The binding of adenosine to polyuridylic acid in which uracil residues are chemically altered.

The binding of adenosine-¹⁴C to polyuridylic acid (poly(U)) and several modified poly(U)s has been studied by equilibrium dialysis. The poly(U) was modified by addition of appropriate reagents across the 5,6-double bond of the uracil ring to form the photohydrate, photodimer, dihydrouracil, the HOBr addition product and the HSO₃^? addition product. Modification of the uracil rings decreases the amount of adenosine which can be bound to the poly(U); the decrease in binding is a function of the fraction of uracil rings which have been changed. Using the expression S = S₀(1 ? αr)² to relate the fraction of uracil rings modified (r) to the number of binding “sites” remaining (S), it is found that α is about 1 for all the modifications except photodimer where it is about 2. These observations are taken to mean that the loss of binding capacity of the poly(U) resulting from modifications of the uracil ring is caused by loss of planarity of the uracil rings caused by the modifications, and consequent loss of double helix structure, but that for all modifications except photodimer there is no disruption of the poly(U) double helix on either side of the leison. There does appear to be local melting on either side of the photodimer lesion. The sigmoidal binding isotherms (A_b versus C_a) of modified and unmodified poly(U) can be approximated closely by the following equation: ((1)) (1) where A_b = bound A, C_a = free A, n = minimum number of adjacent A′s in complex, S = concentration of sites on poly(U), and K₁ = (K_m)^1/m for all m ≥ n. The stacking energy of adenosine (w) can be calculated accurately using the following equation, where dθ/d ln C_a is obtained from Eq. (1). ((2)) (2) For unmodified poly(U), w is ?2.0 kcal/mole and ΔG° (?;RT ln K₁) is ?3.2 kcal/mole. The difference (?1.2 kcal/mole) is attributed to hydrogen bonding. Heavily photohydrated poly(U) does not bind guanosine or guanosine-5′-phosphate.     

Quartz crystal microbalance investigation of the interaction of bacterial toxins with ganglioside containing solid supported membranes

The binding of cholera toxin, tetanus toxin and pertussis toxin to ganglioside containing solid supported membranes has been investigated by quartz crystal microbalance measurements. The bilayers were prepared by fusion of phospholipid-vesicles on a hydrophobic monolayer of octanethiol chemisorbed on one gold electrode placed on the 5 MHz AT-cut quartz crystal. The ability of the gangliosides G_M1, G_M3, G_D1a, G_D1b, G_T1b and asialo-G_M1 to act as suitable receptors for the different toxins was tested by measuring the changes of quartz resonance frequencies. To obtain the binding constants of each ligand-receptor-couple Langmuir-isotherms were successfully fitted to the experimental adsorption isotherms. Cholera toxin shows a high affinity for G_M1 (K_a = 1.8 ⋅ 10⁸M^–1), a lower one for asialo-G_M1 (K_a = 1.0 ⋅ 10⁷ M^–1) and no affinity for G_M3. The C-fragment of tetanus toxin binds to ganglioside G_D1a, G_D1b and G_T1b containing membranes with similar affinity (K_a∼10⁶ M^–1), while no binding was observed with G_M3. Pertussis toxin binds to membranes containing the ganglioside G_D1a with a binding constant of K_a = 1.6 ⋅ 10⁶ M^–1, but only if large amounts (40 mol%) of G_D1a are present. The maximum frequency shift caused by the protein adsorption depends strongly on the molecular structure of the receptor. This is clearly demonstrated by an observed maximum frequency decrease of 99 Hz for the adsorption of the C-fragment of tetanus toxin to G_D1b. In contrast to this large frequency decrease, which was unexpectedly high with respect to Sauerbrey's equation, implying pure mass loading, a maximum shift of only 28 Hz was detected after adsorption of the C-fragment of tetanus toxin to G_D1a. Received: 14 January 1997 / Accepted: 15 April 1997     

Studying Genetic Code by a Matrix Approach

Following Petoukhov and his collaborators, we use two length n zero-one sequences, α and β, to represent a length n genetic sequence ((a) || (b)){\alpha\choose\beta} so that the columns of ((a) || (b)){\alpha\choose\beta} have the following correspondence with the nucleotides: C ~ (0 || 0)C\sim{0\choose0} , U ~ (1 || 0)U\sim{1\choose0} , G ~ (1 || 1)G\sim{1\choose1} , A ~ (0 || 1)A\sim{0\choose1} . Using the Gray code ordering to arrange α and β, we build a 2 ⁿ ×2 ⁿ matrix C _n including all the 4 ⁿ length n genetic sequences. Furthermore, we use the Hamming distance of α and β to construct a 2 ⁿ ×2 ⁿ matrix D _n . We explore structures of these matrices, refine the results in earlier papers, and propose new directions for further research.     

