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Measurements of speeds of sound uhave been made in binary liquid mixtures of cyclohexanone (C6H10O) and dichloromethane (CH2Cl2), or trichloromethane (CHCl3), or trichloroethene (CHClCCl2), or 1,2-dichloroethane (CH2ClCH2Cl) at T=303.15 K. The values of uhave been used to calculate the apparent excess speeds of sound Δuand the isentropic compressibilities κSfor these mixtures. The excess isentropic compressibilitiesκSEhave also been calculated from the values ofκS. The κSEproperty has been found to be negative throughout the entire composition range for mixtures of C6H10OandCH2Cl2, or CHCl3, or CHClCCl2. For {xC6H10O +(1 −x)CH2ClCH2Cl},κSEhas been found to be positive at low values of xand negative at higher values of x, with an inversion of sign occurring at x≈0.24. The values ofΔuand κSEhave been fitted to representative equations.

In an effort to characterize the cellular antigens recognized by anti- neutrophil antibodies in autoimmune neutropenia, we studied sera, purified immunoglobulin G (IgG) and isolated F(ab')2 from 70 neutropenic patients suspected of this diagnosis. Anti-neutrophil antibodies were found in the sera of 36 of these patients by either 125I-staph A binding or immunofluorescence cytometric techniques that detected increased binding of patients' IgG to normal neutrophils. Anti- neutrophil antibody positive sera were then evaluated for specific binding to electrophoretically separated neutrophil membrane-associated proteins by immunoblotting. A 43-Kd protein was consistently identified by eight anti-neutrophil antibody positive sera. The specificity of binding to this protein was confirmed with affinity purified IgG and F(ab')2 fragments prepared from these sera. Sera from 20 healthy normal controls and from 22 non-neutropenic, anti-neutrophil antibody negative rheumatoid arthritis patients failed to bind this protein. Separate studies identified the 43-Kd protein as actin. Purified Acanthamoeba actin comigrated with the protein and was specifically bound by anti- neutrophil antibody positive IgG. Moreover, two actin-specific monoclonal antibodies bound to the 43-Kd membrane-associated protein in immunoblots. In addition, a rabbit anti-actin antiserum not only bound to this same 43-Kd protein but also expressed anti-neutrophil antibody activity against normal human neutrophils, as did purified human anti- actin IgG prepared by affinity chromatography from the serum of one of the index patients. These studies indicate that the anti-neutrophil antibodies of certain patients with autoimmune neutropenia include autoantibodies specific for actin. The molecules on the surface of neutrophils, which have actin-like antigenic epitopes and are recognized by these anti-actin antibodies, remain to be characterized.

Summary The entropy of melting in inorganic polymers and that of the helix-coil transition has been estimated on the basis of rotational isomeric state model. The values of entropy permol. segment obtained are in good agreement with those reported in the literatures.

Summary Immunofluorescent staining of kinetochores in the micronucleus was used for distinguishing between clastogenic and aneuploidogenic effects of various chemicals. The technique consists of culturing cells on glass slides, chemical treatment, fixing cells in situ, and fluorescent staining. Anti-kinetochore antibody was used as a primary antibody with fluoresceinated goat anti-human IgG as a secondary antibody to locate kinetochores, and 4,6-diamidino-2-phenyl-indole (DAPI) or propidium iodide was used for identifying the nucleus, micronucleus, and cytoplasm. Kinetochores appear yellow/green against the orange/red nuclear material and the cytoplasm appears faint green in slides counterstained with propidium iodide. In DAPI counterstained slides, kinetochores appear apple green and the nucleus and micronucleus appear light blue. Thus, location of kinetochores and nuclear material within the cell boundary can be achieved. The presence/absence of kinetochore(s) in a micronucleus is used as an indicator to determine the origin of the micronucleus. The proportion of kinetochore-positive and kinetochore-negative micronuclei is used in the determination of the clastogenic and aneuploidogenic properties of different genotoxic agents. This method is simple and applicable to a variety of cells.

