Demonstration of neuraminidase activity in human blood serum and human milk using a modified, radioactively labelled alpha1-glycoprotein as substrate.

N-Acetylneuramini acid of alpha1-glycoprotein was oxidized with a small molar excess of periodate and reduced with tritium-labelled borohydride. By this method about 50% of the N-acetylneuraminic acid was converted to its radioactively labelled C8-analog and 25% to its C7-analog. Using this modified alpha1-glycoprotein as substrate, minimum neuraminidase concentrations of 10(-18) units/ml, related to the activity of neuraminidase from Vibrio cholerae, could be determined. Neuraminidase activity was demonstrated in 95% of the sera or blood plasma samples from a series of 417 healthy or ill human individuals and in milk samples from 5 different donors. The neuraminidase in both serum and milk had optimal activity at pH 5.5. On an average, 10(-10) neuraminidase units were found in 1 ml serum and 10(-8) units in 1 ml milk. Although the neuraminidase activities in the sera varied, a correlation with definite pathological states is not yet possible. N-Acetylneuraminate pyruvate-lyase activity could not be detected in human serum.


Metabolic transformation of colchicine, IV[1]. On the interaction of colchicine and colchiceine with sulfhydryl compounds.

The interaction of colchicine and one of its metabolites, O10-demethylcolchicine (colchiceine), with sulfhydryl compounds was studied in several experimental systems. Colchiceine protects reduced glutathione from oxidation by atmospheric oxygen. Inactivation of enzymes with catalytically essential sulfhydryl groups is prevented by colchiceine. In affinity chromatographic experiments, Agarose-bound colchiceine adsorbs sulfhydryl enzymes which may be eluted specifically by SH-compounds like mercaptoethanol. No such effects are shown by colchicine. The data demonstrate a novel type of biochemical reactivity in the metabolite colchiceine with potential biological relevance.


Uptake of cortisol by isolated rat liver cells. A phenomenon indicative of carrier-mediation and simple diffusion.

The uptake of cortisol by isolated rat liver cells was studied. Cortisol was taken up rapidly; the uptake increased with increasing temperature and reached a plateau after 45 s at 37 degrees, C, after 60 s at 27 degrees C, and after 90 s at 22 degrees C; at 5 degrees C the uptake increased linearly with time. The uptake was linear up to 1.5 mg of cell protein. Analysis of uptake as a function of increasing concentration of cortisol in the external medium indicated the presence of two saturable systems: a high-affinity system with an apparent Km value of 190 +/- 25 nM and a low-affinity system with an apparent Km value of 2200 +/- 180 nM. Above 600 nM, the rate of uptake of cortisol increased almost linearly with increasing cortisol concentration. Treatment of cells with KCN or 2,4-dinitrophenol inhibited the two saturable components, leaving the nonsaturable system unaffected. The affinity constants, Ka, were 6 X 10(6) M-1 and 0.6 X 10(6) M-1 for the high and low affinity components, respectively. These values increased approximately two-fold when uptake rates were corrected for diffusion. Cortisone and corticosterone inhibited the uptake of cortisol by liver cells competitively; dexamethasone inhibited cortisol uptake noncompetitively. Similarly, oestrone, oestradiol and testosterone decreased the uptake of cortisol, at a concentration of 2000 nM in the external medium, by 20, 49 and 35 percent, respectively; the inhibition was noncompetitive. p-Chloromercuribenzoate, N-ethylmaleimide and 1-fluoro-2,4-dinitrobenzene decreased the uptake of cortisol. Ouabain did not influence the uptake of cortisol; varying the external sodium concentration also did not affect uptake of cortisol. Cyclic-3',5'-adenosine monophosphate had a stimulatory effect. The results show that the first step, before cortisol is bound to intracellular binding proteins, is the uptake of cortisol by proteins in the plasma membrane. At lower concentrations of cortisol, uptake takes place by saturable processes; at higher concentrations saturation is not achieved, indicating that simple diffusion becomes the major route of transport into the cell. The proteins in the plasma membrane probably function as carriers to transport the glucocorticoid into the cell.


The complete amino acid sequences of both subunits of the sweet protein monellin.

The amino acid sequences of both chains of the sweet protein Monellin have been determined. Since chain separation could not be accomplished easily, cyanogen bromide cleavage at the only methionine residue (in the B-chain) was performed and the three products obtained after cyanogen bromide cleavage were separated. For the identification of amino acid phenylthiohydantoins, high performance liquid chromatography was employed. Thus 37 out of a total of 44 residues of the A chain and 40 out of a total of 42 residues of the large CNBr fragment of the B chain could be determined after Edman degradation of the polypeptides on an automated sequenator.


[Quantitative determination of taurine and glycine in the picomole range using [14C]dansyl chloride (author's transl)].

Taurine, a possible neurotransmitter, and the transmitter substance glycine can be determined quantitatively in the picomole range using [14C]dansyl chloride even in the presence of a several thousand fold excess of other amino acids. However, it is necessary to eliminate the excess of dansyl chloride after complete reaction by adding diethylamine. In corresponding microchromatograms on 3 X 3 cm polyamide sheets the dansyltaurine is clearly separated as a circumscript spot without fluorescing contaminations. But commercial [14C]dansyl chloride preparations regularly contain radioactive impurities which do not fluoresce and therefore can only be detected by autoradiography. Since one of these labelled impurities migrates with the usual solvents together with [14C]dansyltaurine during chromatography a method is described for a chromatographic separation of [14C]dansyltaurine from these impurities. As internal standards for preparing calibration curves, alpha-phenylglycine and sarcosine are used. gamma-Aminobutyric acid as well as glutamic acid can react with dansyl chloride to form the corresponding lactams. These lactam rings can be converted with ammonia into the corresponding amides (dansylglutamine and dansyl-gamma-aminobutyramide).


Storage of mannose-containing material in cultured human mannosidosis cells and metabolic correction by pig kidney alpha-mannosidase.

Skin fibroblasts from healthy individuals and a mannosidosis patient were cultured in the presence of [2-3H] mannose and the cell homogenates were fractionated by trichloroacetic acid precipitation into a precipitable and a non-precipitable portion. In uptake as well as in chase experiments the precipitable fractions show no significant difference in their content of radioactivity, while an increased level of radioactivity is found in the non-precipitable fraction of mannosidosis cells. This higher radioactivity content is due to a higher mannose content and is caused by a slower degradation of this fraction. The differences between the metabolisms of the two cell lines can be expressed by the ratio of radioactivity in the non-precipitable and the precipitable fractions. This value is about three times higher for mannosidosis than for control cells. Pig kidney alpha-mannosidase is taken up by both cell lines and is able to correct the impaired degradation of the non-precipitable material in mannosidosis cells, as shown by the normalization of the above defined ratio of radioactivity for this cell type.


Determination of arginine in the reactive site of proteinase inhibitors by selective and reversible derivatization of the arginine side chain.

Reversible and selective modification of the guanidino group by 1,2-cyclohexanedione or 2,3-butanedione in borate buffer was applied to identify arginine residues in the reactive sites of proteinase inhibitors. The ability of the method to distinguish between arginine and lysine residues was examined. Trypsin inhibitors with arginine at the reactive site were almost completely inactivated within 2 h, e.g. the low-molecular weight inhibitors from soybeans (Kunitz), peanuts and porcine seminal plasma. Inhibitors of the lysine-type were not inactivated by 1,2-cyclohexanedione/borate, e.g. the bovine trypsin/kallikrein inhibitor, the trypsin inhibitors from cow colostrum, porcine pancreas, or snail epidermis (isoinhibitor K from Helix pomatia), and the human alpha1-proteinase inhibitor (alpha1-antitrypsin). However, 2,3-butanedione/borate does not show this high specificity. The method was applied to four high-molecular weight proteinase inhibitors from the albumin gland of the snail (Helix pomatia). All four inhibitors were subject to inactivation by the reagent, indicating an arginine residue at the reactive site. Evidence is given by NMR, UV, and titration data that a complex is formed from 1,2-cyclohexanedione and borate. Due to this complex formation, the amount of free carbonyl component is drastically decreased. Since 1,2-cyclohexanedione without borate lacks the high specificity for arginine residues, the borate complex of the reagent is presumed to be responsible for the high specificity. The new procedure for identification of reactive sites is based on a reversible modification of the guanidium group of the arginine residues.


Occurrence of multiple forms of bull and ram acrosin during proenzyme activation and inhibition of activation by p-nitrophenyl p'-guanidinobenzoate.

Proacrosin was extracted from freshly ejaculated bull and ram spermatozoa with acidic buffer solution pH 3.0, and was partially purified by gel filtration to remove acrosin inhibitors. The occurrence of multiple active forms during proacrosin activation at pH 7.8 was monitored by active enzyme staining of samples after polyacrylamide gel electrophoresis at pH 3.8. For comparison, the protein pattern of such activation samples was also determined after sodium dodecylsulfate-polyacrylamide gel electrophoresis. Proacrosin activation was completely prevented in the presence of 10(-4) M p-nitrophenyl p'-guanidinobenzoate.


[Binding of insulin analogs to partially purified insulin receptor from rat liver membrane (author's transl)].

The insulin binding fraction was solubilized from rat liver membrane vesicles by triton and partially purified up to the specific binding activity of 2.1 pmol/mg. A further characteristic of the partially purified soluble receptor is a decrease in irreversible binding to 0.36% (+/- 0.28) with regard to total iodine insulin and to (+/- 1.8) in comparison to reversible binding. From Scatchard plots a high affinity binding site (KD = 5 X 10(-10)M) with low capacity (5 pmol/mg) and a low affinity binding site (KD = 3 X 10(-8) M) with high capacity (30 pmol/mg) can be seen. With carefully prepared liver membrane vesicles, the dissociation constant of the high affinity binding site from Scatchard plot is only 2 X 10(-9)M. With liver membrane vesicles, isolated for preparative purification procedure, the high affinity binding site could not be demonstrated. Displacement studies with insulin analogs were performed with [A1-D-alanine] insulin, [A1-L-alanine] insulin, [des-Gly-A1, NB1, NB29-(Msc)2]-insulin, proinsulin and [desoctapeptid B23-B30]-insulin. Results of binding measurements are presented in half-maximal iodo-insulin binding, in determination of inhibitor- and dissociation constants from Dixon-, Scatchard- and Lineweaver-Burk plots. There are equal relative binding potencies of analogs, observed with crude membrane vesicles and partially purified soluble receptor, although there is a 50-fold difference in specific binding activity. Biologically active insulins are characterized by strong binding to the high affinity binding site. The binding to the low affinity binding site is not correlated to the biological activity of the insulin analog. With insulin and biologically responsive analogs a non-linear curve in the double-reciprocal Lineweaver-Burk plot can be observed. Analogs with low biological activity show a linear dependency. Functional interactions of insulin with the receptor can be demonstrated in a high affinity binding with the first binding site of the Scatchard plot and in a non-linear hyperbolic Lineweaver-Burk plot.


