Distribution of podolactone-type plant growth inhibitors in the coniferae

Plant growth inhibitors of the podolactone-type have been detected by bioassay in ten further species of Podocarpus. The most active extracts in P. elatus were from root tips, root cortex and very young leaves. Fifty-seven other conifers were examined for this type of activity. It is present in Cephalotaxus harringtonia where it is probably due to the presence of harringtonolide, which, like momilactone B from rice husks, shows podolactone-type inhibition of the growth of etiolated dwarf pea hooks.     

Immune cytolysis of human malignant melanoma by antibody to oncofetal antigen-I (OFA-I)

Summary IgG anti-OFA-I found in melanoma patients was tested for its ability to lyse human tumor cells in antibody-dependent cell-mediated cytotoxicity (ADCC). Sera from 89 stage II melanoma patients which contained non-HLA-related IgG antibody to an OFA-I-positive melanoma cell line (M14) as tested by indirect membrane immunofluorescence (IMI) were originally chosen as possible sources of IgG anti-OFA-I. Of those tested for specific IgG activity to OFA-I by IMI, anti-OFA-I was found only in those patients immunized with OFA-I-positive tumor cells. When the same sera were tested in ADCC, no non-HLA-related activity could be demonstrated. This result was confirmed with purified IgG fractions that could, nevertheless, show anti-OFA-I reactivity in a complement-dependent cytotoxicity assay. The fact that naturally occurring IgG anti-OFA-I antibody was not readily detectable in patients' sera and that induced IgG anti-OFA-I did not participate in ADCC indicates that OFA-I-related tumor cell lysis via ADCC is an unlikely phenomenon in cancer patients.     

Direct assay for creatine kinase by reversed-phase high-performance liquid chromatography

A direct assay for creatine kinase (CK) activity was developed based on the separation and quantitation of adenosine triphosphate (ATP) by high-performance liquid chromatography. The total incubation time is 13 min and the elution time for ATP is 16 min. Using lyophilized CK as the sample, a sensitivity in the range of 8 U/l (units/liter) was obtained. The method presented also has clinical significance in that CK levels in serum can easily be determined with minimal sample preparation. Using serum samples from a healthy patient and a heart attack victim, activities of 26.6 U/l and 609.0 U/l, respectively, were obtained. Because of the direct measurement of ATP, this method eliminates the coupled reactions encountered in the common spectrophotometric and colorimetric methods of analysis resulting in a simpler and inexpensive assay.     

Evidence that calmodulin may be involved in phytohaemagglutinin-stimulated lymphocyte division

Phytohaemagglutinin-stimulated and non-stimulated incorporation of [³H]thymidine into human peripheral blood lymphocytes is inhibited by the calcium antagonist PY 108–068 and by the calmodulin antagonists trifluo-perazine andN-(6-aminohexyl)-5-chloro-l-naphthalene sulphonamide (W7). It is argued that calmodulin may be involved in both non-stimulated [³H]thymidine uptake in lymphocytes and also in the lymphocyte response to phytohaemagglutinin.     

Effect of adenosine deaminase inhibitors on Fcμ receptor expression in human T-cell cultures

Incubation of human peripheral blood T-lymphocyte cultures with 100 μM EHNA or 1 μM pentostatin results in the delayed appearance of Fcμ, receptor-bearing cells. The percentage of Tμ-positive cells approaches control levels by 24–30 hr despite the lack of detectable adenosine deaminase activity. Tγ to Tμ transition was more effectively inhibited than To to Tμ in studies with the individual subpopulations. The inhibitory effect of pentostatin or EHNA on the appearance of Fcμ receptor-positive cells was reversible with exogenous undine or 2′-deoxycytidine. These results indicate that pyrimidine deprivation affects the appearance of Fcμ receptors in T-cell cultures, and that despite the continued inhibition of ADA the effect on Fcμ is reversible by 24 hr without the addition of exogenous pyrimidines.     

