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The interaction of purified human antithrombin-heparin cofactor and human plasmin was studied in the presence and absence of heparin. Antithrombin is a progressive, timedependent inhibitor of the proteolytic and esterolytic activities of plasmin. Incubation of plasmin with antithrombin for 15 to 30 min results in 90 to 100% inhibition of both activities of the enzyme. The presence of heparin dramatically accelerates the rate of interaction of antithrombin and plasmin, with nearly complete inhibition within 30 s of incubation. Sodium dodecyl sulfate gel electrophoresis of reduced and nonreduced proteins indicates that antithrombin functions as a potent antiplasmin by forming an undissociable complex which is stable in the presence of denaturing or reducing agents (or both). This complex represents a 1:1 stoichiometric combination of enzyme and inhibitor. Heparin increases the rate of formation of this complex without affecting its dissociability or stoichiometry.

Oocytes, 80 μm-30 mm in diameter, from ovaries of White Leghorn hens have been examined. Structures are described that appear to represent intermediate stages in lining body formation. The formation (or assembly) of lining bodies occurs in the follicle cell cytoplasm, a few microns distant from the follicle cell plasma membrane. In the very late stages of their formation or after their formation the lining bodies become associated with follicle cell plasma membrane. A parallel process takes place in the oocyte cytoplasm. Structures we have termed oocyte bodies are being formed or assembled. Oocyte bodies are seen forming near Golgi complexes and many oocyte bodies ultimately become associated with Golgi cisternae. The evidence presented in this paper demonstrates that the two structures, lining bodies and oocyte bodies, are distinct and originate in the follicle cell and oocyte cytoplasm, respectively.

Nucleotide triphosphate hydrolase (NTPase, EC 3.6.1.4) is present in the vitelline membrane of the avian egg and increases strikingly, both in total and in specific activity, within a few minutes after ovulation. Protein material found within the lumen of the first part of the oviduct is also high in NTPase activity. Hydrolytic activity toward other phosphate-bearing substrates is minor in the case of both vitelline membrane and oviductal lumen material. Oviductal lumen NTPase is very similar to vitelline membrane NTPase in pH dependency, solubility characteristics, substrate specificities, and electrophoretic mobility. Furthermore, oviductal lumen material has no activating effect on preovulatory vitelline membrane NTPase. It is concluded that the source of vitelline membrane NTPase is material (probably secretory in nature) present in the first part of the oviduct.

The ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity of the vitelline-plasma membrane complex markedly increases at the time of ovulation. A fraction, high in ATPase activity, that remains in the supernatant after centrifugation at 100000 × g for 60 min, can be isolated from this membrane complex. The method for solubilizing this fraction utilizes repeated extraction in buffered KCl and differential centrifugation, a procedure analogous to that used to solubilize muscle. ATPase. The solubilized fraction provides three bands on gel electrophoresis and demonstrates a 30-fold increase in activity over the bound fraction. It is stable during storage at 4° for many days. The fraction can be isolated from the unfertilized egg and appears to differ from the cleavage-membrane ATPase of the fertilized egg.

Proton magnetic resonance data and 13C chemical shifts of numerous acetylenes are presented and analyzed. For 1-alkynylphosphines and -phosphinoxides also 31P shifts are given. It is concluded, that electronic charge shift from triple bond to substituent occurs in phenylethyne, ethylthioethyne, triethynylphosphine, triethynylphosphinoxide and ethynyltriethylgermanium. In 1-alkynyl ethers and amines a charge shift to the triple bond is present. A relatively high shielding of C, F, and P, attached to the triple bond, is observed. This effect is ascribed to a reinforcement of diamagnetic anisotropy caused by π-overlap between triple bond and substituent. To explain this reinforcement the hypothesis of coupling of ring currents is introduced.

Sputtering yields of various metals have been determined for He+‐, Kr+‐, and Xe+‐ion bombardment in the energy range 100 to 600 ev. Yields at 400 ev show a periodic dependence on atomic number similar to that of the reciprocals of the heats of sublimation.

