Relationship between the RB1 mRNA level and the expression of phosphorylated RB protein in human breast cancers: their relevance in cell proliferation activity and patient clinical outcome.

The aim of the present study was to ascertain the relationship between the level of RB1 mRNA and the expression of phosphorylated RB protein and the relevance of these two parameters in cancer cell proliferation and clinical outcome in human breast cancer. Sixty-eight primary human breast cancers were considered. The amount of RB1 mRNA was evaluated by quantitative RT-PCR analysis. The level of RB phosphorylation was immunohistochemical defined by measuring the phosphorylated (pp) RB labelling index (LI). Cell proliferation rate was measured by calculating the Ki67 LI. No relation was found between the RB1 mRNA level and the ppRB LI (p=0.565). Both RB1 mRNA value and ppRB LI were related (in an inverse and direct manner, respectively) to Ki67 LI. RB1 mRNA expression was more strictly associated with KI67 LI (p=0.001) than the ppRB LI (p=0.013). Regarding the patient clinical outcome, the separately considered RB parameters did not reach the prognostic significance. However, patients with low RB1 mRNA quantity and patients with high ppRB LI, taken together, had a significantly shorter disease free and overall survival than the group comprehending patients with high RB1 mRNA value and low ppRB LI, and this despite the low number of patients considered. Our results demonstrated that the ppRB LI was independent of the RB1 mRNA level; that both RB parameters are related to the cell proliferation rate and, if collectively considered, have a high informative value on breast tumour prognosis.

Concurrent EGFr and Cox-2 expression in colorectal cancer: proliferation impact and tumour spreading.

BACKGROUND: Many reports were produced on single epidermal growth factor receptor (EGFr) and cyclo-oxygenase-2 (Cox-2) evaluation using immunohistochemical techniques (IHC), but very few works considered concurrent expression of these two proteins in the light of their impact on proliferation and tumour spreading. At least three molecular pathways (EGFr, Cox-2, and APC/beta-catenin molecular cascade) may interact in this malignancy giving rise to cross talking effects on proliferation and cancer spreading. PATIENTS AND METHODS: To better detail these two latter aggressive features, we studied 205 sporadic colorectal cancer patients, comparing concurrent expression of EGFr, Cox-2, Ki-67, Cyclins D1-A, and E, with tumour spreading (budding) (BUD) and pN status. RESULTS: Our results point to a different aggressive molecular profile due to Cox-2 expression. Cox-2 High expressing cases showed a clear EGFr proliferation-promoting role. On the contrary, EGFr seems directly involved in cancer cells spreading rather than in promoting cancer proliferation in Cox-2 Low/Negative cases. CONCLUSIONS: Immunohistochemical profiling of colorectal cancer seems to be a promising approach, not only to define prognostic impact, but also to detail proliferation-related molecular interplays between EGFr and Cox-2 pathways, with these two latter proteins, at present, being the hottest pharmacological targets for colorectal cancer (CRC) chemoprevention and therapy.

Seropositivity to HCV in Macaca fascicularis.

Several non-human primate species are used as laboratory animals for research purposes. Non human primates represent a potential hazard for laboratory animal handlers as they exceed all other species in importance as potentiators of disease in laboratory personnel (Quist K.D., 1972). Hepatitis viruses cause some of the prevalent diseases in man which constitute an important public health problem. The first outbreak of the infection was related to non human primates and occurred in 1958-1960 in USA, with more then 200 human cases. Chimpanzee is the main species that has been implicated but others have also been involved. We report a case of seropositivity to HCV antigens in Macaca fascicularis using a third generation RIBA assay. The nature of reactivity of the positive samples could not be resolved as no animal in the breeder colony had been exposed to an HCV source. Furthermore, Macaca spp. did not appear to be a susceptible species in previous studies.

Immunohistochemical and ultrastructural investigation of neural differentiation in Ewing sarcoma/PNET of bone and soft tissues.

