Progesterone concentration in placenta, molar tissue, and ovarian theca lutein cyst.

Progesterone concentrations in 2 full-term normal placentas, molar tissue from 3 cases of hydatidiform mole were measured by competitive protein binding. Sephadex LH-20 column chromatographic separation of extracts from these tissues showed that, in placental and molar tissue, 85 to 95% of the extracts were progesterone and very small amounts of 20 alpha-dihydroprogesterone and 17 alpha-hydroxyprogesterone. The concentration of progesterone in the placenta were 368.8 and 317.2 ng/g tissue, respectively, while in molar tissue the concentration were 2474.5, 1974.6, and 4146.0 ng/g tissue, respectively. Progesterone concentration in ovarian tissue was between 1121;2 and 1440.9 ng/g tissue. It is suggested that the high concentration of progesterone in molar tissue reflects functional capacity of the abnormal trophoblast in progesterone synthesis and accumulation of progesterone due to absence of a fetus. The lower concentration of progesterone in the ovary in molar pregnancy would appear to suggest that the ovary is a secondary source of progesterone in molar pregnancy.


Cytomegalovirus endometritis: report of a case associated with spontaneous abortion.

Maternal cytomegalovirus infection is a relatively common, yet very often silent complication of pregnancy. The association between early gestational wastage and cytomegalovirus endometritis has been documented in recent tissue culture studies without the morphologic demonstration of the virus. This case study presents the clinical and pathologic findings of a young, secundigravida female with cytomegalovirus endometritis and spontaneous abortion. The intranuclear inclusions were restricted to the endometrial glands; Involvement of the endocervix by cytomegalovirus has been the only site in the female reproductive tract, excluding fetal tissues, affected by this virus which has been demonstrated histologically. The data are still incomplete regarding the etiologic role, if any, of cytomegalovirus and the occurrence of spontaneous abortion.


Ultrastructural study of focal epithelial hyperplasia.

Biopsy material from two young Israeli patients with focal epithelial hyperplasia revealed acanthosis, papillomatosis, and the presence of many epithelial cells with a clear cytoplasm in the basale and stratum spinosum. Many of the inclusions seen with the electron microscope had the morphologic appearance of lysosomes, but definite identification must await cytochemical evidence.


Ultrastructure of an odontogenic myxoma.

The ultrastructural findings in a case of odontogenic myxoma are described. The main cell type was characterized by several cytoplasmic processes, intracytoplasmic fibrils, numerous glycogen particles, and salient Golgi complexes. A few mitochondria and a scarce endoplasmic reticulum was noted. The matrix consisted of numerous granules with fibrillar projections, collagen bundles, and smaller fibers. The over-all ultrastructural features of this tumor were similar to those described in the human cardiac myxoma and in Wharton's jelly and could be clearly differentiated from the fine structural characteristics of other connective tissue tumors.


Symposium on sensorineural hearing loss in children: early detection and intervention. Impedance measurement.

Although impedance measurement is a relatively new procedure, at least in this country, it is rapidly being recognized as one of the most significant contributions since the development of the pure tone audiometer. With the basic applications of the procedure it is possible to determine not only the presence but also to a great extent the specific nature of middle ear impairment, even in patients for whom audiometric results are limited and uncertain. Further applications of impedance measurement include the determination of nonorganic hearing loss and site-of-lesion testing to distinguish between cochlear and retrocochlear lesions. Finally it appears that the procedure may even permit the prediction of severity and configuration of sensorineural loss. Although impedance measurement has great value in all patients with hearing disorders, it is particularly appropriate in pediatric evaluation. Its objectivity, its ease of application, and the information that can be obtained with it make impedance measurement a significant and valuable addition to the battery for evaluation of hearing disorders in children.


Bioregulation of lysosomal enzyme secretion from human neutrophils: roles of guanosine 3':5'-monophosphate and calcium in stimulus-secretion coupling.

The biologic role of calcium and guanosine 3':5'-monophosphate (cyclic GMP) in the immunologic secretion of lysosomal enzymes from human neutrophils was studied. Contact of neutrophils with zymosan-treated serum or the divalent cation ionophore A-23187, in the presence of extracellular calcium, resulted in beta-glucuronidase (beta-D-glucuronide glucuronosohydrolase, EC 3.2.1.31) secretion and a concomitant accumulation of cyclic GMP without any loss of cell viability. Acetylcholine (0.1 muM), in the presence of calcium, enhanced the immunologic stimulation of cyclic GMP accumulation and lysosomal enzyme discharge. A marked and rapid association of 45CaCl2 with neutrophils occurred during cell surface contact with zymosan-treated serum, and this effect on calcium association was enhanced with 0.1 muM acetylcholine. The precise mechanism of the neutrophil-calcium interaction is presently not well understood. However, the finding that 0.5-1.0 muM A-23187 also provoked a rapid association of extracellular calcium with neutrophils suggests that calcium mobilization into the intracellular environment may account, at least in part, for this association between cells and calcium. The close temporal relationship between beta-glucuronidase secretion, cyclic GMP accumulation, and calcium mobilization during cell contact with membrane active agents such as immune reactants, acetylcholine, and ionophores suggests that these three cellular events bear a cause and effect relationship. On the basis of our findings to date, we propose that the immunologic secretion of lysosomal contents from human neutrophils is signaled by intracellular cyclic GMP and that extracellular calcium, by gaining access to the intracellular processes responsible for cyclic GMP accumulation, serves as the link to stimulus-secretion coupling.


Inhibition of replication of normal adrenocortical cells in culture by adrenocorticotropin.

Adrenocorticotropic hormone (ACTH) inhibited [3H]thymidine incorporation in normal adrenocortical cells of adult rats in culture, with a concomitant increase in corticosterone production and a characteristic retraction of cells. Both dibutyryl cyclic AMP and an analog of ACTH, which produces virtually no cyclic AMP, inhibited DNA synthesis and stimulated steroid production. ACTH inhibited the proliferation of adrenocortical cells obtained from suckling rats as well as the cells obtained from the capsular tissue of adult rat adrenal glands, whereas insulin caused a stimulation of DNA synthesis. These results suggest that the major role of ACTH is to induce the transformation of the undifferentiated cells of the adrenal gland into functional fasciculata cells and that the proliferation of adrenocortical cells may be under control of factors other than ACTH.


An electron microscopic study of virus particles in rhesus monkey placenta.

Examination of chorionic villi from rhesus monkey placenta revealed the presence of C-type virus particles budding from syncytial trophoblast, pericytes, Hofbauer cells, and mesenchyme. In addition, particles, were found budding from cells of the cytotrophoblastic cell column and decidual basalis. They measured 30 nm in diameter, had a dense central core, surrounded by a narrow, electron-lucent zone, and were enclosed by an outer unit membrane.


Biosynthesis of polypeptides of cytochrome c oxidase by isolated mitochondria.

Yeast mitochondria, incubated with radioactive amino acids in a "protein-synthesizing mixture" containing an oxidizable substrate and an ATP regenerating system, have been shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to incorporate label into polypeptides equivalent in molecular weight and relative amount ot those made in vivo in the presence of cycloheximide. The ability of these isolated mitochondria to synthesize "native" polypeptides was assessed by examining the incorporation of label into subunits of cytochrome c oxidase (EC 1.9.3.1). An analysis of immunoprecipitates formed by incubating cholate extracts of labeled mitochondria with an antiserum against holocytochrome c oxidase revealed that label was incorporated into three polypeptides of sizes equivalent to those of cytochrome c oxidase subunits I, II, and III, shown from earlier studies in vivo to be translated on mitochondrial ribosomes. Further evidence that these polypeptides made in vitro are "native" and identical to subunits I, II, and III was provided by the observation that labeled polypeptides equivalent in size to subunits I-III- ARE ALSO IMMUNO-PRECIPITATED BY ANTISERUM AGAINST SUBUNITS V plus VII, an antiserum that can precipitate subunits I, II, and III only when they are complexed to the cytoplasmically synthesized subunits, V and VII, of the enzyme. These results suggest that isolated mitochondria are capable of synthesizing three subunits of cytochrome c oxidase and assembling them into a holoenzyme.


Cyclic AMP-dependent endogenous phosphorylation of a microtubule-associated protein.

Microtubules prepared from chick brain homogenates by successive cycles of assembly-disassembly were found to contain two high-molecular-weight proteins, designated microtubule-associated protein1 and microtubule-associated protein2. Microtubule-associated protein2 (apparent molecular weight 300,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was the preferred substrate for an endogenous cyclic AMP-dependent protein kinase which appeared to be an integral component of the microtubules. The initial rate of phosphorylation of microtubule-associated protein2 was enhanced 4- to 6-fold by cyclic AMP, with half-maximal stimulation occurring at 2 times 10-7 M cyclic AMP. Under optimal conditions, a total of 1.0 and 1.9 mol of phosphate was incorporated per mole of microtubule-associated protein2, in the absence and presence of cyclic AMP, respectively. Cyclic AMP also stimulated the phosphorylation of tubulin, but the rate of phosphate incorporation per mol of tubulin was only 0.15% that of microtubule-associated protein2. The data raise the possibility that the cyclic AMP-dependent phosphorylation of microtubule-associated protein 2 may play a role in microtubule assembly or function.


Interferon action II. Membrane-bound alkaline ribonuclease activity in chick embryo cells manifesting interferon-mediated interference.

Membrane fractions from chick embryo cells manifesting viral interference mediated by interferon or poly(I)-poly(C) contain high levels of an alkaline ribonuclease. Enhanced RNase activity is not observed when inhibitors of cell protein or RNA synthesis are present during interferon treatment, or when heterologous interferon is used. The RNase associated with comparable membrane fractions from cells treated with mock-interferon is about 1/10 as active, and shows qualitative differences. In principle, divergent views of interferon action may be reconciled to a common mode of action by postulating that viral interference results from a newly induced or activated RNase of cellular origin and proper specificity that acts to reduce the accumulation and functional capacity of newly synthesized viral RNAs, particularly mRNA. Previous data in support of interferon's acting to inhibit virion-derived transcription in vivo are now interpreted as demonstrating enhanced degradation of viral transcripts (mRNA).