We evaluated a 3-year-old female patient with leukocytosis, recurrent infections, severe periodontal disease, and a history of delayed separation of the umbilical stump. This patient's polymorphonuclear leukocytes (PMNs) had normal membrane depolarization responses, normal oxygen metabolism, normal granule secretion responses, normal bactericidal activity, and normal C3b rosetting. However, by fluorescent cell analysis and C3bi rosetting, it was determined that her cells lacked the C3bi receptor. In addition, the patient's PMNs showed markedly abnormal chemotaxis, adherence, and aggregation responses, and partial abnormalities were detected in PMN spreading and phagocytosis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the subject's neutrophil cytoplasts were missing a 180,000-dalton moiety. Her monocytes also had defective chemotaxis and failed to adhere and grow normally in culture. Epstein-Barr virus- transformed B cells from the patient lacked an aggregation response to phorbol myristate acetate. Laboratory and clinical evaluations of this patient's mother showed no abnormalities. These studies demonstrate that C3bi receptor deficiency can be associated with functional abnormalities in multiple myeloid cells and that the absence of C3bi receptor is associated with abnormal adherence-related functions of these cells.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal-elution chromatography were employed to identify the diploid wheat species ancestral to the B genome of Triticum turgidum. 3H-T. turgidum DNA was hybridized to the unlabeled DNAs of T. urartu, T. speltoides, T. sharonensis, T. bicorne, T. longissimum, and T. searsii. 3H-Labeled DNAs of T. monococcum and a synthetic tetraploid AADD were hybridized with unlabeled DNAs of T. urartu and T. searsii to determine the relationship of the A genome of polyploid wheat and T. urartu. The heteroduplex thermal stabilities indicated that T. searsii was most closely related to the B genome of T. turgidum (AB) and that the genome of T. urartu and the A genome have a great deal of base-sequence homology. Thus, it appears that T. searsii is the B-genome donor to polyploid wheat or a major chromosome donor if the B genome is polyphyletic in origin.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal chromatography were employed to help identify the species ancestral to the B genome of the polyploid wheats. We hybridized 3H-Triticum aestivum DNA to the unlabeled DNAs of T. urartu, T. speltoides, T. sharonensis, T. bicorne, T. longissimum, and T. searsii. 3H-Labeled DNA of T. urartu was hybridized with the DNA of a synthetic tetraploid, AADD. The heteroduplex thermal stabilities indicated that T. searsii was most closely related to T. aestivum (ABD) and that the genome of T. urartu was more closely related to the A genome than the B genome. The degree of reassociation which may have occurred between the six diploid species and the D genome of T. aestivum was evaluated by hybridizing 3H-T. tauschii DNA with the DNAs of the diploids. The results indicated that T. urartu had the least sequence homology to T. tauschii, the D-genome donor lending additional support to the conclusion that T. urartu is related to the A genome. Thus, it is highly probable that T. searsii is the B-genome donor to the polyploid wheats or a major chromosome donor if the B genome is, in fact, polyphyletic in origin.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal-elution chromatography were employed to identify the diploid Triticum species ancestral to the B genome of T. turgidum. Unique and repeated sequences from the various Triticum species were separated by hybridization and thermal elution on hydroxyapatite. Unique- and repeated-sequence fractions of labeled T. turgidum var. durum DNA were hybridized to the corresponding fractions of unlabeled DNAs of T. searsii, T. speltoides, T. longissimum, T. sharonensis, and T. bicorne. Thermal stability profiles were constructed to evaluate base-sequence complementarity between T. turgidum var. durum and the diploid Triticum species. The heteroduplex thermal stabilities indicated that, of the five species examined, T. searsii was the most closely related to the B genome of T. turgidum var. durum. The thermal stability profiles further indicated that the repeated DNA fractions from the Triticum species are more similar than the unique-sequence fractions. This indicates that all of the Triticum species are very closely related and, in all probability, have diverged from a single progenitor species.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal-elution chromatography were employed to identify the diploid Triticum species ancestral to the G genome of Triticum timopheevii. Total genomic, unique-sequence, and repeated-sequence fractions of 3H-T. timopheevii DNA were hybridized to the corresponding fractions of unlabeled DNAs of T. searsii, T. speltoides, T. sharonensis, T. longissimum, and T. bicorne. The heteroduplex thermal stabilities indicated that, of the five species examined, T. speltoides was the most closely related to the G genome of T. timopheevii. Thus, T. spelotides appears to be the G-genome donor to T. timopheevii. The thermal stability profiles further indicated that the repeated DNA fractions from the five diploid species and the tetraploid T. timopheevii are more similar than the unique DNA fractions. This indicates that all of these species are closely related and that the sequences which comprise the current repeated fractions in the various species have not undergone any significant change since the formation of various species.

Individual variability of scoring foci positive for transformation presents a difficult problem in assessing the transformation assay. In this study, an attempt was made to identify five morphologically distinct types of transformed foci based on size (2–3, 3–4, and ≥4 mm in diameter), invasiveness (smooth vs. invading margins), and other properties (piling vs. spread) induced by 3‐methylcholanthrene in Balb/c‐3T3 cells. The transformed focal cells were used in in vitro studies including anchorage‐independent analysis, focal reconstruction, gene transfection using NIH‐3T3 host cells, and Southern blotting to assess amplification of five proto‐oncogenes (K‐ras, H‐ras, c‐fos, c‐jun, c‐myc) and a tumor suppressor (p53) gene. Results showed that 1) there was a significant increase in anchorage‐independent growth of all five types of foci ranging from 7–12%; 2) all five morphological types of transformed foci showed 8–15% focal reconstruction; 3) DNA from all five types of transformed foci induced transformation in NIH‐3T3 cells at a level significantly above the control DNA; 4) gene amplification studies indicated amplification in both K‐rasand H‐rasproto‐oncogenes; however, c‐fos, c‐jun,and c‐mycdid not show DNA amplification. The tumor suppressor gene (p53) was activated and the increase was up to 3‐fold over the normal Balb/c‐3T3 DNA. These findings are consistent with our hypothesis that all five morphologically different foci have preneoplastic potential and that any foci of size ≥ 2 mm regardless of invasiveness and piling should be scored as positive during the transformation assay. Environ. Mol. Mutagen. 32:369–376, 1998 © 1998 Wiley‐Liss, Inc.