On the biosynthesis of plasmalogens during myelination in the rat. VIII[1]. Incorporation of 1-[1-14C] alkyl-2-acyl-3-sn-glycerophosphoethanolamine with different fatty acids.

The uptake of 14C labelled alkyl chains into the alkenyl chains of plasmalogens was estimated 12, 24, 48 and 72 h after intracerebral administration of different substrates to 14 day-old rats  We used as substrates 1-[1-14C] alkyl-3-sn-glycerophosphoethanolamines, containing in the 2-position of the glycerol residue a stearoyl (substrate I), an oleoyl (II), an archidonoyl (III), a 4, 7, 10, 13, 16, 19-docosahexaenoyl (IV) or no acyl residue (substrate V, so-called lysoether phosphatide). 1) If the fatty acid in the 2-position of the substrate is a saturated one (as in experiment I), the recovery of radioactive alkyl chains is relatively high, but their desaturation is very slow and seems not to reach its maximum even after 72 h. 2) If a substrate with an unsaturated fatty acid in the 2-position is applied, the number of radioactive chains (alkyl plus alkenyl) depends on the chain length and/or the number of double bonds in the fatty acids. The desaturation of the alkyl chains, however, reaches its maximum about 48 h after application (except for substrate IV). 3) Not only is the recovery of radioactivity in the glycerophosphoethanolamine lipids by far the highest in experiment V at any time, but also the total activity of the alkenyl chains has already reached a maximum 24 h after application.


On the mode of action of eucaryotic initiation factor 1.

A factor isolated from the human tonsillar ribosomal wash specifically stimulated the poly (U)-dependent binding of Phe-tRNA to 40S subunits at low Mg2+ concentration and without any requirement for GTP. The stimulated binding of Phe-tRNA to 40S particles was inhibited in proportion to added deacylated tRNA. The factor was inactivated by N-ethylmaleimide, but in the presence of 40S subunits a considerable protection was observed. 40S subunits, incubated with the factor and isolated by centrifugation, carried significant factor activity. The results imply that the human tonsillar factor, which shows a great functional analogy to the eucaryotic initiation factor 1 from other sources, exerts its effect by an initial interaction with the 40S subunit.


[Aminolysis and racemization of activated esters with the use of high temperatures (author's transl)].

The reactivities in aminolysis of the benzyl, phenyl, p-nitrophenyl and pentachlorophenyl esters of benzyloxycarbonylphenylalanine were ascertained at different temperatures up to 90 degrees C and the occurrence of racemization investigated. It turned out that the protective function of the benzyloxycarbonyl group in relation to the racemization was conserved, even at the high temperatures used in the peptide synthesis.


A rapid method for dry weight determination of proteins on a micro scale with an electrobalance.

A method to determine the dry weight (0.25-2 mg) of aqueous protein solutions within 1 h, using an electrobalance, is described. The drying of 50-200 mul solution pipetted onto a glass fiber disc is carried out in vacuo at 70 degrees C until the recorded dry weight becomes constant (within 25-40 min). It has been shown that the dried residue can subsequently be used for other purposes, such as quantitative amino acid analyses. The method is also suitable for the determination of moisture content in lyophilized protein samples.


Operational dimensions of one multihospital system.

The pressing realities of health care delivery in the 1970s call for new solutions and fresh approaches to health care administration. The Lutheran Hospitals and Homes Society of America was founded to assist communities that were encountering difficulties in providing for the health care needs of their people. This article describes how the society meets these needs through a multihospital approach in which the "parent" organization totally operates its member facilities rather than fragmenting its services by allowing member units to utilize one or more of its services.


Effectiveness of hospital administrators: a study.

A survey study of the effectiveness of hospital administrators was conducted by Graduate Studies in Health Services Management, University of Missouri, Columbia. The purpose of the study was to determine whether or not hospital administrators and others knowledgeable about hospital administrators believe that these professionals are effective in their work. In general, administrators, chiefs of staff, and board chairmen rated the administrator's effectiveness higher than did directors of health agencies.


Fat distribution in the adrenal cortex as an indication of the mode of intrauterine death.

The presence of fat in the fetal zone of the adrenal cortex in the stillborn macerated fetus is indicative of the mode of intrauterine death. Three basic patterns are recognized. In the type I pattern there is only a scant amount of fat, close to the medullary zone. In the type II pattern fat is more widespread and is present in many more cells of the fetal zone. The type III pattern reflects a massive fatty change of almost all cells throughout the fetal cortical zone. Correlation of the three fat patterns with the pathology of the placenta and clinical data relevant to the etiology of intrauterine death has led to the following conclusions. The type I fat pattern is indicative of an acute mode of death, the type II pattern is indicative of a more prolonged period of intrauterine deterioration prior to death, whereas the type III pattern is indicative of a chronic mode of death. Determination of the fat patterns in the fetal zone of the adrenal cortex can be helpful when the clinician is confronted with the problem of fetal death, especially when maceration may hamper more detailed pathologic studies.


Malignant fibrillo-caveolated cell carcinoma of the human intestinal tract.

The ultrastructural features of a peculiar midgut carcinoid, containing cytoplasmic filaments, fibrils, and caveolae, are presented. Because of the morphologic similarities between the tumor cells and the recently described intestinal caveolated cell, it is proposed that the cells of the reported carcinoid represent the malignant counterpart of this new type of cell. The name suggested for this variant of a tumor of the enterochromaffin group of intestinal endocrine cells is malignant fibrillocaveolated cell carcinoma.


Focal nodular hyperplasia of the liver: a clinicopathologic study and review of the literature.

We received the clinical records and pathologic material of 20 patients with biopsy proven hepatic focal nodular hyperplasia. The majority of the patients were females of child bearing age, five of whom had a history of oral use of contraceptives. In every instance focal nodular hyperplasia was an incidental finding; liver function tests were always normal. Focal nodular hyperplasia is a distinct histopathologic entity, distinguishable from liver cell adenoma. Specifically it consists of nodular aggregates of cytologically normal hepatocytes with foci of intranodular bile duct proliferation. Focal nodular hyperplasia appears to be a benign entity, even in patients in whom the lesion was not excised. The association between focal nodular hyperplasia and oral use of contraceptives may be coincidental, although hormonally related vascular changes may be responsible for rupture of the lesion.


Structural basis of ventricular arrhythmias in human myocardial infarction: a hypothesis.

The present study was undertaken using light and electron microscopic techniques to determine whether Purkinje fibers survive in the subendocardial region of anteroseptal infarcts in humans. Tissue was obtained for this purpose from 11 patients with 12 documented infarctions at the time of autopsy; six patients died within 72 hours of the infarction and five had healed infarcts. Seven of the 11 patients had ventricular arrhythmias. Light microscopic study indicated that intact cells with a normal appearance remained on the subendocardial surface, although the underlying ventricular muscle either was necrotic or was replaced by fibrous tissue. Electron microscopy demonstrated that these intact surviving cells over the surface of the infarct had few randomly oriented myofibrils, abundant glycogen, and other characteristics of Purkinje fibers. These cells could be readily distinguished from normal or infarcted ventricular muscle cells. Purkinje fibers, the most peripheral part of the conduction system, survive in extensive anteroseptal infarcts and may be the site of origin of ventricular arrhythmias.


Multiple sebaceous adenomas and internal malignant disease: a case report with chromosomal analysis.

A case of multiple sebaceous adenomas associated with internal malignant disease is further documented. Thirteen similar cases reported in the literature are also reviewed. We support the thesis that this association represents a unique syndrome. It can be seen in both sexes and usually is manifested during the fifth and sixth decades. A family history of malignant disease was revealed among seven of the nine cases mentioned. Of the other skin lesions, keratoacanthoma is also frequently seen. The internal malignant tumors most commonly afflict the gastrointestinal tract, especially the large intestine. With proper treatment the internal tumors appear to be of a low degree of malignancy. However, these patients have a propensity to develop muliple visceral tumors and require periodic follow-up. Some patients develop skin tumors prior to the discovery of internal malignant disease, an event that offers some prognostic value. Chromosomal study of the cultured blood lymphocytes by the Giemsa banding technique failed to reveal any abnormality.


A new case of trisomy for the distal part of 13q due to maternal translocation, t(9;13)(p21;q21).

The first child of a mother with a balanced translocation (9;13) revealed a trisomy for the distal third of 13q. Clinical signs were microcephaly, hemangiomata, long incurved eyelashes, strabismus, enlarged bridge of the nose, abnormally long philtrum, high-arched palate, low set ears, hexadactyly of the four extremities, umbilical and inguinal hernias, neonatal respiratory distress, psychomotor and growth retardation. The proband presented also male pseudohermaphroditism and trigonocephaly. This last trait is the object of a discussion in which cases of partial trisomy 13q cited in the literature are considered for study of the incidence of this dyscephaly in this partial syndrome.


Familial C/D translocation t(9;13)(9p23.13q21) in a male associated with recurrent abortion.

A familial reciprocal translocation, established by R-banding as t(9;13) (9p23;13q21), is described in a phenotypically normal male carrier, whose father is also a balanced carrier and wife had four consecutive spontaneous abortions. The role of translocation in reproductive failure through production of chromosomally unbalanced gametes or by impairment of the spermatogenesis is briefly discussed.


Normal meiosis in two 47,XYY men.

Details of testicular histology and meiosis are given for two 47,XYY men, one an oligospermic childless individual, the other a fertile man with near-normal spermatogenic activity in his testes. Examination of the chromosomes at meiosis, with "Q" and "C" staining, gave no evidence for the occurrence of the second Y chromosome in the germ line of either individual.


Follow-up till age 3--4 of unselected children with sex chromosome abnormalities.

Follow-up examination of 15 unselected children with aneuploid sex chromosome abnormalities has been made till between the age of 2 1/2 and 5 years. The mental development of the 15 children was in all cases within the normal range, but there was a tendency to some differences compared with their siblings.


Fine structure of 33 258 H-treated chromosomes.