Natural occurrence of poly(ADP-ribosyl) histones in rat liver.

Poly(ADP-ribose) bound to histones has been isolated from rat liver. When [14C]ribose was administered intraperitoneally to rats at a dosage of 300-750 mug (100-250 muCi)/10o g, approximately 1% of the radioactivity was recovered in the acid (5% CLCCCOOH)-INSOLUBLE MATERIAL OF THE LIVER NUCLEI 2 HR AFTER INJECTION. Of the acid-insoluble radioactivity, 4.5-9% was extractable with 0.25 N HCL. Carboxymethyl-cellulose column chromatography of the HCl-extracted material revealed that the radioactivity cochromatographed with histone subfractions f1 and, to a lesser extent, f2 and f3. Part of the protein-bound radioactivity was rendered acid-soluble by treatment with either snake venom phosphodiesterase or neutral NH2OH. From the enzyme digest, 5'-AMP and psiADP-ribose [2'-(5"-phosphoribosyl)-5'-AMP] were recovered, while the NH2OH treatment yielded ADP-ribose monomer and, presumably, oligomer. These observations indicate that ADP-ribose is attached to histones in vivo and is present both as a monomer and a polymer.


The antagonistic effect of an inhalation anesthetic and high pressure on the phase diagram of mixed dipalmitoyl-dimyristoylphosphatidylcholine bilayers.

Several workers have shown that phase transition-related changes in membrane lipids have a profound effect on membrane-solvated protein function. This phase transition temperature dependence has been explained as resulting from the formation of lateral phase separations within the membrane bilayer. The present study demonstrates that a clinical concentration of an inhalation anesthetic produces changes in both the phase transition temperature of pure lipid bilayers and the lateral phase separation temperature of mixed dipalmitoyl- and dimyristoylphosphatidylcholine bilayers of a magnitude sufficient to influence protein function. It is further shown that pressure is able to antagonize the effect of the anesthetic on these transition temperatures. It is proposed that anesthetic action within nerve membranes may be the result of changes in the lateral phase separation-controlled environment of the membrane-solvated proteins essential to nerve function.


Translation and identification of the mRNA species synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus.

Vesicular stomatitis virus messenger RNA has been transcribed in vitro from the viral genome by the virion-associated RNA polymerase in quantities suitable for translation. Wheat germ cell-free extracts programmed with the isolated in vitro 12-18S RNA fraction synthesize polypeptides similar to the viral N, NS, and M proteins, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and tryptic peptide mapping of the in vitro products and the viral marker polypeptides. In addition, the RNA synthesized in vitro also codes for a protein of molecular weight 63,000 which may be a nonglycosylated form of the viral glycoprotein G. The 12-18S RNA has been partially separated into individual messenger species and these have been identified by the proteins for which they code. There are four monocistronic messenger species in the in vitro 12-18S RNA and the coding capacity of three of these molecules agrees with the estimated molecular weight of the polypeptide assigned to it.


Methylated nucleotides block 5'-terminus of vaccinia virus messenger RNA.

Studies on the nature and location of the methylated nucleotides in mRNA synthesized in vitro by vaccinia virus particles revealed an unusual 5'-terminal structure. Evidence that the pyrophosphate group is blocked by 7-methylguanosine and that both 2'-O-methyl-adenosine and 2'-O-methylguanosine occupy penultimate positions was presented. According to this model, the 5'-termini of vaccinia virus mRNAs are: 7MeG-5'ppp-5'GMepNp and 7MeG-5'AMepNp.


Competent Bacillus subtilis cultures synthesize a denatured DNA binding activity.

A protein activity has been detected in soluble extracts of competent cultures of bacillus subtilis which protects denatured DNA from nuclease solubilization. Both binding activity and competence development occur simultaneously but not coordinately, since the peak of binding activity appears 3 hr after the peak of competence. The binding activivity is not detected in physiologically noncompetent cultures, in an isogenic noncompetent cultures, in an isogenic noncompetent mutant grown through the competence regimen, or in early sporulating cells. The activity is included on Sephadex G-100 gel filtration columns but elutes in a broad nonhomogeneous peak. This binding activity may play a role in uptake of DNA and/or integration of exogenous DNA into the genome of competent B. subtilis cells.


Mechanism of action of cholera toxin and the mobile receptor theory of hormone receptor-adenylate cyclase interactions.

Rat liver membrane adenylate cyclase (EC 4.6.1.1) that has been stimulated more than 10-fold by cholera toxin (choleragen) has a 3-fold greater sensitivity to stimulation by glucagon. Choleragen similarly increases the sensitivity of cyclase to other peptide (ACTH, vasoactive intestinal polypeptide) and nonpeptide (catecholamines) hormones in this and other tissues. The rate of 125I-labeled glucagon-membrane dissociation is decreased about 2-fold in toxin-treated liver membranes. Toxin-activated cyclase activity of fat cell membranes is retained upon solubilization with Lubrol PX. Provided 125I-labeled choleragen is first incubated with cells under conditions resulting in enzyme activation, the solubilized cyclase activity migrates with a component of 125I-labeled choleragen on gel filtration chromatography. Agarose derivatives containing the "active" subunit (molecular weight 36,000) of the toxin can specifically adsorb solubilized adenylate cyclase. Toxin-stimulated cyclase can be immunoprecipitated with antitoxin or anti-"active" subunit antibodies. There is a large excess of membrane receptors (ganglioside GM1) which, with the use of choleragenoid, can be shown to be functionally equivalent with respect to cyclase activation. Choleragenoid, an inactive competitive antagonist of toxin binding, can occupy and block a large proportion of toxin receptors without affecting toxin activity. A scheme of toxin action is proposed that involves lateral membrane diffusion of the initially inactive toxin-receptor complex with subsequent direct interaction with and modulation of adenylate cyclase. The basic features of this scheme may be pertinent to the mechanisms by which hormone receptors normally modulate adenylate cyclase.


Effect of a low-molecular-weight DNA binding protein, H1 factor, on the in vitro transcription of the lactose operon in Escherichia coli.

H1 protein, a heat-stable low-molecular-weight DNA-binding factor previously described by Cukier-Kahn et al. [Proc. Nat. Acad. Sci USA (1972) 69, 3643-3647] markedly stimulates in vitro synthesis of lac-specific RNA directed by bacteriophage lambdah80 dlac or phi80 dlac DNA templates in the presence of purified E. coli RNA polymerase holoenzyme. The extent of stimulation obtained by addition of H1 alone is usually greater than that observed with the cAMP receptor protein-cAMP combination. H1 effect varies quite appreciably (from 4- to 16-fold) with the functional state of the promoter, being much larger with lambdah80 dlac p-s, a transducing DNA carrying a superpromoter mutation, than with lambdah80 dlac p+. H1 and cAMP receptor protein effects are nearly additive, although interpretation of the data obtained at high H1 concentration is complicated by the appearance of some inhibitory property. While the cAMP-receptor-protein-mediated synthesis is asymmetrical ("I" strand almost exclusively copied), the degree of asymmetry observed with H1 is less pronounced, suggesting asymmetrical copying from the lac promoter and symmetric transcription from other regions of the DNA. Synthesis of lac-specific RNA from lambdah80 dtrp/lac or phi80 dlac p-r uv5 templates, in which lac promoters are insensitive to cAMP receptor protein, either as a result of lac fusion to the trp operon or mutation in the lac promoter, is totally H1-insensitive. Glycerol (10-15% w/w) can fully substitute for H1 in stimulating lac RNA synthesis in a fashion analogous to that reported for the cAMP receptor protein-cAMP system. The possibility that H1 acts by causing conformational modifications at the promoter level in a way that increases its functional state, and that this effect is more pronounced with operons sensitive to cAMP receptor protein, is discussed.


Fate of viral RNA of murine leukemia virus after infection.

[3H]Uridine-labeled Rauscher leukemia virus was used to infect mouse embryo fibroblasts. After the infected cells were separated into nuclear and cytoplasmic fractions nucleic acid was extracted by sodium dodecyl sulfate-phenol-chloroform treatment and analyzed by Cs2SO4 and sucrose density gradient centrifugation. Between 45 and 70 min after infection a transient and synchronized shift of the acid-insoluble radioactive peak toward the RNA-DNA hybrid region occurred in both the nuclear and cytoplasmic fractions. The density of the cytoplasmic hybrid shifted to 1.56 g/ml (RNA equals about 50%), while the sedimentation rate decreased from 36 S to 14 S; however, the density of the nuclear hybrid shifted to 1.58-1.48 g/ml (RNA equals 57-17%, respectively), while its sedimentation rate remained about 65 S. The hybrids in both the nuclear and the cytoplasmic fractions still showed hybrid density after heat denaturation. The processes of the early stages of RNA tumor virus infection are discussed with regard to the functions of viral RNA-dependent DNA polymerase (reverse transcriptase) and a possible integration of viral genetic information into the host chromosome.


The structure of a yeast hexokinase monomer and its complexes with substrates at 2.7-A resolution.

From a 2.7-A resolution electron density map we have built a model of the polypeptide backbone of a monomer of yeast hexokinase B (EC 2.7.1.1). This map was obtained from a third crystal form of hexokinase, called BIII, which exhibits space group P212121 and which contains only one monomer per asymmetric unit. The 51,000 molecular weight monomer has an elongated shape (80 A by 55 A by 50 A) and is divided into two lobes by a deep central cleft. The polypeptide chain is folded into three structural domains, one of which is predominantly alpha-helical and two of which each contain a beta-pleated sheet flanked by alpha-helices. Both glucose and AMP bind to these crystals and produce significant alterations in the protein structure. Glucose binds in the deep cleft, as was observed previously in the BII crystal of the dimeric enzyme. AMP, however, binds to a site that is different from the major intersubunit ATP binding site observed in the crystalline dimer. The AMP is found near one of the beta-pleated sheets. From our current interpretation of this electron density map we conclude that neither of the two nucleotide binding regions has the same structure as has been observed for the nucleotide binding regions of the dehydrogenases, adenylate kinase, and phosphoglycerate kinase, although some similarities exist.


