Japanese encephalitis virus RNA detected in Culex pipiens mosquitoes in Italy.

Mosquitoes collected in northern Italy were screened for flavivirus RNA. Positive amplicons were sequenced and found most similar to insect flavivirus (ISF), Usutu virus (USUV) and surprisingly also to Japanese encephalitis virus (JEV). The sequence (167 bp), obtained from one pool of Culex pipiens, was found identical to JEV strains from bats in China. Unfortunately additional sequence data or virus isolations were not obtained in this study. Confirmation of potential introduction of JEV to Italy and other European countries is urgently needed.

The role of fetal autopsy and placental examination in the causes of fetal death: a retrospective study of 132 cases of stillbirths.

PURPOSE: To investigate the most plausible cause of stillbirth by evaluating clinical records and postmortem examination findings including placental analysis. METHODS: A retrospective cohort study concerning 132 stillbirths from 124 pregnancies occurred in the Mother-Infant Department of the University Hospital of Modena, Italy, from January 2000 to December 2004. Collected data were reviewed and classified according to the Gardosi ReCoDe system. RESULTS: A reasonable cause of fetal death was identified in 99/124 pregnancies (79.84%). No associated relevant factors were disclosed in 25 fetuses (20.16%) classified as unexplained stillbirths. A succeeding scrupulous analysis of the placenta and an accurate clinical record review were useful to detect other conditions in 82 cases, including 5 cases of unexplained stillbirth. The major relevant conditions associated to stillbirths were feto-placental infection especially in the early fetal gestation age, under the 24th week of gestation, and placental insufficiency occurred both in early and late gestation age fetuses and mainly associated with a IUGR (<10th customized percentile). The main frequent secondary conditions were represented by placental anomalies including cluster of avascular villi with stromal fibrosis associated to thrombosis in minor and/or major vessel(s). Through the further analysis of the placenta, we were able to reduce the unexplained stillbirth rate from 20.16 to 15%. CONCLUSION: Accurate fetal autopsy and placental examination related to meticulous clinical collecting data are requisites in the valuation of stillbirth and could play an important role in reduction of unexplained stillbirth rate.

Contrast-enhanced MRI and PET-CT in the evaluation of patients with suspected local recurrence of rectal carcinoma.

PURPOSE: This study aimed to evaluate the role of contrast-enhanced magnetic resonance imaging (MRI) and positron emission tomography-computed tomography (PET-CT) in the assessment of local recurrence of rectal cancer. MATERIALS AND METHODS: Among 200 patients scheduled for CT follow-up, 60 (48 low risk; 12 high risk) were selected due to CT findings suspicious for or suggestive of local recurrence. Patients underwent contrast-enhanced MRI and PET-CT within 2 weeks. Biopsy was considered the gold standard in 39 cases and follow-up at 6 and 12 months in the remaining 21. RESULTS: Local recurrence was confirmed by histology in 15 cases (7 low risk; 8 high risk) and was excluded in 21 cases by long-term follow-up and in 24 by histology. Sensitivity, specificity, positive and negative predictive value and accuracy were 86.7%, 68.9%, 48.1%, 93.9% and 73.3% for contrast-enhanced MRI and 93.3%, 68.9%, 50%, 96.9% and 75% for PET-CT. CONCLUSIONS: Contrast-enhanced MRI and PET-CT can help in the detection of local recurrence of rectal cancer, even though their roles in early detection remains debatable, as the value of these techniques in current surveillance protocols is still to be defined.

Expression of estrogen receptor in hemangioma of the uterine cervix: reports of three cases and review of the literature.

The occurrence of hemangioma in the female genital tract, particularly in uterine cervix, is rare. The majority of them show asymptomatic behavior. Surgical excision remains curative in most of the cases. Conservative therapies such as sclerosing agents, cryotherapy, and CO(2) laser excision may be alternatively applied. We present three cases of hemangiomas of the cervix in asymptomatic women, diagnosed as cavernous hemangioma in two cases and capillary hemangioma in one. All tumors were immunoreactive for CD31, CD34, factor-VIII-related antigen. Focal expression of estrogen receptors was detected. No positivity was obtained with progesterone receptor antibodies. The presence of estrogen receptor in the endothelial cells of the hemangioma of the cervix suggests a direct role of this hormone in the hemangioma development. A possible target therapy is discussed.

Cataract development in a young patient with lathosterolosis: a clinicopathologic case report.

