Characterization of a transferrin-diphtheria toxin conjugate

We report here the synthesis and properties of a hybrid toxin prepared by covalently coupling diphtheria toxin to transferrin. The purified material contained two major hybrid protein species and was highly cytotoxic to mouse LMTK- cells in culture, reducing protein synthesis by 50% in 24 h at a concentration of 1 ng/ml. Cytotoxic activity was completely abolished in the presence of exogenous transferrin or anti-transferrin or anti-diphtheria toxin, thus demonstrating that the hybrid toxin was intoxicating cells via their transferrin receptors and that both the diphtheria toxin and transferrin components of the conjugate were necessary for activity. NH4Cl, a drug that elevates the pH within acidic intracellular vesicles, also blocked cytotoxic activity, suggesting that a low intravesicular pH was required for activity. The inhibitory effect of NH4Cl could be abolished by exposing toxin-treated cells to acidic culture medium, further implicating an acid-dependent step in the mechanism of the hybrid toxin action. Studies on the kinetics of intoxication also implied that endocytosis and exposure to a low pH within vesicles were necessary for cytotoxicity. Altogether, the results suggest that the transferrin-diphtheria toxin conjugate binds to transferrin receptors and is internalized into acidic endocytic vesicles. The enzymatic moiety of diphtheria toxin then apparently enters the cytosol in response to the low pH and subsequently arrests protein synthesis.     

Immunocytochemical detection of triphosphoinositide in vestibular hair cells

Triphosphoinositide (TPI), an aminoglycoside receptor and a possible regulator of cationic permeation through its ability to bind with Ca++, was localized by the protein-A gold technique in vestibular sensory epithelia using an antibody highly specific to TPI. TPI was detected on the stereocilia, kinocilia, and cuticular plate of hair cells, and in the reticular membrane of supporting cells. The cilia of hair cells are damaged by aminoglycosides at a relatively early stage of toxicity. Ca++-regulated bioactivity in this area is probably involved.     

Modulating effect of cerulein on benzodiazepine receptors

Subcutaneous administration of caerulein (100-500 micrograms/kg) significantly reduced the development of picrotoxin (8 mg/kg) seizures in male mice. The same doses of caerulein inhibited 3H-flunitrazepam binding in in vivo experiments. Proglumide, an antagonist of cholecystokinin receptors, in low dose (5 mg/kg) potentiated the effects of caerulein (100 micrograms/kg), whereas the administration of proglumide in high dose (25 mg/kg) reduced the action of caerulein on 3H-flunitrazepam binding and picrotoxin seizures. Caerulein (5-1000 nM) decreased 3H-flunitrazepam binding in in vitro experiments only after supplementation of the binding medium with 120 mM NaCl and 5mM KCl. The results suggest the possible interaction of caerulein with chloride ionophor. It seems probable that the direct interaction of caerulein with chloride ionophor in involved in the inhibitory effect of caerulein on picrotoxin seizures and 3H-flunitrazepam binding.     

Segregation Distortion of Marker Nuclear Genes in Alloplasmic and Isoplasmic Lines of Barley

Inheritance of barley nuclear genes responsible for various morphological marker traits was studied in hybrid populations F₂ and F_a. Nine marker genes showed deviation from Mendelian monogenic inheritance depending on the cross direction and maternal cytoplasm. Segregation biases to both recessive mutant and dominant normal phenotypes were observed. Mechanisms of the segregation bias related to cytoplasm substitution in iso- and alloplasmic lines are discussed.     

The phospholipid-bound fatty acids of fowl and turkey spermatozoa

The composition of the phospholipid-bound fatty acids in the spermatozoa of the turkey, Meleagris gallopavo, and fowl, Gallus domesticus, was studied. The types of fatty acids were similar in the two birds. The ratio of polyunsaturated : saturated fatty acids was generally low but slightly higher in the turkey than in the fowl. The significance of the findings in relation to the origin of the semen collected in these gallinaceous birds and the greater difficulty of freezing turkey spermatozoa was discussed.     

Distribution of calcareous nannoplankton in surface sediments from intertidal and shallow marine regimes of a marginal sea: Jason Bay,South China Sea

The surface sediments collected from the intertidal and shallow marine (0–20 m) regimes of Jason Bay, South China Sea contain calcareous nannoplankton assemblages in whichGephyrocapsa oceanica comprises 99% of the assemblage. The calcareous nannoplankton diversity is very low and the abundance of species tends to increase with both depth and distance offshore and becomes abundant in samples from water depths of 18 m and 20 m.Emiliania huxleyi is absent from all studied samples. The sediments from the intertidal regime contained rare calcareous nannoplankton.     

Ultrastructure of a colonial radiolarian Collozoum inerme and a cytochemical determination of the role of its zooxanthellae

Collozoum inerme (Müller) is a colonial Radiolarian containing numerous cells bound in a common gelatinous matrix. The cells do not possess a skeleton as observed in many unicellular Radiolaria, but the cytoplasmic organization is similar. The cells are multinucleate and a complex system of cellular processes containing mitochondria, Golgi, and numerous vacuoles radiate out from the nuclear region. The endoplasm is connected to the ectoplasm across a double membrane boundary by thin cytoplasmic strands called fusules whose structure resemble those in unicellular Radiolaria. The ectoplasm contains a lacy network of vacuoles containing an osmiophilic substance. Rhizopodia emerge from the ectoplasmic sheath. Some are thin and densely granular. Larger diameter rhizopodia, containing less dense cytoplasm, sequester the zooxanthellae which present a typical dinoflagellate fine structure. Some of the zooxanthellae are apparently cultivated since they are sometimes observed dividing and persist in large numbers when colonies are cultivated under illumination for several weeks in the laboratory. However, colonies maintained in the dark have a decline in number of zooxanthellae and light microscopic examination shows they are being drawn into the ectoplasm of the radiolarian cells. Electron microscopic examination of zooxanthellae drawn into the ectoplasm sheath indicates they are digested. C. inerme is a remarkable example of a simple cellular aggregate that has exploited its colonial habit to culture algae and use them as food thus possibly enhancing the viability of the colony.     

Intercentral correlations in the cerebral cortex according to the EEG coherence index after restoration of consciousness and speech following prolonged coma

Power spectra and coherence function of EEG of various cortical areas of both hemispheres were analyzed in 9 patients with extremely protracted loss of consciousness. Five patients were in the state of posttraumatic apallic syndrome lasting for more than 4 years in one patient, and 4-9 months with successive lethal outcome in 4 patients. One patient for more than 2 years was in a state of areactivity to external signals. In 3 patients the process of recovery of consciousness and speech began in 1-2 months. At the apallic syndrome, only low-frequency EEG components were present in spectrograms, and the values of coherence function were sharply decreased. With recovering consciousness and speech, a gradual appearance of alpha-activity was observed as well as an increase of coherence values at the frequency of the alpha-rhythm. The recovery of intercentral EEG relations in the motor-verbal cortical area was shown to play a special role in further normalization of connections in the cerebral cortex.