The effect of sodium dodecyl sulphate, urea or guanidinium hydrochloride on the sedimentation velocity, viscosity, ultra violet spectra and fluorescence spectra of the 11S protein of guar seed has been determined. Sodium dodecyl sulphate dissociates the protein directly to the 2S protein, whereas urea or guanidinium hydrochloride produces an intermediate 7S protein. These reagents denature the protein also. Both the dissociative and the denaturation effect follow the order, sodium dodecyl sulphate > guanidinium hydrochloride > urea when the concentration are expressed as mols per litre. The denatured states in the three cases probably differ.

Wave force coefficient variability for cylinders, from wave to wave in a train of periodic waves, has been shown to be dependent on the phase of the force record relative to the ambient flow. The phase varies due to vortex shedding, but the maximum force is approximately constant as seen from this work and the work of other investigators. Thus, the maximum force coefficient is tightly organized according to the Keulegan-Carpenter number and scatter is seen in the phase angle versus Keulegan-Carpenter number. On the other hand, both Cd and Cm have scatter due to these phase differences from wave to wave. For unknown reasons, even when averaged over several wave cycles there is scatter in the results for Cd and Cm. This investigation shows that the maximum force coefficients for a heavily roughened vertical cylinder are tightly arranged according to the Keulegan-Carpenter number and the period parameter. Furthermore, the phase angle is similarly much more organized than for the smooth cylinder.

Estimating the force coefficients Cd and Cm for the two‐term Morison equation for a cylinder in waves depends on accurately measuring the amplitudes and phase shifts between the kinematics of the ambient wave and the wave‐induced force. A regression analysis is used to develop an algorithm that illustrates both the parametric dependency of the data on the Dean eccentricity parameter and the variability of these force coefficients if incorrect amplitudes or phase shifts are introduced into the analysis. The algorithm assumes that the two‐term Morison equation represents the “true” forces exactly and that linear wave theory represents the wave kinematics exactly. A least‐squares analysis of the time‐averaged, mean‐squared error between a “true” and a “computed” force is used. Dimensionless variations in the coefficients depend on two parameters: (1) A dimensionless force amplitude ratio W (proportional to the Dean eccentricity parahieter E); and (2) a dimensionless velocity amplitude ratio V, which is a function of the vertical elevation in the water column z. A new way to illustrate the importance of data conditioning is presented that combines the parametric dependency of the Dean eccentricity parameter with the wave phase/amplitude dependency. Good agreement is obtained with laboratory data of wave forces on a vertical, sand‐roughened cylinder wherein the force measurements were synthetically phase shifted with respect to the wave phase in small increments, up to ±33.8 degrees (±3π/16 radians).

Free and bound acid phosphatase were investigated biochemically and electrophoretically in male sterile and fertile plants. Scutella at the 72-hr seedling stage and anthers with enclosed reproductive tissue at the premeiosis, meiosis, and mature developmental stages were tested. Biochemical data show that while the total amount of free acid phosphatase behaved similarly in fertile and sterile plants, specific activities decreased in fertile plants and remained unchanged or increased in sterile plants. Total amounts of bound acid phosphatase increased significantly in fertile plants while specific activities decreased. In sterile anthers both specific activity and amount of bound acid phosphatase decreased significantly (except cms-S). Electrophoretic results indicated that the basic form of the enzyme was very similar in each location.

The mechanism of cytoplasmic male sterility was investigated in maize by isolating mitochondria from seedlings and various anther stages and analyzing cytochrome oxidase and succinic dehydrogenase biochemically and electrophoretically. Sterile anthers exhibited a lack of biochemical activity and fewer isozymatic bands for cytochrome oxidase. No apparent differences were detected biochemically or electrophoretically between fertile and sterile anthers for succinate dehydrogenase.

Summary Procedures for preparing and culturing Chinese hamster bone marrow and spleen cells for cytogenetic studies are described. Animals are killed by cervical dislocation, then the bone marrow is flushed from femora and tibia with Ham's F12 medium into centrifuge tubes. Bone marrow cells are collected by low-speed centrifugation (285 ×g). Approximately one million cells are cultured in a 30-ml Falcon flask with 5 ml complete medium containing 3.95 ml Ham's F12 medium, 1.0 ml fetal bovine serum (FBS), and 0.05 ml penicillin-streptomycin (5000 U/ml and 5000 µg/ml stock). Spleens are obtained aseptically, transferred to centrifuge tubes containing 2 ml of RPMI 1640 and smashed with a sterile spatula. The debris is removed, cells are collected by low-speed centrifugation (285 ×g), and washed three times with phosphate buffered saline supplemented with 2% heat inactivated FBS. Approximately one million cells are suspended in a 30-ml Falcon flask with 5 ml complete medium containing 3.70 ml RPMI 1640. 1.0 ml heat inactivated FBS, 0.05 ml penicillin-streptomycin, 0.05 ml of 200 mMl-glutamine solution, 1 × 10-5M 2-mercaptoethanol, and 0.20 ml lipopolysaccharide. For sister chromatid exchange analysis, 20 µM of 5-bromo-2' -deoxyuridine is also added to the culture medium for 24 to 28 h for bone marrow cells and 36 to 40 h for spleen cells. These culture systems can also be used for chromosomal aberration studies.