PURPOSE: To report a unique case of cataract in a young patient with lathosterolosis, a singular defect of cholesterol biosynthesis, and to report the clinical results and histopathologic findings after cataract surgery. METHODS: A 7-year-old patient with lathosterolosis, a rare defect of cholesterol biosynthesis, presented with a complex phenotype, including severe mental retardation, liver disease, multiple congenital anomalies, and bilateral posterior subcapsular cataracts. After a severe metabolic decompensation, a worsening in the lens opacity of the left eye occurred. The authors thus performed cataract surgery and made a histopathologic analysis of aspirated lenticular samples. The following examinations were performed at 1 day, 1 week, 3 months, 6 months, 12 months, and 24 months: refraction, biomicroscopy, and fundus evaluation. Visual acuity was not assessable due to lack of patient collaboration. The postoperative follow-up period was 24 months. RESULTS: Histopathologic findings on lenticular fragments revealed the presence of fibers disposed in a honeycomb, samples with homogeneous eosinophilic lens fibers, and other fragments characterized by bulgy elements referable to cortical fibers with degenerative characteristics. After surgery, biomicroscopic evaluation revealed no significant inflammation and good intraocular lens centration at the various control visits. No intraoperative or postoperative complications occurred. No posterior capsule opacification occurred 2 years after surgery. CONCLUSIONS: Lathosterolosis may lead to dysmetabolic cataract development; this unique case of cataract in such a patient has been successfully managed with surgery. Clinical results were excellent, and no complications occurred either intra- or postoperatively.

Conjugated poly(D,L-lactide-co-glycolide) for the preparation of in vivo detectable nanoparticles.

Cellular localization of nanoparticles (Np) represents an important target in the understanding of their distribution after endovenous injection. The need of suitable devices and methodologies capable to detect Np in tissues or in cellular districts can be satisfied by Np which have to be easily recognizable by simple methods. Conjugations of poly(D,L-lactide-co-glycolide) with fluorescein and biotin allow fluorescent and immuno-histochemically active Np to be obtained. The fluorescein Np are detectable using fluorescent microscopy whereas biotin Np can be detected by optical microscopy after streptavidin-biotin-peroxidase complexation. In vivo experiments confirm the ability of these particles to be easily detected in the brain parenchyma or in the liver cell population according to the infusion pathway.

Anatomy of Tunga trimamillata Pampiglione et al., 2002 (Insecta, Siphonaptera, Tungidae) and developmental phases of the gravid female.

This paper deals with some internal anatomical features observed in histological sections and freshly dissected mounts of Tunga trimamillata, a Siphonaptera recently discovered in Andean regions of Ecuador from several mammals, including man. It was possible to study in males and also non-gravid and gravid females, the location and anatomy of several organs not previously described for this species: the testes, epididymis, ganglia, Malpighian tubules, eyes, rectal ampulla with one of its pads and structures which could be interpreted as midgut diverticula, whose presence has not been recorded in the Siphonaptera. The process of neosomy in the female during pregnancy is illustrated by photographs of the consecutive developmental phases, taken at the stereomicroscope. Furthermore, some details of the exoskeleton, spermatheca during different phases of pregnancy of the gravid female and the presence of a foreign body (parasite?) within the haemocoel have been displayed in specimens cleared with Hoyer's medium.

Gastric localization of Leishmania in a patient with acquired immunodeficiency syndrome. A case report.

We report a case of gastric localization of Leishmania in a 29-year old man affected by AIDS. Gastric biopsies revealed macrophages infected with intracytoplasmic organisms attributable to Leishmania amastigotes. The authors emphasize the importance of performing random biopsies in the absence of endoscopic abnormalities.

Histoplasma capsulatum var. capsulatum occurring in an HIV-positive Ghanaian immigrant to Italy. Identification of H. capsulatum DNA by PCR from paraffin sample.

Histoplasmosis, which is highly endemic in the United States, is rare in Europe, usually imported but sometimes autochthonous. In Africa, histoplasmosis capsulati coexists with "African histoplasmosis", a characteristic skin infection caused by H. capsulatum var. duboisii. Histoplamosis due to H. capsulatum is one of the 12 secondary infections listed in the surveillance definitions of AIDS. We report the case of a 36-year-old black man with acquired immunodeficiency syndrome (AIDS) who was living in Italy but originally came from Ghana. Histoplasmosis was disseminated with fever and cutaneous manifestations. The diagnosis was demonstrated morphologically based on the presence of yeast, observed by light microscopy, in skin lesions and by identification of H. capsulatum var. capsulatum DNA by nested PCR from a paraffin sample. No clinical reports of histoplamosis capsulati in Ghana have been published until now. The present case stresses the role of immigration of subjects from outside Europe who have been infected in their native country.

Histopathological evidence of North American blastomycosis in Italy: report of two cases.

No clinical reports of blastomycosis in Italy have been published until now. We here report two cases of histologically diagnosed, unexpected cutaneous involvement in patients, aged 78 and 52 years, living in North Italy and never having been abroad. The histological differential diagnosis between blastomycosis and other fungal pathogens is discussed. Even in the absence of culture the present cases can confidently be considered as genuine examples of Blastomyces dermatitidis infection in Italy.

