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Biomedical subjects

M H Johnson

Publications and source records attributed to M H Johnson.

At least 271 records · Page 15Linked to original sources
PubMed1984

Hemoglobin Shelby [beta 131(H9) Gln----Lys] a correction to the structure of hemoglobin Deaconess and hemoglobin Leslie.

Hemoglobin Shelby, detected in two unrelated black families, has an electrophoretic mobility like Hb F on cellulose acetate (pH 8.4) and a mobility between Hbs S and C on citrate agar (pH 6.2). Globin chain analysis in acid and alkaline buffers revealed an abnormal chain migrating between beta A and beta S. Tests for unstable hemoglobins were positive. Hematologic data on both families indicated carriers have mild anemia. The variant showed a slightly lower affinity for oxygen with normal cooperativity and Bohr effect, and its reactions with 2,3-diphosphoglycerate and inositol hexaphosphate were similar to those of Hb A. Sequence analysis indicated the substitution of lysine for glutamine at position 131 in the beta-chain. In a previous report (1) we described a variant, Hb Deaconess, in which this residue was deleted. On reexamination of the data, we find that Hb Deaconess is identical to Hb Shelby.

Adolescent↗
PubMed1984

The distribution of cytoplasmic actin in mouse 8-cell blastomeres.

Three non-homogeneous patterns of cytoplasmic actin distribution have been demonstrated in pairs of 8-cell blastomeres. Newly formed blastomeres showed an actin distribution associated with the remnant of the previous mitotic spindle. Subsequently blastomeres showed a zonal clearing of actin from regions of intercellular contact, the extent of the clearing increasing with the extent of contact. A polarized distribution of actin was evident from the early to mid 8-cell stage and coincided with the movement of nuclei towards the point of intercellular contact. The detection of polar actin preceded by 2-4 h the detection of a surface polarity as assessed by the FITC-Con A binding pattern and the distribution of cortical microvillous actin. However, once a surface pole of microvilli had formed, it persisted under conditions which led to loss of polar cytoplasmic actin. Incubation in cytochalasin D (CCD) resulted in a dispersed homogeneous pattern of actin distribution but did not prevent the formation of surface poles as assessed both by the Con A binding pattern and detection of polar microvilli. However, the poles formed were less clearly defined and the density and length of microvilli within them was variable. Moreover, when CCD was added early during the 8-cell stage the position of the poles was frequently not on an axis perpendicular to the point of intercellular contact. Cytochalasin D also affected the movement of the nucleus that occurs during the process of polarization. On the basis of these experiments, we conclude that actin is likely to be involved in the events of polarization, but that its precise role remains to be determined.

Actins↗
PubMed1984

Trophectodermal processes regulate the expression of totipotency within the inner cell mass of the mouse expanding blastocyst.

Mouse blastocysts, aged 0, 2, 6 and 12 h from the onset of cavitation, were examined by transmission (TEM) and scanning (SEM) electron microscopy. In TEM sections, trophectoderm cells (TE) differed morphologically from those of the inner cell mass (ICM) by their flattened shape, paler cytosol staining and polarized disposition of both junctional complexes (apicolateral) and intracellular secondary lysosomes (SL; basal). Throughout this period of development, cytoplasmic processes, characterized by abundant SLs, cover approximately 80% of the juxtacoelic face of the ICM. These processes are shown to be derived from the basal surface of TE cells intermediately placed between polar and mural regions. In SEM preparations of the juxtacoelic ICM surface, revealed by 'cracking open' blastocysts, the processes appear as tongue-shaped, centripetally oriented structures which terminate collectively at a central area on the ICM surface. The potential of cultured ICMs to generate TE was demonstrated following their immunosurgical isolation from blastocysts aged up to 12 h post cavitation and by examining the sequence of ultrastructural changes associated with TE generation by ICMs from 2 h blastocysts. In contrast, the juxtacoelic cells of similarly aged ICMs observed in situ in ultrasections of intact embryos showed little or no evidence of totipotency expression as judged by the absence of TE characteristics. Since TE expression within presumptive ICM cells is thought to be generated by an asymmetry of cell contacts (Johnson & Ziomek, 1983), we propose that the juxtacoelic TE processes, by providing a cellular cover to the ICM, function in suppressing the expression in situ of ICM totipotency.

Animals↗
PubMed1984

Changes in actin distribution during fertilization of the mouse egg.