Summary A procedure for preparing and culturing mouse bone marrow cells for cytogenetic studies is described. Animals are killed by cervical dislocation, the bone marrow is flushed from femora and tibia with Ham's F12 medium into centrifuge tubes. Bone marrow cells are collected by low-speed centrifugation (285 ×g). One million cells are suspended in a 30-ml Falcon flask with complete medium containing 3.45 ml Ham's F12 medium, 1 ml fetal bovine serum, 0.05 ml penicillin-streptomycin, and 0.5 ml whole uterus extract from pregnant mice. For sister chromatid exchange analysis, 20 µM of 5-bromo-2' -deoxyuridine is also added to the culture medium for 24 to 28 h. The cell cycle is approximately 12 to 14 h, in culture. This culture system can also be used for chromosomal aberration studies.

Excess molar volumes, %@mt;sys@%%@ital@%V%@rsf@% %@sx@%m%@be@%E%@sxx@%,%@mx@% have been measured for binary mixtures of butanol (n-C4H9OH) + pentane (n-C5H12), + hexane (n-C6H14), + heptane (n-C7H16), + octane (n-C8H18), + 2,2,4-trimethylpentane (2,2,4-TMP), and + carbon tetrachloride (CCl4) at 288.15 K and 298.15 K, and refractive indexes, nD, have been measured for these mixtures at 298.15 K. At both temperatures %@mt;sys@%%@ital@%V%@rsf@% %@sx@%m%@be@%E%@sxx@%%@mx@% has been found to be positive throughout the entire range of composition for xn-C4H9OH + (1 − x)n-C7H16 and + (1 − x)n-C8H18. At both temperatures 288.15 K and 298.15 K, %@mt;sys@%%@ital@%V%@rsf@% %@sx@%m%@be@%E%@sxx@%%@mx@% is positive at low mole fractions of n-C4H9OH and negative at its higher mole fractions in the case of mixtures of n-C4H9OH with n-C5H12, n-C6H14, 2,2,4-TMP, and CCl4. Values of %@mt;sys@%%@ital@%V%@rsf@% %@sx@%m%@be@%E%@sxx@%%@mx@% and nD for the various mixtures of n-C4H9OH have been fitted in smoothing equations.

Excess molar volumes, %@scp@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%,%@mx@% relative permittivities, εr, and refractive indexes, nD, have been measured for binary mixtures of 1,1,2,2-tetrachloroethane (CHCl2CHCl2) with pyridine (C5H5N), anisole (C6H5OCH3), methyl ethyl ketone (CH3COC2H5), and 1,4-dioxane (1,4-C4H8O2) at 303.15 K. %@scp@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%%@mx@% has been found to be negative throughout the entire mole fraction range for all these mixtures. The values of the deviations of εr and nD from a mole fraction average, which are represented respectively by Δεr and ΔnD have been calculated. The results of %@scp@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%,%@mx@% ΔnD, and Δεr for the various mixtures have been fitted by the method of least squares to smoothing equations.

Abstract.: Antimitotic agents administered at the time of synapsis (leptotene/zygotene) have been shown to induce synaptic abnormalities visible during pachytene in the male mouse. The object of this study was to test the hypothesis that cells with relatively large amounts of colchicine-induced damage to the synaptonemal complex (SC) are eliminated from prophase whereas cells with relatively small amounts of SC damage proceed through to the end of prophase. Male mice were injected with tritiated thymidine to mark a cohort of spermatocytes at premeiotic S-phase for tracking through pachytene. Forty-eight hours later, when those cells were at leptotene/zygotene, colchicine was administered intratesticularly. Whole-mount SC spreads were made from animals sacrificed at various times following colchicine administration, and prepared for autoradiography. The marked cells were examined by light and electron microscopy and the kind and number of synaptic abnormalities were scored throughout pachytene. Colchicine-induced SC damage included single axial elements (univalents), together with partially synapsed and nonhomologously synapsed SCs. The amount of SC damage (amount and type per cell and frequency of cells with damage) scored at early pachytene exceeded by three- to fivefold the amount at late pachytene. This is consistent with spermatogenic cell loss from the seminiferous tubule via colchicine-induced destruction of Sertoli cell microtubules. The presence of spermatocytes with no more than four autosomal univalents at late pachytene indicates that some cells with low amounts of synaptic damage progress to the end of pachytene. The loss of the most severely damaged cells may represent a meiotic checkpoint at early pachytene in the male mouse.