Aedes albopictus (Diptera:Culicidae) is incompetent as a vector of hepatitis C virus.

In laboratory experiments, Aedes albopictus females were fed hepatitis C virus (HCV)-infected human blood with a unit-feeding apparatus. Pools of engorged females were tested at various time intervals after their blood meal for the presence of HCV RNA using two methods in parallel: reverse transcription-nested polymerase chain reaction (PCR) and PCR with the Amplicor HCV system. Comparable results were obtained with both methods. Mosquitoes were found to be positive only immediately and 24 h after feeding. Moreover, female progeny produced negative results. Our results indicate that the Ae.albopictus strain used is incompetent as a vector of HCV.

Placental candidiasis: report of four cases, one with villitis.

Four cases of placental candidiasis, an uncommon complication of rupture of the membranes, are presented. In addition to chorioamnionitis, in one of these cases villitis was also observed. Villitis is a rare occurrence in Candida infection and this represents only the second case in the literature. The involvement of villi may be suggestive of blood-borne infection. However, since neither the mother nor the foetus presented any signs of systemic dissemination, the authors suggest a hypothesis of contamination of the villi from foci of chorioamnionitis.

Atypical (bizarre) leiomyoma of the nasal cavity with prominent myxoid change.

A 72 year old woman presented complaining of nasal obstruction, rhinorrea, and epistaxis. At examination, a polypoid mass on the right posterior choana was revealed and subsequently removed. Light microscopic findings consisted of a diffuse proliferation of spindle shaped, pleomorphic cells with eosinophilic cytoplasm and blunt ended nuclei in a prominent myxoid background. The presence of numerous plurinucleate, bizarre cells made it very difficult to determine the malignant potential. Immunohistochemical evidence for leiomyogenic markers coupled with the low mitotic rate, the lack of an infiltrating growth pattern, and the indolent clinical course led to the diagnosis of atypical leiomyoma with prominent myxoid change. A literature survey confirmed that such a tumour is extremely rare at this site, but the biological behaviour seems to be similar to its uterine counterpart. Clinicians should be aware of this occurrence to prevent misdiagnosis because a conservative therapeutic approach is necessary in this disease.

Pitfalls and difficulties in histological diagnosis of human dirofilariasis due to Dirofilaria (Nochtiella) repens.

The authors have conducted a histologic study of over 90 cases of Dirofilaria infection in man due to Dirofilaria repens, the most extensive study of its kind in the world. On the basis of their findings, they illustrate the diagnostic difficulties that may arise when regressive phenomena render the parasite's morphologic characteristics largely or completely unrecognizable. These phenomena are due to the death of the filaria inside the inflammatory nodule weeks or months prior to surgical removal, and to the consequent invasion of inflammatory cells. They start at the nematode's natural orifices (mouth, vulva, anus, cloaca) and gradually spread throughout its body. In cases of difficult diagnosis, therefore, it is recommended that the nodule be sectioned at different points comprising caudal, cephalic and middle body aspects, so as to reveal the few morphologic features that are still recognizable and which may afford a correct diagnosis. The authors present examples of histologic sections in color plates in which the nematode was in an excellent state of preservation and others illustrating the full range of regressive phenomena, from slight to total disintegration of the bodily structure.

The Italian experience of a Pap test and speculoscopy based screening programme.

OBJECTIVES: The main objective of the study was to evaluate if speculoscopy, a magnified chemiluminescent examination, combined with a Pap smear, could improve the detection of early cervical lesions compared with the Pap smear alone. SETTING: Pap tests and speculoscopies were performed in two family planning centres located in the surrounding areas of Modena. Colposcopic investigations and biopsies of the uterine cervix were performed in a second level centre (Gynaecological Prevention Centre of Modena Policlinico). Histological specimens were analysed at the Section of Pathological Anatomy of Modena Policlinico. SUBJECTS: The study population comprised 1000 women aged 25-64 years, invited to undergo a Pap smear every three years in accordance with the screening programme for cervical cancer started in Modena in 1997. METHODS: Midwives performed the Pap smear and speculoscopy in succession. Women with a positive Pap test and/or positive speculoscopy underwent colposcopy and, if colposcopic findings were positive, targeted punch biopsies were performed. RESULTS: A total of 1000 patients were subjected to cytology and speculoscopy examinations. Among these women, 10 had abnormal Pap smear findings whereas 144 had an abnormal speculoscopic pattern. Only three of 59 patients with a histological diagnosis of cervical intraepithelial neoplasia grade I (CIN I)/human papillomavirus and only three of seven patients with CIN II/CIN III had a positive Pap test. CONCLUSIONS: Speculoscopy combined with a Pap test can significantly increase the detection of cervical lesions when included in a screening programme.