Unfertilized mouse oocytes and eggs 1-8 h after fertilization in vitro were examined at the light microscope level for structural changes, distribution of actin (as assessed by both antiactin antibodies and NBD-phallicidin), surface binding of concanavalin A (Con A) and chromosomal distribution and condensation. The influence of cytochalasin D on these events was also assessed. Changes in actin distribution were associated with rotation of the second anaphase spindle, formation of the second polar body, the events following incorporation of the sperm nucleus, and formation and migration of the pronuclei. Cytochalasin D prevented spindle rotation, polar body formation, pronuclear migration and the restoration of Con A binding over the site of sperm entry and the site of polar body formation, but did not affect sperm fusion and entry, the loss of Con A binding at the site of sperm entry and pronuclear formation.

Actins↗
PubMed1984

The relationship between cleavage, DNA replication, and gene expression in the mouse 2-cell embryo.

The 2-cell stage of mouse embryogenesis is characterized by two phases of alpha-amanitin-sensitive polypeptide synthetic activity, which appear to mark the first major expression of the embryonic genome, as assessed by examination of in vitro translates of mRNA. Using populations of embryos synchronized to the first cleavage division, we have established that DNA replication takes place over the period 1 to 5.5 h after the first cleavage division; the two bursts of putative transcription take place before and immediately after DNA replication, and the translation products are detectable in each case within 3-4 h. In addition, we have shown that suppression of cytokinesis and the second round of DNA replication does not affect synthesis of the alpha-amanitin-sensitive polypeptides, and that neither DNA replication nor the loss of maternal mRNA that take place during the 2-cell stage are dependent upon synthesis of the alpha-amanitin-sensitive polypeptides.

Amanitins↗
PubMed1984

The nature of intercellular coupling within the preimplantation mouse embryo.

The changing nature of intercellular coupling during the 4- and 8-cell stages of mouse early development has been investigated by iontophoretic injection of carboxyfluorescein, horseradish peroxidase and current into individual blastomeres in either the intact embryo or after their disaggregation and reaggregation into pairs. Coupling junctions that allowed only molecules of low molecular weight (putative gap junctions) were found not to appear until 2-5 h beyond the 3rd cleavage division (8-cell stage). However, intercellular junctions that were not size selective were detected in intact embryos only throughout the 4- and 8-cell stages. It is proposed that this junctional communication results from the persistence of midbodies through all or part of the two, and in a few cases the three, cell cycles following their formation at the first and second cleavage divisions. We conclude that the cells of the early mouse embryo may be linked in a more extensive syncytial network than was hitherto suspected.

Animals↗
PubMed1983

Hemoglobin Evanston: alpha 14(A12) Trp leads to Arg. A variant hemoglobin associated with alpha-thalassemia-2.

Hb Evanston (alpha 14 Trp leads to Arg) was detected on cellulose acetate at pH 8.4 as a band with an electrophoretic mobility similar to that of Hb S. In addition, a band migrating cathodic to Hb A2 suggested the presence of a variant Hb A2 with a substitution in the alpha-chain, a fact that was later confirmed by structural analysis. An unusual feature of Hb Evanston is its low percentage; less than 10% occurs in the hemolysate. Studies indicate that the variant is not unstable, but there appears to be a defect in globin-chain synthesis. Gene mapping also shows that it is associated with the alpha-thalassemia-2 gene. The variant has high oxygen affinity with normal cooperativity and a normal Bohr effect. The combination of Hb Evanston with alpha-thalassemia-2 produced anemia in this black family.

Female↗
PubMed1983

Cell interactions influence the fate of mouse blastomeres undergoing the transition from the 16- to the 32-cell stage.

Newly formed polar and apolar 1/16 blastomeres were isolated and cultured singly, or in various combinations, through division to form 32-cell blastomeres. The morphology of the resulting cell cluster appeared to depend upon the nature and composition of the cell combination used. In most polar + apolar couplets, the polar cell enveloped the apolar cell, and following division, a 4/32 cluster was thereby generated containing two trophectoderm-like external cells derived from the polar cell and two ICM-like internal cells derived from the apolar cells. A polar cell cultured in isolation divided to give either two trophectoderm-like external cells or a trophectoderm-like cell and an ICM-like cell. Two polar cells cultured together generated clusters in which the ratio of trophectoderm-like:ICM-like cells was 4:0 or 3:1. Most apolar cells cultured together in couplets polarized, and generated 4/32 clusters containing either purely trophectoderm-like or a mixture of trophectoderm- and ICM-like cells. The results are consistent with the notion that continuing interactions between polar and apolar cells are necessary to maintain their respective fates as trophectoderm and ICM, and that in the absence of these interactions polar cells can generate ICM cells by a differentiative division and apolar cells can generate trophectoderm cells by polarizing in response to asymmetric cell contacts.

Animals↗
PubMed1983

Effect of cervical vagal stimulation on chicken heart rate and atrioventricular conduction.