The effect of gamma rays on parameters such as chlorophyll mutation frequency and spectrum, mutagenic effectiveness and efficiency and viable mutation frequency and spectrum were studied in M2 generation of Rhodes grass employing nine doses of gamma rays. The chlorophyll mutation frequency increased in a linear fashion at low and medium doses and was erratic at higher doses. The chlorophyll mutant spectrum ineluded albina, xantha, chlorina, viridis, tigrina, striata, albo-xantha and albo-viridis. The frequency of these mutants varied with treatments. A dose rate of 60 krad was found to be effective as well as efficient both on M1 and M2 plant basis in inducing chlorophyll mutations. The highest frequency of viable mutations was found in 50 krad treatment. Viable mutants found were: nodal variant, peduncle variant, lax ears, coxcomb ears, earhead proliferation, level of fingers, incurved fingers, gappy ears, apical sterility, partial sterility and high sterility.

Non-arrowing and broad-leaved mutants of Rhodes grass have been isolated following gamma irradiation. The green forage yield in non-arrowing and broad-leaved types were respectively 67 and 171 percent more than the control. Crude protein, nitrogen and calcium fractions nearly doubled in the non-arrowing type and remained approxymately the same in the broad-leaved type over controls.

Many workers in the highway construction and roofing industries are potentially exposed to asphalt fumes. However, little is known regarding the carcinogenic hazards of these fumes to the exposed workers. Previous studies have shown that condensates of asphalt fumes are weakly mutagenic to bacteria and are capable of inducing micronucleus formation in cultured mammalian cells. In this study, the induction of DNA adducts in vivo in lung and white blood cells (WBCs) of rats by fume condensates of type I and type III roofing asphalts was investigated using ³²P-postlabeling analysis. Male CD rats (3/group) received 3 intratracheal instillations of fume condensates in a 24-h period. DNA from both lung cells and WBCs were isolated and used to detect DNA adducts. Condensates of both roofing asphalt fumes caused DNA adduct formation in rat lung cells in a similar dose-related manner. Under the conditions studied, however, neither type I nor type III fume condensate induced DNA adducts in WBCs. These results indicate that 1) condensates of fumes from both type I and type III have similar genotoxic activity, 2) chemicals in the condensates of roofing asphalt fumes can covalently bind to the DNA of rat lung cells, and 3) WBCs may not be a suitable surrogate for lung cells in DNA adduct studies of workers exposed to roofing asphalt fumes. Teratogenesis Carcinog. Mutagen. 18:131–140, 1998. © 1998 Wiley-Liss, Inc.

Cellular tubulin is subject to a posttranslational modification involving the reversible addition to tyrosine through peptide linkage to the C-terminal glutamate of the alpha-chain. The synthetic peptide chemoattractant, N-formyl-methionyl-leucyl-phenylalanine, causes a specific, dose-dependent stimulation of tubulin tyrosinolation in rabbit leukocytes. This stimulation is prevented by carbobenzoxy-phenylalanyl-methionine, benzoyl-tyrosine ethylester, and nordihydroguaiaretic acid, which are all inhibitors of chemotaxis presumed to act via membrane-associated events. The combination of 3-deazaadenosine and homocysteine thiolactone, which inhibits phospholipid methylation, and quinacrine, an inhibitor of phospholipase A2, also abolishes the response to the peptide. Colchicine, however, which causes a marked disassembly of cellular microtubules in these cells and also inhibits chemotaxis, does not have any inhibitory effect on the basal or peptide-stimulated rate of tubulin tyrosinolation. In contrast, taxol, a microtubule-stabilizing agent, has an inhibitory effect on both the basal and peptide-stimulated tyrosine incorporation. Taxol also inhibits chemotaxis in rabbit leukocytes. The results strongly suggest the role of closely linked membrane-cytoskeleton interactions in leukocyte chemotaxis, in which tyrosinolation of tubulin may be functionally involved.

Excess molar volumes, %@mt;sys@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%%@mx@% , at T = 298.15 K, and kinematic viscosities, ν, at T = 303.15 K, have been measured for binary liquid mixtures of cyclohexanone (C6H10O) + dichloromethane (CH2Cl2), + trichloromethane (CHCl3), + l,2-dichloroethane (CH2ClCH2Cl), + trichloroethene (CHClCCl2), + 1,1,1-trichloroethane (CCl3CH3), and + cyclohexane (c-C6H12). The values of ν and %@mt;sys@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%%@mx@% have been fitted in appropriate equations using a least-squares method. %@mt;sys@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%%@mx@% has been found to be negative throughout the entire range of composition for mixtures of C6H10O + CH2Cl2, + CHCl3, + CHClCCl2, and + CCl3CH3, and positive for C6H10O + c-C6H12. For C6H10O + CH2ClCH2Cl, %@mt;sys@%%@ital@%V%@rsf@%%@sx@%m%@be@%E%@sxx@%%@mx@% has been found to be positive at lower mole fractions of C6H10O, and negative at higher mole fractions. The analysis has shown that McAllister's approach correlates ν of the present mixtures throughout the entire composition range to a significantly high degree of accuracy.

