Cooley G. Pantazis

Persistent tachypnea in children: keep pulmonary embolism in mind

Purpose: Tachypnea in children is associated with respiratory disorders and nonrespiratory disorders such as cardiac disease, metabolic acidosis. fever, pain, and anxiety. Pulmonary embolism is seldom considered by pediatricians as a cause of tachypnea. Patients and Methods: Three children of various ages with persistent tachypnea are described: a girl after orthopedic surgery for kyphoscoliosis, a boy with nephrotic syndrome, and a neonate with Hirschsprung disease. Other causes of tachypnea were diagnosed and treated before pulmonary embolism was considered. Results: Ventilation-perfusion scanning appeared to be highly probable for pulmonary embolism in these patients. Anticoagulant therapy was started. Conclusion: Pulmonary embolism should be kept in mind in children with tachypnea, especially when other risk factors for venous thromboembolism are present, to avoid delay in anticoagulant treatment and a fatal outcome.Purpose: This article describes an infant with a large abdominal mass and hypertension. Patient and Methods: A 5-month-old infant girl with diarrhea of 1 weeks duration and a large right-sided abdominal mass was brought for treatment. Computed tomography of the abdomen revealed a large, generally homogeneous, hypodense mass, which compressed the right kidney, resulting in dilatation of the right renal collecting system. At surgery, the mass was adherent anteriorly to the transverse colon and attached by a stalk to the mesentery near the origin of the right colic artery. Results: Examination of the mass showed an encapsulated lipoblastoma. Cytogenetic analysis revealed a 46,XX karyotype with a reciprocal translocation between chromosome 2 and chromosome 8 with breakpoints at q23 and q11.2, respectively. Conclusion: Lipoblastoma is a rapidly growing but benign tumor, which can cause severe medical problems by compressing major organs. Cytogenetic analysis can reveal translocations involving chromosome 8 band ql 1.2, which appears to be a specific chromosome marker for lipoblastoma.

Ganglioglioma, a malignant tumor? Correlation with flow deoxyribonucleic acid cytometric analysis.

This study describes the flow cytometric deoxyribonucleic acid (DNA) analysis of a resected ganglioglioma. The initial histopathological analysis revealed a benign tumor characterized by a predominance of mature ganglion cells. The flow cytometric DNA analysis of the necrotic areas, however, demonstrated an aneuploid population of cells. Further examination by histological analysis of the tumor revealed both benign and atypical foci. The retrospective DNA analysis performed from paraffin sections of tissue with benign-histological findings demonstrated euploid populations of cells consistent with a benign, slow-growing lesion. In contrast, DNA analysis performed from tissue with atypical histological findings revealed aneuploid populations of cells consistent with a malignant phenotype. Our analysis provides additional data supporting the existence of tumor progression in some gangliogliomas. Results support the concept of tumor cell heterogeneity and the importance of adequate tumor sampling. The finding of aneuploid populations with unfavorable histology further supports the use of flow cytometry as an adjunct method in assessing tumor biology.

Immunocytochemical study of ras and myc proto-oncogene polypeptide expression in the human menstrual cycle

The human endometrium is a unique and dynamic tissue model system characterized by cyclic processes of cellular proliferation, differentiation, and menstrual desquamation. Both the glandular epithelial and stromal mesenchymal components work in synchronous response to the mitogenetic effect of estradiol and the antimitogenetic effect of progesterone. Mechanisms whereby estradiol and progesterone exert their effects are not completely understood. This study was undertaken to evaluate the expression and localization of the polypeptide products of the ras and myc proto-oncogenes sequentially during the menstrual cycle. Sixteen endometrial biopsy specimens were evaluated. Immunocytochemical quantitation of ras and myc expression was done by use of color image analysis (CoreScan). There was no cyclic variation in the ras polypeptide product, but the expression of myc polypeptide was low in the secretory phase and high in the proliferative phase.

Inhibition of estrogen stimulated mitogenesis by 3-phenylacetylamino-2,6-piperidinedione and its Para-hydroxy analog

3-Phenylactetylamino-2,6-piperidinedione (A10) inhibited estradiol stimulated cell growth in the MCF-7 (E3) human breast tumor cell line in vivo and in vitro. While high concentrations of A10 were needed to inhibit cell proliferation (IC50 = 3 x 10(-3) M in vitro), the compound demonstrated little toxicity. The effect appeared specific since a hydrolysis product of A10, phenylacetylglutamine, demonstrated no growth inhibitory activity at similar concentrations in MCF-7 (E3) cells in vitro. A computer designed analog, p-hydroxy A10, was more potent than A10 in inhibiting activity in MCF-7 (E3) cells in vitro. The IC50 for p-hydroxy A10 was 7 x 10(-6) M which was comparable to that of the antiestrogen, tamoxifen (IC50 1 x 10(-7) M). All three compounds caused a decline in estrogen receptor levels in a dose-dependent fashion. A10 also inhibited estradiol induction of progesterone receptors. Examination of protein kinase activity following an acute exposure to a 10(-11) M growth stimulatory dose of estradiol revealed a 168% increase in protein kinase activity over that of untreated control cells. A10 in a dose-responsive fashion inhibited the estradiol stimulated increase in protein kinase activity. The protein kinase activity was also inhibited by p-hydroxy A10. These activities of A10 and p-hydroxy A10 coupled with the low toxicity and novelty of the basic A10 structure provide an exciting possibility of developing a new class of clinically useful antineoplastic drugs with minimal side effects.

