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Genetic diversity among a complex of cereal cyst nematodes inferred from RFLP analysis of the ribosomal internal transcribed spacer region.

This study examined the restriction polymorphism (RFLP) of the nuclear ribosomal DNA in Heterodera avenae, H. filipjevi, H. mani, H. latipons, and the taxonomically unclear Gotland strain in order to establish a molecular characterization and phylogenetic relationships in the complex of cereal cyst nematodes (CCN). The internal transcribed spacer (ITS) and 5.8S rDNA were amplified by PCR from a single female or a cyst of 27 different geographic isolates of the CCN complex and one population of H. schachtii, used as outgroup. The amplified product was 1.2 kb long and 14 of 15 enzymes produced restriction fragments for each isolate. Relationships between populations were determined from UPGMA analysis based on distance values calculated from RFLP data. Digestions with TaqI clearly differentiated H. avenae, H. latipons, and a group composed of H. filipjevi and the Gotland strain. Six endonucleases (HaeIII, HinfI, ItaI, PstI, TaqI, and Tru9I) produced the same restriction pattern with H. filipjevi and the Gotland strain, and both were clearly separated from H. avenae with PstI. Restriction sites have revealed a mixture of the species H. latipons and H. avenae, and possible infraspecific variation in H. avenae. The inferred phylogenetic relationships of species in the CCN complex are in agreement with their morphological characterization.

Animals↗

MRI in characterization of focal liver lesions: comparison of T2 weighting by conventional spin-echo and turbo spin-echo sequences.

Forty-one patients with 61 proved focal liver lesions underwent MRI of the liver at 1.0 T, with the aim of evaluating the usefulness of turbo spin-echo (TSE) sequences in characterizing focal liver lesions, by comparing them with conventional spin-echo (CSE) sequences. Two different TSE protocols were employed. with constant echo time and varying repetition time: TSE-S (3000 msec) and TSE-L (5100 msec). All images were evaluated quantitatively (signal-to-noise ration 'SNR') and qualitatively: because benign lesions were all liquid (12 cysts and 10 hemangiomas), they were well characterized morphologically on the basis of signal intensity. Mean SNR was significantly different between metastases and benign lesions (P < .0001) with all T2 sequences. Among the single T2 sequences tested, logistic regression analysis showed TSE-L to have the best predictive ability of the nature of focal lesions, with a G value of 42.02, compared to 29.87 of TSE-S and 25.55 of CSE second echo (SE II). The combination of TSE-L with TSE-S did not modify these results, whereas the combination of TSE-L with CSE only resulted in slight improvement (G = 46.95). Comparison of the receiver operating characteristic (ROC) curves showed only SE II (area under the ROC curve of .8312) to be significantly inferior to the best single sequence, or TSE-L (area under the ROC curve of .9176; P = .027). All sequences were equivalent in qualitative evaluation with good reproducibility, sensitivity ranging from .94 to 1.0 and specificity ranging from .86 to .93. This study confirms the value of TSE sequences in characterization of focal liver lesions. Time of acquisition is strongly reduced with these sequences, whereas results are fairly similar to those obtained with CSE. TSE sequences could therefore replace CSE for the study of focal liver lesions.

Adult↗

[Organization of the Pediatric Tumor Cell Bank of the Society of Pediatric Oncology and Hematology (GPOH)].