NMR relaxation processes of 31P in macromolecules

³¹P-Nmr relaxation parameters (spin-lattice relaxation time, linewidth, and nuclear Overhauser effect) were obtained at three different frequencies for poly(U) and a well-defined (145 ± 3 base-pair) fragment of DNA in solution. Data sets for the two samples were analyzed by theories which included relaxation by the mechanisms of ³¹P chemical shift anisotropy as well as by ¹H-³¹P dipole–dipole interaction. Neither data set could be satisfactorily described by a single correlation time. A model of a rigid rotor most nearly fits the data for the DNA molecule. Parameters obtained from the least-square fit indicate (1) that the DNA undergoes anisotropic reorientation with a correlation time τ₀ = 6.5 × 10^?7 sec for the end-to-end motion, (2) the ratio of diffusion constants D_∥/D_⊥ is 91, and (3) that the linewidth is due to chemical shift dispersion to the extent of 0.5 ppm. Some deviations of the calculated from the observed values suggested that significant torsional and bending motions may also take place for this DNA. Another model which contains isotropic motion but with a broad distribution of correlation times was required to fit the data for poly(U). A log ? χ² distribution function of correlation times [Scheafer, J. (1973) Macromolecules 6 , 881–888] described well the motion of poly(U) with the average correlation time τ = 3.3 × 10^?9 sec and a distribution parameter p = 14.     

Purification and Characterization of Chlorophyllase from Alga (Phaeodactylum tricornutum) by Preparative Isoelectric Focusing

Chlorophyllase from a diatom alga (Phaeodactylum tricornutum) was obtained and the partially purified extract has been further purified using preparative isoelectric focusing on a Rotofor cell. Three fractions, FI, FII, and FIII, were separated from the Rotofor cell and salt and ampholytes were removed to give fractions FI′, FII′, and FIII′, respectively. Enzyme fractions FI′, FII′, and FIII′, respectively. Enzyme fractions FI′, FII′, and FIII′ showed specific activities of 15.2 × 10^?4, 226.7 ×10^?4 and 33.8 × 10^?4 µmol/mg protein/min, respectively. Most of the enzyme activity (84%) was in fraction FII′. The optimum pH for chlorophyllase activity was 8.0 for FI′ and 8.5 for both FII′ and FIII′. Apparent K_m values for enzyme fractions FI′, FII′, and FIII′ were 2.1nM, 2.3nM, and 2.0 nM, respectively. Enzyme fractions FII′ and FIII′ showed higher chlorophyllase activity towards the partially purified chlorophyll when it was compared to that with the crude chlorophyll as well as with both chlorophylls a and b. However, the enzyme fraction FI′ had higher activity towards the crude chlorophyll when it was compared to that with both chlorophylls a and b, but with a preference for chlorophyll a over chlorophyll b. The inhibitory effect of diisopropyl flurophosphate (DIFP) on chlorophyllase activity demonstrates a noncompetitive inhibitor kinetics with K_i values of 1.29mM, 2.14mM, and 0.71mM for FI′. FII′, and FIII′, respectively.     

Distribution of hereditary blood groups among Indians in South America. I. In Ecuador

Blood specimens were procured from 658 Quechua, 36 Colorado, 233 Jivaro, 244 Cayapa, and 48 Secoya Indians of Ecuador. These were examined for antigens in the A-B-O, M-N-S-s, P, Rh-Hr, Lutheran, K-k, Lewis, Duffy and Kidd systems and for Diego (Di^a), Wright (Wr^a), and Berrens (Be^a) agglutinogens as well. Hemolystes were prepared and studied for hemoglobin types and the serum samples were tested for haptoglobins and transfserrins. Gene frequencies are high for O, M, s, R¹, (CDe), R² (cDE), Lu^b, k, Kp^b, Le^b and Fy^a; and low or absent for A, B, N, S, Mi^a, Vw, Mt^a, R⁰ (cDe), V (ce^s), Lu^a, K, Kp^a, Le^a, Fy^b, Js^a, Wr^a and Be^a. The Diego (Di^a) gene is present but its frequency varies greatly from tribe to tribe. Gene frequency Hp¹ is well within the range previously reported for Indians in Middle America excepting the Colorado in which population the frequency of 0.889 is unusually high. All 723 serum specimens tested for transferrins were C or CD. No D or BC types were found. All Ecuadorian Indian bloods tested electrophoretically contained only hemoglobin (A) as a major component.     