The effects of supramaximal stimulation of the right and left cervical vagi on heart rate, pacemaker localization, and atrioventricular (AV) conduction were investigated in 15 anesthetized open-chest chickens before and after atropine sulfate. Epicardial bipolar electrograms were recorded from selected atrial sites and right ventricle. A back lead electrocardiogram was also recorded. The effect of stimulation on atrioventricular conduction was evaluated during pacing from one of the right atrial recording sites. Supramaximal stimulation of either cervical vagus produced bradycardia but not cardiac arrest. Heart rate was reduced from an average spontaneous rate of 282 +/- 13 (SE)/min to 161 +/- 13/min with stimulation of the right and left cervical vagus. Pacemaker shifts occurred in over 50% of the vagal stimulations. The most frequent shift occurred to the lower AV node or ventricles. Pacemaker shifts to the AV junctional region producing almost simultaneous activation of the atria and ventricles were not observed. Vagal stimulation during atrial pacing produced minimal prolongation in AV conduction time [right vagus, 13 +/- 3 (SE) ms; left vagus, 8 +/- 2 ms]. Second and third degree heart blocks were not observed during pacing. Vagal stimulation after atropine indicates that the cervical vagi do not contain sympathetic fibers going to pacemaker or AV conduction tissues.

Animals↗
PubMed1983

Hemoglobin Cheverly: an unstable hemoglobin associated with chronic mild anemia.

The evaluation of a family with chronic mild anemia led to the identification of a new unstable hemoglobin (Hemoglobin Cheverly). Modest anemia and reticulocytosis, normal to slightly increased mean corpuscular volume (MCV), and normal mean corpuscular hemoglobin concentration (MCHC) were present in the affected family members. Electrophoresis of blood samples on cellulose acetate and on citrate agar revealed normal patterns. Globin chain analysis and isoelectric focusing data were also normal. After incubation for 3 h at 41 degrees C, Heinz bodies were detected in 95-100% of erythrocytes from affected individuals. Positive heat and isopropanol tests confirmed the initial observation of the Heinz body preparation and indicated that an unstable hemoglobin was present. Structural analysis showed an amino acid substitution of Phe-Ser at position 45 (CD4) in the beta chain. Hemoglobin Cheverly has a reduced affinity for oxygen and a reduced Bohr effect, properties that can be rationalized on the basis of the x-ray crystallographic structure of normal hemoglobin. Despite structural and functional similarities between Hb Cheverly and Hb Hammersmith, beta 42 (CD1) Phe-Ser, the clinical manifestations of Hb Cheverly are mild in contrast to the severe disease observed with Hb Hammersmith. Reasons for the apparently silent clinical expression of Hb Cheverly are not known. We discuss the implications of unstable hemoglobins in the evaluation of chronic anemia in pediatric patients.

Anemia↗
PubMed1983

Echocardiography of cardiac metastases.

Metastatic disease to the heart is a relatively common entity that may involve the pericardium, myocardium, or endocardium singly or in combination. Echocardiography, particularly two-dimensional imaging, is the most sensitive tool for detecting metastatic disease to the heart. Simple, safe, and noninvasive, it can provide better anatomic detail than can more invasive studies. This information is helpful not only in diagnosis and treatment planning, but in differential diagnoses of unusual cardiac configurations and guidance for pericardiocentesis of loculated effusions.

Aged↗
PubMed1983

The fifth cell cycle of the mouse embryo is longer for smaller cells than for larger cells.

Newly-formed pairs of 16-cell blastomeres were collected by periodic observation of isolated 8-cell blastomeres. Any pairs formed by division were recovered and classified as being composed of 1/16 blastomeres that differed in size or were of similar size. All of the latter and some of the former were then cultured for periods of up to 20h. The remaining pairs of different-sized blastomeres were disaggregated to larger or smaller cells. Some of these were reaggregated as smaller: smaller or larger: larger pairs, and these, together with the remaining isolated smaller and larger blastomeres were also cultured for up to 20h. At hourly intervals, cultured cells were sampled and analysed for incidence of division. It was found that larger cells divided on average after 12h whereas smaller cells divided on average after 14h.

Animals↗
PubMed1982

Identification of protein bands in polyacrylamide gel by protein printing.

Previously described methods for identification of proteins separated in cylindrical polyacrylamide gels have been found to be costly in time and antiserum and difficult to apply to small amounts of protein as are found in cerebrospinal fluid. We describe a method which involves printing of the proteins on the cut surface of the gel onto nitrocellulose paper. The protein bands of the imprint can then be identified using labelled antibodies. We have found this to be economical and quick, and it has permitted sensitive and reliable identification of proteins in unconcentrated cerebrospinal fluid and aqueous humour.

Blood Proteins↗