Characterized cell lines are absolutely necessary in applied research of cell biology and medicine. For the completion of diagnosis and therapy especially in pediatric oncology we are establishing a Cell Bank for Pediatric Tumors. The Cell Bank for Pediatric Tumors collects tissue samples of different types of solid malignant tumors from children and young adults. The specimens are transferred to in vitro culture (guidelines of the American Type Culture Collection-ATCC), the resulting cells are characterized to assure accordance with the histogenesis of the original tumor and stored in liquid nitrogen. The cell cultures are characterized morphologically (phase contrast microscopy) and immunocytochemically (ABC-method). To prove the malignancy of cells in primary culture the amount of hypertetraploid cells was determined (DNA-Scanning-Cytophotometry). Cell lines are checked to find out whether they develop tumors in nude mice followed by an analysis of the karyotype. Additional investigations (e.g. in vitro test of cytostatic drug resistance) are carried out on request by the sender. Part of the tumor tissue which is used to start the cell culture is in parallel diagnosed histopathologically at the Children's Tumor Register, Kiel and/or at the Charité. By the end of the year 1995 the Cell Bank for Pediatric Tumors had received 183 different specimens including 123 solid tumors (e.g. 24 neuroblastomas, 18 osteosarcomas, 12 Wilms' tumors, 13 rhabdomyosarcomas), 44 tissue specimens without any malignant cells, 8 probes without vital cells and 8 leukemias and lymphomas. We were able to establish primary cell cultures of 50% of the sterile tumor tissue probes, to cultivate them for a minimum of 5-10 passages, to characterize and freeze them. Six out of these tumor cell lines were already cultivated for one year and are available to the scientific community.

Adolescent↗

Morphological classification and retinal distribution of large ganglion cells in the retina of Bufo marinus.

The retrograde transport of horseradish peroxidase (HRP) and cobaltic-lysine complex (CLC) was used to morphologically characterize large ganglion cells (GCs) and to determine their distribution in retinal wholemounts and in sectioned material in the retina of Bufo marinus. Large GCs, amounting to about 0.5% of total GC population, were defined to be those with very large dendritic field sizes varying between 0.1 mm2 to 0.6 mm2 and cell soma sizes of between 100 microns 2 to 400 microns 2. These cells were subdivided into 3 major groups, Types I, II and III, on the basis of their dendritic field sizes, aborization patterns and the strata of dendritic branching within the inner plexiform layer (IPL). The majority of large neurons (about 90%) were classified as Type I GCs with symmetrical dendritic arbor. These cells had either bistratified branching in the scleral and vitreal sublamina of the IPL (65% of Type I Cells) or unistratified branching in the scleral (26%) or in the vitreal (9%) sublamina. Their dendritic field sizes increased linearly from the retinal centre from 0.13 mm +/- 0.02 mm2 (mean and S.D.) to 0.58 +/- 0.11 mm2 in the retinal periphery. Type II GCs (about 9% of large GC population) were characterized by an asymmetrical dendritic aborization directed towards the ciliary margin with unistratified branching in the scleral sublamina of the IPL. The mean dendritic field sizes of these cells were 0.26 +/- 0.09 mm2. Type III GCs, the least frequent (about 1%) category of large GCs had sparsely branching, elongated dendritic branching aligned approximately parallel with the nasotemporal axis of the retina. The unistratified dendritic branches of these neurons were located in the vitreal sublamina of the IPL with a mean dendritic field size of 0.42 +/- 0.11 mm2. The dendritic field sizes of Types II and III GCs did not increase with retinal eccentricity. Type I GCs were distributed unevenly across the retina, the density being greatest in the visual streak, along the nasotemporal meridian of the retina. The dendritic field sizes of these cells increased towards the retinal periphery, resulting in a constant dendritic field coverage factor across the retina. Each retinal point was covered by the dendritic fields of 4-5 adjacent GCs. In contrast, Types II and III GCs had only discontinuous dendritic coverage. The identification of morphological types of large GCs with previously described functional classes of GCs in the anuran retina is discussed.

Animals↗

Immunogold electron microscopic demonstration of glutamate and GABA in normal and deafferented cerebellar cortex: correlation between transmitter content and synaptic vesicle size.

