Exploring the mesenteric lymphatic apparatus: A morphological and immunohistochemical investigation with clinical correlations.

Modern immunohistochemical techniques allow a detailed study of the lymphatic system in many organs and areas of the body. We performed an in-depth study on lymphatic vessels of the ileal and colonic mesenteries, together with the greater omentum where they appear particularly numerous and mainly represented by capillaries interconnected among themselves and with lymph nodes. The capillary wall consists of a fine single sheath of endothelial cells wrapped around by a subtle collagen membrane and deprived of valves. The progression of lymph flow is promoted by external forces acting on the capillary walls. Only at the mesenteric roots can pre- and post-lymph nodal collector vessels be observed. Our observations help to explain different patho-physiological correlations and the possible presence of skip lymph node metastases.

Deep inside of gastric signet-ring cell carcinoma.

The histology of signet-ring cell carcinoma (SRC) of the stomach has been revisited with the support of current immuno- histochemical techniques in order to explain particular features of this tumor; its great capacity of local diffusion and lymph node metastasis, also through a neo-lymphoangiogenesis. An observational retrospective study on 50 cases of SRC in stage II and III has been performed with the addition of histochemical (Alcian Blue, DDD-Fast Blue B, Mercury Orange) and immunohistochemical (cytocheratin, CD3, CD4, CD8, CD10, CD56, CD68, perforin, granzyme B, podoplanin, collagen type IV) investigations for each case. The signet ring cells, typical for this tumor, show abundant content of electro-negative sialomucins and demonstrate a great capacity of diffusion through the gastric wall. They evoke production and deposition of collagen type IV in the sub-mucosa layer through the local action of fibroblasts. The immunological response to this tumor in the gastric wall and in the metastatic lymph nodes is represented by an increase of B and T-helper lymphocytes, but not of T-killers or natural killers. The neoplastic cells are curiously able to avoid these newly formed 'lymph nodules'. An extended neo-lymphangiogenesis has been observed around the primary tumor and in metastatic lymph nodes. A careful immunohistochemical characterization has allowed a better knowledge of SRC, regarding especially the peculiar behavior of local diffusion of its cells, the associated neo-lymph angiogenesis, and poor immunological reaction.

New advances on placental hydrops and related villous lymphatics.

Fetoplacental hydrops is the final stage of several pathological conditions in which the placenta and umbilical cord become edematous and the fetus develops an anasarcatic state characterized by an excessive accumulation of extravascular fluids in at least two serous cavities of the body. It is a common histological finding of stillbirth, characterized by the appearance of markedly edematous villi, suggesting an increased interstitial fluid accumulation. The recent improved knowledge of lymphangiogenesis and the availability of monoclonal antibodies selectively labeling lymphatic endothelium lead to the hypothesis that villous edema is essentially a lymphedema from defective lymphatic function following inadequate villous blood circulation. Lymphedema is a morphologic phenotype found by our research group in a 24-case series of stillbirths from different morbid conditions such as chromosomal aberrations, congenital malformations, inherited hemoglobinopathies, and prolonged perinatal severe anoxia. Unlike long-lived organs, the placenta is devoid of innervation by the autonomic nervous system; therefore, the vascular tone regulation and the peripheral perfusion are modulated by the expression of the angiotensin converting enzyme (ACE) in the vascular endothelia. This finding may suggest to the clinician to search for a more suitable therapy in case of mother's hypertension during pregnancy.

Acquired immunodeficiency similar to Gulf War illness in a dead former serviceman.

A 38-year-old non-commissioned officer was certified unfit for military duty several months before his death. The forensic autopsy revealed a severe bone marrow aplasia and a pulmonary angioinvasive aspergillosis. Moreover, the presence of inorganic foreign particles in the pulmonary macrophages and intestinal endothelia was observed. The microanalysis implemented on these last selected specimens revealed the presence of silica particles microimpregnated by lanthanides and steel. The patient's acquired immunodeficiency appears comparable with that of Iraqi civilians suffering from Gulf War illness. This is the first report in the literature of the presence of intestinal endothelia engulfed by foreign war particulates; the silica particles may have entered the intestinal endothelia via the blood stream or by ingestion of impregnated fruit and vegetable foodstuffs. This finding provides new perspectives in the assessment of war-associated diseases and includes electron probe microanalysis among the new techniques of military and forensic medicine.

Neurotrophins and neurotrophin receptors in human lung cancer.

