An overview of the genus Radiospongilla (Porifera, Spongillida) with description of a new species from Australia.

A new species of freshwater sponges is described from Australia. The new species Radiospongilla fungosa is characterized by the following traits 'megascleres microspiny stout oxeas with scattered, small, simple spines except tips', and by having 'gemmuloscleres long, slim, spiny strongyles with scattered simple spines and hooked spines clustered at tips', 'gemmular theca with 2 layers of gemmuloscleres radially and tangentially arranged', 'pneumatic layer irregularly filamentous network (near inner layer) and thin laminae irregularly arranged (towards outer layer)'. We discuss the relationships of the new species to other members of the genus both from Australia, particularly North-Western Australia, and from other biogeographic regions on the basis of diagnostic morphotraits. An overview on the taxonomy, biogeographic pattern and phylogeny of the genus is supplied and updated to present, together with an updated identification key for Australia species. The diagnosis of the genus Radiospongilla is emended.

Macroalgal forest vs sea urchin barren: Patterns of macro-zoobenthic diversity in a large-scale Mediterranean study.

The study aimed at contributing to the knowledge of alternative stable states by evaluating the differences of mobile and sessile macro-zoobenthic assemblages between sea urchin barrens and macroalgal forests in coastal Mediterranean systems considering a large spatial scale. Six sites (100 s km apart) were selected: Croatia, Montenegro, Sicily (Italy), Sardinia (Italy), Tuscany (Italy), and Balearic Islands (Spain). A total of 531 taxa, 404 mobile and 127 sessile macro-invertebrates were recorded. Overall, 496 and 201 taxa were found in macroalgal forests and in barrens, respectively. The results of this large-scale descriptive study have met the expectation of lower macrofauna complexity and diversity in barrens rather than in macroalgal forests, and have allowed estimating the differences in levels of diversity and the consistency of variability across Mediterranean sites. Some peculiar patterns in barrens, related to both abundance of specific taxa and to high values of beta diversity, have been evidenced.

Australian freshwater sponges with a new species of Pectispongilla (Porifera: Demospongiae: Spongillida).

This paper focus on the biodiversity assessment of Australian inland water. Checklists of Australian Spongillida are also provided with biogeographic notes together with the geographic range of all species of freshwater sponges in Australia. New discoveries on freshwater sponges are reported from ephemeral freshwater habitats in Kakadu National Park (Australia Northern Territory). Morphological analyses show that the sponges belong to Radiospongilla and Pectispongilla in the family Spongillidae. Radiospongilla cfr. philippinensis shows a single layer of radial gemmuloscleres and the absence of tangential gemmuloscleres in the gemmular theca. Pectispongilla gagudjuensis n. sp. diverges from the diagnostic traits of the four species currently assigned to the genus i.e. skeletal megascleres are dominant acanthostrongyles and less frequent acanthoxeas shorter than in the other species, microscleres are absent, and gemmules are larger than in the other species of the genus.

Effectiveness of a rapid immunohistologic method for tumor cell origin analysis.

A rapid immunohistologic method is described to analyze the possible cell origin of a given neoplasm during surgery by using selected examples of monoclonal and polyclonal antibodies (anti-cytokeratins, anti-vimentin, anti-leukocyte common antigen, and anti-keratin). This rapid (26-28 min) procedure, consisting of a two-step immunoperoxidase method, did not differ in terms of intensity and specificity of the immunoreaction from the control procedure for which conventional longer times of incubation and washing were used. The results demonstrate the feasibility and effectiveness of the procedure.

Heterogeneous immunostaining patterns of follicular dendritic reticulum cells in human lymphoid tissue with selected antibodies reactive with different cell lineages.

