Silver staining combined with alcian blue and hematoxylin-eosin for the detection of Lawsonia intracelullaris in swine proliferative enteropathy.

Fragments of ileum from 663 pigs were collected in abattoirs, prepared with the use of standard histological methods and stained with a novel sensitive histochemical method for the detection of porcine proliferative enteropathy. The method is a combination of the following 3 well-known methods, the Warthin-Starry method, alcian blue and hematoxylin-eosin. In 11 cases, mucus-producing cells were completely absent, severe adenomatous proliferation was observed and intracellular bacteria were found in enterocytes. Disappearance of goblet cells and the presence of adenomatous proliferation without any detectable intracellular bacteria were observed in 16 cases. In the remaining 636 cases, histological changes and intracellular bacteria were not found. When comparing the conventional Warthin-Starry method with the modified staining method presented here, the same 16 cases were found. However, the method presented here enables examination of large numbers of sections in a relatively short period of time.

Meiotic mutants of Medicago sativa show altered levels of alpha- and beta-tubulin.

We have analysed the level of accumulation of alpha- and beta-tubulin polypeptides in flowers collected from different meiotic mutants of alfalfa (Medicago sativa L.). The H33 mutant previously identified as a producer of male and female gametes with the somatic chromosome number (2n gametes) as a result of defective spindle orientation or, more rarely, abnormal cytokinesis, showed a higher level of alpha- and beta-tubulin compared to control diploid plants and approximately the same level as control tetraploid plants. A higher level of tubulin was likewise observed in diploid plants displaying abnormalities in spindle orientation and cytokinesis, which had gone through 3-4 cycles of phenotypic recurrent selection to increase 2n gamete production. A similar analysis was performed on another class of Medicago meiotic mutants characterized by production of 4n pollen (jumbo pollen, due to the absence of cytokinesis at the end of meiosis) and 2n eggs. Again, the level of alpha- and beta-tubulin was found to be higher in the mutants than in diploid controls. We conclude that meiotic defects, such as abnormal spindle orientation or cytokinesis leading to the formation of 2n gametes, determine an increased level of tubulin, the main constituent of plant microtubules (MTs).

Palatal myoclonus and unusual MRI findings in a patient with membranous lipodystrophy.

We describe an Italian male patient, deceased at 29 years of age, affected with a syndrome characterized by childhood-onset seizures, mental disorders, motor dysfunction and bilateral palatal myoclonus. Skeletal X-ray examination showed diffuse osteopenia of the tubular bones, and cyst-like lesions in the carpal, metacarpal and tarsal bones bilaterally and in the proximal end of the right femur. Skin biopsy showed subcutaneous and adipose tissue containing membranocystic structures. Cerebral MR and CT scans showed fronto-temporal atrophy, altered signal of the white matter and mineralization of the caudate and dentate nuclei. These findings strongly recall polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy, but in the present case, bone alterations were not prominent; moreover, palatal myoclonus has never previously been described in this syndrome.

Warm heart surgery eliminates diaphragmatic paralysis.

Since January 1992, we adopted a new method of myocardial protection: warm blood cardioplegia with continuous ante-retrograde combined delivery during normothermic cardiopulmonary bypass, (CPB) instead of cold blood intermittent cardioplegia plus topical ice slush in hypothermic CPB. We have compared postoperative chest X-rays of 50 patients who underwent elective coronary artery bypass with normothermic CPB to postoperative chest X-rays, of 50 patients operated upon with hypothermia. In the cold group transitory diaphragmatic paralysis, as well as pleural effusions and thoracentesis related to the hypothermia, and topical cooling, were statistically increased over that of warm group. The data suggest that topical cooling with slush ice is responsible for phrenic nerve injury and that warm heart surgery has no associated incidence of diaphragmatic injury.

Molecular cloning of two novel rice cDNA sequences encoding putative calcium-dependent protein kinases.

We have isolated, from a cDNA library constructed from rice coleoptiles, two sequences, OSCPK2 and OSCPK11, that encode for putative calcium-dependent protein kinase (CDPK) proteins. OSCPK2 and OSCPK11 cDNAs are related to SPK, another gene encoding a rice CDPK that is specifically expressed in developing seeds [20]. OSCPK2 and OSCPK11-predicted protein sequences are 533 and 542 amino acids (aa) long with a corresponding molecular mass of 59436 and 61079 Da respectively. Within their polypeptide chain, they all contain those conserved features that define a plant CDPK; kinase catalytic sequences are linked to a calmodulin-like regulatory domain through a junction region. The calmodulin-like regulatory domain of the predicted OSCPK2 protein contains 4 EF-hand calcium-binding sites while OSCPK11 has conserved just one canonical EF-hand motif. In addition, OSCPK2- and OSCPK11-predicted proteins contain, at their N-terminal region preceding the catalytic domain, a stretch of 80 or 74 residues highly rich in hydrophilic amino acids. Comparison of the NH2-terminal sequence of all three rice CDPKs so far identified (OSCPK2, OSCPK11 and SPK) indicates the presence of a conserved MGxxC(S/Q)xxT motif that may define a consensus signal for N-myristoylation. OSCPK2 and OSCPK11 proteins are both encoded by a single-copy gene and their polyadenylated transcripts are 2.4 and 3.5 kb long respectively. OSCPK2 and OSCPK11 mRNAs are equally abundant in rice roots and coleoptiles. A 12 h white light treatment of the coleoptiles reduces the amount of OSCPK2 mRNA with only a slight effect on the level of OSCPK11 transcript. With anoxic treatments, OSCPK2 mRNA level declined significantly and promptly while the amount of OSCPK11 transcript remained constant.