The authors evaluated the role of immunohistochemistry and electron microscopy in defining neural differentiation in 28 cases of Ewing sarcoma/PNET. The panel of primary antibodies used included vimentin, MIC-2, NSE, S-100 protein, leu7, neurofilaments, GFAP, and chromogranin A. Cases were considered undifferentiated when neural markers were absent, poorly differentiated if one neural marker was present, and well differentiated if two or more markers were observed. Cases were also evaluated for the presence of cytoplasmic processes, microtubules, and neurosecretory granules as ultrastructural features of neural differentiation: the tumor was classified as well differentiated if two of these features were present; and poorly differentiated if one was evident; all other cases were considered undifferentiated. According to immunohistochemistry, 10 cases (35.7%) were undifferentiated, 12 cases (42.9%) were poorly differentiated, and 6 (21.4%) were well differentiated. According to the ultrastructural analysis, 10 tumors were undifferentiated (35.7%), 14 poorly differentiated (50%), and 4 well differentiated (14.3%). The overall concordance between the two techniques was low (35.7%), and both modalities were concordant in classifying only 1 well-differentiated, 5 poorly differentiated, and 4 undifferentiated tumors. In conclusion, the authors suggest that investigations devoted to test the prognostic significance of neural differentiation in these neoplasms should employ both immunohistochemistry and electron microscopy, separately and in combination, to assess what is the most effective choice for predicting the clinical course.

Desmoplastic small round-cell tumor: a case report on the large cell variant with immunohistochemical, ultrastructural, and molecular genetic analysis.

examination. The patient died 10 months after surgery. Histologically, the tumor was composed of predominantly large epithelioid cells with foci of anaplasia mimicking metastatic carcinoma. Immunohistochemically, the tumor cells stained with anti-cytokeratin, EMA, desmin, and NSE antisera. Electron microscopy showed secretory lumina, desmosomes, cell processes with microtubules and electron-dense granules, and focal whorls of intermediate filaments. Reverse transcriptase-polymerase chain reaction performed on paraffin block-retrieved tissue demonstrated the EWS/WT-1 fusion transcript characteristic of the t(11;22)(p13;q12). This case illustrates a less common histological pattern of DSRCT, i.e., diffuse large cells, thus supporting the view that this tumor presents a wider morphological spectrum than that previously recognized.

Silent oophoritis due to cytomegalovirus in a patient with advanced HIV disease.

A case of isolated necrotizing cytomegalovirus (CMV) oophoritis disclosed only by necropsy studies in a patient with AIDS, is described. This unusual case report is discussed with a review of the literature dealing with CMV involvement of genital organs in the immunocompromised host, and in patients with HIV infection and AIDS.

Splenic mucinous cystadenocarcinoma arising in heterotopic pancreatic tissue.

A mucinous cystadenocarcinoma along with remnants of normal pancreatic tissue was discovered in the spleen of a 21-year-old woman. Extensive clinical, radiological, ultrasound and computed tomography investigation revealed that the patient's pancreas and ovaries were unremarkable. We presume an origin from heterotopic pancreatic tissue for this neoplasm and present a review of the literature in this regard.

[Clear cell sarcoma of soft tissues. Clinico-pathological and ultrastructural analysis of a case in the head-neck region and a literature review]. / Il "sarcoma a cellule chiare delle parti molli". Analisi clinico-patologica e ultrastrutturale di un caso a localizzazione nella regione testa-collo e revisione della letteratura.

BACKGROUND: Clear cell sarcoma of soft tissues (CCSST), even synonymically termed malignant melanoma of soft tissues, is a peculiar malignant tumor with both a prolonged clinical course and a final poor prognosis. Whilst its neuroectodermal histogenesis is most likely, its nosological position is still a matter of debate. Light microscopy (LM), immunocytochemistry (ICC), and electron microscopy (EM) investigations have shown both a melanocytic and schwannian differentiation. Recent findings from cytogenetic analysis documenting a reciprocal translocation between the long arms of chromosomes 12 and 22 [t(12:22)(q13;q12.2)] not seen in cutaneous melanomas nor in other neurogenic sarcomas support CCSST as a distinct entity. MATERIALS AND METHODS: A case of CCSST arising in the preparotid region of a young man with a clinical history of two previous surgical interventions was admitted in 1994 and is herein reported. This patient was previously cured twice in different institutions: the first time he was diagnosed with a "melanoma, NOS" of the face and surgically treated 10 years earlier. The second time--meaning one year earlier the admission--this patient was again surgically treated for a local recurrence which was interpreted as a myoepithelioma (clear cell type) of salivary gland origin. A morphological, immunocytochemical, and ultrastructural analysis performed on the last recurrence is herein presented. RESULTS: On LM the tumor exhibited the classic morphological appearance comprised both of clear and eosinophilic cells with prominent nucleoli, focally containing cytoplasmic granules of melanin pigment. At ICC expression of vimentin and S-100 protein was diffusely demonstrated as well as the melanoma associated antigen HMB-45 was even albeit focally documented.

Primary endocervical extraosseous Ewing's sarcoma/PNET.