A comparative immunohistologic study of the cell density and distribution pattern of follicular dendritic reticulum cells (DRCs) within their follicular microenvironments (germinal centers and mantle zones) was performed by immunoperoxidase technique with a selected panel of antibodies either operationally specific for DRCs (DRC-1) or reported as having additional immunoreactivity with DRCs (antibodies to B- and T-cells, leukocytes, monocytes/macrophages, desmosomal components, and S-100 protein). Twenty-five biopsy specimens, including reactive lymph nodes and tonsils as well as normal spleen tissue, were analyzed. Serial frozen sections were tested either with single antibodies or paired monoclonal reagents in double-labeling procedures. Our results consistently indicated that DRCs positive for Leu M3 and BA-2 antibodies were confined to the central portion of germinal centers, whereas DRCs immunoreactive for S-100 protein and desmoplakin 1 and 2 were localized mostly in the central and pericentral portion of germinal centers. All the DRCs extending from the central portion of germinal centers to the mantle zones were labeled with DRC-1 and, unexpectedly, with OKB7 antibodies. Other immunostainings, such as those for HLA-DR antigens, common leukocyte antigen, and Leu 3a, were not contributory in defining topographic differences of DRCs within the follicle. The consistent heterogeneity of the labeling patterns appears to suggest a possible in situ immunophenotypic grouping of DRCs, and the concept of their possible heterogeneity appears to be corroborated.

Double labeling immunohistologic and flow cytometric analysis of human B cells with particular reference to Leu-8 expression.

Previous results have shown that human lymphocyte subpopulations are heterogeneous as to Leu-8 expression. In the present study, we performed a heretofore unreported immunohistologic analysis and flow cytometric double labeling investigation focused on Leu-8+ and Leu-8- human B cells, with special reference to their expression of other B cell lineage antigens (Leu-14, B1, OKB7 or B2, OKB2, BA-1, BA-2, IgD, and IgM) or of a functional marker of cell proliferation (Ki-67). Immunohistologic analysis was performed on frozen sections of nine normal or reactive lymph node and tonsil biopsy specimens tested with either single or paired antibodies, the latter procedure (double labeling) being directed at revealing positively subtracted Leu-8+ or Leu-8- cells expressing a given marker depending on the antibody staining sequence used. Dual flow cytometry with paired antibodies was performed on cell suspensions from four normal or reactive lymph nodes and tonsils. As to the follicular district, immunohistologic analysis suggests that germinal center (proliferating) B lymphocytes, identifiable in cell suspension as Leu-8-B1+ and representing 80% of the B1+ cells, are Leu-8- and Leu-14+, BA-2+, OKB7+, and IgM+, being rarely IgD+. Most lymphocytes located in the mantle zone (resting cells), identifiable in cell suspension as Leu-8+B1+ and representing the remaining 20% of the B1+ cells, are Leu-8+, Leu-14+. OKB7+, OKB2+, BA-1+, IgD+, and IgM+. Furthermore, from our results, there is indirect evidence to support the existence of a Leu-8+BA-2+ lymphocyte subset located in the mantle zone. Immunohistologic study of four lymph nodes by Leu-8 and Ki-67 antibody (that recognizes a nuclear, cell proliferation-associated antigen absent only in the Go phase of the cell cycle) showed that these markers are mutually exclusive and added further evidence that Leu-8 antigen is a marker of resting lymphocytes.

Inaccuracy of death certificate diagnoses in malignancy: an analysis of 1,405 autopsied cases.

In 1,405 patients who died at the General Hospital of Trieste in 1974 and 1978, malignant neoplasm was revealed at autopsy. Clinical diagnosis was accurate in 54 per cent of these patients. The tumor was clinically suspected in 19 per cent and was undiagnosed in 27 per cent. The accuracy of the clinical diagnoses varied significantly according to the primary site and type of tumor; accuracy was inversely related to the age of the patient and varied also according to the department of the hospital to which the patient had been admitted. This latter variation is age-dependent, too. In the past decades clinical diagnosis of malignancy has not greatly improved, although the autopsy rate has almost everywhere strongly decreased, representing a heavy handicap in the epidemiologic research on cancer as a cause of death.

Demonstration of S-100 protein distribution in human lymphoid tissues by the avidin-biotin complex immunostaining method.