Purified thrombins are isolated from Parke-Davis thrombin topical and from bioactivated crude prothrombins obtained from single animal plasmas. Six thrombin components are present in the former by gel electrophoresis at pH 8.9, ionic strength 0.16, and 30°, but interband protein is high. Ultracentrifugation and gel filtration studies show that, as either ionic strength or temperature is decreased, reversible thrombin self-association and interaction with the gel matrix increase. Electrophoretic mobility and resolution decrease, the latter to give a single diffuse band. Partial fractionation of the six components, numbered according to increasing anodal mobility, was accomplished by differential elution from cellulose phosphate. T2 and T6 were isolated essentially in pure form. Specific activities and component abundances of fractions revealed that T1, T2, and T3 have significantly higher activities than T4, T5, and T6. Thrombins from 60 single animal plasmas contained only T1, T2, and T3, in two distribution types: I, having T1, T2, and T3 in a ratio of 1:2:1, or II, having T1 and T2 as a 1:1 ratio, with T3 absent or present at about 5% of total thrombin. Distribution type for pedigreed animals correlates (a) with age: below 8 months, only I was observed (5 animals); between 8 and 30 months, either I or II was observed (16 animals); and above 30 months, only II was observed (11 animals); and (b) with the two-stage clotting time of the original plasma, with the use of the corresponding serum to supply activators. Other correlations were not found. Component distribution is not a simple, genetically determined characteristic. During activation to produce Distribution I, T1 and T2 may first appear at a ratio of 1:2. An attractive model is as follows. Two prothrombins, P1 and P2, are present in equal amounts. P2 yields T2, and P1 yields T1 and T3, but not sequentially. As the animal ages, the pathway to T3 is lost. Reduction and alkylation of thrombin from thrombin topical results in a dispersion of five fragments between molecular weight of ∼32,000 and ∼5,000. Thrombins from single animal plasmas yield only these two. Probably T4, T5, and T6 are derived from T1, T2, and T3 as a result of peptide cleavages in the larger fragment.

Sputtering yields for 14 metals from the 4th, 5th, and 6th periods were measured for normally incident Hg+ ions at energies between 4–15 kev. The sputtering rate was determined by measuring the time required for the ion beam to pierce a target foil. Results for the various metals show that yields at high energy behave similarly to yields previously found at lower ion energy; i.e., are closely linked to the position of the metal in the periodic chart. Yields increase in the 4th period from Ti through V, Fe, Co, Ni to Cu and in a similar fashion in the 5th period to Ag and in the 6th period to Au. It is believed that Cu, Ag, and Au atoms with their more closely filled d shells behave more nearly like hard spheres and always produce the highest sputtering yields.

Goffman (1961) and others have hypothesized that the experience of being a patient in the traditional type of mental hospital leads to negative consequences for the personality system of the inmate. Rather than being seen as purely a manifestation of internal psychopathological processes, patient behavior is seen, from this perspective, as a response to a peculiar sort of social environment. This environment is seen as disruptive of a positive sense of self in the individual and also as conducive to regressive behavior in several dimensions, including the adoption of a passive, dependent orientation towards the institution. — From a population of psychiatric crisis patients seen at the Emergency Room of Yale-New Haven Hospital, some are sent to the Emergency Treatment Service of the Connecticut Mental Health Center. A small, short term treatment facility, the E.T.S. was designed to avoid many of the negative features of the traditional hospital environment. The small size of the unit leads to a situation where some patients, considered by the reviewing resident to be appropriate E.T.S. candidates, cannot be accommodated because the unit is frequently filled to capacity. A high percentage of these cases are sent to State, a traditional type of public mental hospital. By taking measures on a sample from each of these groups at admission and discharge, it would seem possible to determine the impact of the hospital environment on certain aspects of patient personality and self-image. — The major findings of the study indicate that the traditional hospital environment does, in fact, engender and encourage dependency but that there is a complex interrelation between institutional demands and personality predispositions. Individuals who become long-term mental patients seem to be characterized by a high degree of dependency and a negative definition of self. The hospital environment is thus congruent with certain of their personality demands while it also may serve to emphasize these particular demands from a broader possible range of personality components. Conversely, autonomous individuals seem not to be easily “molded” by the hospital demands and seem able to vacate the patient role very quickly.

The planar waveguide holding planar obstacles is a prototype structure--its fields correspond to some sets of lowest modes in circular and coaxial line waveguides that use coaxial iris loading and in inductive iris loaded rectangular guides. An analysis of a periodically thin-iris loaded planar waveguide is offered, the procedure for which can also be applied to the other waveguides mentioned. A particular example for small iris separation is considered. Among the results of this study are adjustments to some formulas of Brillouin so as to allow consideration of large phase shifts per cell of periodic guide. The procedure, which can be recommended as a formula deriving technique, can find use with such other planar obstacles as the thick periodic iris, the interdigitally placed irises, or the single iris in a multimode guide. Some consideration is given to interdigital loading.

