epidermal nevus syndrome is an heterogeneous entity. In our opinion, this is the first case report with a severe vascular involvement.

[Prenatal diagnosis of hemoglobinopathies in the 1st and 2nd pregnancy trimesters]. / Diagnosi prenatale di emoglobinopatie nel 1. e 2. trimestre di gravidanza.

Fetal diagnosis of haemoglobinopathies, mainly thalassaemia, is widely used for the prevention of these diseases. Since 1975 fetal diagnosis has been carried out in the second trimester of pregnancy (18th - 22th week) on fetal blood samples obtained by fetoscopy or placentocentesis. The biochemical technique most commonly employed has been represented by carboxymethylcellulose chromatography and more recently in our center by isoelectric focusing of haemoglobins which is a faster and cheaper technique. First-trimester fetal diagnosis is now feasible by DNA analysis on chorionic villi. The Milan center experience in second trimester fetal diagnosis of haemoglobinopathies on more than 300 cases and the preliminary data on first trimester diagnosis are reported.