Immunoreactivity for S-100 protein was investigated immunohistochemically in a series of 49 fixed and paraffin-embedded normal, reactive, and neoplastic human lymphoid tissue specimens. The avidin-biotin complex immunoperoxidase method was used, with overnight (12-hour) incubation with a commercially available antiserum to S-100 protein. In addition, cryostat sections were tested with DRC 1 monoclonal antibody to dendritic reticulum cells (DRCs) in three cases and with OKT6 antibody to interdigitating reticulum cells (IRCs) in nine cases. All tissues, including lymph nodes, tonsils, adenoid, spleens, appendices, thymuses, and tissues containing nodular reactive lymphoid infiltrates, demonstrated a consistent immune staining pattern. A striking network composed of dendritic processes that showed finely granular S-100 protein immunoreactivity was observed in most of the follicular germinal centers; a similar dendritic pattern was observed in the follicular centers when the corresponding frozen sections were immunostained with DRC 1. In the extrafollicular areas, the S-100-positive cells topographically and morphologically resembled the IRCs that were demonstrated by OKT6 antibody in the corresponding frozen sections. The results seem to indicate that cells topographically and morphologically similar to IRCs and DRCs in human lymphoid tissues from different sites share immunoreactivity for S-100 protein. The present study confirms the unexpected presence of S-100 protein in dendritic cells of follicular germinal centers by a simple and currently available method.

Differentiation, proliferation and apoptosis levels in human leiomyoma and leiomyosarcoma.

A comparative analysis of the differentiation pattern, the proliferative behaviour, and the level of apoptosis between human benign and malignant neoplasms of smooth-muscle (SM) tissue is lacking. The clinical, histopathological, immunochemical, and immunocytochemical features of leiomyomas (LM) and leiomyosarcomas (LMS) were investigated by a panel of monoclonal antibodies specific for some differentiation markers of SM tissue (SM myosin and alpha-actin, desmin, and SM22) and for markers of non-muscle tissue (vimentin and non-muscle myosin). Proliferating normal and neoplastic cells were identified by proliferating-cell nuclear antigen (PCNA)/Ki67 immunostainings and the apoptotic cells were revealed by means of the terminal-deoxynucleotidyltransferase-mediated dUTP nick-end labelling technique. Gel electrophoresis and Western blotting, performed with anti-(SM1/SM2 myosin isoform) antibody, indicated quantitative differences between LMS and LM, which mirrored higher positive to negative nuclear ratios for PCNA, Ki67 and apoptosis in malignant as opposed to benign neoplasms. With LM, however, a similar SM1 to SM2 ratio could be associated with different proliferation levels. Uterine, gastric and intestinal LMS displayed specific patterns of SM1/SM2 and/or non-muscle myosin expression that were not paralleled by different levels of proliferation/apoptosis. While the level of PCNA/Ki67 correlated with the level of apoptosis in normal SM tissues and LM, that of LMS did not. In vivo at the cellular level, LM and uterine LMS displayed a near-uniform SM tissue differentiation, whereas the other LMS displayed a lesser or a heterogeneous immunoreactivity. In vitro, cultured LMS cells showed a limited and peculiar expression of SM myosin. In conclusion, there is no reciprocal relationship between degree of differentiation and the level of proliferation, as exemplified by the finding that the less differentiated intestinal LMS displays the lowest proliferative behaviour and that the relatively more differentiated gastric LMS/metastasis is more proliferative.

Respiratory and cardiovascular responses to static handgrip exercise in humans.

We studied the time course of respiratory and cardiovascular responses by evaluating changes in the breathing pattern, mean blood pressure (MBP), and heart rate elicited by 3 min of static handgrip at 15, 25, and 30% of the maximum voluntary contraction (MVC) in 15 healthy volunteers. Muscle tension and integrated electromyographic activity remained fairly constant during each trial. During 15% MVC bouts, initially only mean inspiratory flow increased; then, tidal volume and minute ventilation (VI) also rose progressively. No significant changes in MBP and heart rate were observed. During 25 and 30% MVC bouts, not only did mean inspiratory flow, VT, and VI increase but MBP and heart rate increased as well. A slight and delayed rise in respiratory rate was also observed. Unlike 15 and 25% MVC handgrip, 30% MVC handgrip caused a small decrease in end-tidal PCO2. Changes in the pattern of breathing occurred more promptly than those in cardiovascular variables in the majority of subjects. Furthermore, we found a positive correlation between changes in VI and those in cardiovascular variables at the end of 25 and 30% MVC trials. This study indicates that respiratory and cardiovascular responses to static handgrip exercise are controlled independently.

