Phenylalanine ammonia-lyase immobilized in microcapsules for the depletion of phenylalanine in plasma in phenylketonuric rat model

Microencapsulation of the enzyme phenylalanine ammonia-lyase was developed for in vivo depletion of systemic phenylalanine in phenylketonuric rats. Compared to normal rats, systemic phenylalanine blood levels in phenylketonuric rats was increased by 15-20-fold. Daily oral administration of 1 unit of phenylalanine ammonia-lyase-loaded artificial cells to phenylketonuric rats lowered the systemic phenylalanine level to 58% +/- 18% (mean + S.D.) in 7 days (P less than 0.010), while 5 units lowered the systemic phenylalanine level to 25% +/- 8%. 5 units of the immobilized enzyme lowered the systemic phenylalanine level to normal levels within 6 days. Phenylketonuric treated rats showed no signs of abnormal behavior and weight loss compared to phenylketonuric non-treated rats. The immobilized enzyme within artificial cells is therefore protected against low gastrointestinal pH and proteolytic enzymes.     

Syngeneic monoclonal internal image anti-idiotopes as prophylactic vaccines

A syngeneic monoclonal anti-idiotope that behaves as an internal image of the mammalian reovirus type 3 cellular attachment protein (viral hemagglutinin) was used in the syngeneic host for the induction of a prophylactic anti-viral antibody response. These studies were performed without the aid of co-stimulation by viral antigens. The high stringency of this system enables us to define the maximum constraints on the use of anti-idiotopes as anti-viral vaccines. We have used the murine BALB/c monoclonal IgM anti-idiotope 87.92.6 to study the idiotope and antigen specificity, kinetics, dose dependence, adjuvant, carrier, and valency requirements of anti-idiotope-induced anti-viral antibody responses. These studies show that the production of high titer neutralizing antibody requires a lengthy (60 day) immunization protocol, which includes the use of adjuvant and multivalent anti-idiotope, and is dependent on anti-idiotope concentrations of greater than 50 micrograms. When administered in this manner anti-idiotope can stimulate serotype-specific antibody responses across species barriers at levels comparable with those obtained after inoculation with virus. The practical efficacy of these reagents and procedures is documented by the ability of maternal immunization with anti-idiotope to confer complete protection in neonates from a potentially lethal reovirus type 3 viral infection.     

Early ontogeny of Adinia xenica (Pisces,Cyprinodontiformes): 3. Comparison and evolutionary significance of some patterns in epigenesis of egg-scattering,hiding and bearing cyprinodontiforms

Synopsis Developmental patterns as seen in cyprinodontiforms fishes with different reproductive styles are compared, and discussed in relation to ecology and evolutionary significance. The discussion centres around Adinia xenica (its detailed ontogeny presented in two previous sequels to this paper), and, from the existing literature, Fundulus heteroclitus (closely related), Austrofundulus myersi (an annual) and Platypoecilus maculatus (a livebearer). The embryonic resting interval is present in various forms in the first three species, and differences in it and the overall patterns of development are shown to be consistent with ecological conditions. Termination of the resting interval leads immediately to hatching, a process in A. xenica, as in F. heteroclitus, apparently initiated by the appropriate summation of internal and external factors. These factors include any or all of: metabolic changes and increased oxygen requirements, response to light, reduced environmental oxygen, agitation, and increased hydrostatic pressure. They all can cause increased movement by the embryo which is credited with rupturing hatching gland cells and releasing the enzyme(s). Annual fishes experience 3 pronounced resting intervals, termed diapauses. These are discussed in the context of apparent steps and thresholds, and evolutionary ecology. A possible evolutionary sequence, from a simple fractional spawning pattern to diapause, is presented. Morphological differences in primary embryonic respiratory surfaces, as seen in the four species, are related to environmental conditions. The above illustrate ways in which the same basic structures and events are modified to cope with different habitats.     

Purification and characterization of yeast topoisomerase I

Yeast topoisomerase I (Mr = 76,000) has been purified to 80% homogeneity using a combination of ion exchange, gel filtration, and DNA-cellulose chromatography. The enzyme was characterized with respect to its ability to relax supercoiled DNA and to catenate nicked circular DNA. Yeast topoisomerase I will remove both positive and negative turns in DNA supercoils in the absence of ATP and magnesium ion. The products of the catenating activity of the enzyme were examined on agarose gels and in the electron microscope. These analyses indicate that yeast topoisomerase I will generate large catenated DNA networks which appear to rearrange to multimeric linear structures upon long incubation time.     

The corrected nucleotide sequence of yeast leucine transfer ribonucleic acid

The nucleotide sequence of “Renaturable” leucine transfer RNA from Baker's yeast has been re-investigated. The results showed that (i) this tRNA has a sequence of DCD at positions 19–21, (ii) it has an anticodon m⁵CAA and (iii) it has a pseudouridine at position 40.