Endonucleolytic cleavage of murine leukemia virus 35S RNA by microsome-associated nuclease.

Moloney murine leukemia virus 35S RNA (molecular weight 3 to 3.4 × 10⁶) is cleaved by nuclease activity present in microsomal fractions from MLV infected or uninfected mouse embryo cells to two RNA species of approximate molecular weights 1.8 × 10⁶ and 1.5 × 10⁶. Microsomal fractions from MLV infected and uninfected cells also contained nucleolytic activity that solubilized [³H]poly(A)·poly(U) but not [³H]poly(C) or [³H]poly(U); the cleavage of poly(A)·poly(U) was inhibited by ethidium bromide. The cleavage of MLV RNA was also inhibited by ethidium bromide, suggesting double stranded regions in 35S RNA as the site of cleavage.     

Reliability of molecular weight determination of proteins by polyacrylamide gradient gel electrophoresis in the presence of sodium dodecyl sulfate

Thirty-four proteins with molecular weights (MWs) between 1.3 × 10⁴ and 9.5 × 10⁵ and seven polypeptide chains of some of these proteins were incubated in sodium dodecyl sulfate and submitted to electrophoresis in a polyacrylamide gradient gel (3 to 30%; relative percentage of bisacrylamide: 8.4%). Analysis of the results confirmed the reliability of the relationship: log(MW) = alog(T) + b, where T is the polyacrylamide concentration reached by a protein, a is the slope, and b is the Y intercept of the regression line. In fact the mean deviation between the expected and the observed MW was ±5.5%. In addition we have shown that MWs of unreduced and reduced proteins can be estimated with the same accuracy and tha MWs of glycoproteins can also be determined with the same accuracy as other proteins.     

Characterization and Localization of Adenosine A2 Receptors in Bovine Rod Outer Segments

Abstract: Previous work from this laboratory has shown that retinal adenosine A₂ binding sites are localized over outer and inner segments of photoreceptors in rabbit and mouse retinal sections. In the present study, adenosine receptor binding has been characterized and localized in membranes from bovine rod outer segments (ROS). Saturation studies with varying concentrations (10–150 nM) of 5′-(N-[2,8-³H]ethylcarboxamido)adenosine ([³H]NECA) and 100 μg of ROS membrane protein show a single site with a K_D of 103 nM and a B_max of 1.3 pM/mg of protein. Cold Scatchards, which used nonradiolabeled NECA (concentrations ranging from 10 nM to 250 nM) in competition with a fixed amount of [³H]NECA (30 nM), demonstrated the presence of a low-affinity site (K_D, 50 μM) in addition to the high-affinity site. To confirm the presence of A_2abinding sites, saturation analyses with 2-p-(2-[³H]-carboxyethyl)phenylamino-5′-N-ethylcarboxamido adenosine (0–80 nM) also revealed a single population of high-affinity A_2a receptors (K_D, 9.4 nM). The binding sites labeled by [³H]NECA appear to be A₂ receptor sites because binding was displaced by increasing concentrations of 5′-(N-methylcarboxamido)adenosine and 2-chloroadenosine. ROS were fractionated into plasma and disk membranes for localization studies. Receptor binding assays, used to determine specific binding, showed that the greatest concentration of A₂ receptors was on the plasma membranes. Therefore, adenosine A₂ receptors are in a position to respond to changes in the concentration of extracellular adenosine, which may exhibit a circadian rhythm.     