Selective labeling of mossy fiber terminals and parallel fibers was obtained in rat cerebellar cortex by a glutamate antibody produced and characterized by Hepler et al. The high-resolution electron microscopic immunogold demonstration of this amino acid offered the possibility of determining the size and shape of synaptic vesicles in glutamate-positive mossy endings. Mossy terminals that stained with the glutamate antibody formed two distinct populations, one with spherical synaptic vesicles with an average diameter of 34.0 nm (more than 90% of all mossy fiber endings) and one with pleomorphic and smaller synaptic vesicles which had an average diameter of 28.5 nm. We present experimental evidence that the mossy terminals with large round vesicles are of extracerebellar origin, whereas those with small pleomorphic synaptic vesicles are endings of nucleocortical fibers. The presence of two distinct classes of gamma-aminobutyric acid (GABA)-containing axon terminals within cerebellar glomeruli has also been demonstrated; those originating from the cerebellar nuclei contain large (36.2 nm) synaptic vesicles, whereas the majority of GABA-stained axon terminals that are of local (cortical) origin contain small (29.1 nm) synaptic vesicles. It therefore appears that, at least in the case of glutamate and GABA, morphological characterization of the axon terminals based on the size and shape of synaptic vesicles is not a reliable indicator of their functional nature (i.e., whether they are excitatory or inhibitory); convincing evidence for the identity of the transmitter can be obtained only by electron microscopic immunostaining procedures. Our results also suggest the existence of both inhibitory and excitatory feedback from cerebellar nuclei to cerebellar cortex.

Animals↗

[DNA content and survival in subserous gallbladder carcinoma].

BACKGROUND: The clinical and morphological characterization of the subserous gallbladder carcinomas is controversial. AIM: To study the prognostic importance of DNA content of subserous gallbladder carcinoma. MATERIAL AND METHODS: We studied 104 females aged 60+/-12 years old and 16 men aged 70+/-13 years old. In all of them diagnosis was established after mapping of cholecystectomy sample and had a complete clinical follow up. DNA content was measured by flow cytometry. RESULTS: All tumors were adenocarcinoma, and only 16% were well differentiated. Aneuploidy was observed in 29 cases (26%) with DNA index fluctuating between 1.1 and 1.8. Lymphatic vessel tumor involvement was present in 16 of 22 cases with aneuploidy and in 22 of 46 diploid tumors (p=0.05). Eighty nine percent of aneuploid tumors were detected macroscopically and 11% were unapparent. Five years survival was non significantly better among patients with diploid tumors than in patients with aneuploid tumors (45 and 28%, respectively, p=0.2). The histological differentiation was the only variable significantly associated with survival. CONCLUSIONS: Aneuploidy is present in 26% of subserous gallbladder carcinoma. It is not related with any of the morphological or clinical variables studied in this series of patients.

Adenocarcinoma↗

The Purkinje neuron: I. A Golgi study of its development in the mouse and in culture.

The development of the Purkinje neuron was studied in organotypic culture and compared to that occurring in the intact animal, using a modified Golgi-Cox method. The post-natal sequence of development in the intact animal occurred in five distinct stages beginning with (1) an immature state (0-3 days), (2) a stage of perisomatic dendritic processes (4-6 days), and then (3) a stage characterized by the presence of spines on the soma region (7-10 days). This stage of somatic spines has not been delineated previously in Golgi studies of the Purkinje cell during its development. There was no evidence that the lateral somatic processes resorb; rather they continue to grow and develop into dendritic branches. It is proposed that by a process of perikaryal translocation, the soma region becomes transferred "downward," resulting in an elongation of the primary, apical dendrite (stage 4, 11-14 days). Beyond 15 days (stage 5) the dendritic branches grow to the pial surface and the neuron has its full complement of secondary, tertiary, and spiny branches. In culture, the development parallels that occurring in the intact animal during the first 10 days (stages 1, 2, 3) despite the absence of extracerebellar afferents and the special conditions of the culture. However, there is an overall absence of lamination of the cortex, the Purkinje neurons do not align, and the developmental process is modified because of the failure of the process of perikaryal translocation in culture. The resultant mature neuron has an altered morphology characterized by the presence of several dendrites and spines attached to the soma, and also lacks the complete development of the smaller dendritic branches.

Animals↗

The feasibility of gene expression profiling generated in fine-needle aspiration specimens from patients with follicular lymphoma and diffuse large B-cell lymphoma.