Kinematic viscosities ν have been measured for binary liquid mixtures of butanol (n-C4H9OH) + pentane (n-C5H12), + hexane (n-C6H14), + heptane (n-C7H16), and + octane (n-C8H18) at T = 298.15 K. Values of ν have been used to calculate dynamic viscosities η, using available density data. The values of the quantity Δη, which refers to the deviations of η of mixtures from those arising from a mole fraction average, have also been calculated. The values of ν, η, and Δη have been fitted in appropriate equations using a least-squares method. The analysis has shown that McAllister's approach correlates the liquid-mixture viscosity data of the present systems throughout the entire composition range to a significantly high degree of accuracy, whereas Lobe's approach predicts ν of mixtures to a lower degree of accuracy.

Abstract: Studies on aneuploidy have shown that the X is the most frequently lost chromosome in females, and that the number of X chromosome-positive micronuclei increases with age in women. Recently, we showed that the inactive X chromosome is incorporated preferentially in micronuclei. The objectives of the current study were, firstly, to determine the incidence of X chromosome incorporation into micronuclei in males and, secondly, to determine the incidence of X chromosome incorporation into micronuclei of females with Turner syndrome. Blood samples were obtained from 18 male newborns and 35 normal adult males ranging in age from 22 to 79 years and from seven women with non-mosaic Turner syndrome aged 11–39 years. Isolated lymphocytes were cultured in the presence of cytochalasin B and 2000 binucleated cells per subject were scored for micronuclei. Cells were then hybridized with the biotinylated X centromere-specific probe, pBamX7, and visualized with fluorescein-conjugated avidin. All micronucleated cells were relocated and evaluated for the presence or absence of the X chromosome. Of the 335 micronuclei observed, 6.6% (22/335) contained an X chromosome. Analysis of variance shows a statistically significant increase, for both males and Turner females, in the number of X chromosome-positive micronuclei with age (P < 0.001). These data also show that the X chromosome is included in micronuclei from males more often than would be expected by chance (P < 0.005; χ2 analysis, 15 df). Here we show that there is a tenfold difference in the frequency of X chromosome-positive micronuclei in 46,XX females compared to 46,XY males and 45,X females, providing further support to our previous finding that the X chromosome in micronuclei is the inactive chromosome.

We have recently reported a specific dose-dependent stimulation of posttranslational incorporation of tyrosine into tubulin alpha-chains of rabbit peritoneal leukocytes as induced by the synthetic peptide chemoattractant formyl-methionyl-leucyl-phenylalanine (FMLP). The present study reports a similar, specific stimulation of tubulin tyrosinolation in human polymorphonuclear leukocytes (PMN). When compared to normal PMN, both the resting and FMLP-stimulated levels of posttranslational tyrosine incorporation were two- to threefold higher in PMN of three patients with the Chediak-Higashi syndrome (CHS). The concentration of cellular tubulin and the specific activity of tubulin tyrosine ligase were similar in PMN of CHS patients and normal donors and resembled that of other non-neuronal cells. The high levels of tyrosine incorporation in PMN of CHS patients were normalized by the administration of ascorbate, both in vitro and in in vivo experiments. In vitro addition of ascorbate also inhibited the FMLP-induced stimulation of tyrosine incorporation in both normal and CHS cells. Normalization of higher levels of tyrosine incorporation in PMN of CHS patients and the inhibition of FMLP-induced stimulation of tubulin tyrosinolation in normal and CHS cells as observed with ascorbate could also be affected by other reducing agents such as reduced glutathione, cysteine, or dithiothreitol. These results suggest a possible relationship between cellular redox and tubulin tyrosinolation in PMN.

The ability to identify the empirical force coefficients used in the Morison wave force equation depends on the condition of the wave kinematic data and the measured force data. Herein the condition of these data is evaluated by three methods: two geometric and one numerical. The Dean error ellipse method is shown to depend explicitly on the Keulegan-Carpenter parameter, K, for simple harmonic kinematics via the Dean eccentricity parameter (E=3K/2π2). For simple harmonic data, the alignment of the semi-minor axis of the Dean error ellipse is shown to be parallel to the Cd–Cm-axes and to depend on K. When E<1.0, then K<11.40 and the semi-minor axis of the error ellipse is parallel to the Cm-axis. When E>1.0, then K>11.40 and the semi-minor axis is parallel to the Cd-axis. When E=1.0, then K=11.40 and the error ellipse is a circle with zero eccentricity. The Dean error ellipse method is compared geometrically with the amplitude/phase error analysis in which the condition of the data is demonstrated geometrically by the slopes of contours of dimensionless coefficient ratios passing through a zero phase error. These two geometric methods are shown to be related by the Dean eccentricity parameter, E. The matrix A condition number is also shown to depend on E. The matrix for simple harmonic kinematics is Hermitian, unitary and becomes a unit matrix with a unit condition number when K=13.16 and E=1.15. There is only a 15-percent difference between K=11.4 and K=13.16, which is usually well within the data scatter band for wave force tests on vertical smooth cylinders near these values of K. Three sets of physical data show that at K values where the data are equally well conditioned to determine both Cd and Cm, a stable condition of periodic, repetitive vortices are formed. For such stable vortex shedding in periodic flow, the force on the cylinder is stable periodic, yielding repetitive values for Cd and Cm from cycle to cycle.