Spontaneous and induced mutagenesis in Western Equine encephalomyelitis virus in chick embryo cells with different repair activity.

Evidence was obtained indicating differences in the survival rate of Western equine encephalomyelitis virus after exposure to ultraviolet radiation and methyl methanesulfonate in commercial and leukosisfree chick embryo cells that differed in repair activity. The levels of spontaneous mutagenesis (on the basis of the yield of small palque variants of the encephalomyelitis virus) did not essentially change when the virus was passage in leukosis-free chick embryo cells, whereas an increase in the number of small palque variants was observed in the cells of commercial chick embryos. A 10-fold increase in the number of induced virus variants was observed in commercial chick embryo cells in experiments with methyl methanesulfonate as compared with the contorl, whereas the induction of virus variants was not noted in leukosis-free cells.


Light-stimulated phosphorylation of rhodopsin in the retina: the presence of a protein kinase that is specific for photobleached rhodopsin.

A protein kinase has been extracted from bovine rod outer segments by a mild procedure. The enzyme acts specifically on photobleached, not unbleached, rhodopsin and will not catalyze the phosphorylation of histones, phosvitin, or casein. We propose the name "opsin kinase" for the enzyme, which is not affected by cyclic nucleotides but which is inhibited by theophylline. Preparations of purified rod outer segments, however, appear to contain only low concentration of opsin phosphatase activity.


Cobalt(III), a probe of metal binding sites of Escherichia coli alkaline phosphatase.

To facilitate the study of individual metal binding sites of polymeric metalloproteins, conversion of exchange-labile Co(II) in E. coli alkaline phosphatase (EC 3.1.3.1) to exchange-inert Co(III) was examined. Oxidation of Co(II) alkaline phosphatase with hydrogen peroxide results in a single absorption maximum at 530 nm and loss both of the characteristic electron paramagnetic signal and of enzymatic activity. Zinc neither reactivates this enzyme nor displaces the oxidized cobalt atoms. Metal and amino-acid analyses demonstrate that oxidation alters neither cobalt binding nor amino-acid composition of the enzyme. Al data are consistent with the conclusion that hydrogen peroxide oxidizes Co(II) in alkaline phosphatase to Co(III). Polymeric metalloenzymes can contain different categories of metal atoms serving in catalysis, structure stabilization, and/or control and exerting their effects independently or interdependently. The in situ conversion of exchange-labile Co(II) to exchange-stable (Co(III) offers a method to selectively and differentially "freeze" cobalt atoms at their respective binding sites. The accompanying spectral changes and concomitant retardation in ligand exchange reactions may be used to differentiate between specific metal binding sites that serve different roles in polymeric metalloenzymes.


Changes of tissue water proton relaxation rates during early phases of chemical carcinogenesis.

Water proton spin lattice relaxation rate (T1) was determined on tissues of rats experiencing early phases of chemical carcinogenesis. Rats were fed a fast acting carcinogen, 3'-methyldimethylaminoazobenzene, and a slower acting carcinogen, 2-acetylaminofluorene, for up to 4 weeks. T1 of blood serum and liver tissue was significantly higher than those of controls after 4 weeks of 3'-methyldimethylaminoazobenzene feeding. This was not the case for 2-acetylaminofluorene. The blood serum T1 increase reflected the onset of liver nodulation (assumed to be preneoplastic). Liver T1 values increased as the degree of nodulation increased. Blood serum T1 correlated inversely with protein content and directly with water content. Liver T1 values correlated with water content, but this was not true for spleen T1 values. Spleen T1 values were significantly lower than controls at the earliest sampling date for each carcinogen: one week for 3'-methyldimethylaminoazobenzene and 4 weeks for 2-acetylaminofluorene. The spleen T1 decrease paralleled an increase of iron detectable by electron spin resonance in this tissue. Spleen T1 decreases are probably not unique to chemical carcinogenesis.


Genetic heterogeneity in xeroderma pigmentosum: complementation groups and their relationship to DNA repair rates.

Fibroblast strains from 12 patients with xeroderma pigmentosum had lower than normal rates of DNA repair, as determined by autoradiographic studies of ultraviolet-induced unscheduled nuclear DNA synthesis. The nuclei in binuclear cells, obtained by fusing fibroblasts from certain pairs of these strains, had a greater rate of DNA repair than the nuclei of either strain's unfused mononuclear cells. These results indicate that complementary corrections of the strains' repair defects had occurred in the fused cells. Four complementation groups were found, indicating that at least four mutations caused decreased DNA repair among these 12 strains. The unfused mononuclear cells of each group had a characteristic rate of repair that differed from the rates of the other groups.


Identification of a calcium-binding protein as a calcium-dependent regulator of brain adenylate cyclase.

An activating factor of adenylate cyclase (EC 4.6.1.1) HAS BEEN OBTAINED FROM DETERGENT-DISPERSED PREPARATIONS OF PORCINE CEREBRAL CORTEX BY COLUMN CHROMATOGRAPHY ON ECTEOLA-cellulose. The factor was identified by acrylamide gel electrophoresis and by enzyme activation studies as the Ca2+-binding protein that regulates the activity of a brain cyclic nucleotide phosphodiesterase. This Ca2+-binding protein confers a Ca2+-dependent activation upon the adenylate cyclase, which is reversed by the subsequent addition of egta in excess of the free Ca2+. It is proposed that this Ca2+-dependent regulator controls enzymatic activities responsible for the synthesis of adenosine 3':5'-monophosphate and for the hydrolysis of guanosine 3':5'-monophosphate.


A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules.

A protein-free nucleic acid preparation method for electron microscopy is described. The basic procedure is very similar to the classical protein monolayer spreading techniques. The carrier protein (usually cytochrome c) is replaced by benzyldimethylalkylammonium chloride. Both the hypophase method and the microdiffusion or droplet method can be applied with this compound. Unlike cytochrome c, benzyldimethylalkylammonium chloride does not lead to any apparent thickening of the nucleic acid strands. Partially denatured DNA spread with this reagent shows a loosened structure with a foamy appearance in the regions previously considered to be "unmelted," which open up locally into melted loops of different size. Specifically bound proteins, such as RNA polymerase on bacteriophage T7 DNA, can be detected unambiguously.


Differentiation in erythroleukemic cells and their somatic hybrids.

Clones of erythroleukemic cells differ in the extent to which they (1) undergo differentiation spontaneously and (2) can be induced to differentiate in the presence of dimethylsulfoxide. Here we demonstrate that relative differences in globin gene expression within and between clones largely reflect differences in the proportion of cells participating in differentiation rather than uniform differences in the extent to which all cells in these clones undergo differentiation. We call this phenotype of a clone its characteristic probability of differentiation, a property that reflects the likelihood that a cell of this clone will undergo erythrodifferentiation under given conditions. We have examined somatic hybrid cells formed between similar erythroleukemic lines, between phenotypically different erythroleukemic lines, between phenotypically different erythroleukemic lines, and between erythroleukemic cells and mouse fibroblasts. Results of these experiments demonstrate that the spontaneous and induced probabilities of differentiation may be altered in an uncoupled fashion, suggesting that each is determined at different steps leading to a common pathway of globin gene expression.


Neuroradiology of the sphenoidal region.

The neuroradiological techniques employed in diagnosing a series of 54 lesions involving the superior orbital fissure and cavernous sinus (sphenoidal region) were reviewed in order to compare the value of sphenoidal tomography, pneumoencephalography, cerebral arteriography and sphenoidal venography. The patients included in the series exhibited clinical syndromes of sphenoidal region cranial nerve deficits. Confirmation of the sphenoidal region disorder was obtained in all cases. Of the four procedures, sphenoidal venography and basal tomography yielded the highest rate of positive studies.


Polytomoencephalography of the optic chiasm and adjacent structures.

The addition of polytomography considerably increases the sensitivity of pneumoencephalography. The chiasmatic area lends itself particularly well to investigation by polytomoencephalography. In a series of 28 patients, the transverse dimension of the optic chiasm ranged from 4 to 20 mm, with a mean of 18 mm. The posterior portion of the chiasm tended to be U-shaped; its anterior portion's shape was that of a dumbbell. Intrachiasmatic tumors distort and enlarge the chiasm. The diaphragma sella is also particularly well evaluated by this technique. The study is safe, reliable, and can be repeated as needed.


Results of postoperative megavoltage radiotherapy of malignant surface epithelial tumors of the ovary.

The results of postoperative radiotherapy in 151 patients with malignant epithelial tumors of the ovary are presented. Actuarial survival at five years for patients in FIGO Stage I was 76%; in Stage II, 56%; Stage III, 16%; and Stage IV, 0%. Survival also varied with histopathological type, the best prognoses occuring in the serous (malignant) and endometrioid types (NED survival of 35% and 50%, totalling all stages, respectively). The presence of ascites or intraperitoneal surgical spillage did not adversely affect survival. The role of lymphangiography in staging is discussed. New approaches to diagnosis, staging and treatment are required for improving the prognosis of patients with ovarian carcinoma.


Conditioned medium from plateau-phase cells. Effect on growth of proliferative cells and on repair of potentially lethal radiation damage.

In vitro cell cultures grown into plateau phase may be more suitable than exponentially growing cells as a model for tumor cell growth in vivo. Conditioned medium extracted from cells in plateau phase inhibits growth of log-phase cultures of either the same or a different cell line. Attempts to demonstrate that this inhibition is due to a heat-labile diffusible substance have failed. Conditioned medium extracted from plateau-phase cultures was effective in enhancing repair of potentially lethal damage following x irradiation. Cell survival is significantly increased following a six-hour postirradiation culture in conditioned medium as opposed to complete fresh medium.


Idiopathic congenital hemihypertrophy with associated ipsilateral benign nephromegaly.

Two cases of idiopathic congenital hemihypertrophy with ipsilateral benign nephromegaly are presented. Excretory urography usually differentiates benign renal enlargement from Wilm's tumor in these patients. Angiography is indicated in equivocal cases. The clinical features, differential diagnosis, associated conditions, and importance of radiologic reevaluation in patients with hemihypertrophy are discussed.