Simultaneous analysis of phenylglycols and phenylethanols in human urine by gas chromatography—mass spectrometry

A method is described for the determination of the neutral metabolites formed from catecholamines and various other structurally related phenylethylamines by using gas chromatography—chemical ionization—mass spectrometry. These metabolites (phenylglycols and phenylethanols) were extracted from urine specimens and converted to pentafluoropropionyl derivatives which were separated on either 3% OV-1, 3% SP-2250, or 3% QF-1 packed columns. Our results demonstrate the presence in human urine of p-hydroxyphenylglycol, a metabolite of octopamine. One patient excreted 13 and 91 μg/day of free and total (free + conjugated) p-hydroxyphenylglycol, respectively. Treatment with a monoamine oxidase inhibitor reduced the excretion of total p-hydroxyphenylglycol to 30% of baseline level.     

Molecular characterization of Ascaris suum DNA and of chromatin diminution

A technique for the extraction of pure somatic (post-diminution) and germ line (pre-diminution) DNA from the parasitic nematode Ascaris is described. Uncontaminated post- and pre-diminution DNAs were sheared and reassociated to different C₀t values. Computer analysis of the complete reassociation kinetics determined that 33% of the germ line genome is eliminated during the process of chromatin diminution. The eliminated DNA is comprised of repetitive and unique sequences in an approx. 1:1 ratio.     

Viral transformation of rat myoblasts: effects on fusion and surface properties.

Lines of rat myoblasts infected by avian sarcoma viruses have been isolated, cloned, and used to study the effects of viral transformation on myogenic differentiation and the surface changes associated with differentiation. The lines transformed by sarcoma viruses failed to fuse into myotubes and did not show the increase in myosin synthesis normally associated with fusion. The parental nontransformed line showed, subsequent to fusion, a surface alteration detectable by external labeling methods. This alteration, an increase in the level of an external protein of MW > 200 × 10³, is similar to that observed in fibroblasts arrested in the G₁ phase of the cell cycle. This protein was absent or greatly reduced on the surfaces of the myoblast lines that had been transformed by sarcoma viruses. Therefore, viral transformation causes loss of several properties normally associated with arrest of myoblasts in G₁.     

Effects of sodium periodate modification of lymphocytes on the sensitization and lytic phases of T cell-mediated lympholysis.

Sensitization of mouse splenic lymphocytes in vitro with sodium borohydride, suggesting that the biologic effects of sodium periodate are-treated autologous spleen cells stimulated a one-way mixed lymphocyte reaction and led to the generation of thymus-derived cytotoxic effector cells. These effectors were capable of lysing in 4 hr periodate-treated syngeneic and, to a lesser extent, periodate-treated allogeneic target cells. These results suggest that sensitization by periodate-treated autologous cells could result either from a specific reaction to modified self components or from a nonspecific mitogenic stimulation. Effector cells generated by allogeneic sensitization were detected on periodate-modified targets, irrespective of the H-2 antigens expressed by the targets. The effects of periodate modification on both stimulator and target cells were reversible by sodium periodate are dependent on the formation of a free aldehyde group on cell surface glycoproteins. Pretreatment of stimulator cells with neuroaminidase prevented the effect of periodate treatment, suggesting that the sensitization involves oxidized sialic acid residues. During the 4-hour 51Cr-release assay periodate-treated targets could be used to detect cytotoxic effector cells of any specificity. Fresh spleen cells and lymphocytes cultured for 5 days without antigen or in the presence of lipopolysaccharide did not lyse periodate-treated targets. An increasing level of cytotoxicity was detected on periodate-treated targets when the effector cells were generated, respectively, by stimulation with concanavalin A, by sensitization with periodate-modified autologous cells. Although the lysis of periodate-treated targets is itself nonspecific, effector cell specificity could be determined by selective blocking of the lytic phase with cells syngeneic to the stimulators. These results indicate that a nonspecific interaction can occur between lymphocytes and periodate-treated target cells, but that this interaction leads to lysis only when the lymphocytes were activated to become cytotoxic effectors.     

Phenolic O-methyltransferase from Lentinus lepideus (basidiomycete)

The fungus, Lentinus lepideus, produces crystalline methyl p-methoxycinnamate in stationary cultures. O-methylation and methyl ester formation of hydroxycinnamic acids were examined with enzyme preparations of the fungus. Using S-adenosylmethionine-¹⁴CH3, it was found that only the methyl esters of the hydroxycinnamic acids are substrates for O-methylation and not the free acids. Benzoic acids and their methyl esters are not substrates. The activity of the enzyme is p-specific and its specific activity decreases with increasing number of hydroxyl groups in the substrate. The same enzyme preparations catalyze the formation of the methyl ester of cinnamic acid from the free acid.