The expression of neurotrophins (NTs) and related high- and low-affinity receptors was studied in surgical samples of histologically diagnosed human tumors of the lower respiratory tract. The experiment was conducted with 30 non-small cell lung cancer specimens and in eight small cell lung cancer specimens by Western blot analysis and immunohistochemistry to assess expression and distribution of NT and NT receptor proteins in tissues examined. Immunoblots of homogenates from human tumors displayed binding of anti-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and NT-3 antibodies as well as of anti-tyrosine-specific protein kinase (Trk) A, TrkB, and TrkC receptor antibodies, with similar migration characteristics than those displayed by human beta-NGF and proteins from rat brain. A specific immunoreactivity for NTs and NT receptors was demonstrated in vessel walls, stromal fibroblasts, immune cells, and sometimes within neoplastic cell bodies. Approximately 33% of bronchioloalveolar carcinomas exhibited a strong membrane NGF and TrkA immunoreactivity, whereas 46% adenocarcinomas expressed an intense TrkA immunoreactivity but a weak immunostaining for NGF within tumor cells. Moreover, squamous cell carcinomas developed an intense TrkA immunoreactivity only within stroma surrounding neoplastic cells. A faint BDNF and TrkB immunoreactivity was documented in adenocarcinomas, squamous cell carcinomas, and small cell lung cancers. NT-3 and its corresponding TrkC receptor were found in a small number of squamous cell carcinomas within large-size tumor cells. No expression of low-affinity p75 receptor protein was found in tumor cells. The detection of NTs and NT receptor proteins in tumors of the lower respiratory tract suggests that NTs may be involved in controlling growth and differentiation of human lung cancer and/or influencing tumor behavior.

[Parosteal rib lipoma: description of a case]. / Lipoma parostale costale: descrizione di un caso.

Parosteal lipoma is a usually indolent, rare benign tumor, characterized by clinicopathological findings similar to those of the commonly occurring subcutaneous lipoma, except for its intimate relationship with the connective tissue of the subjacent periosteal region. Parosteal lipoma commonly affects the diaphysis of long bones of the upper and lower limbs. We report an exceedingly rare case of parosteal lipoma of the rib, which presented in a 59-year-old man experiencing previous multiple traumas in this site.

Correlation between high intake of glycyrrhizin and myolysis of the papillary muscles: an experimental in vivo study.

The ingestion of large quantities of glycyrrhizin, whether as a drug or a sweetener, is known, in susceptible subjects, to induce a syndrome similar to hypermineralcorticoidism, with bouts of hypertension, hypokaliaemia and rabdomyolysis, sometimes associated with severe renal failure and hypokaliaemia-induced arrythmias. Glycyrrhizin is also known to isomerize into the glycyrrhetic (or glycyrrhetinic) acids 18alpha- and 18beta-. In previous works, we reported that these metabolites cause bouts of hypertension and reduction in diuresis at low doses in the rat. In particular, the alpha isomer causes significant elimination of the calcium ion in the urine. The present findings confirm that 18alpha-glycyrrhetic acid is more toxic than either glycyrrhizin or the beta isomer. Histopathological study of tissue samples taken from rats treated with the alpha isomer also reveal selective damage to the myocardium with oedema, myolysis, apoptosis and blistering of the sarcoplasm. These effects begin to appear in the course of subchronic treatment, they manifest themselves in acute treatment and correlate closely with the electrocardiographic changes recorded in rats acutely treated with 18alpha-glycyrrhetic acid.

A case of lung myelolipomatosis in a patient with bronchial carcinoid.

Myelolipomas are very rare benign tumours composed of an admixture of mature adipose tissue and normal haematopoietic cells. Although they are most commonly found in the adrenal glands, extra-adrenal myelolipomas are documented. We described a case of myelolipoma arising in the lung in a 52-year-old man. The lesion was found incidentally in association with a carcinoid. To our knowledge, this is the second instance of this neoplasm presenting as a lung lesion, and the first case associated with bronchial carcinoid. Pathogenesis and aetiology of myelolipomas are referred to in this paper with special regard to the clinical and pathological findings.

Nucleolar protein p120 expression in oral carcinoma.