Lymphoma of germinal center cell (GC) origin generally is an indolent malignancy that transforms progressively into a more aggressive disease. According to the World Health Organization classification, lymphomas of follicular center cell origin are classified as either large B-cell lymphoma (LBCL) or follicular lymphoma (FL). The authors tested the feasibility of performing gene expression profiling using amplified RNA from fine-needle aspirates (FNA) obtained from lymph nodes. Twenty-four samples from patients with a diagnosis of FL or LBCL were obtained after Institutional Review Board-approved informed consent was obtained. The diagnoses were confirmed by 2 pathologists and were classified into 2 groups (10 LBCL samples and 14 FL samples) by using conventional morphology and immunophenotyping. One hundred nanograms of total RNA were subjected to 2 cycles of standard, double-stranded complementary DNA synthesis and in vitro transcription for target amplification using a small-sample target-labeling protocol. The biotinylated cRNA from each sample was hybridized to gene chips. Gene expression profiling results were analyzed first by principal-component analysis (PCA) by using a list of 146 probe sets that represented 62 genes that are characteristic of an activated B-cell (ABC) signature or a GC signature. The analysis identified 5 LBCL samples with an ABC cell signature. Using a list of 207 probe sets that represented 113 genes involved in FL transformation, PCA analysis identified 2 overlapping clusters corresponding to FL and GC-diffuse LBCL. To improve this classification further, the authors generated a list of 72 genes that were expressed differentially between FL and GC-LBCL. Using this list of genes, PCA analysis demonstrated a clear separation between FL and GC-LBCL. However, five FL samples clustered as an intermediate group between FL and GC-DLBCL. These samples were characterized morphologically by a mixed cell pattern with relatively fewer large, noncleaved lymphocytes and more small, cleaved lymphocytes. The results support the feasibility of FNA-based transcription profiles in patients with FL or LBCL, which, in combination with morphology and immunophenotyping, can help in the subtyping of these entities.

Biopsy, Fine-Needle↗

Nephrotoxicity of a novel antineoplastic platinum complex, nedaplatin: a comparative study with cisplatin in rats.

The present study was designed to characterize the nephrotoxicity induced by the antineoplastic platinum complex nedaplatin (NDP) in rats of different ages in comparison with cisplatin (CDDP). A single dose of 15 mg/kg NDP or 7.5 mg/kg CDDP was administered intravenously to 8-, 11-, or 15-week-old male and female SD rats, which were then sacrificed after ten days. Body weight decreases were observed for both drugs, in direct relation to age. CDDP treatment markedly increased urinary excretion of NAG, gamma-GTP, LDH and protein, with peaks on day 4 and complete or partial recovery on day 7; NDP increased NAG, LDH and protein excretion, but to a lesser extent, and these elevations were generally more marked for females. CDDP increased plasma creatinine and BUN in males and females of all age groups at necropsy. No apparent changes were seen following NDP treatment except in the 15-week-old rats. These results also show that NDP is less nephrotoxic than CDDP. CDDP-treated rats showed remarkable proximal tubular lesions in the renal cortex and corticomedullary region, and the papillary lesions were minor. On the other hand, the NDP-induced nephrotoxicity was morphologically characterized by hyaline droplet changes (electron microscopically, hyperplasia of lysosomes), necrosis or hyperplasia of the collecting duct epithelium in the renal papilla and the epithelium covering the papilla. Cortical lesions, indicated by slight tubular dilatation, were found only in the animals with papillary lesions. In summary, NDP is a promising second-generation platinum complex with reduced nephrotoxicity.

Animals↗

Specialized olfactory receptor neurons mediating intra- and interspecific chemical communication in leafminer moths Eriocrania spp. (Lepidoptera: Eriocraniidae).

