IgG-specific helper activity of t lymphocytes from mice lacking theIr-IgG gene

Cooperative interactions between T and B cells from the congenic inbred mouse strains B10.A(2R) and B10.A(4R) in antibody responses controlled byIr genes have been studied. Within theI region of the MHC, these strains share only theI-A subregion. TheIr gene controlling responsiveness to IgA maps in theI-A subregion, both strains being responders to IgA. T cells from 2R mice collaborate effectively with B cells from 2R or 4R mice for antihapten antibody responses to DNP-IgA. TheIr gene controlling responses to IgG maps in theI-B subregion, and 2R mice are nonresponders for this antigen. Nevertheless, 2R T cells primed with IgG can help responder (4R) B cells -but not syngeneic nonresponder (2R) B cells -in responding to DNP-IgG. These results indicate that mice lacking theIr-IgG gene nonetheless may develop helper T lymphocytes specific for myeloma proteins. In addition, they indicate that cells from congenic mice sharing only theA subregion of theI region can collaborate efficiently.     

A Cold-Sensitive Zygotic Lethal Causing High Frequencies of Nondisjunction during Meiosis I in DROSOPHILA MELANOGASTER Females

The X-linked, cold-sensitive zygotic lethal, l(1)TW-6(cs), both in homozygous and heterozygous females, induces nondisjunction of all four chromosomes at Meiosis I at both 25 degrees and 17 degrees . Nondisjunction frequencies approaching 0.5 for the X and fourth chromosomes have been observed at 16 degrees -18 degrees . The disjunction of the X chromosomes in males is not affected. The mutant causes mitotic irregularities in zygotes at both 25 degrees and 17 degrees . Mortality of all zygotes produced by the crosses 6(cs)/6(cs)x6(cs)/B(s)Y and FM7/6(cs)x6(cs)/B(s)Y is respectively 86% and 67-74% at 25 degrees and 99.8-99.9% and 94% at 17 degrees . The mortality of 6(cs) hemizygotes derived from females carrying no doses of 6(cs) (C(1)DX,y f/yx6(cs)/B(s)Y) is 45-55% at 25 degrees and 98% at 17 degrees . The length of the temperature-sensitive period for 6(cs) homo- and hemizygotes is affected by the maternal dosage of 6(cs); the shortest TSP is for zero and the longest is for two maternal doses. Mortality takes place primarily during embryogenesis with some larval and little pupal mortality. Analysis of sectioned embryos indicates that the large array of different patterns of damage observed could have arisen from abnormal cleavage divisions and the incomplete population of the blastoderm with nuclei.     

Effect of dibutyryl cyclic AMP on glucose-6-phosphatase activity in human fetal liver explants

Glucose-6-phosphatase (EC 3.1.3.9) activity in human fetal liver remains constant at 8–28 nmoles/min per mg protein from the 8th week of gestation to at least week 28 and this value is approximately 25–35% of that found in the adult. This enzyme activity was well maintained for 2–3 days in organ culture of fetal liver explants. Incubation with dibutyryl cyclic AMP (0.1 mM) and theophylline (0.5 mM) increased glucose-6-phosphatase activity 4–8-fold within 24 h. Theophylline alone was ineffective, but markedly potentiated the effects of dibutyryl cyclic AMP. This increase in enzyme activity was completely abolished by simultaneous incubation with cycloheximide or actinomycin D. Insulin clearly decreased glucose-6-phosphatase activity in control tissues after 24 h incubation and tended to diminish the elevated glucose-6-phosphatase activity which resulted from pre-incubation with dibutyryl cyclic AMP.The smallest specimen obtained (36 mm crown-rump length = 6 weeks gestation) was capable of elevating glucose-6-phosphatase activity more than 3-fold in response to dibutyryl cyclic AMP incubation, suggesting that the human fetal liver has the competence to respond to hormonal agents at a very early stage of development.     

Contamination of Sputum Induction Equipment During Patient Usage

Sputum induction equipment was evaluated for its capacity to become contaminated by patients harboring Mycobacterium tuberculosis. The mouthpiece, goose-neck, and 10% NaCl solution were found capable of being contaminated by tuberculous patients. Two per cent glutaraldehyde was shown to be an effective means of decontamination. This study indicates that the entire induction apparatus must be cleaned and decontaminated between patients.     

Electron Microscopy During Release and Purification of Mesosomal Vesicles and Protoplast Membranes from Staphylococcus aureus

The mesosomes of log-phase Staphylococcus aureus ATCC 6538P and Staphylococcus aureus phage-type 80/81, as seen in situ in ultrathin sections, were of the vesicular type. The constituent vesicles ranged from 35 to 50 nm in diameter when the glutaraldehyde-osmium-uranium-lead sequence of fixation and staining was used. During protoplasting in hypertonic buffer containing a muralytic enzyme, vesicles of the same size were extruded and required magnesium ion to maintain structural integrity. The vesicles, purified from the protoplasting supernatant medium by density gradient centrifugation, maintained size and configuration in a homogeneous preparation. Cytoplasmic membranes, produced by osmotic shock and nuclease treatment of protoplasts, were similarly concentrated in gradients. However, they were not free of membrane-associated ribosomes nor of mesosomal vesicles except when prepared in the absence of magnesium.     