Nucleolar protein p120 is a proliferation-associated antigen expressed by cells in early G1 phase, identified by the monoclonal antibody FB-2. Its expression has been evaluated in breast and prostate cancer, and proved to be significantly correlated with other prognostic parameters. In oral pathology, p120 protein content is able to distinguish between non-neoplastic and malignant lesions. In the present study, the immunohistochemical expression of p120 protein was evaluated in fifty cases of oral squamous carcinoma and compared with histological grading, pTNM staging, DNA ploidy status and follow-up of the patients, in order to establish its prognostic value, p120 mean area was significantly correlated to all these parameters, apart from lymph node involvement, indicating the strong predictive potential of this marker. In conclusion, quantitative immunohistochemical analysis of p120 protein represents an easy and reliable method for the assessment of clinical outcome and the definition of risk groups in oral carcinoma.

[AIDS splenomegaly and related iron problems]. / Splenomegalia da AIDS e problematiche siderotiche connesse.

Spleens collected from 85 consecutive autopsies of AIDS patients (mean age 37 years) were studied. Splenomegaly, observed in 59 cases (69.4%), does not statistically correlate with life style and blood transfusions. Eleven very large spleens (over 890 g) were associated with opportunistic infections (i.e.: mycobacteria, true fungi and rochalimaea). The histological pattern was characterized by marked lymphoid depletion of the white pulp and--in 67 cases (89.4%)--packing of the pulp cords by macrophages engulfed of brown pigment which was strongly positive to the Perls reaction for ferric iron. The contemporary presence of Perls positive and p24 immunoreactive material was diffusely observed in the cytoplasm of splenic macrophages, also positive to the alkaline tetrazolium reaction. The same was observed in the cytoplasm of monocytes/macrophages of lung and brain (in 5 out of 5 patients with splenomegaly > 600 g, randomly selected). We believe that our findings deal with the formation of a haemoglobin--p24 complex and are in keeping with recent data which suggest the formation of disulphide bonds between viral proteins and haemoglobin.

p120 and AgNOR nucleolar protein expression: a comparison with nuclear proliferation markers in oral pathology.

To find a better method for predicting the biological behavior of certain oral cavity lesions, the expression of nucleolar protein p120 and nucleolar organizer region counts (AgNOR) was compared with that of nuclear proliferation markers MIB-1 and PCNA in 10 cases of keratotic epithelial hyperplasia (KEH), 10 cases of epithelial dysplasia (ED), and 15 cases of squamous cell carcinoma (SCC). Significant differences in p120 and AgNOR mean area values and PCNA labeling index (LI) were recorded between KEH and ED, as well as ED and SCC (Student-Neumann-Keuls test). All markers significantly differed between SCC grades I and III. Significant differences were also noted in AgNOR mean area values between grade I and II SCC and in p120 mean area values. MIB-1 and PCNA LI differed significantly when grade II and III SCC were compared (SNK test). There were significant correlations between p120 and AgNOR (Pearson correlation coefficients) and between both of them and the proliferative indexes. AgNOR correlated with tumor grade, stage, and lymph node status (Spearman correlation coefficients), suggesting a prognostic role for that marker.

Immunohistochemical detection of nucleolar protein p120 in paraffin-embedded tissues.

The monoclonal antibody FB-2 recognizes the antigen p120-kDa protein (p120), associated with the nucleolar matrix. p120 has originally been reported as expressed and detectable in malignant and non-neoplastic proliferating cells, but not in most normal resting tissues and benign tumours. In the present study, a reliable immunostaining method was used to detect p120 on formalin-fixed, paraffin wax-embedded tissue, testing it on 148 samples from different neoplastic and non-neoplastic tissues from different organs (breast, colon, lung, prostate, bladder, lymph nodes, skin, tongue and liver). The immunostaining was performed after the application of a specific antigen-unmasking protocol based on six consecutive cycles of microwave oven heating. Under these retrieval conditions, p120 antigen was clearly detectable, not only in hyperplastic and malignant cells, but also in stromal and normal non-proliferating cells of all the tissues evaluated. Our results show that the nucleolar protein p120 can be detected by routine immunohistochemistry in formalin-fixed, paraffin-embedded tissue and is expressed in all nucleated cells under any biological condition.

The changing histopathology of primitive lung cancer.