A 36-year-old woman presented with intermenstrual spotting and was found to have a cystic mass involving the uterine cervix on a pelvic ultrasound examination. A necrotic and hemorrhagic tumor was excised by hysterectomy and processed for light and electron microscopic investigation and molecular analysis. Microscopic examination revealed a small round cell tumor that immunohistochemical studies (including staining for the highly restricted surface antigen p30/32MIC2) and ultrastructural studies indicated was an extraosseous Ewing's sarcoma (EES)/primitive neuroectodermal tumor (PNET). This diagnosis was established by detection of EWS/ERG fusion transcript through reverse transcription polymerase chain reaction (RT-PCR) with nested primers. Full body computed tomography failed to detect any extrauterine tumor, and the patient is clinically free of disease 18 months after hysterectomy. This case represents the first report of a primary EES/PNET arising in the uterine cervix.

Ultrastructural characterization of oligodendroglial-like cells in central nervous system tumors.

Cells with uniform, small-round nucleus and clear cytoplasm (oligodendroglial-like cell, OLC) are commonly observed in central nervous system (CNS) neoplasm of glial and neuronal lineage, such as oligodendroglioma, clear-cell ependymoma, and central neurocytoma. Immunohistochemistry does not always contribute to the characterization of OLC because of (1) loss of antigen expression; (2) lack of specific markers for oligodendrogliomas; and (3) occasional coexpression of neuronal and glial antigens. An ultrastructural analysis associated with an immunohistochemical study of 20 cases of CNS tumors largely constituted by OLCs has been performed. Neurocytomas (12 cases), medullocytomas (2 cases), cerebral neuroblastoma (1 case), and ganglioglioma (1 case) showed OLCs with ultrastructural features of neuronal differentiation (neuritic processes, dense-core granules, synaptic structures). Oligodendroglioma (3 cases) OLCs were characterized by mitochondrial-rich cytoplasm, and ependymoma (1 case) OLCs showed microrosettes and scattered cilia. The electron microscopic analysis can provide a more precise diagnosis of these OLC-containing tumors despite their uniform morphological appearance.

Microcolposcopic topographic endocervical assessment before excisional treatment of cervical intraepithelial neoplasia.

OBJECTIVE: To evaluate whether microcolposcopic topographic endocervical assessment reduces the failures of excisional treatment of cervical intraepithelial neoplasia (CIN). METHODS: Three hundred fifty patients with colposcopic and histopathologic findings of endocervical CIN were recruited for excisional treatment. Three hundred forty-eight of these were randomized to have or not have microcolposcopy before excisional treatment. Measurement of endocervical lesion was the only aim of microcolposcopic evaluation. When an endocervical extension was available, the cone biopsy was cut according to microcolposcopic measurement. Excision status was evaluated and related to presurgical management on operative specimens. After excision, patients were followed-up for at least 5 years after treatment. Three hundred thirty (171 and 159 with and without preoperative microcolposcopy, respectively) patients completed the study. Disease persistences were defined by cytologic, colposcopic, and histologic results. Microcolposcopic value was defined as completeness of excision and/or lack of persistent disease. RESULTS. On surgical specimens, involved margins were detected in 19 (5.4%) cases. Presurgical microcolposcopy was performed in only one of these cases. The difference of incomplete excision between cases with or without microcolposcopy was statistically significant (P < .001). In patients who were followed-up, persistent disease was detected in one (0.6%) woman in the microcolposcopy group and in 16 (10%) women in the control group. Comparison between the two groups showed a significantly lower risk of persistent disease when presurgical microcolposcopy was performed (P < .001). CONCLUSION: By measuring endocervical extension of the lesion, preoperative microcolposcopy allows individualized cones, thus improving the prognosis after excisional treatment of CIN.

Filamentous inclusions in nonneoplastic and neoplastic pancreas: an ultrastructural and immunogold labeling study.

Filamentous inclusions (FI) are unusual, irregularly shaped cytoplasmic inclusions, which are mostly found in acinar cell carcinomas of the pancreas and are consequently thought to be an abnormal zymogen granule type. This study describes identical inclusions in acinar, centroacinar, and small duct epithelial cells from nonneoplastic pancreas, as well as those found in tumor cells from a mixed acinar-endocrine pancreatic carcinoma. An ultrastructural and immunogold labeling demonstration indicates that these inclusions are aggregates of intermediate filaments immunoreacting with the anti-cytokeratin AE1/AE3 mixture and with V9 clone anti-vimentin monoclonal antibodies. Their pleomorphic appearance, variable immunoreactivity, and frequent association with lipid droplets and secondary lysosomes, mostly of the angulate type, led to the hypothesis that the FI undergo a degenerative remodeling pathway similar to that proposed for hepatic Mallory bodies. A survey of the literature on FI and human tumors suggests that they are a variably expressed ultrastructural feature of tumor cells originating from exocrine cell-containing tissues, namely the pancreas and gastrointestinal tract.