Significance of S-100 protein immunostaining in the immunohistological analysis of normal and neoplastic lymphoid tissues--an appraisal.

S-100 protein is a heterogeneous fraction of dimeric polypeptides (alpha and beta subunits) that can exist in different combination forms within the various tissues. Concerning the S-100 protein immunodetection within lymphoid tissue, the heterogeneity of the S-100 antigen, the tissue quality (frozen or paraffin-embedded after treatment with different fixatives) and the treatment of the tissue with different immunostaining methods and antibodies of different nature, all make for inconsistent results obtained in the immunohistological studies reported in the literature. Most of the S-100-positive cells of the lymphoreticular system are dendritic cells involved in the immune response (interdigitating reticulum cells, Langerhans cells, and follicular dendritic reticulum cells), other S-100-positive cells belonging to the mononuclear/phagocytic system. S-100 protein immunostaining may be used as a helpful immunohistological diagnostic clue to certain malignancies of the immune system (follicular center cell lymphomas) on the basis of their specifically related dendritic cell microenvironment. In addition to monoclonal antibodies for the immunophenotypic characterization of dendritic cells and macrophages and to enzyme reactions, the combined use of anti-S-100 antibodies specific for each of the S-100 protein subunits, tested with sensitive procedures, would be a very useful tool in the attempt to classify the proliferative disorders of dendritic cells and macrophages.

S-100 protein detection in "follicular macrophages" of mouse lymphoid organs by ABC immunoperoxidase method.

The distribution of S-100 protein was investigated in normal or reactive lymphoid organs from adult mice, rats and humans by the avidin-biotin complex ABC) immunoperoxidase method. In mouse lymph nodes and spleen the protein appeared to be confined to the "tingible-body macrophages" and occasionally in sinus histiocytes in the lymph nodes. No immunoreaction product was detected in the other cell types present inside or outside the follicles. In the thymus the immunostained cells were located in the medullary area and corresponded morphologically to mononuclear PAS-positive cells with histiocytic appearance. By contrast, in rat and human organs S-100 protein was detected in interdigitating reticulum cells and dendritic reticulum cells. The present findings indicate that S-100 protein is located differently in mouse lymphoid organs and in other mammalian species.

Enzyme and immunohistochemistry of follicular hyperplasia in AIDS-related lymphadenopathy.

We used a panel of monoclonal and polyclonal antibodies to analyze frozen and paraffin-embedded lymph node biopsy specimens from 25 intravenous drug abusers (IVDA) with acquired immunodeficiency syndrome (AIDS)-related lymphadenopathy histologically characterized by follicular hyperplasia. Our aim was to obtain diagnostic clues to this commonly occurring pattern. Double-labelling immunohistological studies were also performed on selected frozen sections and 13 plastic-embedded specimens were tested by a number of enzyme reactions. Consistent features in IVDA included abnormally high numbers of intrafollicular T-cells, positive for acid phosphatase and beta-glucuronidase, most of which had Leu-2a-positive phenotype; a marked reduction or loss of mantle zone B-cells (positive for surface IgD-IgM and alkaline phosphatase); and disarray of the network of follicular dendritic reticulum cells (DRCs), as revealed with DRC-1 and anti-S-100 protein antibodies or with reaction for 5'-nucleotidase. When present, distinctive intrafollicular clusters of Leu-2a-positive T-cells and mantle zone B-cells were nearly always associated with areas lacking DRCs in some patients. The intrafollicular hypervascularity invariably found in IVDA proved to be of a true capillary nature, as demonstrated by alkaline phosphatase, 5'-nucleotidase, and ATPase reactions. In control tissues, all showing absence of Leu-2a-positive intrafollicular T-cells, most of the above individual changes could be detected, although they were occasional, mild, and never associated within the same follicle. By contrast, combined immunohistological and enzyme histochemical findings in IVDA indicated that in most follicles such changes were marked and very often associated within the same follicle in each case.(ABSTRACT TRUNCATED AT 250 WORDS)

Hemangioendothelioma of bone. A report of an unusual case with lymph node metastasis.

