Synthesis and biological activity of adipokinetic hormone analogues modified at the C-terminus

A series of Locusta adipokinetic hormone I (AKH-I), C-terminal threonine residue using a combination of solid- and liquid-phase methodology and evaluated in Locusta migratoria, in a lipid mobilization assay in vivo and an acetate uptake assay in vitro. Modifications at Thr¹⁰ of AKH-I involved replacement of its C-terminal amide by the groups -OH, -OCH₃, -NHCH₃, -N(CH₃)₂, and -NHC₆H₅; the last three groups were also applied to the amide of AKH-I-[Thr(Brl)¹⁰]. The methyl ester, monomethyl, and dimethyl analogues were all of lower activity than the parent in the lipid mobilization assay, but lost less than two orders of potency. In the acetate uptake assay, again the methyl ester analogue showed the greatest retention of biological activity of all modified peptides. A cyclic analogue, cyclo (PLNFTPNWGT), was active in both assay, but only at very high concentrations. Almost all analogues were more active in the acetate uptake assay than in the lipid assay, but unusually, AKH-I-NHCH₃ and AKH-I-N(CH₃)₂, together with cyclo (PLNFTPNWGT), were more active in the lipid mobilization assay. In addition, the acid AKH-I analogue did not suffer as large a loss in potency in the lipid mobilization assay as in the acetate uptake assay, although it was less potent in the former. The relative potencies of these two methyl analogues contrast with those for AKH AKH-I-[Thr(Bzl)¹⁰]-NHCH₃ and AKH-I-[Thr(Bzl)¹⁰]-N(CH₃)₂, which, together with both phenyl analogues, were significantly more active in the acetate uptake assay. We conclude that the acetate uptake assay has a greater preference for a hydrophobic C-terminus, compared with the lipid mobilization assay.     

THE DEPENDENCE OF CELL DIVISION IN CHLORELLA ON TEMPERATURE AND LIGHT INTENSITY

The effects of temperature and light on cell division were studied in synchronized suspensions of the high-temperature strain Chlorella 7–11–05. It was found that the time for incipient cell division, the progress in the process after it started, and the number of cells produced are influenced by temperature and light intensity. Within limits, cell division is generally favored by the increase in temperature. The increase in light intensity first favors cell division then, after the optimal light intensity is attained, a further increase in light intensity inhibits cell division. Observations are discussed in connection with the findings of other investigators. The limitations of cell division by temperature and light intensity are considered to be separate from the effects of these factors on growth.     

Van Slyke's buffer values for cell secretions

Summary Van Slyke's buffer indexes () were determined for secretions of synchronized cells of the green, high-temperature alga, Chlorella 7-11-05. The change in pH and the value of the buffer index were found to depend on initial pH, duration of observation, and population density. The major component of the secreted buffer system was identified as bicarbonate, and the major cation, as potassium.This work was supported by funds from the National Aeronautics and Space Administration.Scientific Article A 1129, Contribution No. 3580 of the University of Maryland Agricultural Experiment Station.     

Telemetric Electrocardiography

By means of a satellite transmitter nearby the subject an electrocardiogram can be obtained by telemetry in a hospital electrocardiographic (ECG) laboratory from an area of approximately 20 miles in diameter within the city of London, Ontario. The technique employed in this preliminary trial produced excellent tracings, quite comparable to tracings obtained using the same leads with direct recording. Ninety-four trials were performed on 63 subjects, 37 of whom were patients with a suspected or known myocardial abnormality. A normal group of 26 healthy subjects, 16 of whom were strenuously exercised on a bicycle ergometer to the point of physical exhaustion, were also studied. Monitoring patients during ordinary daily activities has proved extremely useful in the detection and diagnosis of cardiac arrhythmias, intermittent conduction disturbances, latent coronary artery disease, and asymptomatic heart disease.     

Renal parathyroid hormone-dependent adenylate cyclase activity after repletion of vitamin D-deficient rats with vitamin D-2

Rats fed a diet deficient in both vitamin D and Ca²⁺ exhibited a greater depression of the renal parathyroid hormone (PTH)-dependent adenylate cyclase than was observed in rats fed diets deficient in either vitamin D or calcium. Total serum Ca²⁺ was decreased from a control level of 11.2 mg/dl to 8.5 mg/dl in rats fed the diet deficient in calcium alone, and to 5.4 mg/dl in rats fed the diet deficient in vitamin D. Serum calcium was decreased further to 4.3 mg/dl in rats fed the diet deficient in both vitamin D and Ca²⁺. Serum immunoreactive PTH was significantly elevated over control levels when rats were fed the test diets; however, there were no significant differences between the elevated levels in the three experimental groups. Repletion of rats deficient in vitamin D only with a single oral dose of 3200 I.U. vitamin D-2 resulted in restoration of serum calcium to normal levels, a return of serum PTH to the control state, and an associated increase in PTH-dependent adenylate cyclase activity to the control level by 72 h. Repletion of rats deficient in both vitamin D and Ca²⁺ with the same dose of vitamin D-2 raised serum Ca²⁺ to 7.2 mg/dl by 72 h, but did not cause a reduction in circulating PTH, nor did it result in any significant improvement in the responsiveness of the membrane adenylate cyclase to PTH. These results suggest that elevated PTH is a factor in the down regulation of the PTH-dependent adenylate cyclase, but do not rule out a role for calcium as a regulatory factor.     

Antigenic differentiation of the kinetoplastids Leishmania braziliensis and Trypanosoma cruzi by means of monoclonal antibodies

BALB/c mice were hyperimmunized with non-infectious extracts of either Leishmania braziliensis promastigotes or Trypanosoma cruzi epimastigotes. When spleen cells from these mice were fused with P3X63Ag8 plasmacytoma cells, the resultant hybridomas synthesized monoclonal antibodies which displayed specific reactivity by indirect immunofluorescence with distinct subcellular components of the parasites. These studies revealed that antigens associated with the flagellum and with a nongranular component of the cytoplasm would account for much of the serologic cross-reactivity observed between the two species. Conversely, antigens associated with surface and/or cytoplasmic granules and with an intracellular organelle believed to be the kinetoplast appeared to be species-specific.