Cellular localization of müllerian inhibiting substance messenger ribonucleic acid during human ovarian follicular development

Müllerian inhibiting substance is expressed in the human reproductive system and has been associated with oocyte meiotic arrest. In situ hybridization was used to selectively localize ovarian cells containing high levels of müllerian inhibiting substance messenger ribonucleic acid, a müllerian inhibiting substance precursor, during different stages of human follicular development. Müllerian inhibiting substance transcript was noted in the granulosa cells of primordial, primary, and antral follicles. Surprisingly, transcript was also identified within the cytoplasm of oocytes and throughout the ovarian stroma. Controls included sense oligoprobe, positive and negative tissue controls, and treatments minus the detection antibody. Localization of transcript within the cytoplasm demonstrates that active transcription of müllerian inhibiting substance messenger ribonucleic acid occurs within both fetal and adult human female gonads. The presence of müllerian inhibiting substance messenger ribonucleic acid within oocyte cytoplasm could implicate an autocrine role for müllerian inhibiting substance-derived peptides in the establishment of oocyte competence.

Acute megakaryocytic leukemia. Flow cytometric analysis of DNA content at diagnosis and during the course of therapy.

Flow cytometry (FCM) for the determination of DNA content and cell cycle analysis was performed on multiple bone marrows from a case of acute megakaryocytic leukemia in a 71‐year‐old woman. Two aneuploid (multiploid) peaks were present at diagnosis. This multiploidy was a stable characteristic when studied temporally. The disappearance of the peaks in the bone marrow specimens directly correlated with complete remission morphologically. These findings further support the concept of FCM as an adjunct to morphologic study in identifying residual tumor burden.

Intermediate structures in nuclear morphogenesis following metaphase from HeLaS3 cells can be isolated and temporally grouped

Previously nuclear reformation following metaphase in HeLaS3 cells was conceptualized in terms of a stepwise process which was continuous throughout anaphase and telophase. This concept was based on a three-dimensional visualization by scanning electron microscopy (SEM) of individual, organically prepared chromatid structures (prenuclei) which could be sequentially arranged. Morphologic analysis revealed unique topographies and morphometric properties which suggested that it should be possible to isolate populations of prenuclei aqueously. Such an isolation using detergents and density centrifugation is presented which yields metaphase plates and two populations of prenuclei with distinctive morphology. Essentially, prenuclei are freed from late mitotic cells in suspension cultures of synchronized HeLaS3 cells by treatment with 0.1% Nonidet-P40 followed by treatment with a mixture of Tween 40-desoxycholate (0.5%). Critical for the isolation is the presence of a divalent cation (5 mM Mg+ +) and an acid pH (~ 5.8). After density centrifugation, 2N decondensing structures (late intermediates) are recovered from 42% Percoll, and a mixture of 2N predecondensing (early intermediates) and 4N metaphase plates are recovered from 52% Percoll. The latter intermediates can be further separated into highly enriched populations (>94% pure) by fluorescence-activated sorting. Predecondensing structures are of the same overall morphology as prenuclei isolated previously by organic means, can also be ordered sequentially to demonstrate nuclear morphogenesis, and retain centromere/kinetochore loci. These chromosomal loci based on immunostaining of individual structures appear to be positioned centrally during chromatid reassociation and then appear to be dispersed prior to structural rearrangements leading to formation of a disc-like prenucleus. The significance of grouping intermediates temporally and of two protocols of isolation yielding the same structures is discussed with regard to a study of the requirements for nuclear morphogenesis in late mitosis.

Down's syndrome and acute myelofibrosis. Time study of dna content during the progression to leukemia

Flow cy tometry (FCM) for the determination of cellular DNA content was performed on multiple bone marrow biopsy specimens from a 3‐year‐old boy with Downs syndrome and myelofibrosis. A rapidly fatal acute nonlymphocytic leukemia developed within 3 months after initial bone marrow evaluation. The clinical and morphologic changes corresponded to the development of aneuploidy as determined by FCM and cytogenetic analysis. These findings support the clinical observations of the premalignant potential of myelofibrosis in Downs syndrome.

Immunocytochemical study of insulin - like growth factor I and insulin - like growth factor I receptor in human endometrium during the normal menstrual cycle

Our aims were to detect, using immunocytochemistry, IGFI and IGFI R in the human endometrium and to assess semiquantitatively their levels in the phases of the normal menstrual cycle. Twelve normal proliferative and 10 normal secretory endometrial samples were studied. Each specimen was subjected to an immunocytochemical peroxidase antiperoxidase protocol. The antibodies used to detect IGFI and IGFI R were, respectively, 3D1/2/1 and alpha 1R3. Analysis of variance (ANOVA) was performed to evaluate mean IGFI and IGFI R levels in the glandular epithelium and stroma of each sample while correcting for intra- and interobserver variation. These experiments show the presence of IGFI and IGFI R in human endometrium. There are significant variations in the IGFI and IGFI R levels from patient to patient within each cycle phase, and between glands and stroma within each sample. These findings highlight the importance of the use of in situ studies to clarify endometrial IGFI and IGFI R physiology.

Teratocarcinqma Cells Cultured on Embryonic Substrates Show Accelerated Migrating Behavior in Vitro and Increased Metastatic Activity in Vivo

Abstract Teratocarcinoma cells (402AX) were grown on feeder layers of whole mouse embryos (Day 4) or mouse embryonic fibroblasts and then were either examined in vitro or transplanted in vivo. After twenty-four hours of coculture, teratocarcinoma cells demonstrate accelerated cell migration in vitro- Furthermore, transplantation of teratocarcinoma cells with embryonic substrates into syngeneic hosts produces grossly detectable lymph node metastases. These effects appear to be due to soluble factor(s) produced by embryonic substrates which enhance tumor cell proliferative/migratory activity. This suggests that tumor cell invasion and metastasis may be stimulated by soluble factors produced by host tissues.