ATPase activity has been demonstrated to be present in the vitelline membrane complex of the hen's egg. This activity increases markedly after ovulation. The vitelline membrane complex has been isolated by dissection and studied in an Ussing chamber such that the inner and outer surfaces can be examined independently. Coupled activity has been detected whereby the presence of substrate on one side affects activity on the other side. Measurements demonstrate that the vitelline membranes of eggs laid in the fall have low enzymatic and no coupled activity. The vitelline membranes of eggs laid in the summer have high enzymatic and marked coupled activity. The coupled activity is reversed in the fertilized compared to the unfertilized egg. This study sheds light on both the physiological character of the ATPases and the roles played by the vitelline membrane in the development of the ovum prior to and after fertilization.

Similarities and differences between “soluble” and ‘membrane-bound’ ATPase (ATP phosphodyhrolase, EC 3.6.1.3) have been found for the membrane complex of the hen's egg. Under conditions of varying ionic strength at pH 9.0, both “soluble” and “membrane-bound” ATPase activities are enhanced by Na + and are suppressed by Ca 2+ . At both pH 6.0 and 9.0 the “membrane-bound” and “soluble” ATPases exhibit opposite patterns of K + enhancement or suppression of activity. The “soluble” and “membrane-bound” fractions exhibit similar cold lability and biphasic pH dependencies. The presence of similar pH differentials in vivo suggests a functional correlation between activation of “membrane-bound” ATPase and changes in the ionic environment. The ATPases appear to represent a family of enzymes with marker characteristics of enzymes associated both with “transport” and “contractility” depending on variations in the species, absolute concentrations and ratios of ions, as well as variations in pH and ionic strength of the reaction.

The finishing of fabrics made from blends of Acrilan and wool is described. The process includes a heat-treatment and a washing-off process, easily carried out on commercial machinery. The mechanical properties of a selection of wool, Acrilan and Acrilan/wool cloths were measured.

SummaryBovine thrombins (Gel-T) isolated from bioactivated crude prothrombin, obtained from Holstein, Jersey and Hereford single animal plasmas, occur in two distributions: either T1, T2 and T3 as 1:2:1 (Type I), or T1 and T2 as 1:1, with T3 absent or present to the extent of 5% of total Thrombin (Type II). Since components differentially denature during procedures used to fractionate, examinations of Gel-T were conducted so that statistical procedures could be used to evaluate particular variances. The variance due to the Gel-T preparative procedure and Gel-T intrinsic specific activity yield a coefficient of variance (Sc) of 0.55%. Thus 95% of single animal Gel-T specific activities fall within 1.1% of the average value. There are no significant differences between animals of different breed, or of bloods drawn from the same animal at different times. Evidently, Gel-T represent a reproducible reference standard. Analysis of distributions shows that the intrinsic specific activities of thrombin components are identical to within 1 %, and are approximately 1.0 NIH unit per 6.0 × 10−4 absorbance unit (3,250 ±450 NIH units per mg). The clotting time (τ) - thrombin concentration (T) relation, log τ = — ã log T + C, is examined. Over the range of τ from 15 to 60 sec it is shown that a T series can be reproduced to within 1 % in thrombin concentration. The variances in the assay system are then due almost entirely to variance in the assay substrate, fibrinogen. These are an Sc of 1.7% for τ̄ between sets of tubes containing dilute fibrinogen (τ-tubes) from one lot of Stock-F, and Sc of 12% between τ̄ for sets of τ-tubes from Stock-F prepared from different lots of Armour Fraction-I, and an Sc of 1.9% for ã̄( =0.679) for sets of T-tubes, obtained either from one Stock-F or Stock-F obtained from different lots of Fraction-I. Errors in several procedures for determining unknown thrombin concentrations are examined.