Metaphase chromosomes of mouse strain L cells show strikingly uncondensed pericentric heterochromatic regions after treatment of living cells with the benzimidazol-derivative 33 258 Hoechst. In electron micrographs of total preparations after G-band staining the chromosomes are seen to be made up of irregularly folded fibrils of 200-400 A in diameter. In the uncondensed regions only very few fibrils laid loose loops are present, making it probable that only one fibril forms one chromatid.


The human leukocyte test system. IX. DNA synthesis and mitoses in PHA-stimulated 2-day cultures set up with Eagle's mimimal essential medium (MEM).

In human leukocyte cultures set up with Eagle's MEM and stimulated with Difco's PHA M, DNA synthesis and mitotic indices were analyzed by means of 3(H)-thymidine autoradiography and cell counting from 23 up to 52 h after culture initiation. Considerable amounts of DNA synthesis and mitoses were found in this time span. This resembles the results found with Ham's F-10 medium. However, the DNA synthesis pattern and the distribution of mitotic indices a higher yield of asynchrony in Eagle's MEM as compared with Ham's F-10 cultures. Proportions of first, second, and third mitoses at 72 h culture time were determined with different methods.


Bf polymorphism: study of a new variant (F0.55).

A variant band in the Bf polymorphism has been found in the serum of a healthy women, her mother, her sister, and two brothers. By direct comparison the band was found to migrate faster than F but slower than F1. It is highly probable that the band represents a new allele at the Bf locus, Bf 0.55. The new allele was transmitted in this family together with the HLA haplotype A11, B27.


Heterogeneity of Dyggve-Melchior-Clausen dwarfism.

Sibs with apparent Dyggve-Melchior-Clausen (DMC) dwarfism and normal intelligence are described. Three other familial and 3 sporadic cases with DMC dwarfism and normal intelligence are known. Twelve familial and 9 sporadic cases are known with the usual combination of DMC dwarfism and severe mental retardation. Since the two conditions appear to breed true they seem to be genetically different. We propose to name the former "Smith-McCort dwarfism" to clearly distinguish it from the DMC syndrome in which mental retardation is a constituent part. Both conditions are inherited as autosomal recessive traits. Spinal cord compression due to atlantoaxial instability is a serious and preventable complication of both disorders.


Polymorphism of properdin factor B in South African Negroid, Indian and Colored populations.

Properdin factor B phenotypes were determined in 7 Bantu speaking Negroid populations, 1 Indian, and 1 Colored population of South Africa in a total of 1258 individuals. In the Negroid populations allele frequencies were: BfF 0.655, BfS 0.282, BfRARE 0.063, in the Indian population: BfF 0.322, BfS 0.645, BfRARE 0.033, and in the Colored population: BfF 0.513, BfS 0.435, BfRARE 0.052. In addition, 2 so far unknown F alleles and possibly 1 new S allele were discovered.


A familial tetraphocomelia syndrome involving limb deformities, cleft lip, cleft palate, and associated anomalies--a new syndrome.

This paper reports a rare malformation syndrome which is observed in two sibs (brother and sister) of a family. It consists of nearly symmetric reductive defects of the limbs, flexon contractures of various joints, cleft lip and cleft palate, multiple minor abnormalities including capillary hemangioma of the forehead, hypoplastic cartilages of ears and nose, micrognathia, intrauterine growth retardation, and possibly mental retardation. Chromosomes of both parents and propositi are normal. Genetic data suggest autosomal recessive inheritance.


Unbalanced 13q/21q translocation: a revised study of the case previously reported as 21-monosomy.

Reexamination was made on a male infant previously reported as 21-monosomy. Extensive chromosome banding analyses in the patient and parents disclosed an unbalanced de novo translocation between chromosomes 13 and 21. The patient's karyotype was interpreted as 45,XY,--13,--21+der(13),t(13;21) (q2 or 3;q1 or 2)pat. The patient showed many clinical features characteristic of 13q--syndrome.


Tertiary trisomy, 47,XX,+14q--, resulting from maternal balanced translocation, 46,XX,t(14;16)(q11;q24).

A 3-year-old child with tertiary trisomy (14 (+14q--), daughter of a mother with a balanced reciprocal translocation [46,XX,t(14;16)(q11;q24) is presented. Craniostenosis and developmental retardation were the primary presenting features in this patient.


Center for Barr body condensation. A case of Turner's syndrome with 45,X/46,X,dic(X) (Xqter leads to p22::p22 leads to qter).

An 11-year-old girl with karyotype 45,X/46,X,dic(X) (Xqter leads to p22::p22 leads to qter) is presented. The abnormal X is always found to be the inactive and late replicating X, and according to previous investigations by Therman et al. (1974) part of the cells are seen to have bipartite Barr bodies.


Partial trisomy 4q syndrome: case report and review.

This communication contributes an additional case of partial trisomy for the long arm of chromosome 4[46,XX,t(X;4)(Q27;Q25)]. Three generations of the patient's family were karyotyped and her mother and brother were found to be balanced translocation carriers. From the patient's clinical examination and from review of 13 similar cases reported in the literature, the following phenotype emerged: constant features were psychomotor retardation, muscle hypotonia at birth, undescended testes in all males and abnormal auricles, typically with a prominent antitragus. Anomalies of kidneys and other structures of the urinary system were present consistently as probably the most characteristic feature of the syndrome. Over 200 structural anomalies of other systems were reported, many of the common to other autosomal chromosomal syndromes.


A 46,XY, del(18)(pter leads to p1 100:) cebocephalic child from a 46,XX,t(12;18)(18pter leads to 18 p 1100:: 12qter leads to 12pter) normal parent.

A normal female with an 18p--chromosome but without any feature of the 18p--syndrome produced a cebocephalic child whose karyotype included an 18p--chromosome. Evidence is presented that the normal female is a nonreciprocal translocation heterozygote resulting from the short arm of one chromosome 18 becoming attached to the long arm of a chromosome 12.


Acromesomelic dwarfism: description of a patient and comparison with previously reported cases.

A woman is described who has a severe bone dysplasia confined to the limbs. The degree of involvement is more marked in the distal bones. Her forearms and legs are short; the fibulae are represented by distal triangular remnants; the metacarpals are short, several phalanges in the fingers are absent; and the toes are represented by "ball-like" remnants containing a single bone. The findings in our patient are compared to previously reported cases of acromesomelic dwarfism.


Nucleolus organizer and satellite association in a variant D-group chromosome.

A No. 15 chromosome with a short arm longer than usual is observed in two phenotypically normal brothers. This chromosome appears to have no visible satellite, shows no N-band staining, and is never involved in satellite association. These results have led us to the conclusion that this chromosome lacks the nucleolus organizer region.


A new, high frequency variant of alpha1-antitrypsin.

A new variant of alpha1-antitrypsin has been detected with the aid of isoelectric focusing on polyacrylamide slab gels. In contrast with many other variants this new alpha1-antitrypsin allele is found in 10-15% of the phenotypes examined. The electrophoretic properties of the new alpha1-antitrypsin variant in isofocusing polyacrylamide gels differ only slightly from the most common alpha1-antitrypsin allele M. On the basis of its electroforetic behaviour we propose the term MN to indicate this new protease inhibitor variant. The isofocusing technique employed, provides an easy to handle, very reproducible method for determining the alpha1-antitrypsin phenotype and can be employed for large scale screening.


Assignment of a gene for human mitochondrial isocitrate dehydrogenase (ICD-M, EC 1.1.1.41) to chromosome 15.

Segregation analysis of human biochemical markers and chromosomes in human-mouse somatic cell hybrids allowed to demonstrate synteny of ICDM with the genes for phosphomannose isomerase and pyruvate kinase and to assign the linkage group to human chromosome 15.


Rapid detection of glyoxalase I (GLO) on cellulose acetate gel and the distribution of GLO variants in a Dutch population.

A rapid electrophoretic procedure is described for detecting the human red cell glyoxalase I variants (GLO 1, GLO 2-1, and GLO 2) on cellulose acetate gel (cellogel) on which the sites of enzymed activity are visualized as purple bands against white background. The frequency of GLO1 gene in a Dutch population living in and around Leiden was found to be 0.4544.


Persistence of high intestinal lactase activity (lactose tolerance) in Afghanistan.

Two hundred and seventy apparently healthy adult subjects from Afghanistan, mainly from the central and eastern parts of the country, were subjected to a lactose tolerance test. The change of blood glucose from the fasting concentration at 20 min after the administration of lactose showed a bimodal distribution. Forty-seven subjects had a rise of blood glucose concentration of more than 1.1 mmol/l and were classified as persistence of high intestinal lactase activity (PHILA), a term which lays emphasis on the fact that high lactase activity in the adult is an unusual state whose prevalence in some populations requires explanation. In the Afghan sample there were no significant differences of the frequency of PHILA in different ethnic groups.


ESD polymorphism in Norway.

ESD phenotype distribution and allele frequencies in 217 Norwegians and 196 Norwegian Lapps are presented. There is good accordance with Hardy-Weinberg distribution, ESD1 allele frequencies are 0.887 in the Norwegians and 0.872 in the Lapps.


Premature chromosome condensation in irradiated man.

Micronuclei and premature chromosome condensation previously described in mammalian cells exposed to different types of irradiation and chemicals in vitro or in vivo could be demonstrated in cultured lymphocytes derived from chronically low-dose irradiated persons. The usefulness of these findings for biological recording of irradiation effects is suggested.


[A further chromosome analysis in induced abortions (author's transl)].

The cytogenetic data of 500 unpublished cases of induced abortions are reported. The incidence of gross chromosome abnormalities was 7.4% (37/500). The frequencies of chromosome abnormalities after 6 gestational weeks decreased gradually with advancing the gestational weeks, while the frequency in the group of 5 gestational weeks remained considerably high as 25.0%.


Leukocyte peroxidase in Spielmeyer-Vogt's disease.

Leukocyte peroxidase activity was estimated in 5 patients with the juvenile form of neuronal ceroid lipofuscinosis (Spielmeyer-Vogt's disease) and in 15 healthy controls. In contradiction to recent reports normal activity of p-phenylene diamine mediated peroxidase was found in the patients. The possible role of contamination of the white cell preparation with hemoglobin is discussed.


A case of trisomy for the short arm of chromosome no. 9(+9(p)).

A new case of "free" trisomy for the short arm of No. 9 chromosome identified by Giemsa staining and "Giemsa-11 technique" is reported.


Familial 13p+ chromosome with mental retardation and dysmorphic features of two children.

Chromosomal studies of two brothers with mental retardation and dysmorphic features showed a 13p+ chromosome. The same 13p+ chromosome was found in the father and the other sib, who were both phenotypically normal. The relationship between the physical abnormalities and the chromosomal findings are discussed.