An unusual case of hemangioendothelioma of bone with regional lymph node metastasis in a 54-year-old woman is reported. The patient died nine weeks after admission of circulatory collapse. Six possibly similar cases were found in reviewing the literature. It is concluded that lymph node metastases from hemangioendothelioma of bone appear to be an extraordinary event and may represent an extremely unfavourable prognostic finding.

A histopathologic study of persistent generalized lymphadenopathy in intravenous drug abusers.

Persistent generalized lymphadenopathy (PGL) has been reported in groups at risk for the acquired immunodeficiency syndrome including intravenous (iv) drug abusers. We investigated the histology of the lymph node biopsies obtained from 18 iv heroin abusers. All the patients (13 males and 5 females; median age 23.5) had generalized lymphadenopathy. This was associated with one or more systemic symptoms in 72% of the cases. Immunologic abnormalities were present in most of the patients. The overall histological picture consisted of a fairly repetitive pattern dominated by an exuberant and diffuse hyperplasia of frequently misshapen and poorly outlined follicles, associated with a striking proliferation of capillary vessels and a very consistent amount of plasma cells both extending in the follicular and interfollicular areas. Multinucleated giant cells of the Warthin-Finkeldey type ("mulberry cells") were encountered in 22% of the cases. It would appear that in the follicular district the most indicative diagnostic clues of PGL can be obtained if one considers the abnormal morphology of the follicles associated with a consistent presence of plasma cells and a scattered fine network of proliferating capillary vessels.

Amyloidosis of specialized mesodermal tissues in non-myelomatous monoclonal gammopathy. A pathogenetic hypothesis.

The follow-up and the pathological findings of a previously described case of non-myelomatous monoclonal gammopathy with relapsing localized amyloid masses of the shoulder joints are reported. The peculiar "mesodermal" distribution of amyloid deposits, strictly limited to tissues lined by mesothelial or endothelial cells emphasizes the role of local factors in the pathogenesis of amyloidosis thus supporting the hypothesis that amyloid deposition can be related also to some differentiated tissues with peculiar local susceptibility. The morphological findings of the present case enable the Authors to suggest a pathogenic mechanism for amyloid deposition.

Lymph node immunohistology in intravenous drug abusers with persistent generalized lymphadenopathy.

Lymph node biopsy specimens from 16 intravenous drug abusers with persistent generalized lymphadenopathy were evaluated by immunohistochemical methods using a panel of antisera to detect different cell populations. The 11 cases that we tested on cryostat sections showed an increased number of Leu-2a-positive cells (cytotoxic-suppressor phenotype) in the follicular centers and a significantly reduced helper-to-suppressor T-cell mean ratio when compared with control tissues. In these 11 patients the peripheral helper-suppressor ratio was at the lower normal limit or inverted. Ten cases tested for anti-human T-cell lymphotropic virus type III antibodies were positive. In all 16 cases, immunohistology of paraffin-embedded sections demonstrated a polyclonal B population; 12 of 15 patients tested had polyclonal hypergammaglobulinemia, mostly IgG. The mixed-cell population of the lymph node sinuses was composed mostly of Leu-M1-positive and lysozyme-positive cells and, to a lesser extent, by alpha 1-antichymotrypsin-positive and S100 protein-positive cells. It seems that many of the immunologic dysfunctions found in these patients appear to be reflected in a fairly repetitive immunohistologic pattern.

Practical importance of routine paraffin-embedded bone marrow biopsy in multiple myeloma.