Macrocyclic zinc(II) complexes for selective recognition of nucleobases in single- and double-stranded polynucleotides

 As an extension of our earlier discoveries that Zn^II-cyclen complex (1) (cyclen=1,4,7,10-tetraazacyclododecane) and Zn^II-acridine-pendant cyclen complex Zn^II-N-(9-acridin)ylmethyl-cyclen (3) are the first compounds to selectively recognize thymidine and uridine nucleosides in aqueous solution at physiological pH, the interaction of these and a relevant complex, bis(Zn^II-cyclen) (7), has been investigated with a series of polynucleotides, single-stranded poly(U) and poly(G), and double-stranded poly(A)·poly(U), poly(dA)·poly(dT) and poly(dG)·poly(dC). These Zn^II-cyclen complexes interact with the imide-containing nucleobases in the single-stranded poly(U), unperturbed by the presence of the anionic phosphodiester backbone. The affinity constant of 1 for each N(3)-deprotonated uracil base in poly(U) is determined to be log K= 5.1 by a kinetic measurement, which is almost the same as log K=5.2 for the interaction of 1 with uridine. Thus, they disrupt the A-U (or A-T) hydrogen bonds to unzip the duplex of poly(A)·poly(U) or poly(dA)·poly(dT), as demonstrated by lowering of the melting temperatures (T _m) of poly(A)·poly(U) and poly(dA)·poly(dT) in 5 mM Tris-HCl buffer (pH 7.6, 10 mM NaCl) with increase in their concentrations. The order of the denaturing efficiency is well correlated with that of the 1 : 1 affinity constants for each complex with uracil or thymine;7>3>1. The comparison of circular dichroism (CD) spectra for poly(A)·poly(U), poly(A), and poly(U) in the presence of 3 has revealed a structural change from poly(A)·poly(U) to two single strands, poly(A) and poly(U), caused by 3 binding exclusively to uracils in poly(U). On the other hand, the acridine-pendant cyclen complex 3, which earlier was found to associate with guanine by the Zn^II coordinating with guanine N(7), in addition to the π-π stacking, interacts with guanine in the double helix of poly(dG)·poly(dC) from outside and stabilized the double-stranded structure, as indicated by higher T _m. Received: 31 December 1997 / Accepted: 23 February 1998     

Multivariate spectrochemical analysis of interactions of three common Isatin derivatives to calf thymus DNA in vitro

Interactions of Isatin and its derivatives, Isatin-3-isonicotinylhydrazone (IINH) and Isatin-β-thiosemicarbazone (IBT), with calf thymus DNA (ctDNA) have been investigated to delineate pharmaceutical-physicochemical properties using UV–Vis/fluorescence/circular dichroism (CD) spectroscopy, viscosity measurements, and multivariate chemometrics. IINH and IBT molecules intercalate between base pairs of DNA, hypochromism in UV absorptions, increase in the CD positive band, sharp increase in specific viscosity, and the displacement of the methylene blue and Neutral Red dye in complexes with ctDNA, by the IINH and IBT molecules, respectively. The observed intrinsic binding constants (K_{b[IBT–ctDNA]?}=?1.03 × 10⁵ and K_{b[IINH–ctDNA]?}=?1.09 × 10⁵ L mol^?1) were roughly comparable to other intercalators. In contrast, Isatin binds with ctDNA via groove mode (K_{b[Isatin–ctDNA]?}=?7.32 × 10⁴ L mol^?1) without any significant enhancement in ctDNA viscosity. The fluorescence quenching of Isatin by ctDNA was observed as static. CD spectra indicated that Isatin effectively absorbs into grooves of ctDNA, leading to transition from B to C form. Thermodynamic parameters like enthalpy changes (?H < 0) and entropy changes (?S > 0) were calculated according to Van’t Hoff’s equation, indicating the spontaneous interactions. The common soft/hard chemometric methods were used not only to resolve pure concentration and spectral profiles of components using the acquired spectra but also to calculate Stern–Volmer quenching constants, binding stoichiometry, apparent binding constants (K_a), binding constants (K_b), and thermodynamic parameters. The K_b values for Isatin, IINH, and IBT were calculated as 9.18 × 10³, 1.53 × 10⁵, and 2.45 × 10⁴ L mol^?1, respectively. The results obtained from experimental-spectroscopic analyses showed acceptable agreement with chemometric outlines.     

Power of assaying inbreeding through sampling of phenotypes and mating types

Summary Several enzyme polymorphisms and hemoglobin variants were typed in a sample of n=219 non-related Greek blood-donors. The following gene frequencies were observed: p^a=0.201, p^b=0.701, p^c=0.098; PGD^A=0.985, PGD^c=0.015; AK¹=0.942, AK²=0.058; Hb^A=0.988, Hb^S=0.012. No polymorphic variation was seen in LDH, s-MDH, PHI, or SOD. The population genetical aspects of these results are discussed.