The Genetics of Dopa Decarboxylase in DROSOPHILA MELANOGASTER . II. Isolation and Characterization of Dopa-Decarboxylase-Deficient Mutants and Their Relationship to the α-Methyl-Dopa-Hypersensitive Mutants

Of 84 lethals isolated over the dopa decarboxylase (DDC) deficiency Df(2L)50, 8 have been identified as DDC-deficient alleles on the basis of their effect on DDC activity when heterozygous over the CyO balancer chromosome with activities ranging from 28% to 53% of controls. Some of the Ddc-deficient alleles exhibit intracistronic complementation. Most of the complementing pairs of alleles are much reduced in viability, e.g. < 5% of expected, and express a common syndrome of mutant phenes which can reasonably be inferred to derive from inadequately sclerotinized cuticle. Individuals heterozygous for the noncomplementing allele, Ddcⁿ⁷, over the 12-band DDC deficiency, Df(2L)130, die at the end of embryogenesis as unhatched larvae with unpigmented mouth parts.

The Ddc alleles and the l(2)amd α-methyl dopa (αMD) hypersensitive alleles are both located within the 11 band region 37B10-C7. The l(2)amd locus is immediately to the right of hk(2–53.9).Ddc has been mapped within 0.004 Map Units to the right of l(2)amd with a maximum estimated recombination frequency of 0.01%. None of the Ddc/CyO strains are sensitive to the dietary administration of α-methyl dopa (αMD), and complementation occurs between the Ddc deficient alleles and the l(2)amd alleles both on the basis of viability and DDC activity. No effect on DDC by the amd alleles has been found to date. Even in the complementing heterozygote, amd^H1/amd ^H89, the level of activity, thermostability, and in vitro αMD inhibition of DDC remains unaffected. Although no biochemical phene has yet been established for the αMD hypersensitive amd alleles, it seems likely that the two groups of mutants are functionally related.

     

Differences in terrestrial velocity in Macaca and Presbytis

Over 150 sequences of terrestrial movement were timed and measured in the toque macaque, the gray langur, and the purple-faced langur to determine the relation between velocity, gait, and psychosocial context. Species differences were found in mean velocity, “favored” gait, and surface preference. All three species used the walk and the gallop at the slowest and fastest speeds respectively. The macaques and gray langurs walked frequently, but the purple-faced langurs were never seen to do so. At intermediate speeds, the macaques used either the “fast” walk or the “slow” gallop, whereas the gray langurs used irregular patterns of walk-gallop-walk-gallop. The purple-faced langurs were faster (about 20 fps) and less variable than the other species, regardless of distance. These data suggest that motor expression varies among the Cercopithecoid monkeys: the correlation between locomotion and anatomy is not as close as it is among wholly terrestrial or arboreal forms; slowness in the macaque is an expression of social confidence, not of biomechanical inability; high speed in the purple-faced langur is due to psychosocial factors rather than to “terrestrial” adaptability.     

Ultrastructure of human endometrial epithelium in monolayer culture with and without steroid hormones

Summary Colonies of cells of epithelioid appearance were identified in monolayer cultures grown up to 50 days from normal human endometrial cell suspensions obtained by a method designed to insure a maximum harvest of glandular cells. Groups of these cells were separated from stromal cells by means of cloning cylinders. Studies comparing the ultrastructure of cells of this type to fresh endometrial tissue revealed a number of similarities. The morphological characteristics common to both types of samples included junctional complexes, perinuclear microfilaments and microvilli with glycocalyx. Other common features were prominent nucleoli, well developed Golgi, rough endoplasmic reticulum and membranebound electron-dense bodies in the cytoplasm. A stripping technique applie to the fetal bovine serum used in the nutrient medium made it possible to initiate cultures in a steroidfree environment and to maintain them in the presence of the specified concentration of estradiol and/or progesterone. Isolation of epithelial cells of endometrium in monolayer culture may provide a useful model system in which to study the specific effects of steroid hormones on cellular function and differentiation. Supported by grants from the National Institutes of Health (CA 18678 and CA 07368).     

Engineering factors in single-cell protein production. II. Spray drying and cell viability

One important economical method for producing singlecell protein is to spray dry the cultured cells. This study presents some preliminary data on the effects of spray drying on cell viability. Under conditions similar to those for the production of spray-dried milk, 4–5 log cycles destruction occurred. The results indicate that, the activation energy for thermal destruction of yeast was reduced from the normal heat treatment value of 84 kcal/°K mole to about 38 kcal/°K mole.     

Isolation and characterization of six human hepatic isometallothioneins.

Human brain contains one cationic (pI8.3) and two anionic (pI5.5 and 4.6) forms of glutathione S-transferase. The cationic form (pI8.3) and the less-anionic form (pI5.5) do not correspond to any of the glutathione S-transferases previously characterized in human tissues. Both of these forms are dimers of 26500-Mr subunits; however, immunological and catalytic properties indicate that these two enzyme forms are different from each other. The cationic form (pI8.3) cross-reacts with antibodies raised against cationic glutathione S-transferases of human liver, whereas the anionic form (pI5.5) does not. Additionally, only the cationic form expresses glutathione peroxidase activity. The other anionic form (pI4.6) is a dimer of 24500-Mr and 22500-Mr subunits. Two-dimensional gel electrophoresis demonstrates that there are three types of 26500-Mr subunits, two types of 24500-Mr subunits and two types of 22500-Mr subunits present in the glutathione S-transferases of human brain.