Deletion of the short arms of chromosome 20.

A 46, XX, del(20) (p11) karyotype (Paris Conference, 1971) was identified in an 11-month-old French-Canadian girl with a dysmorphic syndrome, multiple congenital anomalies, psychomotor and growth retardation. Both parents had normal phenotype and karyotype.


The different actions of normal and supranormal calcium concentrations on the proliferation of BALB/c 3T3 mouse cells.

In the presence of a normal (1.25 to 1.80 mM) calcium concentration, addition of fresh bovine calf serum or completely changing the medium induces proliferatively quiescent BALB/c 3T3 mouse cells in dense cultures to start a growth division cycle and initiate DNA synthesis about 12 hr later. Fresh, low-calcium (0.02 mM physiologically available) medium also causes cells to start a growth-division cycle. Howver, the development of such stimulated, calcium-deprived cells tops just before the expected time of initiation of DNA synthesis, which can then be rapidly induced by restoration of the normal calcium concentration. Simply raising the calcium concentration to nonphysiologically high levels (without otherwise altering the medium) can mimic the action of fresh serum or fresh whole medium by inducing some of the cells in proliferatively quiescent confluent concluent cultures to start a growth-division cycle and initiate DNA synthesis 22 hr later.


The role of the gas phase in the greening and growth of illuminated cell suspension cultures of spinach (Spinacia oleracea, L.).

The gas phase developed above spinach suspension cultures critically affected their growth and greening. Ethylene accumulation inhibited greening; this effect of ethylene was antagonised when the culture gas phase was enriched with carbon dioxide.  Greening was enhanced by reducing the partial pressure of oxygen below the air level; this effect was observed when oxygen supply did not restrict growth.


Changes in soluble proteins in callus cells of hypersensitive tobacco inoculated with tobacco mosaic virus.

Protein changes occurred in callus cells of hypersensitive tobacco (Nicotiana tabacum var. Xanthi-nc) 72 hr after inoculation with tobacco mosaic virus and incubation on a minimal growth medium. Two protein bands, serologically related to viral coat protein, were obtained from extracts of infected cells following electrophoresis on 7% and 10% polyacrylamide gels. An additional, slower migrating protein, perhaps due to virus-induced stimulation of a host protein, also was detected. Although local lesions appeared on callus after 40 hr of incubation, four proteins previously reported in lesion-bearing hypersensitive tobacco leaves were not found. The possible significance of this and the usefulness of a callus-TMV system as a tool to study virus-induced protein changes are discussed.


[Measles and its complications in Hamburg from 1960 to 1973 (author's transl)].

A study of measles was undertaken from the case histories of 3 093 children under 15 years of age, treated in the children's clinics of Hamburg between 1960 to 1973. 562 children were suffering from pneumonia (18,2%), 230 had otitis (7,4%) and 229 (7,4%) showed neural affections (143 cases with fits and 86 patients with encephalopathy or encephalitis), Various parameters were analysed: age, sex, seasonal distribution, fatality, age disposition to convulsions and measles encephalitis, "incubation period" of neural affections, sequelae. Permanent sequelae due to measles encephalitis were seen in 14% of the cases. A plea is made for an universal immunisation against measles.


[Seasonal patterns of coronavirus infections in Hamburg].

During a serological surveillance programme between October 1974 and September 1975 in Hamburg, 1624 sera from all age groups of the population were examined for the presence of coronavirus antibodies (OC 43 agent). 61,51% of the sera showed HI-titres between 1:8 and 1:128. The geometric mean antiboty titre (GMT) for the whole group was 1:12,2. The serological seasonal pattern suggests that coronaviruses are circulating the whole year in the urban population with a prevalence in the cold and a decrease in the warm season (June). The highest incidence of seropositive reactions (82,9%) was found in the 15-24 years old individuals (GMT 1:13,8). These data indicate an increased spread of coronaviruses in this segment of the population and a marked susceptibility of the group.


[Identification of dextrans using rapid fluid chromatography].

It is reported about an analytical method for routine control of clinical dextrans, a method which is considerably quicker than the previously used ones. It is based on the application of HPLC (high pressure liquid chromatography) to permeation chromatography of dextrans on controlled pore glass beads (CPG). The time required for a permeation chromatogram of a dextran sample could be reduced to less than one hour. Calibration curves, necessary for the evaluation of the chromatograms, were established by use of Pharmacia-Testdextrans.


[New system for no-wait continuous infusion of several different solutions using an additional apparatus].

This article introduces a simple coordination system for two or more infusion liquids. Solutions drip down one after the other in a continuous flow, with no attendance required, two or more infusion containers being connected in a way described below. There is no intermingling of solutions when using non-rigid containers, and only a slight one when using glass or plastic material. The check valves work through the pressure exerted by the columns of liquid, the containers being placed on different levels. The essential advantages are: less discomfort and less danger for patients and staff. The system can be used for a washing drainage.


Effect of glucose on the activity of feeding centres before and after parasagittal cuts between medial and lateral hypothalamus.

Intracarotid administration of isotonic glucose (0.5 ml of 5.4%) in the starving albino rats produced an increase in the multiunit activity (MUA) of ventromedial hypothalamus (satiety centre) and a decrease in the MUA of the lateral hypothalamic area (feeding centre). Intracarotid infusion of normal saline did not change the MUA of any of these centres. Parasagittal knife cuts placed in between the satiety and feeding centres did not affect the responsiveness of these centres to glucose administration as indicated by the recorded multiunit activity.


[Osmolality problems].

This is a survey about all factors influencing the osmotic pressure. The actual osmotic pressure in the plasma of healthy individuals is regulated essentially by the natrium ions and the corresponding anions. The one exception is the dysfunctioning diabetes mellitus with distinctly advanced hyperglycaemia. Syndromes with an accompanying azotemia demonstrate a seeming hyperosmolality as long as the osmotic pressure is measured or calculated by the customary method. The real and present osmotic condition can only be calculated when the blood urea concentration is no longer considered. It is proposed that the term "corrected osmolality" be used for the values determined in this manner.


A comparative study of several methods for recording spontaneous motor activity in mice under drug effects.

Various existing methods of recording motor activity in animals are briefly reviewed. An electronic adaptation of an old one, the Jiggle cage, is described. The relative advantages and demerits in recording behaviour of mice under drug effects are compared with those of the traditional Jiggle cage, an electromagnetic activity counter and observational ratings.


In vitro anthelmintic action of some indigenous medicinal plants on Ascardia galli worms.

Different parts of ten indigenous medicinal plants were screened for their in vitro anthelmintic activity against Ascaridia galli worms of the birds. Preparations from Carica papaya, Sapindus trifoliatus, Butea frondosa and Momordica charantia were more effective than piperazine hexahydrate.


[Nutrition problems in kidney diseases].

The practice of dietetic therapy is unusual today for patients suffering from renal failure without hypertension and reduction of glomerular filtration rate. Specific treatment is needed, however, for arterial hypertension, uremia, calculus and uralith disease. Experiments in rats showed, that a lot of uremic symptoms following poorly functioning kidneys are partly at least caused by disturbances in amino acid metabolism. Uremia patients with dysfunctioning plasma protein metabolism (transferrin, complement, cholinesterase, prealbumin and retinolbinding protein) need oral, respectively parenteral substitution of essential amino acids. This substitution is very important under catabolic stress conditions in uremic syndrome with and without vividialysis treatment.


[Effect of canrenoate potassium on intraoperative disturbances of electrolyte equilibrium].

In 61 patients the influence of canrenoate potassium on the shift of electrolytes during the intraoperative phase is studied. Without medication of canrenoate potassium depletion of potassium, magnesium and zinc occurs, a sign of transitory intraoperative hyperaldosteronism. On the other hand, preoperative and intraoperative infusion of 1000 mg canrenoate potassium prevents an increased excretion of electrolytes. Causes and consequences of the electrolyte depletion as well as the effect of canrenoate-potassium medication are discussed.


Ability of temperature-sensitive mutants of the recombinant influenza S/N (H2N1) virus to induce immunity to parental (H0N1 and H2N2) viruses.

The behavior in mice of two thermosensitive (ts) mutants (denoted ts217 and ts700) of the recombinant influenza virus S/N (H2N1) was studied. The parental thermoresistant (tr) virus and both of the mutants were capable of inducing protection against pneumotropic A/Singapore (H2N2) and A/WS (H0N1) challenge viruses. Immunity against the Singapore virus, with which the S/N virus shared the hemagglutinin, developed earlier than against the WS virus, with which the S/N virus shared the neuraminidase. The tr and ts217 viruses were immunologically more active than the ts700 virus. The first two viruses grew markedly better in mouse lungs than did the latter. In the course of ts217 virus replication in vivo, revertants capable of growing at 39 degrees C appeared readily. On the other hand, the ts700 virus proved to be genetically stable. These data seem to provide evidence of a linkage between the stability of the ts phenotype, reproductive capacity in mouse lungs, and immunogenicity in the viruses examined.


Comparison of replication of adenovirus type 2 and type 4 in human lymphocyte cultures.

Adenovirus type 2 was capable of replicating in purified lymphocyte cultures from human adenoid specimens. Phytohemagglutinin stimulation enhanced the replication of virus. Viral titers of 103 to 104 50% tissue culture infective doses per ml were reached after 4 to 8 days. Only 1 to 3 per 106 cells were found to produce virus. In contrast, there was no evidence that lymphocyte cultures could support the replication of adenovirus type 4. The life span of cultures infected with type 2 or 4 was not reduced. The possibility that lymphocytes infected with virus play a role in initiating natural, persisting adenovirus infections of human adenoids is discussed.


Isolation of a purified skin test antigen from Blastomyces dermatitidis yeast-phase cell wall.

Preparative polyacrylamide gel electrophoresis was used to isolate individual components of an alkaline-soluble-water-soluble fraction of the cell wall of Blastomyces dermatitidis yeast phase. One component isolated demonstrated exceptional specificity and reactivity when tested on guinea pigs infected with B. dermatitidis. This component displayed no cross-reactivity when tested on animals infected with Histoplasma capsulatum. The significance of isolation of a purified, specific antigen is discussed.


Significant contribution of spleen cells in mediating the lethal effects of endotoxin in vivo.