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Zusammenfassung An einer Stichprobe von n=219 erwachsenen griechischen Blutspendern wurden verschiedene Enzympolymorphismen und Hämoglobinvarianten untersucht. Folgende Genfrequenzen konnten beobachtet werden: p^a=0,201, p^b=0,701, p^c=0,098; PGD^A=0,985, PGD^c=0,015; AK¹=0,0942, AK²=0,058; Hb^A=0,988, Hb^S=0,012. Keine polymorphe Variation wurde bei LDH, s-MDH, PHI und SOD gefunden. Die populationsgenetischen Aspekte dieser Befunde werden diskutiert.

     

Evolution of 1, 3, 5-trisubstituted bipyrazole scaffold based platinum(II) complexes as a biological active agent

Abstract

Square planar mononuclear platinum(II) complexes having general formula [Pt(Lⁿ)Cl₂], (where, Lⁿ?=?L^1–4) were synthesized with neutral bidentate heterocyclic 1,3,5-trisubstituted bipyrazole based ligands. The synthesized compounds were characterized by physicochemical method such as TGA, molar conductance, micro-elemental analysis and magnetic moment, and spectroscopic method such as, FT-IR, UV–vis, ¹H NMR, ¹³C NMR and mass spectrometry. Biological applications of the compounds were carried out using in vitro brine shrimp lethality bioassay, in vitro antimicrobial study against five different pathogens, and cellular level cytotoxicity against Schizosaccharomyces pombe (S. Pombe) cells. Pt(II) complexes were tested for DNA interaction activities using electronic absorption titration, viscosity measurements study, fluorescence quenching technique and molecular docking assay. Binding constants (K_b) of ligands and complexes were observed in the range of 0.23–1.07?×?10⁵?M^?1 and 0.51–3.13?×?10⁵?M^?1, respectively. Pt(II) complexes (I–IV) display an excellent binding tendency to biomolecule (DNA) and possess comparatively high binding constant (K_b) values than the ligands. The DNA binding study indicate partial intercalative mode of binding in complex-DNA. The gel electrophoresis activity was carried out to examine DNA nuclease property of pUC19 plasmid DNA.     

Na+ and H+ dependent Mn2+ binding to phosphatidylserine vesicles as a test of the Gouy-Chapman-Stern theory

Summary Mn²⁺ binding to phosphatidylserine (PS) vesicles was measured by EPR as a function of [Na⁺] and pH. At nearly physiological monovalent salt concentration the apparent Mn²⁺ affinity (K _a) increased monitonically over the pH range 5.7–8.35, withK _a roughly [H⁺]^–1 above pH 7.3. It was found, moreover, thatK _a fell off more rapidly with added NaCl at pH 6.1 than at pH 7.87. Qualitatively, these results are consistent with two types of Mn²⁺-PS binding: (i) simple adsorption and (ii) adsorption with the release of an amino proton from PS. The existence of Mn²⁺-induced H⁺ displacement from PS was verified through titration measurements, employing a pH electrode.When H⁺ displacement is taken into account, the variation inK _a with [Na⁺] observed at pH 6.1 is found to be in reasonably good agreement with that expected from the Gouy-Chapman-Stern theory of ionic binding to charged surfaces.     

The species–area relationship derived from species-specific incidence functions

The species–area relationship (SAR), describing the increase in species number (S) with increasing area (A), is one of the most robust patterns in ecology with great significance for conservation. The SAR is generally formulated as a power function, S = kA^z, although the semilogarithmic form S = a + b log A has often been used by botanists. Here we unite the two forms by deriving SARs from the incidence functions of the species that make up the community. We show how the decisive scaling parameters z and b relate to the properties of individual species, and highlight why the biological interpretation of SARs has been so enigmatic.     