Fever, lethargy and rash complicating treatment for Wilms' tumor-a new syndrome?

A syndrome of fever, lethargy, irritability and rash occurred in patients undergoing surgery, irradiation and chemotherapy for Wilms' tumor. The incidence, course and severity of these findings are described. Potential etiologic factors are analyzed and a hypothesis advanced.


The hazards of myelography.

The history of myelography and reactions to various contrast media is reviewed. Air is completely absorbed without producing long-term side effects. Lipiodol and Pantopaque are not absorbed and residual medium in the spinal canal can cause arachnoiditis. Other agents as Diodrast and Thorotrast never gained widespread acceptance. The water-soluble Dimer-X and Conray 60 have come into prominence but they are not totally without reactions. Recently a new water-soluble nonionic contrast medium has been developed in Norway. Metrizamide (Amipaque) has the same overall density as other water-soluble media but produces fewer reactions.


Computed tomography of liver specimens.

Fourteen liver specimens were studied by computed tomography with pathological correlation to evaluate the possible application of this technique to the diagnosis of hepatic lesions. Lesions were clearly demonstrated in 9 of 10 abnormal specimens, but differentiation among hepatomas, metastases, and cysts was not possible in vitro.


Angiographic signs of corpus callosal tumors: a reappraisal.

Angiographic findings of eleven corpus callosal tumors are analyzed. Three new angiographic signs are demonstration of the lateral ventricular roof veins, spreading of the septal veins, and actual demonstration of corpus callosal thickening. In contrast to traditional concepts, diagnostic abnormalities were angiographically demonstrated in all eleven cases. Biplane angiotomography is of vital importance in the angiographic diagnosis of this frequently subtle tumor.


Zwoegerziekte virus, the causative agent for progressive interstitial pneumonia (maedi) and meningo-leucoencephalitis (visna) in sheep.

The final results of experimental infections with virus recovered from the lungs of sheep suffering from progressive interstitial pneumonia (=zwoegerziekte=maedi) are reported. The virus could be reisolated from blood samples of all experimentally infected sheep. Every animal produced antibodies against the virus. The neutralising, complement-fixing and precipitating antibodies remained present in the blood for six years. Fourteen out of 21 intrapulmonarily infected sheep developed clinical and/or histopathological lung lesions and in three a meningo-leucoencephalitis was detected in addition. One of these three developed the clinical and pathological signs of 'visna' 14 months after inoculation. Signs of visna were seen in eight of 10 sheep that had been inoculated intracerebrally. Furthermore, nine of these sheep suffered from progressive interstitial pneumonia. Hence the name maedi-visna virus is proposed for the agent which causes both disease entities. Three sheep that yielded virus after infection and in which antibodies were detected, did not develop histopathological lesions.


Some aspects of lipid metabolism in fatty liver and kidney syndrome in chicks.

Various aspects of lipid metabolism were examined in broiler chicks affected with fatty liver and kidney syndrome (FLKS). Plasma free fatty acid concentrations were invariably elevated. Plasma triglyceride concentrations were increased amounts of triglyceride-rich lipoproteins. Lipoprotein lipase activity in adipose tissue was considerably reduced, but in heart tissue the enzyme activity was increased. Hepatic lipogenesis was reduced. Rates of oxidation of palmitic and succinic acids by liver, heart and kidney were normal. The increased oxidation rate of palmitic acid following the addition of carnitine was also normal. These findings indicate that elevated blood lipid levels are likely to be an important factor contributing to the development of fatty deposition, particularly in extrahepatic tissues.


The relation between the rabbit potency test and the response of sheep to sheep clostridial vaccines.

Six commercially available clostridial vaccines comprising one oil-emulsion, two alum-precipitated and three aluminum hydroxide adjuvanted preparations, each containing between two and seven antigenic components, were administered to groups of 10 rabbits and eight sheep in accordance with manufacturers' recommendations. Serum antitoxic values to Cl welchii beta, Cl welchii epsilon, Cl septicum, Cl oedematins and Cl tetani toxins were determined 14 days after completion of each vaccination course. The overall pattern of mean antitoxic values was found to be similar in sheep and rabbits, a vaccine eliciting a comparatively high antibody titre to any given antigen component in sheep also inducing a comparatively high titre in the corresponding group of rabbits. Similarly, comparatively poor responses in sheep were associated with poor responses in rabbits. The degree of variation in response within groups of animals was greater in sheep than in rabbits for all five antigenic components assayed. Sheep consistently developed higher titres than rabbits to Cl oedematins component but consistently lower titres to both Cl welchii beta and epsilon components irrespective of the type of vaccine used. The response of both species to Cl tetani antigen was similar in terms of serum antitoxic values. It was concluded that rabbits provide a suitable model for the assessment of potency of sheep clostridial vaccines.


Insulin receptor: role in the resistance of human obesity to insulin.

Large adipocytes from obese subjects have similar receptor numbers and affinities for insulin as small adipocytes from subjects of normal weight. These results indicate that the insulin insensitivity of large fat cells from obese humans occurs after the insulin-receptor interaction and might be explained by either a dilution of receptors over the cell surface or by alterations in intracellular metabolism.


Induction of neural differentiation in cultures of amphibian undetermined presumptive epidermis by cyclic AMP derivatives.

Induction of neural differentiation in cultures of undetermined presumptive epidermis from three amphibian species was achieved by the addition of 1 millimolar dibutyryl adenosine 3',5'-monophosphate, 8-bromadenosine 3',5'-monophosphate, or adenosine C',E'-monophosphate together with theophylline. Adenosine 5'-monophosphate, adenosine 2',3'-monophosphate, dibutyryl guanosine 3',5'-monophosphate, and butyrate at 1 millimolar are ineffective. These results suggest that the action of the primary inductor or inductors may be mediated via adenosine 3',5'-monophosphate.


The lamellar substructure of osmiophilic inclusion bodies present in rat type II alveolar pneumonocytes.

When rat pulmonary tissues are processed as outlined in this paper, the type II cell's surfactant containing osmiophilic inclusion bodies possess a periodic substructure. This substructure consists of electron-opaque lamellae, 26 A in thickness alternating with 16 A wide electron-translucent zones.


The inhibitive effects of steroid analogues in the binding of tritiated 5alpha-dihydrotestosterone to receptor proteins from rat prostate tissue.

The relationship between molecular structure and the binding potential of steroids to receptor proteins was investigated. Twenty-four selected steroids were studied in minced incubations of rat prostate tissue. Measurements of the inhibitory effects of the steroids on the binding of tritiated 5alpha-dihydrotestosterone to the receptor proteins were obtained in the 100,000 times g dialysed supernatant and the purified nuclear component of the prostate cells. The steroids that achieved the highest degree of inhibition were those compounds that exhibited a generally planar geometric shape and were known to possess potent androgenic activity. Several of the compounds were shown to possess a higher degree of inhibition than that of testosterone or 5alpha-dihydrotestosterone. The data is further supportive of the two step theory that necessitates the complexing of the free steroids to the receptor proteins in the cytosol before transport to the nuclei. Evidence is also suggestive of the presence of 17-esterase activity. The inhibitory effect of the steroids apparently involves the binding to the intracellular receptors and is not related to the uptake of 5alpha-dihydrotestosterone into the cell.


The urinary metabolites of 17alpha-ethynylestradiol-9alpha,11xi-3H in women. Chromatographic profiling and indentification of ethynyl and non-ethynyl compounds.

Metabolites of 17alpha-ethynylestradiol (EE2) were obtained from human urine following ingestion of tritium-labeled EE2. Over 95% of the recovered activity was found as conjugated steroids and these were separated into four groups by chromatography of the urine extract on Sephadex LH-20 with chloroform-methanol (1/1) + 0.01M NaCl. The two major conjugate fractions appeared to be almost exclusively glucosiduronates. Enzymatic hydrolysis liberated at least ten different EE2 metabolites as shown by chromatography on Sephadex LH-20 with benzene-methanol (85/15). After additional separation and purification of these metabolites, positive identification was obtained for nine radioactive compounds by either gas liquid chromatography-mass spectrometry or reverse-isotope recrystallization. Five were ethynyl compounds: EE2, 2-MeO EE2, 16beta-OH EE2, 2-OH EE2 and 6alpha-OH EE2. The other four were de-ethynylated estrogens: estrone, estradiol-17beta, estriol, and 2-Me-O-estradiol-17beta.


Androgen-induced nuclear accumulation of the estrogen receptor.

The effect of androgens on the nuclear uptake of both tritiated estradiol (3H-E2) and the estrogen receptor was studied in immature rat uteri. It was demonstrated that in vitro preincubation of immature rat uteri with various androgens (1 muM to 50 muM) followed by incubation with 3H-E2 (20 nM) resulted in a greatly decreased specific nuclear uptake of 3H-E2. Non-androgenic steroids had no effect. It was also confirmed that 5alpha-dihydrotestosterone (DHT) causes the accumulation of the estrogen receptor in the nuclei of uterine tissue. In vitro incubations of rat uteri with DHT (1muM and 50muM) were found to cause maximal nuclear estrogen receptor accumulation to the same degree as caused by estradiol, i.e. the nuclear uptake of approximately 100% of the estrogen receptor. Antiandrogens, which block the binding of androgens to the testosterone receptor in various tissues, did not inhibit the DHT - induced decrease in the 3H-E2 uptake by the uterine nuclei or the DHT - caused accumulation of the estrogen receptor in nuclei. These results seem to indicate that the uterine testosterone receptor has no role in the androgen - induced nuclear uptake of the estrogen receptor. However, the non-steroidal antiestrogens inhibited the DHT - induced nuclear accumulation of the estrogen receptor. This would seem to indicate that the estrogen - and androgen - induced nuclear accumulation of the estrogen receptor share a common mechanism.


[Membrane antigens of Zajdela ascitic hepatoma].