A procedure is presented for purifying antithrombin-heparin cofactor from human plasma. The final product is homogeneous as judged by disc gel electrophoresis, sodium dodecyl sulfate gel electrophoresis, and immunoelectrophoresis. The final yield averages 12%. A specific antibody directed against pure inhibitor preparations precipitates virtually all of both antithrombin and heparin cofactor activity from defibrinated plasma. The purification, the immunoprecipitation, and other data, indicate that both activities are properties of a single molecular species. The inhibitor and thrombin form a 1:1 stoichiometric complex which cannot be dissociated with denaturing and reducing agents. Addition of heparin, a widely used anticoagulant which specifically accelerates the action of our inhibitor, increases the rate of formation of this complex without altering its stoichiometry or its dissociability. Interaction of thrombin with antithrombin-heparin cofactor requires the presence of the active center serine of the enzyme and arginine residue(s) on the inhibitor, since chemical modification of either of these critical residues inhibits complex formation both in the presence and absence of heparin. We suggest that, in analogous fashion to trypsin-trypsin inhibitor systems, a specific interaction occurs between the active center serine of thrombin and a unique arginine-x reactive site on the antithrombin-heparin cofactor. Furthermore, lysyl residues of the inhibitor probably serve as a binding site for heparin, since chemical modification of these residues virtually eliminates heparin cofactor activity with only minimal reduction of antithrombin activity. We postulate that heparin binds to the inhibitor and causes a conformational change which results in a more favorable exposure of the arginine reactive site, allowing a rapid interaction with thrombin.

The addition of 0.08 M sucrose to a culture medium containing Chang-strain human liver cells causes intense cytoplasmic vacuolation. Electron microscopy of these cells grown inferritin, time-lapse cinematography, and radioautography reveal that the vacuoles arise by endocytosis and that the sucrose is taken into the cell and localized in the vacuoles. Tracer studies demonstrate that sucrose-3H provides a marker for quantitation of endocytosis and that it neither induces nor stimulates endocytosis. Electron micrographs of vacuolated liver cells show microfilaments in close proximity to the inside of the plasma membrane, in the pseudopodia, and to the cytoplasmic side of the membrane surrounding endocytosis vacuoles. Cytochalasin B (CB), a mold metabolite that inhibits various types of cell motility, has a dose-dependent inhibitory effect on the uptake of sucrose-3H by these cells. This inhibition is accompanied by a cessation of the movement of ruffles and pseudopodia on the surface of the cells and the formation of blebs which arise from the cell's surface. These morphological changes are quickly reversible upon removal of CB. Alterations in the appearance and location of microfilaments are also observed in CB-treated cells.

Several new 4-amino-5-arylsulfonylpyrimidines were prepared via the reaction of various α-(ethoxymethylene)arylsulfonylacetonitriles with guanidine or variously substituted amidines (Table II). 2,4-Diamino-5-(p-chlorophenylsulfonyl)pyrimidine (IIIg), a typical example, was prepared from the reaction of 2-(p-chlorophenylsulfonyl)-3-ethoxyacrylonitrile (IId) with guanidine in refluxing ethanol containing sodium ethoxide. With proper substitution of the ethoxymethylene intermediate, the method was found suitable for the preparation of other compounds having either a hydroxy or methyl group at the 4-position of the 5-arylsulfonylpyrimidine. The fluoro group in 2,4-diamino-5-(p-fluorophenylsulfonyl)pyrimidine (IIIx) was successfully replaced by nucleophilic reagents such as sodium ethoxide, N-methylpiperazine and N,N-diethylethylene-diamine. Attempts at direct displacement of fluorine by ammonia at 190° were unsuccessful.

The technique of isolation consists of a combination of homogenization, differential centrifugation, gradient centrifugation and the multiple poolings of isolated fractions. Large numbers of embryos are used to provide 5–10 g of accumulated liver tissue from the younger embryos (7, 11 days). Contamination by components of red blood cells or nuclear envelopes is avoided. The tissues are homogenized as dilute suspensions in either isotonic Krebs-Ringer solution or isotonic sucrose solutions supplemented with magnesium. Intact red blood cells are separated from the homogenate. Initial fractions are separated by multiple differential centrifugations. Fluffy coats and supernatants are pooled sequentially to increase the final yield of membranes. Intact nuclei can be separated by centrifugal rotation at 100 000 × g for 2 h over appropriate sucrose solutions. The separation of the plasma membrane fraction is carried out by density gradient centrifugation at 150 000 × g for 2 h. Fractions isolated in either isotonic sucrose or saline appear to be similar. The cytochrome oxidase and glucose-6-phosphatase activities are negligible. β-Glucuronidase activity is exceedingly low. Mg2+-dependent ATPase activity is enhanced by the presence of Na2+. The isolation of plasma membrane of embryonic cells allows for a more direct assessment of membrane development, cell-contact phenomena and membrane function.

