Fast atom bombardment combined with tandem mass spectrometry for the study of dinucleotides

A study of mono- and dinucleotides by utilizing negative ion fast atom bombardment (FAB), metastable decomposition of (M-H)- species, and collisionally activated decomposition (CAD) of (M-H)- species is reported. Data were obtained for several complete series containing the standard nucleosides (guanosine, adenosine, cytidine, thymidine, and uridine): the 3'- and 5'-monophosphate mononucleotide series for both ribo- and 2'-deoxyribomononucleotides, all possible combinations for the 3'(-)----5'-ribodinucleotides, and all possible combinations of the 3'(-)----5',2'-deoxyribodinucleotides. The metastable and CAD spectra provide more information than the FAB mass spectra. The (M-H)- ions of all dinucleotides decompose either as metastable ions or upon collisional activation to eliminate BH (B = base) preferentially from the 3'- rather than the 5'-terminus. Isomeric dinucleotides can be distinguished on the basis of this fragmentation. To establish the identity of the base at the 5'-terminus, collisional activation is preferred. By comparing relative abundances of BH elimination observed, the inherent basicities of the nucleoside base anions can be inferred to be C- greater than A-, T-, greater than G-.     

Systematics within Gyps vultures: a clade at risk

Background  

Populations of the Oriental White-backed Vulture (Gyps bengalensis) have declined by over 95% within the past decade. This decline is largely due to incidental consumption of the non-steroidal anti-inflammatory veterinary pharmaceutical diclofenac, commonly used to treat domestic livestock. The conservation status of other Gyps vultures in southern Asia is also of immediate concern, given the lack of knowledge regarding status of their populations and the continuing existence of taxonomic uncertainties. In this study, we assess phylogenetic relationships for all recognized species and the majority of subspecies within the genus Gyps. The continuing veterinary use of diclofenac is an unknown but potential risk to related species with similar feeding habits to Gyps bengalensis. Therefore, an accurate assessment of the phylogenetic relationships among Gyps vultures should aid in their conservation by clarifying taxonomic uncertainties, and enabling inference of their respective relatedness to susceptible G. bengalensis.     

Mapping of antibody specificities to V H gene families

V _H gene segments represent the products of the repeated duplication and subsequent diversification of a primordial V gene element. It is widely assumed that natural selection, operating via pathogens, has played the dominant role in this process. Here, we screen some 3.7 × 10⁴ C ⁺ colonies of mitogen-activated B cells for the production of antibodies specific for phosphorylcholine or hen egg lysozyme and expression of the V _H X-24, S107, Q52, or J558 gene families. These gene families were expressed at frequencies proportional to their genomic complexity among both unselected and antigen-specific C ⁺ colonies. Thus, the capacity to encode equivalent antibody-combining sites is dispersed uniformly among V _Hfamilies. This result suggests that individual V _H genes have not evolved to address specific antigens.     

Regulation of idiotope expression. III. H-2 influences the magnitude and the idiotypy of a T-independent antibody response in mice of certain genetic backgrounds

Antibody response to the phosphocholine (PC) epitope on Streptococcus pneumoniae R36a (Pn), a T-independent Ag type 2, was studied in H-2 congenic mouse strains. The PC-specific antibody plaque-forming cells (PFC) were enumerated in the spleen at various intervals after the primary Pn injection, and the proportion of PFC that produced antibody expressing the AB1-2 idiotope (Id) was determined by using the corresponding monoclonal anti-Id. AB1-2 is a cross-reactive Id, detectable on germline-encoded PC antibody of the T15 family, and on most, but not all, somatic variants of that antibody. The specific PFC responses in BALB/c (H-2d) and BALB.B (H-2b) strains were of comparable magnitude and most, if not all, PFC were ABl-1 Id-positive (AB1-2+). This was not the case in the responses of the B10D2 (H-2d) vs C57BL/10 (H-2b) strains and the D1.C (H-2d) vs D1.LP (H-2b) strains (on DBA/1 background). In each of these pairs, the H-2d mice were high responders, and the response was dominated by AB1-2 Id (greater than or equal to 80% AB1-2+ PFC at the peak, on day 5). The H-2b mice were low responders, and only a minor proportion of PFC (less than or equal to 30%) were AB1-2+; an increase of AB1-2+ was seen later in the response (d.10). The results of PFC assays were confirmed by measuring the PC-binding antibody and AB1-2 Id in the sera of D1.C and D1.LP mice immunized repeatedly with Pn. Moreover, D1.LP mice that had very low levels of AB1-2 Id had higher serum levels of antibody expressing two other T15 Id, B36-82, and B24-44. The B36-82 and B24-44 Id have been previously found on somatic variants of PC antibody expressed independently of the Ab1-2 Id. The concentrations of these two Id in D1.LP mice after repeated immunization approached those in D1.C. These results indicate that 1) the H-2 allelism may have a significant effect on TID antibody response in mice of a certain genetic background, but not in the BALB/c; and 2) the idiotypic repertoire of the response may be influenced by H-2 at the level of clonal variants of PC-reactive cells.     

