Epidermal growth factor induces terminal differentiation in human epidermoid carcinoma cells

In this study, spermatogenesis in the adult Djungarian hamster is described. Undifferentiated spermatogonia topographically arranged as Asingle (A(s)), Apaired (Apr), and Aaligned (Aal) spermatogonia were observed, as were six generations of differentiating spermatogonia (A1, A2, A3, intermediate, B1, and B2). The differentiating spermatogonia divided at regular intervals during the cycle of the seminiferous epithelium. Mitosis of these cells was observed at the transition from stage IX to stage X (mitosis of A1 into A2 spermatogonia), at the transition from stage XII to stage I, at the transition from stage II to stage III, at the transition from stage IV to stage V, at the end of stage VI, and at approximately the middle of stage VII. Cellular associations in the cycle of the seminiferous epithelium are described. The seminiferous epithelium was divided into 12 stages, based upon the developmental steps in spermiogenesis, and the frequency of these stages was determined. The duration of the cycle of the seminiferous epithelium, determined by [3H]thymidine incorporation, was shown to be 7.90 +/- 0.01 (mean +/- SEM) days.     

De novo epileptic confusional status in a patient with cobalamin deficiency

A 80-year-old man with cobalamin deficiency and no history of epilepsy developed a partial complex epileptic confusional status (ECS) unresponsive to acute i.v. diazepam. Brain CT scan and MRI investigation ruled out a focal cerebral lesion. Therapy with high doses (10,000 micrograms i.m. daily) of cobalamin alone was started, and the patient fully recovered in the following 72-hour. Control EEGs repeatedly performed days and weeks later showed progressive disappearance of the frontal interictal spiking, while the patient was on monotherapy with cobalamin (5,000 micrograms i.m. weekly). A month later the patient unfortunately discontinued replacement therapy and 13 weeks later he developed a fatal convulsive epileptic status. To our knowledge the association of ECS and cobalamin deficiency has not been previously reported.     

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine as a substrate of cytochrome P450 2D6: allosteric effects of NADPH-cytochrome P450 reductase

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin that produces Parkinsonism symptoms in man, has been examined as a substrate of recombinant human cytochrome P450 2D6. When cumene hydroperoxide is used as an oxygen and electron donor, a single product is formed, identified as 4-phenyl-1,2,3,6-tetrahydropyridine. The K(m) for formation of this product (130 microM) is in agreement with the dissociation constants for MPTP binding to the enzyme determined by optical and nuclear magnetic resonance (NMR) spectroscopy. When the reaction is carried out with nicotinamide adenine dinucleotide phosphate (reduced) (NADPH) and recombinant human NADPH-cytochrome P450 reductase, a second product, identified as 1-methyl-4-(4'-hydroxyphenyl)-1,2,3,6-tetrahydropyridine, is formed in addition to 4-phenyl-1,2,3,6-tetrahydropyridine. The K(m) values for formation of these two products are 19 microM and 120 microM, respectively. Paramagnetic relaxation experiments have been used to measure distances between the protons of bound MPTP and the heme iron, and these have been used to construct models for the position and orientation of MPTP in the active site. For the cytochrome alone, a single mode of binding was observed, with the N-methyl close to the heme iron in a position appropriate for the observed N-demethylation reaction. In the presence of the reductase, the data were not consistent with a single mode of binding but could be explained by the existence of two alternative orientations of MPTP in the active site. One of these, characterized by a dissociation constant of 150 microM, is essentially identical to that observed in the absence of the reductase. In the second, which has a K(d) of 25 microM, the MPTP is oriented so that the aromatic ring is close to the heme iron, in a position appropriate for p-hydroxylation leading to the formation of the product seen only in the presence of the reductase. In the case of codeine, another substrate for cytochrome P450 2D6, the addition of reductase had no effect on the nature of the product formed, the dissociation constant, or the orientation in the binding site. These observations show that NADPH-cytochrome P450 reductase has an allosteric effect on the active site of cytochrome P450 2D6 that affects the binding of some substrates but not others.     

Differential expression of D1 and D5 dopamine receptors in the fetal primate cerebral wall

Previous film autoradiographic studies demonstrated that, during corticogenesis, dopamine receptors of the D1 class are abundant in the embryonic primate cerebral wall. In the present study, we expand these findings by identifying the cellular elements of the fetal occipital cerebral wall expressing D1 and D5 subtypes of the D1 dopamine receptor class. We have examined tissue from monkey fetuses collected at 70, 90 and 120 days of gestation using antibodies directed against C-termini of the D1 and D5 dopamine receptors. At all three embryonic ages studied, we found D1 and D5 receptors expressed by multiple cell types of the embryonic cerebral wall. Both D1 and D5 receptor proteins are produced by pyramidal neurons of the cortical plate and by a variety of interstitial neurons of the subplate and intermediate zones. D1 and D5 receptors are also present in cells of the proliferative ventricular and subventricular zones, some of which were identified as dividing cells. In addition, D1 and D5 receptors are detectable in the protoplasmic astroglial and ependymal cells distinguishable in monkey fetuses collected at 120 days of gestation. Some cellular elements of the embryonic monkey cerebral wall express only one subtype of the D1 dopamine receptor class. For example, embryonic Cajal-Retzius neurons in the marginal zone and migrating neurons in the intermediate zone are immunoreactive only to D5 antisera. In contrast, radial glia can be labeled only with D1 receptor-specific antisera. Finally, only D1 receptors are detectable in the blood vessels penetrating the embryonic monkey cerebral wall. Based on these observations, we propose that dopamine receptors of the D1 class play an important role in regulating cerebral cortical formation and that D1 and D5 receptor subtypes may participate in regulation of different aspects of this process.     

Differential expression of mammalian TRP homologues across tissues and cell lines

Mammalian homologues of the Drosophila trp gene have been invoked as the structural basis for the currents associated with capacitative Ca2+ entry (CCE) in many cell types. Trp homologues are members of a large protein family that may associate as channel subunits providing an explanation for the functional diversity of store-operated channels observed in these cells. However, there is little information as to which of these genes are co-expressed at the cellular level. We have examined the tissue specific expression of five mammalian trp genes and determined which are co-expressed in five different cell lines. The results show tissue- and cell-specific co-expression of multiple trp forms. This implies that the subunit composition of a particular CCE channel may vary depending on the cell type.     

Gastric ECL-cell hyperplasia produces enhanced basal and stimulated gastric acid output but not gastric erosion formation in the rat

Cervical involvement is one of the major prognostic factors in carcinoma of the endometrium confined to the uterus. The purpose of this study was to determine whether intrauterine ultrasound with a high-frequency miniature probe can depict the degree of cervical involvement of the disease. Thirty-two women with endometrial carcinoma underwent preoperative transvaginal and intrauterine sonography. By both scans, the degree of cervical involvement was prospectively evaluated. Sonograms were compared with the findings from histologic examination. Intrauterine sonography was completed in 30 of the 32 patients. In these 30 patients, the degree of cervical involvement (none, endocervical gland, or cervical stroma) based on transvaginal scan was correct in 23 cases (77%), and that based on intrauterine scan was correct in 26 cases (87%). Three tumors with endocervical glandular involvement were correctly diagnosed by intrauterine sonography, whereas they were incorrectly diagnosed by transvaginal scan. The specificity and positive predictive value of intrauterine sonography for the assessment of the presence of cervical stromal invasion are 100% (26/26 and 3/3, respectively). Although this study is preliminary, our experience with intrauterine sonography shows that it has potential for assessing cervical stromal invasion in endometrial carcinoma.