We performed a physiological and morphological characterization of sensilla auricillica in male Eriocrania semipurpurella moths. Each auricillic sensillum contained three olfactory receptor neurons. Responding neurons (87 of 139) could be grouped into five physiological types. Type 1 responded to (R,Z)-6-nonen-2-ol and type 2 to its enantiomer (S,Z)-6-nonen-2-ol, both of which are pheromone components of E. semipurpurella. Type 3 responded to both (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol, which are pheromone components of the sympatric species E. cicatricella. Types 4 and 5 responded to the ketones (Z)-6-nonen-2-one and/or nonan-2-one, which are found in the pheromone glands of female E. semipurpurella. Field-trapping showed that type 3 receptor neurons mediate strongly antagonistic effects of (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol on E. semipurpurella, while nonan-2-one should possibly be included as a synergist in the sex pheromone blend of this species. The attraction of E. cicatricella and E. sparrmannella to compounds mixed with the pheromone blend of E. semipurpurella shows that the pheromone components of E. semipurpurella have little or no antagonistic effects on these species. The morphology and physiology of eriocraniid pheromone sensilla are very similar to those found in the order Trichoptera (caddisflies), suggesting a homology between pheromone detection systems in the two sister orders Lepidoptera and Trichoptera.

Action Potentials↗

In situ classification and image cytometry of pelagic bacteria from a high mountain lake (gossenkollesee, austria).

We describe a procedure to measure the cell sizes of pelagic bacteria after determinative hybridization with rRNA-targeted fluorescently labeled oligonucleotide probes. Our approach is based on established image analysis techniques modified for objects simultaneously stained with two fluorescent dyes. It allows the estimation of biomass and cell size distribution and the morphological characterization of different bacterial taxa in plankton samples. The protocol was tested in a study of the bacterioplankton community of a high mountain lake during and after the ice break period. Cells that hybridized with a probe for the domain Bacteria accounted for 70% of the bacterial abundance (range, 49 to 83%) as determined by 4(prm1),6(prm1)-diamidino-2-phenylindole staining (K. G. Porter and Y. S. Feig, Limnol. Oceanogr. 25:943-948, 1980), but for >85% of the total biomass (range, 78 to 99%). The size distribution for members of the beta subclass of the Proteobacteria shifted toward larger cells and clearly distinguished this group from the total bacterial assemblage. In the surface water layer beneath the winter cover, bacteria belonging to the beta 1 subgroup constituted about one-half of the beta subclass abundance. The mean cell volume of the beta 1 subgroup bacteria was significantly less than that of the beta subclass proteobacteria, and the beta 1 subgroup accounted for less than 30% of the total beta subclass biovolume. Two weeks later, the biovolume of the beta Proteobacteria had decreased to the level of the beta 1 subgroup, and both the biovolume size distributions and cell morphologies of the beta Proteobacteria and the beta 1 subgroup were very similar. We could thus quantify the disappearance of large, morphologically distinct beta subclass proteobacteria which were not members of the beta 1 subgroup during the ice break period. Our results demonstrate that changes in biovolumes and cell size distributions of different bacterial taxa, and eventually of individual populations, reveal hitherto unknown processes within aquatic bacterial assemblages and may open new perspectives for the study of microbial food webs.

Journal Article↗

Adult T-cell lymphoma/leukemia in a Caribbean patient: cytogenetic, immunologic, and ultrastructural findings.

Cytogenetic findings on immunologically and morphologically characterized leukemic cells from a Caribbean patient with adult T-cell lymphoma/leukemia (ATLL) (HTLV+) are reported. Marker studies on peripheral blood lymphoid cells showed a mature postthymic phenotype: TdT-, OKT3+, OKT4+, OKT6-, OKT8-, 3A1-, anti-HLA-DR-. Light and electron microscopic analysis revealed a great cellular pleomorphism with respect to nuclear features. Three main types of leukemic cell were observed: typical multilobed ATLL lymphocytes, Sézary's syndrome (SS) cells, and cells intermediate between those two. Chromosome studies on PHA-stimulated cultures revealed three clones. The predominant clone was hyperdiploid; significant abnormalities were 1q+, 14q+, and 6q- (breakpoint q21), which are known to occur in lymphoid malignancies, together with trisomy 7q and i(17q), which have been reported previously in Japanese ATLL and the small variant of SS, respectively. The 14q+ marker was t(11;14)(q13;q22-24). The incidence of 6q-, trisomy 7q, and i(17q) varied within the main clone, and it is speculated that these chromosome abnormalities might be related to the variation observed in the cell types of this patient. Two minor clones had 6q- (breakpoint q25) and 13q+ markers, respectively. It was not possible to unequivocally establish the relationship between these three clones. The chromosomal, morphologic, and immunologic findings in this case support a close relationship between ATLL in Japan and in the Caribbean basin, as well as between the proliferating cells of ATLL and SS.