The membrane antigens of Zajdela ascitic hepatoma cells were investigated. Living cells were studied by immunofluorescence method, and solublized membrane preparations by the precipitation reacting in agar gel. Testing of the tumor cells with organospecific anti-kidney serum caused a specific fluorescence of tumor cells surface. This can be due to incorporation into the antigenic structure of the Zajdela hepatoma cell membranes of at least one organospecific antigen. Treatment of the tumor cells with organospecific anti-liver serum led to specific fluorescence of tumor cells surface. In solubilizates of the tumor cells one of the three organospecific antigens peculiar for the normal liver cells, was detected.


[Quantitative changes in proteins in the cat parietal zone large pyramids in various sleep stages and during wakefulness].

Protein content in large pyramidal cells of the parietal cortex was shown to be by 28.9% lesser during the slow--wave sleep than during the paradoxical sleep.


[Electron microscopic and electron histochemical study of mitochondrial inclusions in insect intestine epithelial cells].

The electron microscope investigation of the midgut epithelial cells of Chaoborus obscuripes revealed special membrane formations in mitochondria. The mitochondrial inclusions (MI) look like small membranous whorls about 0.2--0.3 microns in diameter. In a certain period of functional activity, MI start leaving the mitochondria and appear in cytoplasm. Later, large multimembrane structures (MS) composed of parallel membranes in concentric or whorl--like arrangement are observed in the cytoplasm. At first MS are situated in the vicinity of mitochondria, then they gather near the base of the villus. The relationship between appearance of MI and MS in the cytoplasm and the digestion process was revealed. Treatment of ultrathin sections with H202 or H5J06 showed MI and MS to be composed mainly of lipids. This suggests that MI and MS,probably, take part in the lipids metabolism of the midgut epithelial cells of insect.


[Benign non-epithelial tumors of the stomach].

Data relative to 36 patients with benign gastric tumors of various histological structure have been studied. The clinical picture, methods of diagnosis and operative technic wwere discussed. Based on the author's personal data a conslusion is drawn on the possibility of a preoperative diagnosis of benign non-epithelial tumors of the stomach and indications to dissection of a tumor instead of gastric resection.


Effects of proteases and neuraminidase on RBC surface charge and agglutination. A kinetic study.

Electrophoretic mobility, membrane sialic acid content and agglutinability by "incomplete" antisera against Rh-o, hr' and k antigens were determined for red blood cells in the course of treatment with trypsin, ficin and neuraminidase. Neuraminidase gradually produces a slight to moderate agglutinability as it reduced surface charge density in proportion to the amount of sialic acid removed. Proteases acted in two distinct steps. The first stage is characterized by the cells rapidly becoming highly agglutinable and by the unmasking of new negative charge as the first half of the sialic acid is removed. In the second stage the cells show a slight gain in agglutinability as surface charge is removed in proportion to sialic acid removal as in the case of neuraminidase. Neuraminidase-treated cells are considerably less agglutinable than cells reduced to the same zeta-potential by protease treatment. The greater efficacy of proteases compared to neuraminidase in making cells agglutinable could be because they not only reduce surface charge density but also increase antigen-antibody bond strength, render antigens more mobile in the membrane to allow clustering in regions of cell to cell antibody bridging and remove glycopeptide chains which may be causing steric hindrance to antigen-antibody binding or to cell-cell contact.


[Determination of carboxycathepsin (peptidyl dipeptidase) activity in human blood serum].

Based on fluorometric determination of a dipeptide histidil-leucine, which is splitted off from a synthetic substrate (Cbz-Phe-His-Leu) by carboxycathepsin (carboxydipeptidyl peptide hydrolase), a method was developed for estimation of the enzymatic activity in human blood serum. The method is characterized by simplicity and high sensitivity; Use of small amount of blood serum (about 0.03 ml) was possible. In normal human blood serum the carboxycathepsin activity varied from 7.5 to 18 nmoles of the dipeptide, liberated per mg of protein per hr.


The relative roles of iodination and iodothyronine content on thyroglobulin stability.

We have independently varied the degree of iodination and of iodothyronine formation over a wide range by acutely administering various doses of perchlorate and/or methimazole to severely iodine-deficient rats 30 min before giving 131-I- with graded quantities (1-100 mug of 127-I-). Thyroids were removed 4 h later and the soluble protein analyzed for labelled iodoamino acid composition and with sucrose density gradient ultracentrifugation. Since the total thyroid iodine content before administering 127I- was less than 1 mug, calculation of the degree of iodination and iodothyronine content of the labelled Tgb could be made from the known specificity of the injected labelled iodide. Newly organified iodine ranged from smaller than 0.1 to 1.4 mug/thyroid and labelled iodothyronines from smaller than 5 to 962 pmoles/thyroid. Both the degree of iodination and iodothyronine content varied directly with Tgb stability in the absence of inhibitors. But when Tgb iodination was kept constant, Tgb stability at pH 10.1 varied directly with iodothyronine content. When iodothyronine content was kept constant, Tgb stability was independent of the degree of iodination. Correlation of stability with iodothyronine content was highly significant (r=0.79, Psmaller than 0.001) but not of stability with iodine content (r=0.49, P larger than 0.05). We conclude that the primary determinant of Tgb stability in mild alkali is the iodothyronine content and not the degree of iodination of the protein. The increased Tgb stability may be induced by coupling between iodotyrosil residues of different 12 S subunits rather than between residues of the same 12 S subunit.


Regulation of plasma oestrogens in normal adult males. II. Response of oestradiol, oestrone, testosterone and cortisol to dexamethasone and ACTH administration.

The response of plasma oestradiol (Oe-2), oestrone (Oe-1), testosterone (T) and cortisol to dexamethasone and ACTH administration in normal adult males was ascertained. After an oral dose of 1.5 mg dexamethasone Oe-1 decreased from 2. 51 to 1.85 ng/100 ml (74% of the control level, mean of 15 subjects) over night and after continuing the dexamethasone administration for two more days (twice daily 1.5 mg) Oe-1 decreased to 1.30 ng/100 ml (52%). Oe-2 (control level 1.71 ng/100 ml) and T (678 ng/100 ml) did not change over night, but at the end of the test Oe-2 decreased to 84% and T to 91% of the control level. After a single im injection of 2 mg beta 1-24 cortrophin Oe-1 increased 1.9 times (meanof 13 subjects) without further increase during two more days of treatment (twice daily 1 mg), while Oe-2 did not change over night and decreased to 75% of the control level at the end of the test. Under ACTH T decreased to 66% of the control level over night and to 50% at the end of the test indicating a suppression of the testicular hormone secretion. From these results and those reported previously it was concluded that the testicular hormone secretion influences mainly Oe-2 in peripheral plasma, while Oe-1 depends mainly on the adrenal hormone secretion.


Progesterone-binding proteins: in vitro binding and biological activity of different steroidal ligands.

Progesterone-binding proteins from human, rabbit, sheep and guinea pig myometrial cytosol, all induced with oestradiol, as well as from pregnant guinea pig myometrium and plasma were investigated. The physico-chemical properties of the oestradiol-induced binding proteins were very similar in all the species studied. In all, 63 steroids were tested for their ability to compete with tritiated progesterone for the binding sites on these six proteins and their relative affinities were determined. The studies reveal that the ligand specificities of oestrogen-induced myometrial binding proteins from human, rabbit and sheep are rather similar, whereas that from guinea pig myometrium has different binding characteristics. The properties of the binding proteins from pregnant guinea pig uterus and plasma differ substantially from all of the induced proteins. It is clear from the different physico-chemical characteristics and binding specificities that the oestrogen-induced myometrial protein of the guinea pig is not the same as that which appears in the myometrium and plasma during pregnancy. The binding energies of the well bound progestational compounds were of the order of -12 Kcal/mole, half of which stems from the interaction of the steroid nucleus with the protein. The specific interaction of the protein with the two functional groups, the 3-keto-4-ene system and the acetyl sid chain each contributed-3 Kcal/mole. In the case of the rabbit, sheep and human proteins a 17alpha-ethynyl-17beta-hydroxyl function could replace the acetyl side chain. For a large number of steroids reasonable agreement existed between the degree of binding to the rabbit myometrial protein and in vivo biological activity (Clauberg-McPhail test) in the same species. The data suggest that as far asthe binding aspect is concerned, the rabbit is an appropriate model for assessing the biological activity of compounds under development for human application. The in vitro binding system is also a useful tool to assess whether steroids need to be bio-activated before eliciting a biological response.


[Synthesis and properties of 5-acetyl-4-methyl-1-(beta-d-ribofuranosyl)-imidazole-5' di-and-triphosphate].

5-Acetyl-4-methyl-1-(beta-D-ribofuranosyl)-imidazole-5'-phosphate reacts with diphenylphospho chloridate forming the asymmetrical pyrophosphate ester. This in turn reacts with tri-n-butyl-ammonium phosphate yielding 5-acetyl-4-methyl-imidazole-riboside-5'-diphosphate and with tri-n-butylammonium pyrophosphate to give the nucleotide triphosphate. 5-Acetyl-4-methyl-imidazole-riboside-5'-pyrophosphate shows in the test with pyruvate kinase a reaction rate three times slower than that of ADP; but the same Km as that of ADP. The ATP analogue is only about 10% as effective as ATP itself in the test with hexokinase, 3-phosphoglycerate kinase and gloconate kinase. Adenylate kinase and NAD" kinase show no activity when ATP is replaced by the nucleotide-triphosphate-analogue. In presence of ATP the analogue strongly inhibits the reaction of adenylate kinase.


[Change in the glycolytic and glucose-6-phosphate dehydrogenase activity in experimental cirrhosis of the liver].

Development of cirrhosis of liver tissue did not influence the intensity of glycolysis, with glucose as a substrate, in supernatant fraction of liver homogenate in chronic intoxication with CCL4. In preparations of cirrhotic liver, as compared with liver from the intact animals, more distinct activation of glycolysis was caused by addition of ATP and NAD at the stage of 3-week intoxication and also by addition of hexokinase, glyceraldehydephosphate dehydrogenase and lactate dehydrogenase at the stage of distinct cirrhosis of liver (6 weeks of CCL4 intoxication). Km values for glucose-6-phosphate dehydrogenase increased over all the periods of intoxication.


Progesterone-binding proteins from endometrium and myometrium of sheep uterus: a comparative study.