Paraffin-embedded bone marrow biopsy specimens obtained prior to (37) or after (25) therapy from 62 patients with multiple myeloma (MM) were analyzed with particular reference to infiltration pattern, extent of infiltration, and myeloid to myeloma tissue percentage ratio (MMR) to verify their mutual relationships and clinicopathologic relevance. Fifty-nine biopsies were evaluable for infiltration pattern (diffuse in 27, interstitial in 25, and nodular in 7). Diffuse and interstitial patterns were more common (P less than 0.025) in stage III and stage I patients, respectively. A higher (P less than 0.001) mean serum paraprotein level was found in patients with the diffuse pattern than in those with the interstitial pattern. The average extent of infiltration by myeloma cells in the residual myeloid tissue was higher (P less than 0.001) and a high extent (75% or more) was more frequently (P less than 0.005) seen in diffuse than in interstitial pattern cases. The average MMR value was lower (P less than 0.001) and a MMR value less than 1 was more frequently (P less than 0.005) seen in the diffuse pattern group than in the interstitial pattern group. All these differences were present also when a separate analysis was performed for treated and untreated patients. It seems that a diffuse histologic pattern, as opposed to interstitial, would significantly predict a bone marrow extent of infiltration of 75% or more, a MMR lower than 1, a higher serum paraprotein level, and a clinical stage III. Bone marrow biopsy appears thus to play a role in providing parameters of prognostic relevance in MM also in the course of the disease. Prospective studies are needed to establish whether histologic pattern has an independent prognostic value.

Role of fine-needle aspiration cytology in the preoperative evaluation of smooth muscle tumors.

A preliminary study was undertaken to assess the feasibility and the diagnostic role of fine-needle aspiration cytology (FNAC) in the preoperative evaluation of eight uterine smooth muscle tumors manifesting as single large masses with signs of growth. Percutaneous FNAC was performed under echographic control with a 22-gauge needle and the material was stained according to conventional techniques. Histology of surgically resected specimens was available for final diagnosis and comparative analysis in all the cases, including five leiomyomas (LM), one smooth muscle tumor of uncertain malignant potential (TUMP), and two low-grade leiomyosarcomas (LMS). Cellularity, as indicated by the density (crowding) of nuclei reflecting the amount of cytoplasmic volume, and the cohesiveness of the tissue fragments in the smears appeared to be the most important diagnostic parameters in the distinction between LM and LMS. LM usually showed few scattered poorly cellular fragments of highly cohesive tapering cells without nuclear crowding and with abundant cytoplasm. LMS usually showed a large number of single cells and fragments of loosely arranged tapering cells with nuclear enlargement and crowding and ill-defined scanty cytoplasm. Borderline forms such as TUMP were hardly distinguishable from LMS and LM. FNAC appears to be a feasible preoperative procedure in uterine smooth muscle tumors and may play a diagnostic role, especially in distinguishing frankly benign from overtly malignant forms.

[Temporal aspects of the function of the renin-angiotensin-aldosterone system. Contribution to the methodological standardization of explorative test (author's transl)]. / Aspetti temporali della dinamica del sistema renina-angiotensina-aldosterone. Contributo alla standardizzazione della metodologia di esplorazione funzionale.

The purpose of this study is to throw further light on the problem of the methodological standardization in the study of the renin-angiotensin-aldosterone system (RAAS). Thus, in 10 normal volunteers and 33 patients with moderate, recent and uncomplicated essential hypertension, divided into the subtypes with normal, high and low renin, it has been performed a timing analysis of the behaviour of plasma renin (PRA) and aldosterone (PA). PRA and PA were measured by radioimmunological methods in blood samples simultaneously collected in steady state conditions (study of circadian rhythmicity) and during the course of manipulative tests (orthostatism, dietary sodium restriction, orthostatism associated with sodium restriction, furosemide administration). The study revealed that each category of the present series of healthy and hypertensive individuals has an own temporal organization in the entire function of RAAS. This finding allowed to standardize the RIA reference indices as chronocorrelated intervals of normality and to establish the optimal criteria for the assessment of biochemical data and methodological study of hypertensive patients.