Antigens, Surface↗

Drug and radiation sensitivity measurements of successful primary monolayer culturing of human tumor cells using cell-adhesive matrix and supplemented medium.

The limitations of the agar suspension culture method for primary culturing of human tumor cells prompted development of a monolayer system optimized for cell adhesion and growth. This method grew 83% of fresh human tumor cell biopsy specimens, cultured and not contaminated, from a heterogeneous group of 396 tumors including lung cancer (93 of 114, 82%); melanoma (54 of 72, 75%); sarcoma (46 of 59, 78%); breast cancer (35 of 39, 90%); ovarian cancer (16 of 21, 76%); and a miscellaneous group consisting of gastrointestinal, genitourinary, mesothelioma, and unknown primaries (78 of 91, 86%). Cell growth was characterized morphologically with Papanicolaoustained coverslip cultures and cytogenetically with Giemsastained metaphase spreads. Morphological features such as nuclear pleomorphism, chromatin condensation, basophilic cytoplasm, and melanin pigmentation were routinely seen. Aneuploid metaphases were seen in 90% of evaluable cultures, with 15 of 28 showing 70% or more aneuploid metaphases. Colony-forming efficiency ranged between 0.01 and 1% of viable tumor cells, with a median efficiency of 0.2%. This culture system uses a low inoculum of 25,000 viable cells per well which permitted chemosensitivity testing of nine drugs at four doses in duplicate from 2.2 X 10(6) viable tumor cells and radiation sensitivity testing at five doses in quadruplicate from 0.6 X 10(6) cells. Cultures were analyzed for survival by computerized image analysis of crystal violet-stained cells. Drug sensitivity studies showed variability in sensitivity and in survival curve shape with exponential cell killing for cisplatin, Adriamycin, and etoposide, and shouldered survival curves for 5-fluorouracil frequently seen. Radiation sensitivity studies also showed variability in both sensitivity and survival curve shape. Many cultures showed exponential cell killing, although others had shouldered survival curves. This method for growing cells from primary human biopsy specimens is more efficient than the agar culture method, enables easier and better biological analysis of the actual cells grown, and permits improved characterization of drug and radiation survival curves.

Biopsy↗

Characterization of secretory and morphologic properties of primary cultured endocrine cells from porcine pancreata.

We characterized morphologic and secretory properties of porcine pancreatic endocrine cells in primary culture obtained by autolytic preparation without any exogenous proteolytic enzymes. The endocrine cells exhibited a neuron-like shape, and insulin granules were accumulated at the terminal of the processes. Thus derived endocrine cells survived in culture medium containing nicotinamide and remained sensitive to glucose for at least 6 weeks after preparations. The cells responded well to physiologic concentrations of glucose, and high K+ depolarization and the antidiabetic sulfonylureas, tolbutamide, and glibenclamide also elicited the release. With high glucose, insulin release was markedly potentiated by forskolin, glucagon, glucagon-like peptide-1, and arginine and inhibited by somatostatin, the Ca2+ channel blocker nitrendipine, and the ATP-sensitive K+ channel opener diazoxide. Epinephrine had dual effects on the release by glucose; enhanced within a low nanomolar range and inhibited at 1 micromol/L. However, the cells were unresponsive to leucine. Such secretory sensitivities to nutrients, hormones, and pharmacologic agents, and long survival rate (as long as 5-6 weeks) of these cells suggest to us therefore that derived endocrine cells may be useful for xenotransplantation of pancreatic beta cells for treatment of insulin-dependent diabetes mellitus.