The binding of tritiated proesterone to the cytoplasmic progestogen receptors from oestrogen-primed sheep endometrium and myometrium has been investigated. The binding characteristics of the progesterone receptors from both sources were determined in the supernatant fractions obtained after high speed centrifugation of the myometrial and endometrial homogenates. High affinity binding proteins with identical association constants for progesterone (1 times 10-9 M-1) were detected in both endometrium and myometrium. The concentration of binding sites was also of the same order of magnitude in both tissues. After centrifugation on surcrose gradients, these binding proteins were shown to have similar sedimentation constants, 7S in a gradient containing no added KC1 and 4S in a gradient containing 0.4 M KC1. The binding peaks from both sources could be abolished by heating the cytosol at 60 degrees C for half an hour. The proteinaceous nature of the binding materials was demonstrated by incubation of the endometrial and myometrial cytosols with pronase, DNase and RNase: the binding was totally eliminated by the proteolytic action of pronase whereas DNase and RNase had no effect. The ligand specificity of the two progesterone binding proteins was studied using a competitive protein-binding technique. Both the endometrial and the myometrial receptor proteins were shown to bind only steroids which are known potent progestogens. In addition, the relative affinities of the binding proteins for 37 steroidal compounds closely resembled each other. Thus, the physiochemical and binding data obtained show that a very close similarity exists between the progesterone-binding proteins in the endometrium and myometrium of ovariectomized sheep after oestradiol treatment.


Plasma testosterone in Klinefelter's syndrome: diurnal variation and response to ACTH and dexamethasone.

The mean basal plasma testosterone level in 28 patients with chromatin positive Klinefelter's syndrome was significantly lower than in 58 healthy male controls. In 12 of the patients the hormone levels were in the normal range. In both the eugonadal males and the Klinefelter patients the plasma testosterone levels spontaneously decreased throughout the day, the relative decrease in both groups being of the same order of magnitude. Short term ACTH infusion and the administration of dexamethasone did not significantly influence the plasma testosterone concentration in the Klinefelter patients. These findings do not support the view that the adrenal cortex plays a major role in contributing to the circulating plasma testosterone levels in this syndrome.


Viral infection and psychiatric disorders.

A psychiatric population of 94 inpatients and 12 outpatients was investigated on referral to a psychiatric unit in a general hospital for serum antibody titres to several viruses by a complement fixation technique. Of the total population studied, only eight were considered to have antibody titres of possible significance. This result would appear to indicate that viral infection does not play a major part in the causation or precipitation of psychiatric disorders.


Characteristics of proliferative cells from young, old, and transformed WI 38 cultures.

Mitotic cells were obtained by a shake off procedure from cultures of normal WI 38 cells at various passage levels, and from SV-40 virus transformed cells. The size of all mitotic WI 38 cells was similar regardless of in vitro age, whereas cells from the monolayer displayed an age-related increase in size. Mitotic transformed cells were similar to normal in size, but no size changes were observed in transformed monolayer cells during serial passage. Ultrastructural studies of mitotic WI 38 cells revealed no consistent change in the numbers of mitochondria or lysosome-like bodies during aging in culture. Mitotic transformed cells displayed numbers of mitochondria comparable to normal cells, but lysosome-like bodies occurred less frequently. Size distribution and structural characteristic are presented in relation to the ability of cells to synthesize DNA and to divide. These results support the contention that aging in WI 38 cultures is characterized by a declining fraction of homogeneous, actively dividing cells, and an increasing fraction of heterogenous nondividers that display senescent changes.


The effect of age on mitochondrial ultrastructure and enzymes.

The ultrastructure of perfused livers and of mitochondrial fractions from 6 months and 30 month-old C57/BL mice were studied. In old mice the liver cell mitochondria were enlarged and rounded with a light "foamy", vacuolated matrix, short cristae and a loss of dense granules. Quantitative studies showed a 60% increase in the mean size and an increased proportion of larger mitochondria in intact 30 month-old perfused livers. Endothelial and Kupffer cell mitochondria were smaller than those of the parenchymal cells. Mitochondria in pellets prepared from 6 and 30 month-old livers were rounded and condensed although there were a few larger and "foamy" mitochondria in the preparations from old mice. Up to 47% of large mitochondria in the old livers were lost during cell fractionation. The levels of cytochrome oxidase and malate dehydrogenase were slightly decreased with age but their cytochemical localization was unchanged.


Viral infection and interferon in cell cultures aged in vitro.

Cells from leucosis-free chick embryos (skin and muscles), grown in vitro 1 and 7 days with no medium change, are used as a model for young dividing and stationary in vitro aged cells, respectively. Aged cultures are advantageous for IF induction by cytopathic and noncytopathic viruses. Decreased virus production in them is ascribed to a back effect of the early formed IF onto the inducing virus. A factor which stimulates viral IF interferon induction also in young cultures, is released from aged cells into the medium. This factor could belong -- according to its properties and action -- into the class of chalones.


Ultrastructure of in vitro aged chick embryo cell cultures in relation to viral infection.

The ultrastructure of young uninfected and infected with herpes simplex virus chick embryo cells was compared with that of aged cells. In contrast to the nuclei of young chick embryo cells 24 hr after infection, the aged chick embryo cells 24 hr after infection presented very expressive blocks of perinuclear chromatin. The nucleoplasm contains forming naked HSV particles. The lesions seen are dilation of the perinuclear cisternae, dilatation of rough endoplasmic reticulum cisternae and formation of multilamellar bodies. The moderate swellings of the mitochondria with violation of cristae arrangement and losing of mitochondrial matrix were observed. In contrast to the cytoplasm of young infected cells no formation of crystal-like arrangement of cytoplasmic ribosome tetrads occurs.


Pathogenetic mechanisms in atherosclerosis.

Four theories of atherogenesis are briefly reviewed and criticized: the degenerative, the thrombogenic, the platelet aggregation and the insudative theory. Evidence is presented in detail to suggest that a modified form of the insudative theory (1) accounts more satisfactorily than the other theories for the known association of risk factors with atherosclerosis and (2) allows one to understand how some of the more important risk factors operate at the level of the arterial wall. It is proposed that atherosclerotic plaques, and also certain extravascular lesions broadly associated with atherosclerosis (corneal arcus, xanthomas), arise because altered endothelial permeability allows certain reactive macromolecular plasma proteins (the plasma low density and very low density lipoproteins and fibrinogen, which are normally largely confined to the circulation) to permeate endothelium and interact with charged components of the connective tissue gel of the arterial wall or other tissues. The effect of hyperlipidemia, hypertension, arterial disease or injury upon this process, and the manner in which these factors interact, is examined in relation to experimental findings and clinical observations.


Mechanisms of cholesterol accumulation in the arterial wall.

The rate of cholesterol accumulation is a function of three separate processes: the transfer of lipid or lipoprotein from blood plasma to the artery, the binding and sequestering of lipids in the arterial wall and the solubilization and removal of lipid from the artery. These processes have been studied with lipids or lipoproteins labeled with radioisotopes by autoradiographic and quantitative chemical procedures. More recently immunochemical procedures have been applied to this problem. Studies have been performed with intact animals, isolated organs and cell cultures. In addition, homogenates have been used successfully to study intraarterial transformations of lipids, (for example, cholesterol esterification). Although epidemiologic and clinical studies, as well as animal experiments, have provided evidence that the concentration of plasma low or very low density lipoproteins parallels the rate of atherogenesis, the nature of the causal chain linking plasma lipoproteins to atherosclerosis is as yet unclear. A possible link between plasma lipoproteins, arterial liproprotein lipase and atherogenesis has been postulated.


Aortic mitochondrial synthesis of lipid and its response to cholesterol feeding.

The rates and products of lipid synthesis from acetate-I-14C were studied in mitochondria isolated from control and atherosclerotic rabbit aorta. More acetate was incorporated into fatty acids in the cholesterol-fed animals. The mechanism was one of chain elongation, and the resultant products were longer in chain length. The newly elongated fatty acids were esterified mostly into phospholipids, presumably those turning over most rapidly. A hypothesis is proposed for the pathogenetic sequence: Cholesterol feeding alters transport functions of the mitochondrial membranes of aortic smooth muscle cells. Respiration and redox state are altered and consequently acetate is incorporated more rapidly into fatty acids as an alternate mechanism for oxidation of the reduced form of nicotin-amide-adenine dinucleotide (NADH). Rapidly turning over phospholipids esterify these fatty acids and may transfer them ultimately to cholesterol. Esterified cholesterol is less exchangeable and commences to accumulate.


Lipid metabolism in perfused human and dog coronary arteries.

The paper represents a summary of our studies in which in vitro perfusion of human and animal coronary vessels was carried out. Formation and uptake of lipids in perfused human coronary arteries were studied under a vairety of experimental conditions, including exposure to carbon monoxide. The effect of collagenase on lipid synthesis and transport in carotid arteries of dogs was also studied. Human plasma with hydrogen-3-labeled cholesterol and carbon-14-acetate was used to perfuse human blood vessels. Autologous plasma was employed. Inhibition of cholesterol uptake was accomplished by the addition of 7-ketocholesterol (concentrations of 0.005 to 1 mum/ml) to the perfusate. Both atherosclerotic and normal human coronary arteries incorporated 14C-acetate into lipids but failed to synthesize either cholesterol of cholesterol esters. Similar results were obtained in human saphenous veins perfused at arterial pressure. Cholesterol uptake from the perfusion fluid was demonstrated in atherosclerotic and normal human coronary arteries as well as in human saphenous veins. Carbon monoxide increased permeability of the arterial wall to cholesterol uptake. In dog arteries exposed to collagenase marked increases in cholesterol uptake were found, but total lipid synthesis was reduced; the relative synthesis individual lipids remained unchanged. The addition of 7-ketocholesterol to the perfusate reduced cholesterol uptake by the vessel by 90 percent. Inhibition of cholesterol uptake was present in all species and was not due to oxidation of cholesterol to 7-detocholesterol in the perfusate. The results illustrate that human coronary arteries as well as human saphenous veins synthesize lipids but not cholesterol. Cholesterol flux into the artery is augmented by carbon monoxide and collagenase. The data also show that active inhibition of cholesterol uptake in the arterial wall can be accomplished by competitive inhibition with 7-ketocholesterol.