Presurgical assessment and therapy of microinvasive carcinoma of the cervix.

Retrievable pathological specimens and clinical data on 70 patients with microinvasive carcinoma diagnosed on surgical specimens from cone biopsy or hysterectomy (Stage IA) were reviewed and compared to pertinent findings in the literature with the intent of evaluating diagnostic criteria and defining pathological features that may influence the outcome by therapy. Emphasis was given to the preoperative assessment emphasizing that both an accurate colposcopic evaluation and a detailed pathological analysis may reliably point to a conservative therapeutic approach. Increasing depth of stromal invasion was associated with lesion width as well as with endocervical extension, as measured on colposcopy, microcolpohisteroscopy, and histology. Lymph-vascular space involvement was significantly related to depth of invasion. Two patients of 28 with dissected nodes had node metastases as well as lymph-vascular space involvement. Both developed a pelvic recurrence. One had a > 1- to < or = 3-mm invasion depth, the other a > 3- to < or = 5-mm lesion invasion. While advocating a conservative procedure for Stage IA1, we suggest discrimination with regard to Stage IA2 because we believe that lymph-vascular involvement should be meticulously evaluated. In fact, > 1- to < or = 3-mm lesions without lymph-vascular space involvement can be conservatively treated, while for any other lesion falling within the Stage IA2 category a modified radical histerectomy plus pelvic lymphadenectomy should be recommended.

Gastro-intestinal stromal tumors: an ultrastructural reinterpretation of the clear cell component.

The histology, immunohistochemistry and ultrastructure of six gastro-intestinal stromal tumors of the stomach (GSTs) showing a focal to diffuse clear cell component are reported. At light microscopy, all GSTs had typical histopathological features with one case additionally displaying stromal myxoid changes and scattered multinucleated giant cells. Immunohistochemically, 6 of 6 GSTs stained positive for vimentin, 2 of 6 for smooth muscle specific actin and 1 of 6 for desmin. At electron microscopy, GSTs showed microfilaments with focal densities as well as other smooth muscle features, such as subplasmalemmal linear densities and foci of external lamina. Ultrastructural appearances of tumor cells with clear cell features showed these not to be an artifact of fixation, but the expression of an unusual cytophagocytic activity. Inclusions of auto- and heterophagocytic nature were found responsible for the origin of the large, mostly lipidic vacuoles which displaced cell nuclei peripherally in a signet-ring fashion. It is concluded that such previously unrecognized features are ultrastructural aspects of GSTs with smooth muscle differentiation.

Smooth muscle differentiation in normal human ovaries, ovarian stromal hyperplasia and ovarian granulosa-stromal cells tumors.

An immunohistochemical and ultrastructural investigation on the presence of "smooth muscle differentiation" in stromal ovarian tissue was carried out in 10 adult granulosa cell tumors, six juvenile granulosa cell tumors, six thecoma/fibrothecomas, six cases of stromal hyperplasia, and in 10 normal ovaries. For immunohistochemistry, formalin-fixed paraffin-embedded tissues were processed using anti-alpha smooth muscle actin (alpha-SM actin) and anti-desmin as primary monoclonal antibodies. All adult granulosa cell tumors and juvenile granulosa cell tumors showed an intense alpha-SM actin immunoreaction, but weaker for desmin. Immunostain was diffuse in six out of 10 and five out of six adult granulosa cell tumors and juvenile granulosa cell tumors, respectively. Ultrastructurally, intermediate filaments focally converging into well developed desmosomes as well as peripheral bundles of myofilaments were documented both in adult granulosa cell tumors and juvenile granulosa cell tumors. In thecoma/fibrothecomas and stromal hyperplasia, alpha-SM actin and desmin expression was minimal or absent; on electron microscopy some "myoid" features and myofibroblasts were also seen. In normal ovaries, alpha-SM actin was found intensely expressed in the theca externa, focally identified in cortex-medulla, and unstained in the theca interna layer. Immunoreaction increased during folliculogenesis, going from a thin positive alpha-SM actin layer around secondary follicles to a strong diffuse stain in mature follicles. Our immunohistochemical and ultrastructural results indicate that a "smooth muscle differentiation" is a typical component of the specialized gonadal stromal tissue. A diffuse and focally alpha-SM actin is constantly present in granulosa cell tumors and thecoma/fibrothecomas, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The morphology of elastin in non-specific and inflammatory abdominal aortic aneurysms. A comparative transmission, scanning and immunoelectronmicroscopy study.