Two closely related, histocompatible mouse strains that have marked differences in both in vitro and in vivo responses to endotoxin were used to evaluate the contribution of lymphoid cells to the lethal effect of endotoxin. C3H/HeJ mice are endotoxin resistant, whereas C3H/HeN mice are endotoxin sensitive. In vitro spleen cell mitogenic responses to endotoxin were similar in untreated mice and in mice that received sublethal irradiation (450 R) followed by reconstitution with autologous spleen cells. Reconstitution with spleen cells from the related strain produced chimeric animals with spleen cell mitogenic activity like that of the donor strain. When chimeric animals were subjected to a lethal challenge of endotoxin, their response was markedly altered by the transferred lymphoid cells. C3H/HeJ animals reconstituted with C3H/HeN cells became more endotoxin sensitive, whereas C3H/HeJ cells became more endotoxin resistant. These results indicate that spleen cells play a significant, detrimental role in endotoxin-induced lethality.


Hepatitis B antigen-associated deoxyribonucleic acid polymerase activity and e antigen/anti-e system.

Serum samples of 403 asymptomatic blood donors carrying hepatitis B surface antigen (HB5Ag) were concentrated threefold and tested for e antigen and antibody to e antigen (anti-e) by immunodiffusion. Hepatitis B antigen (HBAg)-associated deoxyribonucleic acid (DNA) polymerase activity was specifically determined by the difference in incorporation of [methyl-3H]thymidine 5' -triphosphate into DNA by an aliquot of centrifuged serum samples after it had been treated either with normal rabbit serum or with rabbit antibody to HBSAg. All of 58 serum samples containing e antigen revealed HBAg-associated DNA polymerase activity, whereas none of 96 samples containing anti-e did. In the remaining 249 samples in which neither e antigen nor anti-e was found, 62 showed specific DNA polymerase activity, although at lower levels than the samples containing e antigen.


Immunological relationships between glucosyltransferases from Streptococcus mutans serotypes.

Partially purified glycosyltransferase enzymes for Streptococcus mutans GS-5 (serotype c) have been utilized to prepare antibodies directed against the soluble glucan-synthesizing activity, GTF-B, and the insoluble-soluble glucan synthetic activity, GTF-A. Anti-GTF-A inhibited insoluble glucan formation catalyzed by the extracellular enzymes from strains GS-5 and FA-1 (serotype b) to a much greater extent than that of strains HS-6 (serotype a) or OMZ-176 (serotype d). This antibody fraction also inhibited both the cell-associated glucosyltransferase activities as well as the sucrose-mediated adherence of cells to glass surfaces by strains GS-5 and FA-1 but not that of strains HS-6 and OMZ-176. Anti-GTF-B inhibited soluble glucan formation catalyzed by the extracellular enzymes of strains GS-5 but not that of strain HS-6, FA-1, or OMZ-176. However, this antibody fraction did not strongly inhibit either the cell-associated glycosyltransferase activity or cellular adherence of any of the four strains. These results with body antibody fractions were also correlated with the ability  of the antibodies to agglutinate the cells and form precipitin bands after immunodiffusion with the extracellular enzymes. Antibody prepared against the homogeneous soluble glucan-synthesizing enzyme demonstrated similar effects to the anti-GTF-B fraction. These results are discussed in terms of the antigenic relationships existing between the glucosyltransferases from different serotypes of S. mutans.


Requirements for ingestion of Chlamydia psittaci by mouse fibroblasts (L cells).

Ingestion of 14C-amino acid-labeled Chlamydia psittaci (6BC) by mouse fibroblasts (L cells) was inhibited when the host cells were incubated for 30 min at 37 degrees C in Earle salts containing 10 mug of crystalline trypsin per ml. Tryptic digestion also inhibited the ingestion of 1-mum polystrene latex beads. Trypsin-treated L cells almost completely recovered their ability to ingest chlamydiae after 4 h at 37 degrees C in medium 199 with 5% fetal calf serum. Cycloheximide (10 mug/ml) blocked this recovery. Heating 14C-amino acid-labeled C. psittaci for 3 min at 60 degrees C inhibited its ingestion by L cells, whereas inactivating it with ultraviolet light was without effect on the ingestion rate. These results show that efficient ingestion of C. psittaci by L cells involves trypsin-labile sites on the host and heat-sensitive sites on the parasite. The failure of excess unlabeled infectious C. psittaci to promote the ingestion of 14C-labeled heat-inactivated chlamydiae suggests that direct interaction between these two sites must occur for uptake to proceed normally.


Serological and genetic examination of some nontypical Streptococcus mutans strains.

Thirty-four strains of Streptococcus mutans whose antigenic or genetic positions were unclear or unknown with respect to the serological scheme of Bratthall (1970) and Perch et al. (1974), or the genetic (deoxyribonucleic acid base sequence homology) scheme of Coykendall were analyzed to clarify their relationship to previously well-characterized strains. Strain OMZ175 of the "new" serotype f was genetically homologous with strains of S. mutans subsp. mutans. Strains of the "new" serotype g were homologous with serotype d strains (S. mutans subsp. sobrinus). Strains isolated from wild rats constituted a new genetic group but carried the c antigen. Thus, strains within a "genospecies" (subspecies) of S. mutans may not always carry a unique or characteristic antigen. We suggest that the existence of multiple serotypes within subspecies represents antigenic variation and adaptations to hosts.


Molecular forms of neurotoxins in proteolytic Clostridium botulinum type B cultures.

A modified purification method was used to isolate the neurotoxin of proteolytic Clostridium botulinum type B strain Lamanna. The preparation was found to be a mixture of two protein forms. They were of molecular weight 152,000 and could not be separated by ion-exchange chromatography or electrophoresis in polyacrylamide gel. One was a single polypeptide chain, and the other was a dichain molecule (nicked toxin) held together by an interchain disulfide bond(s). Trypsinization increased the toxicity of the toxin preparation and converted the single-chain molecules into dichain forms that were indistinguishable from the endogenously generated nicked toxin. A protease of the type B culture, with substrate specificity similar to that of trypsin, did not change detectably the molecular form of unnicked type E toxin, although toxicity was increased. Higher toxicity was obtained when unnicked type E was trypsinized; the resulting preparation contained only nicked toxin molecules.


Susceptibility of mice treated with cyclophosphamide to lethal infection with Leptospira interrogans Serovar pomona.

Mice not normally susceptible to infection with Leptospira interrogans serovar pomona were rendered susceptible to lethal infections by treatment with a single dose of 300 mg of cyclophosphamide (Cy) per kg administered optimally from 4 days before to 1 day after infection. Cy-treated mice with either passively or actively acquired antibody were protected from death. Blood levels of leptospires in infected untreated and in Cy-treated mice remained similar until 2 days after infection, when untreated mice cleared the leptospires. Soon afterwards, opsonizing and agglutinating antibody appeared. Cy-treated mice given spleen cells from other normal or specifically immune mice were protected from infection. An important factor in the natural resistance of mice to leptospiral infection appears to be their capacity to produce circulating antibody within 48 to 72 h. Applications are suggested for this animal model in vaccination and protection studies.


Use of semisolid agar from initiation of pure Bacteroides fragilis infection in mice.

The development of a pure Bacteroides fragilis infection in mice is described. The infection produces large subcutaneous abscesses at the site of injection which can be observed grossly within 7 days after injection. The infection was initiated by infection of pure cultures grown in semisolid agar medium. Similar infections were also produced with pure cultures of B. distasonis, B. ovatus, B. thetaiotaomicron, and B. vulgatus. However, a distinct deoxyribonucleic acid homology group, formerly classified as B. thetaiotaomicron, did not produce abscesses in any of the mice tested.


Ultrastructural and biochemical alterations effected by viridin B, a bacterocin of alpha-hemolytic streptococci.

Viridin B, a bacteriocin produced by Streptococcus mitis strain 42885, was shown previously to exert a bactericidal effect against a Neisseria sicca strain but only a bacteriostatic effect against a coagulase-negative staphylococcus. In the present report, this dual effect was defined further. Viridin B caused the appearance of mesosomes in both indicator strains early in the course of exposure. Whereas mesosomes were the only noticeable alterations in the staphylococcal strain at all times of exposure to bacteriocin, progressive changes occurred in the N. sicca cells involving the nucleoid and cytoplasmic contents and resulting in the appearance of bacterial ghosts. Biochemical studies indicated a rapid and complete cessation of the incorporation of [3H]thymidine in N. sicca and in the staphylococcus. Whereas the incorporation of [3H]leucine and [3H]uridine in N. sicca was completely stopped, such incorporation was only reduced in the staphylococcus.


Relationship of bacteriophages to alpha toxin production in Clostridium novyi types A and B.

The relationship of specific bacteriophages to the production of the lethal alpha toxin in Clostridium novyi types A and B was investigated. When type A strain 5771 reverted to the phage-sensitive state, it ceased to produce alpha toxin but continued to produce the gamma and epsilon antigens. This "nontoxigenic" culture, therefore, more closely resembled C. botulinum types C and D than the other C. novyi types. Phage-sensitive type B strains also ceased to produce the alpha toxin but continued to produce the beta toxin, and therefore very colesly resembled C. novyi type D (C. haemolyticum). Alpha toxin was again produced when the phage-sensitive cultures were reinfected with the respective tox+ phages. Alpha toxin production could also be induced in the "nontoxigenic" phage-sensitive derivatives from type B strain 8024 by tox+ phages isolated from other strains of type B. tox- phages were also isolated, but they did not affect alpha toxin production. The tox+ phages also caused a marked change in the colonial morphology of type B strains. In this report we present evidence that alpha toxin production by C. novyi type A strain 5771 and type B strain 8024 depends upon the continued presence and participation of specific bacteriophages designated as NA1tox+ and NB1tox+, respectively.


Morphological and antigenic relationships between viruses (rotaviruses) from acute gastroenteritis of children, calves, piglets, mice, and foals.

The reovirus-like particles present in the feces of young pigs and foals with acute enteritis and the virus causing epizootic diarrhea of infant mice were found to be indistinguishable morphologically from each other, from the South African SA. 11 and "O" viruses, and from the rotaviruses of children and calves. The inner capsid layer of each of these viruses reacted seriologically with sera of children, calves, mice, piglets, and foals convalescent from infection with their respective rotaviruses. These sera reacted by immunofluorescence with human, bovine, porcine, and murine rotaviruses, SA.11, and "O" viruses in tissue cultures and with human bovine, procine, nad murine viral antigens by complement fixation and gel diffusion. However, the antisera differed in their ability to react serologically with the outer capsid layer of the viruses investigated and in their ability to neutralize tissue culture-adapted calf virus. These two tests may demonstrate strain or host specificity among rotaviruses. Since the porcine, murine, and equine viruses are closely related serologically to and are morphologically identical to the human and bovine viruses, they should be included in the group of viruses for which the term "rotavirus" has been suggested. All known members of this proposed group of viruses share a common antigen, probably situated within the inner capsid layer; thus, any one of the viruses may be used for the preparation of antigen or antibody for diagnostic tests, and this will aid in the diagnosis of virus infection in those species from which a rotavirus has not been cultured.