Animals↗

Human colostral cells. I. Separation and characterization.

Analyses of the cells present in human colostrum obtained from fifty-four healthy donors during the first four days of lactation revealed that there were 3.3 x 10(6) (range 1.1 x 10(5)--1.2 x 10(7)) cells per ml of colostrum. Based on histochemical examinations, it was found that this population consisted of 30--47% macrophages, 40--60% polymorphonuclear leucocytes, 5.2--8.9% lymphocytes, and 1.3--2.8% colostral corpuscles; epithelial cells were rarely encountered. The identity of various cell types was confirmed by Wright's stain and by a series of histochemical techniques which disclosed the presence of non-specific esterase, peroxidase, and lipids. For further characterization, the different types of cells were separated by various methods, such as Ficoll-Hypaque density centrifugation, isokinetic centrifugation on a linear Ficoll gradient, adherence to glass or plastic, and phagocytosis of carbonyl iron. Immunohistochemical staining with FITC- and/or TRITC-labelled reagents to IgA, IgM, IgG, K- and lambda-chains, secretory component, lactoferrin, and alpha-lactalbumin were applied to unseparated as well as separated colostral cells. Polymorphonuclear leucocytes (staining for peroxidase) as well as macrophages and colostral corpuscles (staining for non-specific esterase) exhibited numerous intracellular vesicles that contained lipids as well as various combinations of milk proteins. Lymphoid cells did not stain with any of these reagents and plasma cells were not detected among the colostral cells. Individual phagocytic cells contained immunoglobulins of the IgA and IgM classes, both K and lambda light chains, secretory component, lactoferrin, and alpha-lactalbumin. The coincidental appearance of these proteins in single, phagocytic cells but not in lymphoid cells indicate that the cells acquired these proteins by ingestion from the environment. Markers commonly used for the identification of B lymphocytes (surface immunoglobulins) and T lymphocytes (receptors for sheep red blood cells) were unreliable for the analysis of colostral cells (unless accompanied by subsequent morphological characterization) because strong fluorescence was observed on the surface of many non-lymphoid cells and because numerous macrophages and colostral corpuscles formed rosettes with sheep red blood cells (SRBC). Lymphocytes, often found in association with colostral macrophages or corpuscles, were classified as T cells.

Cell Count↗

[The pocket infection-erosion of permanent pacemakers: the results of a conservative approach without substitution of the components].

OBJECTIVE: To assess the value of a morphologic classification of pocket lesions in eroded-infected permanent pulse generators and its influence in the efficacy or surgical management without removal and with system reutilization. DESIGN: Retrospective evaluation of patients, presenting with eroded-infected pacemaker, submitted to surgery. SETTING: Pacing Center and Cardiology Department of Faro District Hospital. PATIENTS: 43 consecutive patients treated by the same surgeon between 1985 and 1994. INTERVENTIONS: We considered three groups, based on morphologic characterization of the pocket lesions: G1-partial extrusion from the pulse generator with focal, adherent, discoloured and eroded skin; G2-identical to the previous group but with inclusion of lead segments or only lead extrusion; G3-tender pacing sites with fluctuation and, usually, fistula formation without the skin lesions of the other groups. In every patients a new ipsilateral generator pocket has been created just aside from the previous one with reimplantation of the same pacemaker. Failure was defined as the need for explantation of the system, with a new pacemaker implantation on the contralateral site, after the first attempt in G3 and a second in the other two groups. MEASUREMENTS AND RESULTS: The erosion-infection rate was 6.9%. Recurrences appeared in, 8.3%, 36.3% and 75% for, respectively, G1, G2 and G3. The initial and final success rates of G1 were significantly different from G3 success rates (p, respectively, < 0.07 and < 0.001). At the end of our study, reutilization surgery was successful in 91.4% of G1 and G2 assembled patients compared with 25% of the G3 patients (p < 0.003). Mean hospital stay for G3 patients was 7.4 +/- 1.0 days while G1 and G2 were managed as outpatients. The mean follow-up of patients without recurrences of pocket erosion-infection was 32.8 months. CONCLUSIONS: Our data suggest that a morphologic classification of lesions is very useful for treatment choice. It is possible, then, to select patients that may successfully managed by ipsilateral reimplantation without removal of pacing systems or hospital stay. The differences in the appearance of lesions and management results must reflect different etiopathogenic mechanisms.