lndeIxes of permuted notations constitute one application of the WWswesser Line Notation (WLN) in industrial organizations. The Institute for Scientific Information (ISI) has introduced an index of this type for new compounds being reported in the journal literature-more than 150,000 per year. This new tool, called the C#rwrrica/ Substructure /ndex (CSI), was created solely for substructure searches. The creation, advantages, and limitations of the CSI are discussed along with criteria used for selecting the more than 1,000,000 entries for the 1970 Annual. Also discussed are design features of the CSI that enable chemists unfamiliar with WLN to conduct substructure search~, In recent years, the extremely limited value of traditional indexes for substructure searching has become well recognized, As a result, considerable research effort has been financed for substructure search systems. However, with few exceptions, this research effort has been aimed at developing computer systems, even though most chemists do not have immediate access to computers. Manual substructure searches remain laborious or impossible to perform. The development of “desk-top” manually searchable indexes by permuting line notations was first reported

The angular distribution of material sputtered under normal Hg+‐ion incidence from flat, polycrystalline targets at 100 to 1000 ev was measured and plotted in polar diagrams. The distribution is ``under cosine,'' approaching a cosine distribution, at higher ion energies. Mo and Fe show a more pronounced tendency to eject to the sides than Ni or Pt. Under oblique ion incidence atoms are sputtered preferentially in the forward direction.

ANTITHROMBIN III (heparin cofactor) is known to inhibit thrombin1 and Factor Xa1–3 (activated Factor X). We have purified antithrombin4 from human plasma by a series of chromatographic and electrophoretic separation techniques. The homogeneity of the final product is demonstrated by disc gel electrophoresis, SDS gel electrophoresis and immunoelectrophoresis4. Using this preparation, we have shown that antithrombin forms a 1 : 1 stoichiometric complex with thrombin which cannot be dissociated with denaturing and reducing agents. Addition of heparin dramatically accelerates the rate of formation of this complex. Complex formation is completely dependent on an interaction between the serine active centre of thrombin and an arginine reactive site of antithrombin. Furthermore, e-amino lysyl groups of antithrombin serve as binding sites for the highly negatively charged heparin. Based on these data and other evidence, we have proposed that heparin acts to accelerate inhibitor function by binding to antithrombin and inducing an allosteric modification in it, which renders the arginine in its reactive site more accessible to the serine in the active centre of thrombin4.

Summary. An acquired haemorrhagic syndrome is described in which the failure of haemostasis is due to impaired crosslinking of fibrin. This defect, demonstrated by both monochloroacetic acid clot-solubility test and disc-gel electrophoretic analysis of individual fibrin chains, existed despite normal fibrin stabilizing factor (factor XIII) levels in the patient's plasma. Activation of the factor by thrombin, measured by amine incorporation into casein, was normal. The unique molecular defect in this patient could be fully accounted for by the presence of an inhibitory IgG antibody against the crosslinking sites in fibrinogen-fibrin. No alteration could be seen in either the rate or in the extent of the reversible aggregation of fibrin. However, large amounts of cryoprecipitate were found in the patient's plasma. This phenomenon could be reproduced in vitro by incubating the chromatographically purified IgG of the patient with normal fibrinogen at 4°C. It is conceivable that the antibody in the patient could have arisen in response to the modification of normal fibrinogen by isonicotinic acid hydrazide which the patient had ingested for 8 yr.

Human mouse and avian oviduct mucosa was studied histochemically and with light and electron microscope for the presence and localization of adenosine triphosphatases (ATPases); peritoneal fluid and serum were also studied. ATPase activity was found in oviductal fluid in all 3 species; no ATPase activity was found in sera and only very small amounts in peritoneal fluid. Light microscopy revealed ATPase-secreting cells most heavily concentrated in the lower preampulla of hens oviducts. ATPase secreting cells in human and mouse oviducts varied with age and hormonal states. Infant postmenopausal and follicular phase oviducts showed few active sites; oviducts from secretory phase oviducts had very high activity along the preampullar mucosal surface. Highest ATPase activity was demonstrated in and on the surface of secretory cells bordering the lumen.

Observations have been made on the response, in vitro, of cultured and freshly dissociated cells to mechanical deformation. Large numbers of individual cells were studied by means of a special culture chamber bounded by two parallel glass coverslips whose spacing could be reduced from 140 to 2 microns in steps of roughly 0.5 micron. The degree of deformation required for herniation of the cell surface was measured. These measurements lead to the definition of a statistical index characteristic of the extensibility of cell surfaces. This index has been shown to be distinctive for several types of cells; to alter with certain stages of embryonic development; and to be stable with respect to the culturing of cells and certain alterations in the method of cell culture.