Methylxanthines (MX) inhibit cell division in sea urchin and clam eggs. This inhibitory effect is not mediated via cAMP. MX also inhibit respiration in marine eggs, at concentrations which inhibit cleavage. Studies showed that no changes occurred in ATP and ADP levels in the presence of inhibitory concentrations of MX, indicating an extra-mitochondrial site of action for the drug. Subsequent studies revealed decreased levels of NADP+ and NADPH, when eggs were incubated with inhibitory concentrations of MX, but no change in levels of NAD+ and NADH. MX did not affect the pentose phosphate shunt pathway and did not have any effect on the enzyme NAD+ -kinase. Further studies showed a marked inhibitory effect on the glutathione reductase activity of MX-treated eggs. Reduced glutathione (GSH) could reverse the cleavage inhibitory effect of MX. Moreover, diamide, a thiol-oxidizing agent specific for GSH in living cells, caused inhibition of cell division in sea urchin eggs. Diamide added to eggs containing mitotic apparatus (MA) could prevent cleavage by causing a dissolution of the formed MA. Both MX and diamide inhibit a Ca2+-activated ATPase in whole eggs. The enzyme can be reactivated by sulfhydryl reducing agents added in the assay mixture. In addition, diamide causes an inhibition of microtubule polymerization, reversible with dithioerythritol. All experimental evidence so far suggests that inhibition of mitosis in sea urchin eggs by MX is mediated by perturbations of the in vivo thiol-disulfide status of target systems, with a primary effect on glutathione levels.

A specific stimulation of tubulin tyrosinolation in human neutrophils (PMNs) is induced by the synthetic peptide chemoattractant N-formylmethionylleucylphenylalanine (fMet-Leu-Phe), and this stimulation is closely associated with activation of the NADPH oxidase-mediated respiratory burst (Nath, J., and Gallin, J. I. (1983) J. Clin. Invest. 71, 1273–1281). In contrast, along with tubulin tyrosinolation, a distinctly different respiratory burst-associated random posttranslational incorporation of tyrosine into multiple PMN proteins is observed in PMNs stimulated with the phorbol ester phorbol 12-myristate 13-acetate (PMA) or sn-1,2-dioctanoylglycerol (DAG). In studies exploring the mechanism(s) of signal transduction for these distinct neutrophil responses, we found that the fMet-Leu-Phe-induced stimulation of tubulin tyrosinolation in PMNs and in differentiated HL-60 cells is completely blocked by pertussis toxin, while the PMA-induced random incorporation of tyrosine is not inhibited. We also found that expression of the fMet-Leu-Phe-mediated stimulation of tubulin tyrosinolation in HL-60 cells is correlated with increases in the specific activity of protein kinase C and with the acquisition of respiratory burst activity which occur during induced myeloid maturation of these cells. Furthermore, both the fMet-Leu-Phe-induced stimulation of tubulin tyrosinolation and the PMA or DAG-induced random posttranslational incorporation of tyrosine into multiple proteins in activated neutrophils, were found to be reversibly inhibited (>70%) by the protein kinase inhibitors 1-(5-isoquinolinesulfonyl)piperazine (C-I) and 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), in parallel with inhibition of superoxide (O2−) generation. In related studies, we also found that fMet-Leu-Phestimulated O2−production is comparably inhibited by C-I and H-7, but in a highly temperature-dependent manner. Inhibition was observed only when C-I or H-7 is added to PMNs at physiologic temperature, i.e. 37 °C. Interestingly, inhibition of the PMA-induced generation by C-I or H-7 was not found to be similarly temperature-dependent. Considered together, these findings argue against the suggestion that there is a protein kinase C-independent pathway for activation of the respiratory burst in neutrophils stimulated with N-formyl peptides.

The arabinose-resistant and Ames assay systems of Salmonella typhimurium were used to evaluate the mutagenic potential of azo dyes and their aromatic amine reduction products. Azo dyes, namely direct black 38, direct blue 75, and direct red 2, were mutagenic in the arabinose-resistant and Ames assays with both hamster and rat liver S9 activation. Both assays gave relatively higher mutagenic responses with hamster S9. Reduction products of these dyes, namely benzidine, o-dianisidine, and o-tolidine, were mutagenic in the Ames assay. Benzidine was weakly mutagenic and o-dianisidine and o-tolidine were nonmutagenic in the arabinose-resistant assay. These results indicate that both arabinose-resistant tester SV50 and Ames tester TA98 were sensitive in detecting mutagenicity of azo dyes. The use of the standard plate protocol with Ames tester TA98 is more efficient than the modified azo dye protocol in detecting mutagenicity of aromatic amine reduction products. Additional modifications in either the standard plate or modified azo dye protocols may improve detection of mutagenicity of these compounds in the arabinose-resistant assay system.