Successful bone marrow transplantation in children with severe aplastic anemia using HLA-partially matched family donors.

Bone marrow transplantation (BMT) using HLA-partially matched family donors has produced disappointing results (25-30% of long-term survivors) in patients with severe aplastic anemia. We describe two children affected by severe aplastic anemia, not responsive to immunosuppressive therapy, who underwent allogeneic bone marrow transplantation using a HLA-partially matched family donor. Both cases presented 2 first class HLA-antigens (A and B) disparity between donor and recipient. The pretransplant conditioning regimen consisted of cyclophosphamide, thoracoabdominal irradiation, cytosine-arabinoside, and antilymphocyte globulin. As graft versus host disease (GVHD) prophylaxis, Cyclosporine-A was administered at usual dosages for 6 months. A full marrow engraftment was observed in both cases. Only grade I acute GVHD, promptly responsive to corticosteroid therapy, developed with no chronic GVHD. Five months after transplant, both children progressively developed hypertension, renal function impairment, thrombocytopenia, and severe normochromic anemia, with erythropoietin serum levels lower than expected for the haematocrit. After antihypertension treatment and supportive therapy, the clinical picture progressively improved, while treatment with recombinant human erythropoietin completely corrected the long-lasting anemia. The two children are alive and well 28 months after the transplant, with a Karnofsky score of 100% and a normal peripheral blood count. The authors suggest that, once immunosuppressive therapy has failed, BMT from donors other than HLA-identical sibling is a feasible approach in children affected by severe aplastic anemia, not having an HLA-identical donor.

[Significance of the combination of small intestine enema and ultrasonography in the follow-up of Crohn's disease. Clinical correlations]. / Validità dell'associazione del clisma dell'intestino tenue e dell'ecotomografia nel follow-up della malattia di Crohn. Correlazioni cliniche.

High-resolution US yields important information for the study of Crohn's disease. A more accurate follow-up is possible by the integration of US with radiological and clinical findings. Sixty-two patients with suspected or known Crohn's disease were examined with double-contrast small bowel enema and high-resolution US. The latter allowed the demonstration of such pathologic findings as mesenteric thickness greater than 15 mm, submucosal thickness (third hyperechoic layer greater than 50% of total bowel wall thickness), lymphadenopathies, and fluid collections. The authors suggest that US grading be performed, and radiological correlations made. The combined use of the two techniques yields better results and allows a better assessment of the patients, which is very useful for both prognosis and therapy.

Translocation (8;21) in two cases of refractory anemia with excess of blasts in transformation.

We report two cases of refractory anemia with excess of blasts in transformation (RAEB-T) with the translocation (8;21), which is frequent in ANLL but not in myelodysplastic syndromes (MDS). A review of such cases leads us to conclude that myeloproliferative disorders characterized by the t(8;21) may be preceded by an MDS phase.

Allogeneic bone marrow transplantation in children from other than HLA-identical sibling donor.

Optimal allogeneic bone marrow transplantation (BMT) presupposes the use of a HLA-identical sibling as donor. Unfortunately, only about 30% of patients have an HLA-matched donor, so that the use of alternative donors has been increasingly used. We report an analysis of 13 children transplanted using an HLA-partially matched donor as source of haemopoietic stem cells. They suffered of ALL (3 pts), ANLL (1 pt), SAA (2 pts), Osteopetrosis (1 pt), Wiskott-Aldrich Syndrome (2 pts), Severe Combined Immunodeficiency Disease (2 pts) and Familial Haemophagocitic Lymphohistiocytosis (2 pts). Full engraftment was obtained in all 11 of the patients who survived longer than 14 days and, globally, a moderate incidence of acute GvHD (grade II-IV) was observed in the evaluable patients (3 out of 11 with a percentage of 27%); only a patient of the six survivors more than one hundred days after BMT had severe chronic GvHD (16.6%). Four pts (31%) are actually alive and well (mean follow-up 358 days) with a mean Karnofsky score of 95%. Our data suggest that BMT from HLA-partially matched donors could represent a possible alternative therapeutic strategy in children when a compatible donor is not available. This is especially due to the reduced severity of GvHD in childhood and because of T-cell depleted marrow transplants could obtain more satisfactory results when employed in typical pediatric non-malignant disorders (i.e. immunodeficiencies) rather than in leukemia.

In vivo and in vitro growth of SCLC cells derived from biopsies.