Function of the fat-soluble vitamins.

Much has been learned about the function of the fat-soluble vitamins in recent years. Their mechanisms are divergent ranging from a role in transcription of DNA or differentiation to a direct enzymatic activation of the gamma-carboxylation of glutamate residues of a preexisting protein. Thus the only resemblance so far among these physiologically active agents is their lipid solubility. However, in general the fat-soluble vitamins play important roles in specialized functions carried out by highly differentiated organisms.


On certain electrical phenomena for the interpretation of ch'i in Chinese acupuncture.

The author describes a series of experiments on the "microelectric phenomenon" and tires to give new scientific interpretations of acupuncture. Thus, the concept of "2-M-C" was developed, which may generate new hypotheses concerning the modus operandi of needle treatment.


Limits of applicability of the firefly luminescence ATP assay for the detection of bacteria in clinical specimens.

ATP measurement can be used as an indicator of biological mass, and the extreme sensitivity of the firefly ATP assay has led to its use in bacterial detection systems. Clinical specimens present problems not encountered with cultured isolates of known bacterial species. The lower limit of sensitivity for detecting bacteria using the firefly assay is 100,000 bacteria per ml. Non-bacterial ATP, which is probably present in all clinical specimens, produces false-positive results unless it is completely destroyed, and this destruction must be carried out under conditions that do not affect bacterial ATP. A cause of false-negative results is the presence in all urine specimens of unidentified materials that inhibit the luminescent enzymic reaction. These considerations indicate that application of the firefly ATP assay in bacterial detection systems for clinical specimens is feasible only if a preparatory step separates bacteria from interfering materials and from non-bacterial sources of ATP, and concentrates microorganisms to measurable levels. These limitations sharply curtain the applicability in diagnostic microbiology of this exotic chemical reaction.


Granular cell tumor (myoblastoma) of the orbit.

An 8-year-old white boy had a tumor in the left orbit with clinical manifestations of muscular impairment, exophthalmos, and reduced visual acuity. Five months after exenteration of the orbit the boy was healthy without tumor recurrence. Based on our observations with electron, light, and phase microscopy, and histochemical studies, we conclude that this tumor is benign, of histiocytic origin, and we propose the term granular cell histiocytoma be used to describe this entity.


Occupational therapy in a community aftercare program.

The need for effective community treatment for psychiatric clients has increased in recent years. The aftercare program descirbed in this paper provides a community treatment program for adult psychiatric clients that aims at facilitatin their adequate and independent adjustment to the community. The occupational therapist functions as a case manager of the treaatment team and is also program coordinator for the arts and crafts groups and the skill development groups. The arts and crafts groups are designed primarily to meet basic mental health needs; the skill development groups are aimed at teaching independent functioning in basic living skills. The behavior rehearsal model provides the theoretical framework for the skill development groups and is an effective approach in teaching skill acquisition to clients treated in the community.


Ultrastructure of medullary, intraductal, tubular and adenocystic breast carcinomas: comparative patterns of myoepithelial differentiation and basal lamina deposition.

Samples from medullary, intraductal, tubular and adenocystic breast carcinomas were studied ultrastructurally, with emphasis on the patterns of myoepithelial differentiation and basal lamina deposition. Myoepithelial cells, while prominent in the intraductal and adenocystic carcinomas, were rarely found in the medullary neoplasms and appeared absent in the tubular neoplasms. Parallel to the above, basal laminae were most abundant and evenly deposited in the intraductal and adenocystic tumors, but were infrequent in the medullary tumors and seemingly absent in the tubular carcinomas. Well-defined myoepithelial cells and retention of the capability to synthesize basal lamina are evidence of differentiation in neoplastic cell populations. Thus, their presence in intraductal and adenocystic carcinomas (generally associated with a good prognosis) is not surprising. However, their scarcity or absence in the similarly favorable group of medullary and tubular carcinomas suggests that other factors may also influence the invasive and metastasizing ability of breast carcinomas.


The nuclear membranes in hypertrophied human cardiac muscle cells.

Nuclear membranes of cardiac muscle cells were studied in 134 patients with cardiac hypertrophy of various causes. Abnormalities observed consisted of: a) increased foldings and convolutions; b) nuclear pseudoinclusions formed by cytoplasmic organelles protruding into saccular invaginations of the nuclear membranes, and c) intranuclear tubules. The increased foldings and convolutions of the nuclear membranes and the nuclear pseudoinclusions appear to result from synthesis of nuclear membranes in excess of that needed to accommodate the increase in nuclear volume which occurs in hypertrophy. Intranuclear tubules were found in 6 patients and consisted of tubular invaginations, 400 to 650 A in diameter, of the inner nuclear membranes into the nucleoplasm. Some of these tubules were straight and cylindrical, and were associated with a peripheral layer of marginated chromatin; others were not associated with chromatin, appeared coiled and followed irregular courses. Intranuclear tubules in cardiac muscle cells probably represent an extreme cellular response to the stimulus of hypertrophy.


Ultrastructure of inclusions in peripheral blood mononuclear cells in sarcoidosis.

Bundle-shaped tubular (BST) inclusions were identified by electron microscopy in 1 to 6 percent of peripheral blood mononuclear cells obtained from 10 to 11 patients with sarcoidosis. The frequency of the inclusions within mononuclear cells did not correlate with the clinical status of the patients. The relationship, if any, of BST inclusions to sarcoidosis and the immunologic defects in sarcoid patients remains to be determined.


Effect of ACTH on lipolysis in adipose tissue of normal and adrenalectomized rats in vivo.

Parametrial fat-pads of fed rats were perfused in vivo. Rates of release of glycerol and free fatty acids (FFA) were determined from venous-arterial difference and plasma flow. Adrenalectomy lowered basal release of FFA and glycerol, but not reesterification of FFA within the fat-pad. ACTH (5 mug iv) in normal rats increased release of FFA and glycerol (mumol-g-1-h-1) from basal values of 0.90 and 0.48 to 3.2 and 1.3, respectively, and in adrenalectomized rats from 0.41 and 0.33 to 1.5 and 3.1, respectively. Thus in normal rats ACTH increased the molar ratio of FFA to glycerol released from 1.9 to 2.5, whereas in adrenalectomized rats the ratio fell from 1.3 to 0.5. After stimulation of lipolysis in normal rats 85% of the FFA formed were released but only 20% in adrenalectomized rats; the remainder was reesterified to triglyceride. It is concluded that adipose tissue of adrenalectomized rats is sensitive to the lipolytic activity of ACTH, but increased glucose utilization by adipocytes in the absence of glucocorticoid enhances reesterification and reduces release of FFA by the tissue.


Postnatal development of adenylate cyclase in rat salivary glands: patterns of hormonal sensitivity.

In order to compare the known morphological changes which occur during the postnatal development of the salivary glands in the rat with alterations in membrane function, we measured adenylate cyclase activity and its responses to sodium fluoride (NaF), norepinephrine, and isoproterenol in salivary gland membranes at various times after birth. In the parotid gland, basal enzyme activity did not change significantly during postnatal life, but fluoride-stimulated activity rose on day 15; A similar marked rise in activity stimulated by norepinephrine (0.02 mM) and isoproterenol (0.03 mM) was noted simultaneously. In the submandibular gland, basal adenylate cyclase activity was higher just after birth than at 25 days of life or in maturity. Fluoride-stimulated activity was 7 times higher than basal activity on day 1, greater than 10 times higher on day 25, and 30 times greater in the adult. The gland was as responsive to norepinephrine and isoproterenol on day 5 as it was on day 25 or in the mature animal, showing a two- to threefold increase over the basal enzyme value at each time point studied. Residual phosphodiesterase activity in the membranes was always negligible. The data demonstrate a time-dependent developmental change in the responsiveness of the parotid gland to norepinephrine and isoproterenol, which corresponds to the time when morphological maturation normally occurs. By contrast, in the submandibular gland, membrane-bound adenylate cyclase is fully developed at the time of birth.


Brief hospitalization of patients with families: initial results.

A total of 175 newly admitted inpatients who lived with families were randomly assigned to three treatment groups: standard inpatient care (discharge at the therapist's discretion), brief hospitalization (one week or less) with transitional day care available, and brief hospitalization without day care. Outpatient aftercare was offered to all patients. The three groups showed no significant differences as to amount of improvement in levels of psychopathology at 3 and 12 weeks, but the briefly hospitalized patients were able to resume their vocational roles sooner. There were no significant differences among the groups in readmission rates.


Alternative to the hospital: a controlled study.

The authors describe the evaluation of a treatment model that makes use of the community to help mentally ill patients acquire necessary coping skills. This approach is based on the assumption that deficiency in coping skills and aggressive dependency are primarily responsible for high readmission rates to mental hospitals and that coping skills and autonomy are best learned in the community, where the patient will be needing and using them. The treatment model is evaluated by comparing it with progressive in-hospital treatment and follow-up care. The results suggest that the model described can successfully treat patients with a high level of symptomatology in the community rather than in the mental hospital.


Arteriographic demonstration of intravenous tumor extension.

The arteriographic demonstration of a linear, striated arterial pattern in the anatomic course of the draining vein is reliable evidence of venous extension by a tumor and represents the supply to the intravenous component of the neoplasm. In addition to its previously described occurrence with hypernephromas, this report documents the characteristic angiographic appearance with hepatocellular tumors and a retroperitoneal osteosarcoma as reliable evidence of tumor extension into the inferior vena cava. It is possible that in some cases, the parallel arteries represent markedly enlarged vasa vasorum of the involved venous structure. The frequent invasion of the inferior vena cava by hepatomas and the importance of inferior vena cavography in their angiographic evaluation is also emphasized.


Metastatic disease of the sella turcica.

Eleven cases of known metastatic disease to the sella turcica are reported. The clinical, plain film roentgenographic and angiographic presentations are discussed with reference to the differential diagnosis and possible distinguishing characteristics of metastatic disease.