Pregnancy luteoma with granulosa cell proliferation: an unusual hyperplastic lesion arising in pregnancy and mimicking an ovarian neoplasia.

A pregnancy luteoma (PL) was incidentally found at a term cesarean section in a 27-year-old black woman without any endocrine abnormality. The lesion involved only the left ovary; it had a nodular and focal pseudoalveolar growth pattern and was associated with areas of tubular sertoliform component, consistent with granulosa cell proliferation. Immunohistochemistry revealed a diffuse positivity to Inhibin A, CD99, cytokeratin and vimentin. The ultrastructure was typical of steroid-producing cells. PL is a tumor-like lesion arising in pregnant women and often misdiagnosed as a neoplastic lesion; awareness of this rare entity and its differential diagnoses may avoid unnecessary surgery in young patients.

Gelatin microspheres crosslinked with D,L-glyceraldehyde as a potential drug delivery system: preparation, characterisation, in vitro and in vivo studies.

To overcome the restriction in using crosslinked gelatin in the pharmaceutical field, D,L-glyceraldehyde (GAL), a non-toxic crosslinking agent, was proposed. Gelatin microspheres crosslinked with different concentrations of GAL (0.5, 1 or 2%, w/v) and for different time periods (1 or 24 h) were prepared. The effect of the preparation variables was evaluated analysing the extent of crosslinking, the morphological aspect, the particle size and the swelling behaviour. To evaluate the pharmaceutical properties, an antihypertensive drug, clonidine hydrochloride, was chosen as drug model and loaded into the microspheres. Either the increase of the crosslinker concentration or of the crosslinking time period decreased both the swelling and the in vitro drug release processes of the microspheres. After the subcutaneous injection, the loaded microspheres crosslinked with the lowest GAL concentration (0.5%, w/v) or for the shortest time period (1 h) showed a reduction of systolic blood pressure (SBP) similar to that recorded with a clonidine hydrochloride solution having the same drug concentration. Instead, the microspheres crosslinked for 24 h with concentrations of GAL higher than 0.5% (w/v) produced a more gradual and sustained SBP reduction and the antihypertensive effect was maintained until 52-72 h. The biocompatibility studies showed that the microspheres crosslinked with GAL are well tolerated in vivo. These results suggest the potential application of gelatin microspheres crosslinked with GAL as a suitable drug delivery system for the subcutaneous administration.

Primary high-grade mucosa-associated lymphoid tissue-type lymphoma of the cervix presenting as a common endocervical polyp.

Lymphomas of the uterine cervix are uncommon neoplasms and typically appear as diffuse cervical enlargement. We describe a rare case of primary high-grade lymphoma of mucosa-associated lymphoid tissue of the uterine cervix in a 46-year-old white woman. The tumor, incidentally disclosed at gynecological examination, appeared as a single common polyp. Immunohistochemical investigation found the lesion to consist of a monomorphic CD20-positive infiltrate of large blasts and rare intermingling centrocyte-like lymphoid cells. A dense area of monotypic (lambda light-chain restriction) plasma cells was found beneath the endocervical mucosa; only a few scattered lymphoepithelial lesions were present. The neoplastic cells did not stain for CD5, CD10, CD23, CD43, or cyclin D1. A bone marrow biopsy displayed a paratrabecular, centrocyte-like B-cell infiltration, but no lymphadenopathy was detected by instrumental examination (computed tomographic scan, magnetic resonance imaging). The tumor was successfully treated by multiagent chemotherapy followed by total hysterectomy. To our knowledge, this case represents the second reported example of mucosa-associated lymphoid tissue-type lymphoma occurring in the uterine cervix. We highlight the very unusual gross appearance of this case and emphasize the difficulty of interpreting lymphoid infiltrates in the lower genital tract by microscopy.

Acanthamoeba meningoencephalitis in a patient with acquired immunodeficiency syndrome.

Several cases of Acanthamoeba encephalitis (ie, granulomatous amebic encephalitis) have been reported in patients with acquired immunodeficiency syndrome from the United States. To our knowledge, none so far has been reported from Europe, and this is the first case of amebic meningoencephalitis due to Acanthamoeba in a patient with acquired immunodeficiency syndrome from Italy. The patient was a 24-year-old, human immunodeficiency virus-positive heterosexual man with a 6-year history of intravenous drug use. He was admitted to the hospital because of severe headache, confusion, nuchal rigidity, jaundice, and ascites. He died 5 days later. At autopsy, the brain showed extensive hemorrhagic necrosis with numerous trophic and cyst forms of Acanthamoeba. The amebas were identified as Acanthamoeba divionensis by the indirect immunofluorescence test.