Reovirus-like agent associated with fatal diarrhea in neonatal pigs.

Large numbers of a reovirus-like agent were visualized with electron microscopy in bacteria-free gut homogenates obtained from piglets with a fatal diarrhea resembling transmissible gastroenteritis. The syndrome, of vomiting, diarrhea, dehydration, and death, was reproduced in piglets artificially infected with these bacteria-free gut homogenates. Reovirus-like particles persisted in serial piglet passage and none was seen in uninfected, asymptomatic controls. Hyperimmune sera (made in recovered piglets) aggregated the reovirus-like particles, as judged by immunoelectron microscopy, and neutralized the infectious agent. The cytoplasm in enterocytes on infected intestinal epithelium fluoresced when this hyperimmune sera was used in an indirect fluorescent antibody test. Feeding cow colostrum or diets containing porcine gamma globulin protected infected piglets. No cytopathogenic effect was noted in infected tissue cultures, nor did this agent affect neonatal guinea pigs, hamsters, mice, and rats. The agent did not agglutinate human O or A erythrocytes.


H and M antigens of Histoplasma capsulatum: preparation of antisera and location of these antigens in yeast-phase cells.

Antiserum has been prepared in rabbits against the H and M antigens of H. capsulatum with immunoelectrophoretic precipitin arcs used as vaccines. The antiserum is specific for H. capsulatum in the immunodiffusion test and can be used as reference serum for identifying antibodies to these antigens in sera from suspected cases histoplasmosis. We found that (1) hand m antigens are not located on the surface of yeast-phase cells and (ii) complement fixation releases the antigens reactive in the complement fixation test from yeast-phase cells.


Effect of aeromonas salmonicida (furunculosis) infection on the incorporation of [1-14C]acetate into lipids of trout liver.

[1-(14)C]acetate is incorporated into lipids more rapidly by liver from Aeromonas salmonicida-infected trout than by normal trout liver, both in vitro and in vivo. The increase was, in general, about twofold to less than threefold.


Comparison of the in vitro bactericidal activity of human serum and leukocytes against bacteroides fragilis and Fusobacterium mortiferum in aerobic and anaerobic environments.

In aerobic and anaerobic in vitro environments, Fusobacterium mortiferum was killed by human leukocytes or serum alone or in combination; clinical isolates of Bacteroides fragilis were killed only by leukocytes in the presence of serum.


Binding of morphine by serum globulins from morphine-treated rabbits. II. Antibody nature of the binding globulins.

Data are presented which show that the increased binding of morphine by sera from rabbits which had been treated with morphine in an appropriate manner is mediated by immunoglobulins and is specific. These observations, together with findings reported in the preceding paper, suggest that morphine or one of its derivatives can evoke specific immune responses. It seems most likely that the response is evoked by a complex, formed in vivo, of a tissue or serum protein and morphine or one of its metabolites.


Characteristics of guinea pig IgE.

Anti-dinitrophenyl (DNP) IgE antibody was produced in guinea pigs (GP) by immunization and repeated boosters with 1 mug DNP-Ascaris extract in A1 (OH)3. The isoelectric point was determined to be 6.48 and the molecular weight to be 185,000 daltons. In absorbtion chromatography, rabbit anti-GPIgG1 could not absorb the anaphylactic activity of the IgE. The activity of IgE was destroyed by heating at 56 degrees C for 1 h. IgE persisted in the skin for at least 3 weeks.


Bovine reaginic antibody. I. Rat mast cell degranulation by bovine allergic serum.

A method of utilizing morphological changes in rat mast cells to determine reaginic antibody activity in bovine serum is described. This technique, which has been shown to be useful for the diagnosis of allergies in man, relies on the ability of antigen to degranulate mast cells sensitized with allergic serum. Experiments with radioactively-labelled allergic bovine globulin indicated the specificity of the binding of such proteins to rat mast cells. Cross-reaction between reaginic bovine antibody and human IgE was shown by a binding assay involving the uptake of 125I-labelled anti-human IgE globulin by mast cells incubated with bovine passive cutaneous anaphylaxis positive globulin.


In vitro diagnosis of atopic allergy. V. The value of repeated testing using several batches of the allergen extract.

The radioallergosorbent test (RAST) has been used for in vitro diagnosis of allergy in a selected group of children with asthma and allergic rhinoconjunctivitis. The results were compared with those of provocation tests and also with the results of skin tests. Where the results of RAST and the provocation tests differed, repeated tests were performed, initially in vitro and secondly in vivo. Selected commercial allergens of good quality were used and, for the most part, the same allergen batches were used in RAST, in the provocation tests and in the skin tests. A 76% correlation between RAST and provocation tests was found on preliminary investigation before divergent results were repeated. This degree of correlation is similar to that obtained previously in unselected patients using RAST as a part of the routine diagnostic. On repeated examination, agreement was obtained in an additional 16% of the cases. The major part of the improvement (13%) was due to conversion of preliminary RAST results from negative to positive in children with positive provocation tests. To a minor extent, the improvement was due to the fact that in, some children with positive RAST, negative provocation tests became positive when repeated. Finally, in one case an originally positive RAST in a child with a negative provocation test was found to be negative on re-examination. Comparing the final revised results of RAST and provocation tests there were no cases where a negative provocation test was combined with a positive RAST. Nearly all the children with positive provocation tests in spite of a negative RAST were shown to have low degree allergy. Although circulating reaginic antibodies are not always present, it seems that, on repeated investigations, the great majority of children with asthma and hay fever can be shown to have such antibodies even though the serum concentration may be low or very low. In doubtful cases repeated RAST examinations may be of value.


Passive cutaneous anaphylaxis in the domestic fowl.

Passive cutaneous anaphylaxis (PCA) has been demonstrated in young and adult domestic fowl using antisera obtained from actively sensitized chickens on the day of their maximal susceptibility to systemic anaphylaxis. Young chickens are more sensitive to histamine and PCA reaction than adults. The PCA reaction was inhibited by the H1-histamine receptor blocker, mepyramine. In the PCA reaction of 2 h sensitization period, the main exudating cells were heterophils, mononuclear cells (lymphocytes, monocytes and macrophages) and some basophils. This study suggests a possible role for circulating basophils in the mediation of early acute inflammatory reaction of immunological origin.


Activation and consumption of Hageman factor in the anaphylactic shock of the rat.

In the early stages of anaphylactic shock of rats pretreated with Bordetella pertussis vaccine, a prompt and parallel activation of the factor XIIa-dependent intrinsic coagulation, kinin generation, and fibrinolytic acticity was observed. The coagulation studies, the similarity of anaphylactic results with those produced by a single injection of ellagic acid, and the effective inhibition of the anaphylactic and the ellagic acid-induced activation of these pathways by lysozyme all suggest that factor XII itself becomes activated in rat anaphylaxis. As the reaction proceeded, considerable anticoagulant activities emerged, but the bradykinin and the plasminogen activator levels even further increased. During the first 10 min of anaphylactic shock, factor XII was partly consumed and this was prevented by epsilon-aminocaproic acid infusion. The results show that in pathological conditions such as anaphylaxis there is an intimate in vivo interaction among the three factor XIIa-dependent pathways.


Phytohaemagglutinin (PHA) skin test in predicting prognosis in cancer patients.

The response to intradermal injection of 1 mug of purified PHA was found to correlate with survival of patients with advanced solid tumours treated by chemotherapy, with responders surviving longer than non-responders. In patients with prolonged survival, the reaction was enhanced. Negative intradermal response to PHA was found to be less common in cancer patients than negative responses to tuberculin or streptokinase, and was more easily reversed by successful chemotherapy.


Passive cutaneous anaphylaxis in guinea pigs elicited by gastric absorption of dextran induced by acetylsalicylic acid.

Passive cutaneous anaphylaxis (PCA) was elicited in guinea pigs sensitized with rabbit antidextran by the absorption of dextran macromolecules from the stomach induced by intragastric acetylsalicylic acid. The gastric contents had a pH sufficiently low to maintain the acid mainly in the unionized form since it is the latter which alters gastric permeability. The acid concentration required to induce PCA was below that which caused mucosal cell loss or bleeding. The maximal molecular weight of the absorbed dextran was approximately 25,000. Dextran was chosen as antigen because of its well-characterized physical and immunological properties. It is suggested that ingestion of acetylsalicylic acid may contribute to sensitization and allergic reactions to antigenic food materials by facilitating their absorption from the stomach.


Sugar polymers in rats and man.

Intracutaneous injections of a glucan contaminant of invert sugar solutions and crude cane sugar into human skin produced a localised wheal and erythema reaction. The glucan was also active on intradermal injection into both dextran-sensitive and dextran-resistant rats.


Computerised assessment of cardiac pacemaker performance.

The design and use of a PDP 11/40 based automated on-line cardiac pacemaker assessment system is described. One program has been developed for testing pacemakers on the bench and another for implanted pacemaker tests. Either fixed rate or demand pacemakers can be tested. The on-line system is easy and fast to use and is also highly accurate. Parameters extracted are pulse width, rate and pulse energy. In addition, a graphical representation of the patient's ECG and the pacemaker pulse is obtained on a computer graphics terminal.


Analysis of the human respiratory system in hypoxic and anoxic hypoxia by analogue computer simulation.

The goal of this work is to synthesise a model of an adaptive system of respiration control in man, which could be used for the prognosis of human performance under extreme conditions. The model was partly based on 428 experimental results from hypoxic and anoxic hypoxia. Simulation of the oxygen regulatory circuit was realised on the NADAC 100 and MEDA T 80 analogue computers with satisfactory results. Equations constituting a new version of the model are presented in some detail, together with the results of the simulation of ventilatory mechanics.


[Compartmental model of the adsorption of a dye by proteins. The possible role of adsorption in the hepatic excretion of bromosulphophthalein].

A compartmental model was suggested for the study of the hepatic excretion of bromosulphophthalein (BSP). By using a hypothesis concerning the adsorption of the BSP compound by proteins, one derives from the equations of the model several known experimental results, viz. form of concentration curves that vary in accordance with the strength of the injection, existence of a limiting Tm value, etc. An explicit formula is given for calculating Tm.


Do efferent fibres to hair cells intervene in acoustic stimulus peripheral coding?