This study was performed on 1710 lung epithelial malignant tumours, diagnosed in the biennia 1963/1964 (42 cases), 1973/ 1974 (293), 1983/1984 (637) and 1993/1994 (738). It was aimed at evaluating whether, over a time span that long, the distribution of the histopathological patterns could have changed. The neoplasms were classified according to the WHO criteria (1981). From our data, a striking increase becomes apparent for all histotypes. Squamous cell carcinoma holds the leading position both among males and females, even though its rate of increase tends to slow down among males. Analogous trend is observed for all the other histotypes but adenocarcinoma. Among females, the rate of increase continues to accelerate for all histotypes but large cell carcinoma. The male to female ratio is ranging from 2:6 for adenocarcinomas to 12:1 for squamous cell carcinomas. In conclusion, impressive changes seem to have occurred in the frequency of the different lung oncotypes over this time span, especially for adenocarcinoma (increasing trend) and large cell carcinoma (decreasing trend). It also seems worth underlying that the marked increase among females could make lung carcinoma the number one cause of cancer-related deaths in this sex.

Pulpal neuropeptidergic fibers.

A study was carried out on Met- and Leu-enkephalin, Gastrin/CCK-, SP-, CGRP-, NPY-immunoreactive fibers using paraffin sections of dental pulp taken from 8 apparently normal teeth (wisdom teeth or teeth extracted for orthodontic reasons). Within the limitations of the samples studied, dental pulp is characterized by the presence of sensory (Enkephalin-, Gastrin/CCK-immunoreactive) and pain fibers (SP-immunoreactive) and of fibers with a potent vasodilatory action (CGRP-immunoreactive) and by the absence of fibers with a vasoconstrictor action (NPY-immunoreactive).

[Contribution of echography in the diagnosis of gastro-esophageal reflux in children]. / Apporto dell'ecografia nella diagnosi del reflusso gastro-esofageo nel bambino.

56 children, aged between 0-13 month, affected by chronic vomiting were evaluated both with ultrasound and radiological methods, in a double blind study. 22/56 children resulted to be affected by gastro-oesophageal reflux (GER) with upper GI series versus 18/56 with ultrasound. Four cases, ultrasound negative and X-ray positive, represented a borderline situation in which a mild gastro-oesophageal reflux is to be considered paraphysiological phenomenon. The accuracy and handleness of ultrasound evaluation in GER are emphasized.

[A clinico-statistical study of the anatomico-clinical and immunohistochemical correlations in chronic lesions of the oral cavity]. / Studio clinico-statistico sulle correlazioni anatomo-cliniche e immunoistochimiche di alcune lesioni croniche del cavo orale.

A series of 191 biopsies of the oral cavity (epulides, cysts and granulomas) carried out in 1975-76 (104 cases) and 1985-86 (87 cases) is reported. Data regarding age, sex, site, histopathological diagnosis, dimensions of the lesions and type of operation were processed on an AT mod 280 computer in a file specially created using an Ashton Tate DB 111 Plus. Percentage values relating to epulides proved substantially unchanged. Those regarding cysts showed a particularly evident increase in males, while those relating to granulomas showed a decline in both sexes. From the immunopathological viewpoint, angiomatous epulides, prevalent in the female sex, presented a structural framework different to that of plasma cellular epulides, testified by the presence of vessels lined with endothelia reactive to fucose and with scanty plasmocyte immunoglobulin component.

Immunohistologic analysis of mycobacterial antigens by monoclonal antibodies in tuberculosis and mycobacteriosis.

Four monoclonal antibodies (MoAbs), 60.15, 61.3, 105.10, and 2.16, directed to different proteins of Mycobacterium tuberculosis were used by an indirect avidin-biotin complex peroxidase-antiperoxidase method to detect mycobacterial antigens in lung, lymph node, and joint tissue specimens of tuberculous patients. Using MoAb 60.15, which recognizes a broad range of cross-reactive mycobacterial proteins with a molecular mass of 28 kilodaltons (kD), scattered materials (mycobacterial in origin) were observed, many of which were located within phagocyte cytoplasm. With MoAb 61.3, which reacts with a 35 kD protein present in M tuberculosis, Mycobacterium africanum, and Mycobacterium bovis, many clumped particles similar in size and shape to acid-fast bacilli were observed within the phagocyte cytoplasm (lung tissue) and positive macrophages with lysosomes were distributed throughout the cytoplasm (bronchoalveolar lavage). The specificity of this MoAb (61.3) was confirmed by the negative staining of positive lymph node specimens obtained from a patient infected with Mycobacterium kansasii. MoAbs 105.10 and 2.16 bind to the cross-reactive 65 kD heat shock protein that is present in mycobacteria and stain scattered particles and dark clumps of bacilli within the phagocyte cytoplasm. On the basis of this study, immunohistochemical detection of mycobacterial antigens appears to be useful in establishing the mycobacterial etiology of caseating granulomas and in avoiding the false-negative results obtained by traditional staining methods.