Arbovirological survey in Silica plateau area, Roznava District, Czechoslovakia

The serosurveys conducted in the Silica plateau area of the Slovak karst region revealed the presence of specific neutralizing antibody against tick-borne encephalitis (TBE) virus in 18% of local inhabitants (33 examined, mostly goats and sheep farmers), 54% of goats (26 examined), 18% of sheep (120 examined) and 13% of cattle (60 examined), against Lipovník (LIP) virus in 30% of inhabitants, 88% of goats, 55% of sheep and 45% of cattle, and against Bhanja (BHA) virus in 27% of inhabitants, 46% of goats, 29% of sheep and 23% of cattle. The results of hemagglutination-inhibition tests with TBE and BHA antigens were analogous. A detailed analysis of these serologic data points to a recent enhancement of the circulation of LIP and BHA viruses and to a very low TBE virus activity in this natural focus of arboviral infections. The immunological surveys of the 32 former "Roznava disease" patients, conducted 25 years after an extensive epidemic of a TBE virus infection that originated in Roznava in 1951, revealed the presence of neutralizing (and also hemagglutination-inhibiting) antibodies against TBE virus in as many as 78% of cases. Antibodies against LIP and BHA viruses were also detectable in the sera of 16% and 9%, respectively, of these individuals. Populations of the ectoparasites examined for the presence of arbovirus comprised 231 Ixodes ricinus, 806 Dermacentor marginatus and 204 Haemaphysalis punctata ticks and 117 specimens of the louse-flies Melophagus ovinus. Two strains of arbivirus that were antigenically related to Lipovník and Tribec viruses belonging to a group of Kemerovo viruses were isolated from male and female I. ricinus ticks collected from cattle.     

"Nonspecific" immunoenhancing T cells in tumor-bearing mice include anti-idiotypic subsets

Splenocytes from DBA/2 mice inoculated 3 wk earlier with syngeneic P815 mastocytoma tumor cells produce increased numbers of antibody plaque-forming cells (PFC) when stimulated with either sheep red blood cells (SRBC) or phosphorylcholine (PC) on Streptococcus pneumoniae R36a in vitro. The nature of this nonspecific hyperreactivity was investigated in mixed cultures of purified splenic T and B cells. The addition of T cells from P815 tumor-bearing mice (TP815) into the cultures of normal B cells produced a significant enhancement of the PFC response to both SRBC and PC, when compared with the effect of normal T cells added to control cultures. The idiotypic profile of the enhanced anti-PC response was studied by a PFC-inhibition assay with monoclonal antibodies against two distinct idiotopic determinants (Id) of the T15 family. Normal B cells produced greater than 90% of T15 Id-positive (Id+) PFC. Addition of normal T cells diminished the proportion of T15 Id+ PFC to approximately 60%, whereas the rest of PFC were Id-. Addition of the immunoenhancing TP815 cells into the normal B cells cultures elevated the number of both T15 Id+ and Id- PFC responses, proportionally. However, when TP815 cells were first incubated on T15 protein-coated dishes and the non-adherent fraction was added to B cell cultures, the anti-PC PFC response remained enhanced but consisted of predominently T15 Id- PFC. These observations suggest that the early stage of P815 tumor growth activates various populations of specific helper/amplifier T cells including subsets with anti-idiotypic activity and that the generalized increase of antibody response to various antigens in tumor-bearing mice may be regarded as a polyclonal activation of specific T cells.     

Leonhardi Euleri's “Principia pro motu sanguinis per arterias determinando”

The purpose of this publication as a pedagogical contribution is to illustrate the uniqueness of the method Euler used to determine the unsteady flow of blood in the circulatory system. In his presentation, he accounts for the tapering of the vessels and their elastic behavior. The limitations of this study are based upon the extent of the development of the mathematics of his era. This work may be the first mathematical treatment of circulatory physiology and haemodynamics. Euler perhaps could very well be called the Father of Haemodynamics.The Determination of the Principals of the Motion of Blood through the Arteries.Supported in part by a grant from the American Heart Association (No. 62F4EG).This work was done during the tenure of an Established Investigatorship of the American Heart Association.     

The flow of a viscous liquid in a converging tube

The problem of the viscous flow of an incompressible Newtonian liquid in a converging tapered tube has been solved in spherical polar coordinates. The method of the solution involves the Stokes' stream function and a technique introduced by Stokes in the study of a sphere oscillating in a fluid. The theory for the flow in a rigid tube includes: (1) the pulsatile flow with both radial and angular velocity components; (2) the steady state flow with both radial and angular velocity components and (3) the very slow steady state flow with only a radial velocity component present. For a tapered elastic tube, the velocity of the propagated pulse wave is determined. The solution given is in terms of the elastic constants of the system and the coordinates for this type of geometry. The pulse velocity is then related to the velocity in an elastic cylindrical tube with the necessary correction terms to account for the tapered tube. Supported in part by the American Heart Association (No. 62F4EG). This work was done during the tenure of an Established Investigatorship of the American Heart Association.