In order to increase the availability of SCLC cells derived from biopsies, in vivo and in vitro growth methods were investigated. The cells grown in both conditions were periodically monitored for reactivity with 2 monoclonal antibodies (MAbs): MLuC1 directed against SCLC cells and IM1 which recognizes the class II antigen on activated lymphocytes and macrophages. About 50% of the 28 analyzed SCLC specimens were found to proliferate in one or both systems. The in vitro-grown cells exhibited the same heterogeneity found in the original cell suspensions and moreover, in some cases only normal cells were recovered after several in vitro passages. From the subcutaneous transplanted tumors a large number of MLuC1-positive tumor cells could easily be recovered, thus indicating the validity of the in vivo methodology. The MBr1 MAb, directed against an epithelial antigen, was found to react with about 50% of the 26 tested tumors, mainly those which demonstrated in vivo and/or in vitro growth capacity. These data suggest that only some tumors, presumably with peculiar biological characteristics, can efficiently grow in these artificial systems.

CA 72-4 radioimmunoassay for the detection of the TAG-72 carcinoma-associated antigen in serum of patients.

Monoclonal antibody (MAb)B72.3 has been used to detect the presence of TAG-72 in the serum of carcinoma patients. We have developed new anti-TAG-72 MAbs and have selected one of these, CC49, as the "catcher" MAb with 125I-B72.3 as the detecting antibody in a double-determinant immunoradiometric assay. This combination enabled the development of a sequential assay (designated CA 72-4) that showed optimal quantitative properties as demonstrated by such parameters as linear dose-response, high re-producibility, and lack of serum-matrix and "hook-back" effects. Only 3.5% of 744 normal sera and 6.7% of 134 sera from patients with benign gastrointestinal diseases had TAG-72 levels greater than 6 U/ml. Approximately 40% of 303 patients with gastrointestinal malignancies had serum TAG-72 levels of greater than 6 U/ml (55% of the patients with advanced disease). Thirty-six percent of patients with adenocarcinomas of the lung and 24% of patients with ovarian cancer (53% stage IV patients) also had elevated serum TAG-72 levels. A poor correlation was found between the carcinoembryonic antigen (CEA) and TAG-72 values of sera obtained from gastric cancer patients. Thirty-four percent of CEA negative cases were scored positive in the CA 72-4 assay, suggesting the complementarity of the CA 72-4 assay to CEA assays in the analysis of sera from patients with certain malignancies.

Histopathological characterization of a novel monoclonal antibody, MLuC1, reacting with lung carcinomas.

A monoclonal antibody (MoAb), MLuC1, derived from the fusion of P3-X63-Ag 8-U1 mouse myeloma cells with spleen cells from an HR mouse immunized with the carcinoma cell line SW626, was studied to define its reactivity profile on normal and neoplastic human tissues and its potential clinical applications in lung cancer. Evaluation of paraffin sections using the ABC immunoperoxidase method showed a "pan-epithelial" reactivity; a large majority of epithelial components of organs in the respiratory, digestive and urogenital systems (except liver, rectum and ovary) were immunostained. As regard to neoplastic tissues MLuC1 recognized 84% of lung carcinomas (82% of small cell, 100% of squamous cell, 74% of adenocarcinomas), 86% of breast and 62% of ovarian carcinomas. On the contrary, MLuC1 was non-reactive with the other normal and tumoral non-epithelial tissues. Due to its spectrum of reactivity this MoAb could be useful for different diagnostic purposes such as differential diagnosis and lung cancer cytology.

Chromosomal abnormalities in a primary small cell lung cancer.

A cytogenetic analysis of a fresh primary tumor specimen of small cell lung cancer showed a del(3)(p14p23) in the majority of metaphases. Additional clonal changes were found in the karyotype. No abnormalities for Ha-ras, Ki-ras, N-ras, myb, or myc were detected by Southern blot analysis of the tumor DNA.

[Multi-institution, multidisciplinary radiology and endoscopy trial of the diagnostic accuracy of the double contrast examination of the stomach surface]. / Trial policentrico multidisciplinare radiologia-endoscopia sul rendimento diagnostico dell'esame a doppio contrasto di superficie dello stomaco.

In 12 X-ray and gastroenterology departments 1570 patients, clinically suspected to have a gastric pathology, have been examined by double contrast (surface) examination of the stomach followed by gastroscopy. Diagnostic accuracy was 93% in respect of endoscopic data, assumed correct by hypothesis. On the basis of a cost and benefit evaluation, made according to the well known criteria, a wider routine use of the double contrast technique is proposed.

Skull changes and intellectual status in hydrocephalic children following CSF shunting.

A total of 76 shunt-treated hydrocephalic children were examined at least once between two and 12-years after surgery to determine what skull changes had occurred and to assess the children's intellectual status. The high incidence of skull changes in shunt-treated, non-tumoral, hydrocephalic children was confirmed. A strict relationship existed between skull changes and low intracranial pressure. The results indicate that impairment of mental development parallels the degree of skull changes, suggesting a close relationship between impaired mental development and cerebrospinal fluid hypotension.