The functional roles suggested for Olivo-Cochlear Bundle (OCB) fibres innervating auditory receptors never received a fully convincing demonstration. Assuming that efferent fibres effects, at the level of hair cells, are inhibitory with regard to single auditory nerve fibre activity, a mathematical model of the peripheral auditory system is proposed in which the presence of the descending pathway is taken into account. Computer simulation of the model accounts for most of the salient features of the patterns of activity of single cochlear nerve fibres. In the light of the model predictions, the influence of OCB fibres on peripheral receptors is assumed to act in the direction of improving the signal-to-noise ratio, as a first step of information processing.


Numerical determination of intestinal membrane diffusing constants by a gradient method.

Optimisation problems arising in the identification of kinetic parameters of intestinal membranes are here considered. The dynamic behaviour of the membrane is described by means of a linear compartmental model. Using optimisation techniques of a gradient type, the intestinal kinetic parameters are identified, minimising a quadratic criterion between experimental data of D-histidine transport and model prediction. Numerical results are reported and their physiological implications discussed. The quantitative assessment of the asymmetry of diffusion constants with respect to diffusion direction seems to be an important result of this work.


Methadone treatment: program evaluation and dose response relationships.

Data from a heterogeneous group of 155 clients in a Methadone Treatment Program were analyzed for (A) number of possible methadone administrations since S joined the program, (B) actual number of methadone administrations received, (C) spottiness (A-B), and (D) presence of illicit drugs in urine analysis reports. Data were evaluated for three separate time spans which corresponded with three programmatic changes. Results indicate that initially high blockading doses of methadone are effective in reducing illicit drug use, but as length of time in program increases, the efficacy of low doses of methadone in reducing illicit drug use increases. At least one programmatic change, a more stringent selection procedure, was demonstrated to be highly successful in reducing illicit drug use.


Attitudes toward alcohol as predictors of self-estimated alcohol consumption in college students.

The purpose of the present investigation was to examine the relationship between attitudes toward alcohol use and self-estimated drinking behavior. The results indicated that attitudes can reliably predict variations in self-reported drinking behavior. The implication of these findings for treatment and future research is discussed.


Cognitive functioning under moderate and low dosage methadone maintenance.

The Wechsler Adult Intelligence Scale (WAIS) was administered to two groups of patients in the Santa Clara County Methadone Maintenance Program who were receiving low and moderate daily stabilized dosages of methadone hydrochloride. In the two separate statistical analyses performed, there were no significant differences found between subtest scores or verbal, performance, and full-scale scores of the groups measured. These results, combined with observations regarding the similarities of WAIS profiles of the two groups, indicate that cognitive functioning as measured by the WAIS was not differentially affected by the two dosages.


Case-note abstraction: a methodological substudy as cautionary tale.

A sample of 42 case notes of alcoholic patients were abstracted for 17 items by three different raters. Interrater agreement was generally rather low. Research based on case-note abstraction which does not report on abstraction reliabilities must therefore be viewed with some suspicion. It would be helpful if clinical material could more often be collected in a standardized manner.


The heroin epidemic in San Francisco: estimates of incidence and prevalence.

Much attention has recently been focused on the question of the extent of heroin use in America. In a worthwhile effort to call public attention to the problem, many estimates of its size have evidenced a tendency toward exaggeration and aggrandizement. This paper presents methods of estimation of the extent of heroin addiction which, when carefully employed, should effectively correct such distorted estimates. Two general types of estimation are employed, incidence and prevalence. Incidence estimates are concerned with new cases of heroin addiction that occur in a specific population within a given amount of time. These estimates are based upon self-report data from addicts regarding date of first addiction. When corrected for the "lag phase" (that period of time between onset of first addiction and eventual visit to a treatment program), these data permit accurate retrospective charting of incidence trends. Prevalence estimates focus on all known cases of heroin addiction in a specific population within a given amount of time. Three separate types of prevalence estimates from three separate sources are outlined in this article: estimates based on overdose death data, estimates based on crime statistics, and estimates of "unknown" addicts. In outlining these methods, this article describes the fluctuations in heroin addiction in one major American city, San Francisco, California. After analyzing data gathered from a sample of 2,367 addicts contacted over a 3-year period, this study suggests that the incidence of heroin addiction seems to have declined after 1970. Possible factors underlying this apparent decline in heroin addiction are then discussed, including the post-1970 maturation of the "population at risk," the effectiveness of antidrug media messages, the changing drug fashions in the heroin subculture, and the gradual deterioration of the quality and potency of street heroin.


Readiness for rehabilitation.

High dropout rates of addicts in treatment have long been attributed to a lack of motivation or readiness for rehabilitation. Two groups of subjects were compared on several demographic characteristics and several variables isolated as possible predictors of readiness for rehabilitation.


The validity of self-recording reports made by drug and alcohol abusers in a residential setting.

In this study a self-recording procedure was used to clarify the relationship between urges or cravings for drugs (including alcohol) and actual drug (including alcohol) use within a residential setting. There were two purposes of the study: first, to evaluate the predictive validity of the self-recording procedure; and second, to generate empirical data relevant to the concept of craving, which is crucial to many theories of drug and alcohol addiction. The predictive validity of the urge reports was evaluated using a behavioral measure--actual and suspected drug use incidents. These incidents were reported by staff members on all drug and alcohol abusers in residential treatment during a 2-month period. The results indicate that there is no significant tendency for there to be an increased use of drugs on the same days that drug urges are reported, and that drug use incidents are most likely associated with reports of no urges for drugs. Therefore, the predictive validity of the self-recording reports was not established. The data challenge the notion that drug usage is preceeded by or accompanied by (intense, driving) drug cravings at least within this setting.


Interpersonal behavior in a small group setting during the heroin addiction cycle.

In a study of social interaction during an experimental addiction cycle, male narcotic addicts expressed more hostility after higher doses of heroin. No consistent role variations were observed in dominance, friendliness, or therapeutic-orientation.


Psychopathology, craving, and mood during heroin acquisition: an experimental study.

Six detoxified addict volunteers were allowed to self-administer intravenous heroin on an essentially self-determined schedule. Two periods of heroin acquisition were compared: an unmodified cycle in which patients could become intoxicated and a later cycle in which the effects of heroin were blocked with a narcotic antagonist. In the unblocked condition, patients initially experienced an increase in positive mood, but with chronic administration there was a significant rise in psychopathology and the development of a generalized dysphoric state. Similar changes did not occur when the same patients took heroin while blocked with a narcotic antagonist. Drug craving rose dramatically when "unblocked" heroin was available, but gradually fell during methadone detoxification. Following treatment with a narcotic antagonist, the presence of heroin failed to elicit any sustained rise in craving and drug taking was dramatically reduced.


Drug interactions of the components of Optalidon after oral administration.

An investigation involving seven successive was undertaken on several groups of 10 to 14 volunteers, in order to evaluate any drug interaction between the three active components of Optalidon, namely amidopyrine (A), butalbital (B), and caffeine (C). Each component was investigated after oral administration, alone and in combination either with one of the others (i.e. A+B, B+C, C+A) or with both of the others in Optalidon (A+B+C). The plasma concentration and urinary excretion were recorded for each component as a function of time. For amidopyrine, two metabolites, amino-4-antipyrine and acetamino-4-antipyrine, were also measured in the urine. Based on a pharmacokinetic model, the following conclusions can be drawn: a) There is no change in bioavailability due to the combination of the three components in Optalidon in respect to their single administration. Within each study, there is no significant difference between the elimination rate constants, areas under the plasma concentration/time curve and percentage excreted in urine for the three components administered alone or in any combination with the other components of Optalidon. b) Concerning the absorption half-life, there is no change for amidopyrine. Only caffeine and butalbital show a statistically significant interaction in respect to this parameter and, as a consequence, differences in the time and value of the maximal plasma concentration in Optalidon. However, these differences are scarcely of anyl clinical relevance.


A contribution to the pharmacokinetics of bencyclane (Fludilat) in man.

The quantitative determination of bencyclane from the biological material was carried out with the aid of a combined microchemical method (thin-layer chromatography and measurement of fluorescence) using NBD chloride. The original method [J. Reisch, Z. Analyt. Chemie, 247 (1969) 56; J. Monforte, Clinical Chemistry 18 (1972) 1329; R.S. Fager, Anal. Biochemistry, 53 (1973) 290, etc.] was so modified as to enable attainment of optimal results in respect of sensitivity and accuracy in the determination of bencyclane. The sensitivity of this modified method is 0.1 mug/ml plasma. Volunteer subjects and patients received under standard conditions 2 coated tablets Fludilat (i.e. 200 mg bencyclane hydrogen fumarate) orally as a single dose or repeated 3 times daily over 5 days, or 4 ampoules (= 200 mg) in a single intravenous injection. After a single oral administration, maximum plasma concentrations of approximately 2 mug/ml were attained in about 2 hours. The elimination half-life was about 360-480 min. The appearance of a second peak after about 6-7 hours indicates involvement of several compartments. On intravenous administration, maximum plasma concentrations of above 2 mug/ml were attained. A second peak in the late phase of the elimination was also detected here. The repeated oral administration led to maximum plasma concentrations of above 3 mug/ml without there being any indication of accumulation. Protein binding of about 30% was determined with the aid of the equilibrium dialysis method. A parallel "in vitro" study with 14C-bencyclane (U.R. Kleeberg, 1973, unpublished) showed an approx. 40% protein binding, an approx. 30% erythrocyte binding, and an approx. 10% thrombocyte binding. About 20% bencyclane remain free.


[Elimination kinetics of phenprocoumon (Marcumar) in liver cirrhosis and after premedication with phenobarbital].

In this paper are presented the results of a study on the elimination kinetics of phenprocoumon in patients concomitantly treated with phenprocoumon and phenobarbital as well as in patients with bioptically proved hepatic cirrhosis. The pre-treatment of the patients with phenobarbital (3 X 100 mg), 4 days before phenprocoumon was started, resulted in an accelerated elimination of phenprocoumon. The biological half-life of phenprocoumon was reduced from 150 to 70 hrs when compared to normal controls, and the elimination constant was found to increase to 9,96 X 10(-3). Phenobarbital at plasma concentrations achieved by usual therapeutical dosage schedules of the drug does not interfere with the protein binding of phenprocoumon as could be shown by equilibrium dialysis. Phenobarbital, therefore, does not accelerate the elimination of phenprocoumon. The inducing effect of phenobarbital on the drug metabolizing enzyme system in the liver results in more rapid biotransformation of phenprocoumon. In patients with bioptically proved hepatic cirrhosis, the elimination kinetics did not differ from the results obtained in normal controls.