Cube law, condition factor and weight–length relationships: history, meta-analysis and recommendations

This study presents a historical review, a meta‐analysis, and recommendations for users about weight–length relationships, condition factors and relative weight equations. The historical review traces the developments of the respective concepts. The meta‐analysis explores 3929 weight–length relationships of the type W = aL^b for 1773 species of fishes. It shows that 82% of the variance in a plot of log a over b can be explained by allometric versus isometric growth patterns and by different body shapes of the respective species. Across species median b = 3.03 is significantly larger than 3.0, thus indicating a tendency towards slightly positive‐allometric growth (increase in relative body thickness or plumpness) in most fishes. The expected range of 2.5 < b < 3.5 is confirmed. Mean estimates of b outside this range are often based on only one or two weight–length relationships per species. However, true cases of strong allometric growth do exist and three examples are given. Within species, a plot of log a vs b can be used to detect outliers in weight–length relationships. An equation to calculate mean condition factors from weight–length relationships is given as K_mean = 100aL^b?3. Relative weight W_rm = 100W/(a_mL^bm) can be used for comparing the condition of individuals across populations, where a_m is the geometric mean of a and b_m is the mean of b across all available weight–length relationships for a given species. Twelve recommendations for proper use and presentation of weight–length relationships, condition factors and relative weight are given.     

Dissociation ofH-2 recognition by antibody and cytotoxic T cells of a cloned murine leukemia cell line

The differential expression of H-2 specificities recognized by antibody and by cytotoxic T lymphocytes (CTL) has been studied using a clone (FY7) of the C57BL/6 leukemia cell line FBL-3 (H-2 ^b /H-2 ^b ). Unlike C57BL/10 spleen cells, EL-4 lymphoma cells and Y57-2C leukemia cells (allH-2 ^b /H-2 ^b ), FY7 failed to induce the primary in vitro generation of anti-H-2^b CTL by (B10.A x A)F₁ (H-2 ^a /H-2 ^a or (B10.D2 x BALB/c)F₁ (H-2 ^d /H-2 ^d ) responder spleen cells. In addition, FY7 was not lysed by, and did not competitively inhibit anti-H-2^b CTL. Quantitative absorption tests with H-2K^b and H-2D^b antisera revealed that FY7 expressed these antigens in quantitatively similar amounts to EL-4. The H-2K^b product of FY7 appeared to be identical with that of C57BL/10 spleen cells both in apparent molecular weight and isoelectric point. Yet FY7 failed to inhibit anti-H-2K^b CTL competitively in a cold target inhibition assay. Possible mechanisms are discussed for the lack of T-lymphocyte recognition of the H-2K^b-gene product expressed by FY7.Abbreviations used in this paper CTL cytotoxic T lymphocytes - MHC major histocompatibility complex - MLC mixed lymphocyte culture - PAGE polyacrylamide gel electrophoresis     

Role and strain distribution of genes controlling light chains needed for the expression of an intrastrain cross-reactive idiotype

Several strains of mice were tested for their capacity to provide immunoglobulin L chains required for the expression of the major cross-reactive idiotype (CRI_A) associated with p-azophenylarsonate-specific antibodies of strain A mice. To facilitate testing, mice were bred that were homozygous for Igh-C ^e and Lyt-2 ^a , 3 ^a , i. e., they possessed genes controlling H chains but not L chains required for expression of the CRI_A. Such male mice were mated to females of various strains and their offspring were tested; expression of CRI_A indicated the presence in the female parent of genes controlling the appropriate L chains. All females bearing the Lyt-2 ^a , 3 ^b or Lyt-2 ^b , 3 ^b genotype yielded offspring, most of which were CRI _A ⁺ , whereas all the offspring of females that were Lyt-2 ^a , 3 ^a were CRI _A ^– . The female parents included mice of several strains that are congenic for Lyt-2 ^a , 3 ^b , Lyt-2 ^b , 3 ^b or Lyt-2 ^a , 3 ^b , thus demonstrating very close linkage between the Lyt loci and the expression of CRI_A. In addition, doubly congenic strains of mice with the heavy chain allotype of the CRI _A ⁺ AL/N strain and the Lyt-2 ^a , 3 ^a genotype on a BALB/c background failed to express CRI_A. The data provide further evidence for the similarity of repertoires of L chains in Lyt-3 ^b mice of various strains. When genes were present controlling A/J H chains and L chains of C57BL/6 or BALB/c origin, the quantitative expression of CRI_A was only slightly lower than that observed in A/J mice. Mice possessing genes controlling the H or L chains required for CRI_A expression, but not both, did not express CRI_A but synthesized Ar-specific antibodies which contained low but significant concentrations of the idiotype-associated chain.