Ultrastructural and functional studies of degranulation responses by human neutrophils have suggested that microtubules (MTs) have a role in the intracellular transport of neutrophil granules. We have found that granule-MT complexes can be isolated from disrupted taxol-treated (1.0 microM) neutrophils, visualized by electron microscopy, and quantified in terms of granules per MT length. After incubation of neutrophils with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), granule-MT complex formation was found to be increased two- to threefold. Enhanced binding of granules to MTs was detectable within 30 s of fMLP stimulation and was dependent on the concentration of fMLP. Incubation of cells with dibutyryl cAMP inhibited this fMLP-stimulated granule-MT complex formation in a dose-responsive fashion. These granule-MT interactions could be reproduced in a cell-free system with neutrophil granules isolated by density gradient centrifugation and MTs polymerized from phosphocellulose-purified tubulin. Furthermore, reconstituted granule-MT interactions were found to be modulated by ATPase inhibitors. Sodium orthovanadate increased granule-MT interactions in a concentration-dependent manner, while AMP-PNP, a nonhydrolyzable ATP analogue, and N-ethylmaleimide decreased or eliminated these interactions. In addition, we found that a MT-activated ATPase could be recovered from intact neutrophil granules by salt extraction, and that extracts enriched in this ATPase contained a polypeptide of between 115 and 120 kD which binds ATP and is immunologically related to kinesin. These studies demonstrate that cytoplasmic granules interact with MTs in human neutrophils in a regulated stimulus-responsive manner, and they suggest that such interactions may involve an MT-based, ATPase-dependent, vesicle translocation system as has been demonstrated in other types of cells.

Phleum pratense L., the only species of Phleum that is of economic importance, is found in all temperate regions of the world.

Diploid, tetraploid, and hexaploid plants occurred in foreign plant introductions of P. sp. affin. P. montanum from Turkey and Iran. Within-progeny variations in polyploid levels indicate that divergent races occurred as interspersed but natural populations. One to six B-chromosomes of varying sizes in addition to the normal chromosome complement were present in some plants. Their behaviours appeared essentially similar to those recorded for other species. Certain of the plants examined exhibited cytological features similar to those observed in known hybrids. These included prophase pycnosis, stickiness, split metaphase I plates, unoriented chromosomes at M-1, and low levels of pollen stainability. These features were absent in microsporocytes of other plants. Low open-pollination seed set characterized all plants examined. This may be due to hybridity or possibly to response to ecological conditions of the new environment.

Nielsen, E. L., and J. Nath. (U. Wisconsin, Madison.) Somatic instability in derivatives from Agroelymus turneri resembling Agropyron repens.Amer. Jour. Bot. 48(4): 345–349. Illus. 1961.—Variations in somatic chromosome numbers of 4 to over 80 were observed in root tip cells of Agropyron repens‐like plants derived from Agroelymus turneri.The latter presumably had out‐crossed to Agropyron repens.Split spindles in dividing nuclei and multinucleate cells were observed in the root tip meristem. The nuclear and mitotic behaviors observed can perhaps be attributed to an unbalanced ribonucleoprotein system resulting from the combination of distantly related genomes in the formation of the hybrid.

Nath, J., and E. L. Nielsen. (U. Wisconsin, Madison.) Cytology of plants from self‐ and open‐pollination of Phleum pratense.Amer. Jour. Bot. 48(9): 772–777. Illus. 1961.—Conventional cytological disturbances and several anomalous behaviors such as heavy‐walled coenocytes, uncoiled chromosomes, supernumerary chromosomes, production of cross‐bridges at second division, and the occurrence of globular structures in the microsporocytes were associated with sterility in timothy. These anomalies have not been described previously as contributory to reduced seed set in timothy.

Ten pentaploids were isolated from seedling populations developed from seed of tetraploid plants grown in association with hexaploid timothy. About 80% of the prophase sporocytes exhibited severe or moderate pycnosis. These pycnotic accumulations appear to arrest the meiotic processes and apparently result in sporocyte lethality. Surviving sporocytes at diakinesis possessed nuclei with configurations that varied from 1 VI+14 II+1 I to 9 II+17 I. It is suggested that the conventional sorts of irregularities (univalents, inversions and translocations) cannot account for the low percentages of stainable pollen. Pairing relations indicated that considerable duplication and/or autosyndesis occurred in the genomes combined, and suggest that even though the tetraploid (2n=28) and hexaploid (2n=42) forms of Phleum pratense are closely related they are not identical. A portion of the sterility observed may be due to nuclear-cytoplasmic interaction.