Hemodynamic effects of thiothixene and chlorpromazine in schizophrenic patients at rest and during exercise.

The hemodynamic effects and plasma levels of noradrenaline were studied in schizophrenic patients at rest and during exercise after long-term treatment with chlorpromazine (150-600 mg daily) and thiothixene (60-80 mg daily). The results are compared with those from previous studies in untreated patients and patients receiving very large doses of chlorpromazine. The effects of thiothixene on the different hemodynamic variables were very moderate, and the observed differences between this group and the control group may be due to the different patient materials. In the two groups of patients receiving chlorpromazine, the heart rate at rest and durng exercise tended to be higher than in the control group. There was also a tendency towards a lower stroke volume after this drug and thiothixene during exercise. The noradrenaline levels in plasma were highest after the high dose of chlorpromazine both at rest and during exercise, while they were lower after the moderate chlorpromazine dose. After thiothixene, the values were between those of the group on the low chlorpromazine dose and those of the control group.


Effects of xipamide and triamterene - alone or in combination - on electrolyte excretion in the rat.

The effects of X and T in different doses alone or in combination on renal sodium and potassium excretion were examined in conscious rats. In acute experiments, the combination of both diuretics produced an approximately additive effect on sodium excretion. Addition of T to X in different dose relations (X:T = 1:0.5, 1:1 or 1:1.5) prevented the X-induced potassium loss. These effects were more pronounced at the highest dose of T; the difference to the other combinations, however, was small. In chronic experiments, when the diuretics were administered daily for 13 days, both combinations (X:T = 1:1 and X:T = 1:1.5) resulted in a nearly equilibrated potassium balance at the end of this period. In shorter intervals, however, sometimes considerable oscillations in potassium excretion occured.


Contact allergy to corticosteroids.

Contact dermatitis to topical corticosteroids does occur, although infrequently. Patch testing to the available steroids is recommended for patients with long-standing dermatoses which have responded poorly to topical steroids.


The haem-accessibility in leghaemoglobin of Lupinus luteus as observed by proton magnetic relaxation.

Using the solvent-protons' longitudinal magnetic relaxation rates (p.m.r.) for Lupinus luteus leghaemoglobin derivatives the accessibility of the haem has been evaluated by our "stereo-chemical p.m.r. titration" method with nonexchangeable protons of aliphatic lower alcohols in otherwise deuterated solutions. The haem in leghaemoglobin is more accessible and its protein environment more flexible compared with vertebrate haemoglobins. The correlation time in aquometleghaemglobin aqueous solution has been determined by measuring the frequency dispersion of the p.m.r. rates between 6.1 and 93 MHZ. Taking into account the measured value of tauc = (7.7 +/- 0.5 x 10(-10) s the iron-to-proton inter-spin distances have been calculated. The significance of these distances as well as the electronic g-factor anisotrophy for elucidation of fine structural details of the haem-environment are discussed.


A square pyramidal model for ribonuclease action.

A mechanism incorporating a square pyramidal model (SP) is presented for the action of bovine pancreatic ribonuclease. Its formulation is based on structural principles governing pentacoordinate behavior. The model is compared with a previous trigonal bipyramidal (TP) representation with regard to the geometry of the active site and enzyme constraints. Of two variants of the SP model, an adjacent (cis displacement) and in-line (trans displacement) process, the in-line mechanism, as with the TP model, fits existing model studies. Consideration of the energetics of the SP vs. the TP model leads to an estimated energy difference of about 1-2 kcal/mol. This suggests that the preferred model may be intermediate in geometry between the two idealized representations for the enzymatic hydrolysis. Comparisons are made showing that pseudorotation is an unlikely process in either model.


Retro analogs related to oxytocin.

[D-alle3]-retro-D-deaminotocinamide (I), retro-L-deaminotocinamide (III) and their respective N-formyl derivatives (II and IV) were synthesized by the stepwise active ester method: deaminotocinamide was prepared by the solid-phase method. The retro-analogs of deaminotocinamide, tested at concentrations up to 10)-5)M, were found to be without activity as agonists or antagonists in the oxytocic assay. At 10(-4)M,[D-alle]-retro-D-deaminotocinamide is a weak competitive inhibitor of oxytocin. [D-alle3, Gly7]-retro-D-deaminooxytocin (V) was synthesized either by the active ester method or by a fragment condensation method employing the retro-D-RING, [D-alle3]-retro-D-deaminotocinamide, and D-tail, Boc-Gly-D-Leu-Gly, as the fragments...


Chemical study of the topography of porcine lutropin (LH) using dinitrofluorobenzene and dansyl chloride.

Treatment of porcine lutropin beta-subunit by increasing amounts of dansyl chloride shows that only one fluorescent group can be bound to the free subunit, namely on tyrosine beta-37. This modification prevents reassociation with native chi-subunit. In contrast to dansyl chloride, dinitrofluorobenzene reacts preferentially with tyrosine beta-59. This substitution does not interfere with the reassociation with the native chi-subunit. By using equimolar ratio of dansyl chloride and porcine lutropin chi-subunit it is possible to modify this subunit on a single site, which is found to be a tyrosine residue. The monodansylated X-subunit is still able to recombine with native B-subunit but its fluorescence is found to be markedly quenched upon binding. In addition, the O-dansyl-tyrosyl fluorescence quenching by potassium iodide is more effective on the recombined dimer than on the free chi-subunit. Both reconstituted dimers (native xhi chi dinitrophenylated beta and dansylated chi chi native beta) are without biological activity. It is not clear whether the substituted phenolic groups are essential or whether the added groups prevent the chi beta dimer from taking the active conformation. This alternative is discussed in the light of recent data from this laboratory and others.


The phosphopyridoxyl peptide from the mitochondrial aspartate aminotransferase of beef kidney.

The sequence around the coenzyme-binding lysine of mitochondrial aspartate amino-transferase from beef kidney was determined. The holoenzyme was treated with NaB3H4 and digested with thermolysin; the labelled peptide was isolated and its sequence proved to be identical with sequences around the coenzyme site from the same isoenzyme of different organs and animals (pig heart and sheep liver). The sequences of the phosphopyridoxyl peptides of the mitochondrial aspartate aminotransferases appear to be closely related to the corresponding peptides from the cytoplasmic isoenzymes.


Synthesis of peptides and derivatives of 3- and 4- hydroxyproline.

Synthesis and properties are reported for a number of peptides and related derivatives of 3-hydroxy-L-proline and 4-hydroxyl-L-proline. These were made for several different purposes, namely, sythesis of N-acetyl-O-tosyl-4-hydroxy-L-proline amide as a model for chemical reduction of peptide-bound hydroxyproline, synthesis of tripeptides with the sequence glycyl-4-hydroxy-L-hydroxy-L-prolyl-X, synthesis of the naturally occurring sequence, glycyl-3-hydroxy-L-prolyl-4-hydroxy-L-proline, and monomers for polymerization to yield the sequence -glycyl-prolyl-4-hydroxy-L-prolyl-.


Late results following substitution of colon for the stomach, esophagus, urinary bladder and vagina.

The transverse colon was substituted for the stomach in four patients after total gastrectomy for cancer. Three of them are alive after 21 to 23 years. They are in good condition and fully active. In six patients the esophagus was resected and the colon was interposed transthoracically. Two patients survive 13 to 15 years later. Carcinoma of the urinary bladder was resected three times and colon substituted. In a case of total vaginal aplasia, sigmoid colon was substituted for the vagina. The patient is alive after 17 years and has a normal sexual life. Sixteen patients have had colon transplantations to replace various organs. Seven of them are alive. The average time of survival thus far is 19 years.


A ten-year experience in the evaluation and management of patients with stress urinary incontinence.

The surgical procedures used, the complications encountered and the results obtained in 549 patients with urinary stress incontinence with or without prolapse and 50 patients with prolapse without urinary stress incontinence are presented. Incontinence was cured in 347 patients, improved in 126, unchanged in 66 and worsened in ten. Seven patients operated on for uterine prolapse developed urinary incontinence after surgery. The overall recurrence of SUI was 12.75%. The introduction of suprapubic bladder drainage has practically eliminated postoperative urinary tract infections and reduced the length of hospitalization from 9.1 to 7.2 days. My experience in 214 patients with suprabpubic drainage demonstratedthe superiority of the Ansari method over the cystocath. The addition of Cantor's bladder neck plication improved the results (cured plus improved) from 80% to 100% in the Marshall-Marchetti-Krantz operation and from 81% to 86% when the Marshall-Marchetti-Krantz operation was associated with an abdominal hysterectomy.


Polycystography in the evaluation of carcinoma of the urinary bladder.

Polycystography is a safe, simple, useful, radiological procedure to assess the degree of infiltration of bladder tumors. It broadly classifies bladder tumors into two groups: infiltrating and noninfiltrating. This information is important to plan the treatment. In 96% of the cases, the radiological findings correlated well with other methods of assessing the degree of infiltration. The standard technic, modifications, advantages and disadvantages of this procedure are discussed.


Treatment of lithiasis by the forced injection of liquid and ureteral catheterization by the translumbar route.

We have used translumbar intrapelvic injection of liquid and intraureteral sliding of a catheter to push ureteral calculi and obtain their elimination through the bladder in 32 patients. We have described the technic and the results obtained. We had more success with the intraureteral catheter technic. No complications were observed in our cases. Ureteral physiology in relation to the favorable results obtained is discussed.


Effect of cataract on intraocular pressure.

Cases of immature and mature cataract were selected to study normal values in relation to hypermature cataract. We observed that cases of hypermature cataract had a high intraocular tension which was associated with a high protein level and increase in the number of macrophages in the aqueous. We concluded that a phacolytic mechanism is responsible for a secondary rise of intraocular tension in cases of hypermature cataract.


Candida sepsis successfully treated by parenteral administration of 5-fluorocytosine.

Candida sepsis has become one of the most common and dangerous forms of hospital acquired infection. The recommended drug for parenteral treatment of Candida sepsis is amphotericin B, however, its toxic effects preclude its usage in many patients, particularly in the presence of renal failure. A less toxic antifungal agent is 5-fluorocytosine. A patient with Candida albicans sepsis was treated successfully with 5-fluorocytosine by intravenous administration. The fungal infection developed during the course of acute renal failure, repeated surgical intervention, intravenous hyperalimentation, gastrointestinal bleeding and five months of antibiotic therapy. The clinical symptoms receded rapidly and cultures became sterile after one week of intravenous treatment. The predisposing factors, difficulties in prevention and diagnosis of fungal infection are discussed.