The recent interest in using Buckminsterfullerene (fullerene) derivatives in biological systems raises the possibility of their assay by immunological procedures. This, in turn, leads to the question of the ability of these unprecedented polygonal structures, made up solely of carbon atoms, to induce the production of specific antibodies. Immunization of mice with a C 60 fullerene derivative conjugated to bovine thyroglobulin yielded a population of fullerene-specific antibodies of the IgG isotype, showing that the immune repertoire was diverse enough to recognize and process fullerenes as protein conjugates. The population of antibodies included a subpopulation that crossreacted with a C 70 fullerene as determined by immune precipitation and ELISA procedures. These assays were made possible by the synthesis of water-soluble fullerene derivatives, including bovine and rabbit serum albumin conjugates and derivatives of trilysine and pentalysine, all of which were characterized as to the extent of substitution and their UV-Vis spectra. Possible interactions of fullerenes with the combining sites of IgG are discussed based on the physical chemistry of fullerenes and previously described protein-fullerene interactions. They remain to be confirmed by the isolation of mAbs for x-ray crystallographic studies.

product is not thiocyanate but either the product of its further reaction with BrCN according to eq 1 or a further decomposition product of S(CN)2.BrCN + -SCN -Br + S(CN)2(1)The reactions of ADPaS and ATP0S are not useful for synthesizing adenosine 5'-[180]diphosphates or -triphosphates with chiral Pa or Pd because of the oxygen rearrangements in the polyphosphate systems described herein. These can be prevented by protecting the terminal phosphoryl groups with removable alkyl substituents, as demonstrated by the reactions of the RF and SP epimers of /3-(cyanoethyl)-ADPaS with BrCN in H2180 to produce high yields of the SP and RF epimers of 6-(cyanoethyl)-[a-180]ADP.22 The displacement of thiocyanate by H2180 proceeded with inversion of configuration at Pa, and the /3-cyanoethyl groups were easily removed by treatment with base to produce the SP and Rf epimers of [a-180]ADP.22Eckstein and Lowe and their collaborators have been able to desulfurize nucleoside phosphorothioates with electrophilic brominating agents in acidic solutions with inversion of configuration and without rearrangements in polyphosphates.19c•23 At neutral pHs these reactions also involved rearrangements, suggesting that our observations do not represent an isolated phenomenon observable only in the special case of reactions of BrCN with ADPaS or ATP/3S. This paper and our earlier communication4 provide the first evidence for involvement of cyc/o-diphosphates in chemical reactions. Dimeric phenylphosphonic anhydride is the only fourmembered ring organophosphorus compound reported in the literature that has two P-O-P bonds.24•25 Recently a cyclic phosphoric acid anhydride was postulated as a possible inter- (22)

Ribonuclease P (RNase P) is an essential endonuclease responsible for the 5-end maturation of precursor tRNAs. Bacterial RNase P also processes precursor 4.5S RNA, tmRNA, 30S preribosomal RNA, and several reported protein-coding RNAs. Eukaryotic nuclear RNase P is far more complex than in the bacterial form, employing multiple essential protein subunits in addition to the catalytic RNA subunit. RNomic studies have shown that RNase P binds other RNAs in addition to tRNAs, but no non-tRNA substrates have previously been identified. Additional substrates were identified by using a multipronged approach in the budding yeast Saccharomyces cerevisiae. First, RNase P-dependant changes in RNA abundance were examined on whole-genome microarrays by using strains containing temperature sensitive (TS) mutations in two of the essential RNase P subunits, Pop1p and Rpr1r. Second, RNase P was rapidly affinity-purified, and copurified RNAs were identified by using a genome-wide microarray. Third, to identify RNAs that do not change abundance when RNase P is depleted but accumulate as larger precursors, >80 potential small RNA substrates were probed directly by Northern blot analysis with RNA from the RNase P TS mutants. Numerous potential substrates were identified, of which we characterized the box C/D intron-encoded small nucleolar RNAs (snoRNAs), because these both copurify with RNase P and accumulate larger forms in the RNase P temperature-sensitive mutants. It was previously known that two pathways existed for excising these snoRNAs, one using the pre-mRNA splicing path and the other that was independent of splicing. RNase P appears to participate in the splicing-independent path for the box C/D intron-encoded snoRNAs.RNA ͉ biogenesis R ibonuclease P (RNase P) is a conserved endoribonuclease responsible for removing the 5Ј leader sequence from precursor transfer RNAs (pre-tRNAs) found in bacteria, archaea, eukarya (1, 2). In all cases, with the possible exception of some organelles, RNase P is composed of both RNA and protein subunits. Bacterial RNase P is the simplest form of the holoenzyme, with one large RNA subunit and a single small protein subunit (1). Although the RNA subunit of bacterial RNase P is sufficient for catalysis in vitro at high salt concentrations (3), both the RNA and protein subunits are required in vivo. The protein subunit appears to stabilize the catalytically active conformation of RNase P RNA and assist with substrate binding (4-7). In addition to pre-tRNAs, bacterial RNase P is known to process several substrates that are proposed to contain tRNA-like structures: 4.5S RNA, tmRNA, viral RNAs, mRNAs, riboswitches, ColE1 replication origin control RNAs, and C4 antisense RNA from phages P1 and P7 (8-16). The presence of the protein subunit in the RNase P holoenzyme increases the substrate versatility of the enzyme over the RNA enzyme alone (17).The nature of the eukaryotic nuclear RNase P is much more complex. First, there are two very similar enzymes that are related to bacterial RNase P, termed RNa...

Enhanced recovery after surgery (ERAS) protocols have been shown to be effective at reducing perioperative morbidity and costs while improving outcomes. To date, spine surgery protocols have been limited in scope, focusing only on specific types of procedures or specific parts of the surgical episode. The authors describe the creation and implementation of one of the first comprehensive ERAS protocols for spine surgery. The protocol is unique in that it has a comprehensive perioperative paradigm encompassing the entire surgical period that is tailored based on the complexity of each individual spine patient.