Aged↗

The distribution and morphological characteristics of the intracortical VIP-positive cell: an immunohistochemical analysis.

Using a sensitive immunohistochemical procedure, we have undertaken a detailed morphological characterization of the vasoactive intestinal peptide-positive (VIP-positive) neuron in the cerebral cortex of the rat. VIP-positive neurons are present in all regions of cortex, and are usually strongly bipolar, possessing long, radially directed processes with very limited branching in the tangential plane. The most extensive dendritic branching occurs in layers I and deep IV-superficial V, and the density of axonal varicosities is highest in layers II-IV. In the visual cortex, approximately 50% of the labeled cell bodies are in layer II and III and 80% of the labeled cell bodies are contained in layers I-IV (superficial 600 microns of cortex). In order to determine the density and 3-dimensional distribution pattern of these cells, we prepared serial tangential sections through the rat visual cortex, mapped the distribution of all labeled cells in each section on transparent acetate sheets, and compressed these superimposed maps. This analysis demonstrated that: (1) approximately 1% of the cortical neurons are VIP-positive, (2) their distribution is fairly uniform and statistically random, (3) there are no large areas (i.e. with a diameter greater than 100 microns) that lack a VIP-positive cell, (4) on the average, there is one VIP-positive cell per column of 30 microns diameter, and (5) the average nearest neighbor distance on the compressed display is 15 microns. Given the morphological characteristics of VIP-positive cells, these data indicate that each VIP-containing cell is identified with a unique radial volume, which is generally between 15 and 60 microns in diameter, and overlaps with the contiguous domains of neighboring VIP-positive cells. These morphological data support the notion that VIP-containing neurons play an important functional role within radially oriented columns of cerebral cortex.

Animals↗

Pathology and "molecular histology' of Hodgkin's disease and the border to non-Hodgkin's lymphomas.

HD is characterized by a small amount of tumour cells (the H- and RS-cells) in a background of many, probably reactive, lymphocytes and histiocytes. Lymphocyte predominant HD seems to be a distinct clinicopathological entity of germinal centre cell origin and has a relatively favourable prognosis. This type has to be differentiated from mixed cellularity and nodular sclerosis HD, which show a different marker profile of the H- and RS-cells and their variants. Differentiation of progressively transformed germinal centres from lymphocyte predominant HD may be problematical and is based on the detection of L&H-cells in the latter entity. Diffuse paragranuloma may be difficult to differentiate from classic HD as well as non-Hodgkin's lymphomas, especially T-cell-rich B-cell lymphomas and peripheral T-cell lymphomas. In rare cases a distinction between HD and non-Hodgkin's lymphomas may be impossible. Similar to lymphocyte predominant HD, lymphocyte depletion HD may provide difficulties in its differential diagnosis from non-Hodgkin's lymphomas. The border with anaplastic large cell lymphoma cannot be clearly drawn and may be blurred. The border between HD and non-Hodgkin lymphomas may be especially blurred in composite lymphomas, if single cell studies can demonstrate a common clonal origin of both parts of the lymphoma. Analysis of immunoglobulin or T-cell receptor variable region genes amplified by PCR from single cells picked from histological sections ("molecular histology') combines morphological characterization and identification of H- and RS-cells with a molecular analysis of those cells. This enables one not only to determine the nature of H- and RS-cells and their clonality but also to answer questions about the relationship between Hodgkin and non-Hodgkin's lymphomas.

Diagnosis, Differential↗