The maintenance of the mechanical properties of the vessels results from the correct arrangement of smooth muscle cells and extracellular fibrous proteins (elastin and collagen) in their wall. The morphology of extracellular matrix modifications, particularly of elastin, was investigated in inflammatory (IA) and non specific (NSA) abdominal aortic aneurysms by scanning (SEM), transmission (TEM) and immunoelectron microscopy. Both NSAs and IAs were significantly characterized by extensive extracellular matrix remodelling, including different patterns of elastin degradation. Elastic and collagen fibres distribution appeared to be extensively altered in IAs, while it conformed more to a normal pattern in NSAs. With respect to NSAs, the morphology of elastic fibres in IAs was modified to such an extent that their identification by TEM had a rely on immunocytochemical methods and by SEM on back-scattered electron analysis. The observed ultrastructural changes are indicative of the central role of extracellular matrix modifications in the pathogenesis of IAs and NSAs.

[Wash and non-wash system in the perioperative recovery of blood in prosthetic surgery: ultrastructural assessment of corpuscular blood components]. / Sistema wash e non-wash nel recupero perioperatorio del sangue in chirurgia protesica: valutazione ultrastrutturale della parte corpuscolata del sangue.

AIM: The authors aimed to perform an ultrastructural morphological analysis of blood recovered using wash and non-wash systems in patients undergoing full cement-free hip replacement in order to evaluate the integrity of the various blood corpuscle components. EXPERIMENTAL PROTOCOL: An open prospective study in patients undergoing full cement-free hip replacement at the Orthopedics Division of S. Orsola-Malpighi Policlinico in Bologna. Materials of S. Orsola-Malpighi Policlinico in Bologna. MATERIALS AND METHODS: Blood recovered postoperatively using a non-wash system was studied in 6 patients. In a further 3 patients perioperatively recovered blood was studied after washing using Cell Saver Haemolite 2 before reinfusion. Red globules, white globules and plaelets were isolated from blood collected using these two different recovery systems and analysed by SEM. RESULTS: Study of the ultrastructural morphology of various corpusculated blood fractions. DISCUSSION AND CONCLUSIONS: From the data in our possession it appears that the ultrastructural morphology of the various corpuscle components of blood in subjects undergoing postoperative recovery is better preserved using a non-wash system. There was no sign of "polluting" material in terms of adipose cells or free bone fragments in either group.

Immunocytochemical study of epidermal growth factor receptor, transforming growth factor alpha, and "squamous differentiation" in human endometrial carcinoma.

The expression of epidermal growth factor receptor (EGFr) and transforming growth factor alpha (TGF-alpha) was compared with the presence of "squamous differentiation" (SD) visualized in various histotypes of endometrial carcinoma by using a panel of monoclonal antibodies. The results of the current study demonstrate that EGFr and TGF-alpha are present in routinely processed endometrial carcinoma. The highest positive EGFr and TGF-alpha expression was seen in the group of adenocarcinomas with SD. The more intense EGFr and TGF-alpha immunoreactivity was observed in "squamous" foci both in adenoacanthomas (AA) and in adenosquamous carcinomas (AS). These EGFr- and TGF-alpha-positive squamous areas prevalently displayed a "stratification-related" cytokeratin (CK) immunoprofile characterized by the expression of CKs 1, 4, 5, 10, 13, 14, and 16. No correlation was found between EGFr- and TGF-alpha-positive status and depth of myometrial invasion or surgical stage. These results clearly demonstrate that EGFr and TGF-alpha expression is related remarkably to endometrial carcinoma with "squamous" areas both morphologically and immunophenotypically. This specific association leads us to suggest that EGFr and TGF-alpha expression in endometrial carcinoma may be prevalently involved in the equilibrium of cell differentiation of the "squamous" foci commonly observed in this group of neoplasias.

Nuclear bodies associated with core particles of hepatitis B virus in a healthy carrier.

An unusual association of nuclear bodies (NB) with core particles of hepatitis B virus (HBV) is described in a healthy carrier who histologically showed ground glass hepatocytes with occasional sanded-nuclei. Ultrastructurally, many hepatocytes had typical features of HBV infection. Unexpectedly, a few cells contained previously undescribed complexes of NB studded with a number of HBV core particles. The ultrastructural characteristics of these complexes and their possible relation to HBV morphogenesis are briefly discussed.