Aggressive surgery in treatment for parotid cancer: the role of adjunctive postoperative radiotherapy.

A 7 year follow-up of 120 patients with malignant parotid cancers revealed parotidectomy to be an adequate procedure for low-grade, nonaggressive lesions. The addition of radiation therapy postoperatively will improve local and regional control of high-grade, aggressive lesions. Postoperative radiation therapy may also make the removal of the facial nerve unnecessary in certain clinical presentations.


Metastatic seeding along the small bowel mesentery. Roentgen features.

1. Intra-abdominal malignant seeding along the small bowel mesentery is roentgenologically identifiable. 2. The anatomic features of the small bowel mesentery and the pathways of flow of ascitic fluid along its peritoneal recesses determine the depostion and growth of seeded metastases. 3. Seeded deposits lodge particularly within the lower recesses of the small bowel mesentery and are reflected by typical changes involving distal ileal loops or the cecum. 4. These are characterized by (a) palisading, arcuate scalloped deflections or gross mass displacement of distal small bowel loops in the right lower quadrant, or (b) extrinsic indentations upon the medial and inferior contours of the cecum. Associated desmoplastic reaction may result in tethering of mucosal folds and angulated fixation of bowel loops.


Ecology of respiratory virus transmission: a comparison of three communities in West Bengal.

Respiratory virus transmission in children was studied comparatively in three ecologically different low-income communities in West Bengal: an isolated village, a suburban village, and a crowded urban community. Continued use of contaminated pond water for bathing, irrigation of nasal passages, post-defecation washing of the anus, and washing of food vessels was common to all, as was intense crowding of indoor sleeping quarters during cold and wet seasons. Intensity of infection was highest (26%) in the most crowded urban area, the variety of virus types least in the most isolated village. Sources of drinking water differed but seemed unrelated to virus transmission. Toxigenic diphtheria organisms were found in nonspecific skin lesions in children in each area.


Crimean hemorrhagic fever-Congo (CHF-C) virus antibodies in man, and in domestic and small mammals, in Iran.

A Crimean hemorrhagic fever-Congo (CHF-C) virus antibody survey was carried out in Iran with sera from man and several animal species; this survey was done by means of agar gel diffusion precipitation (AGDP) test with the following results (percent positive of number tested): men, 13% of 351; sheep, 38% of 728; goats, 36% of 135; cattle, 18% of 130; camels, 0% of 157; small mammals, 3% of 274. A number of sera were tested by complement-fixation (CF), neutralization (N), and hemagglutination-inhibition (HI) tests in addition to the AGDP test. A good correlation was found in the results with 105 sera tested by AGDP, HI, and N, with approximately 70% to 75% positive in all three tests; by CF, only 20% were positive. Of 55 human sera, of which 15 could be tested by N test, about half were positive by AGDP and only 10% by HI at low titers; none was positive by N and CF tests. These results suggest that any one of the three serological tests (N, HI, and AGDP) can be used to survey the antibody prevalence in sera from domestic animals; the CF test, not unexpectedly, was less suitable. Our results, however, are inconclusive in regard to the human sera.


Islet cell tumors of the pancreas and the alimentary tract.

Functioning tumors of the pancreatic islets are now recognized as the source of clinical syndromes affecting the gastrointestinal tract which have a wide variety of catastrophic symptoms. Experiences with thirty-six cases suggest at least four separate diagnostic categories in the ulcerogenic tumor syndrome. These include: a typical history, gastric analysis, and roentgenographic findings with boderline fasting serum gastrin levels; ulcerogenic tumor with evidence of hyperparathyroidism; iatrogenic ulcerogenic syndrome associated with failure of a previous operation for duodenal ulcer; and the classic ulcerogenic syndrome associated with a fulminating ulcer diathesis or diarrhea and high serum gastrin levels. The problems presented at operation include: decisions to be make in the presence of a negative exploration; the finding of a solitary tumor in the wall of the duodenum; solitary pancreatic tumors particularly in the body and tail; ulcerogenic tumors in the very young; liver metastases in the elderly; and the wisdom of removing gross metastases in combination with total gastrectomy. The long-term survival in the ulcerogenic tumor syndrome approximated 50 per cent, with 40 per cent of those having proved malignancy living five years. Evidence of hyperparathyroidism is relatively common in association with both the ulcerogenic and the diarrheogenic tumor syndromes. The association may by a result of a congenital abnormality, metabolic alkalosis, or a direct effect of the islet cell tumor. Parathyroidectomy may be indicated when both the serum calcium and parathormone levels are elevated in the presence of borderline fasting gastrin levels. The latter may return to normal after parathyroidectomy. The evidence of hyperparathyroidism closely parallels the episodes of diarrhea in the diarrheogenic syndrome, and hyperparathyroidism may regress spontaneously after total removal of the pancreatic tumor. Just as routine calcium determinations made the diagnosis of hyperparathyroidism more commonplace, it is suggested that the gastrointestinal syndromes associated with islet cell tumor would receive wider recognition if radioimmunoassays for gastrin as well as secretin, and the other secretin-like polypeptides, were carried out routinely.


Immunochemotherapy of hepatoma in rats.

Immunoglobulin G prepared by immunization of rabbits with rat Novikoff ascites tumor cells was prepared and found to by cytotoxic to Novikoff cells and rat lymphocytes. A chlorambucil conjugate was prepared and found to have similar qualities in vitro. Three day culture systems were prepared with various test substances and the immunoglobulin was shown to markedly depress cell multiplication. The chlorambucil conjugate appeared to be more efficacious than the immunoglobulin alone in inhibiting cell growth. In vivo studies revealed a total inhibition of tumor formation in animals treated with the immunoglobulin-chlorambucil conjugate whereas all control groups had 50 to 100 per cent mortality. No systemic toxicity was apparent. These studies indicate that tumor-specific immunoglobulin bound to chlorambucil presents a feasible method of delivering tumoricidal drugs to cancer in body cavities and of treating tumors with an accessible blood supply.


[Clinical considerations and genetics of the lattice dystrophy of the cornea].

Lattice dystrophy of the cornea is reported in 4 families.  For purposes of linkage analysis we did serological tests on pertinent family members, including marker testing of the blood group systems ABO, MNSs, Rh, K, Ky, and P; protein group systems Hp, Ge, Gm, and Inv; and enzyme polymorphisms SEP, PGM, ADA, and AK.


Permeability barriers to cytochrome-c in nerves of adult and immature rats.

Nerves in the tongues of adult and immature rats were examined with respect to their permeability to exogenous cytochrome-c (mol wt 12,000) injected into the tongue. The distribution of cytochrome-c was determined in cryostat sections on the basis of the peroxidase activity of this protein. Nerves of 14-day-old rats were permeable to injected cytochrome-c. The larger nerves of older animals showed only localized accumulations of cytochrome-c reaction product both between and within axons adjacent to endoneurial blood vessels. Reaction product was not found, however, in association with blood vessels penetrating nerves of the tongue that were not within the limits of tracer spread. In the smallest nerve brances, thin linear strands of reaction product filled the interstices between the nerve fibers.


The blood-brain barrier of the rat choroid plexus.

Following intravenous injection, cytochrome c traverses the capillary endothelium of the rat choroid plexus and permeates the perivascular space and the extracellular space between epithelial cells. The tracer is incorporated into pinocytotic vesicles adjacent to the lateral and basal plasmalemmas. Thereafter, cytochrome c is incorporated into multivesicular and dense bodies. Tracerladen vesicles were not found to fuse with the apical plasmalemma and cytochrome c was not discharged into the cerebral ventricles. Acid phosphatase activity of the choroidal epithelium after the administration of cytochrome c was greatly increased and localized in the same intracellular sites shown for cytochrome c. These data suggest that cytochrome c and possibly other proteins that penetrate the choroidal stroma are taken up by the choroidal epithelium and subsequently degraded in lysosomal vesicles. This heterolytic mechanism may be an important means for preventing the entry of certain substances such as proteins into CSF and subsequently into nervous tissue.


Neuromuscular transmission in a mammalian preparation during exposure to enflurane.

The effects of enflurane on motor nerve terminals and muscle endplates were studied in the rat phrenic nerve-diaphragm preparation using standard microelectrode recording techniques. Muscle endplate interaction with enflurane was suggested by depression of the amplitude of miniature endplate potentials (MEPP's) without change in frequency, increased duration of MEPP's and endplate potentials (EPP's) increased threshold for generation of muscle action potentials, and inhibition of the endplate depolarization induced by succinylcholine. Evidence of nerve terminal effects of enflurane was limited to a greater relative decline of EPP amplitude during tetanus, indicative of failure to maintain transmitter output at rapid rates of stimulation. It is concluded that the depressant effect of enflurane on indirectly elicited muscle twitch is primarily due to an impairment of the action of muscle endplate on adjacent membrane; however, the relative importance of prejunctional depression, seen only during rapid nerve stimulation, cannot be assessed from the present data.


Ectopic production of big ACTH in carcinoma of the lung. Its clinical usefulness as a biologic marker.

This report considers the potential usefulness of adrenocorticotropic hormone (ACTH) determinations in diagnosis and in prognosis for therapy of patients with carcinoma of the lung but without clinical Cushing's syndrome. The report is based on radioimmunoassay data from 129 patients, including 62 with lung cancers and 67 with nonmalignant pulmonary conditions. Elevated plasma ACTH was found in 21 of 24 patients with untreated cancer and the hormone was detected in tumor extracts and/or bronchial washings from the remaining 3. Elevation of plasma ACTH was found in only 10 of 38 treated patients. Absence of clinical Cushing's syndrome in spite of high plasma ACTH concentrations is explained by the observation that the predominant form of ectopic ACTH in plasma is immunoreactive but nonbioactive 'big' ACTH. Prolonged survival, for longer than 19 months, was observed in only 5 patients: all patients with low plasma ACTH after resection of the lung tumor and 2 of 3 patients with low plasma ACTH without therapy. ACTH was found in all available malignant tissue, primary and metastatic, from the lung carcinoma group,but not in normal lung or in 5 tumors metastatic to the lung. Of the 39 patients diagnosed initially to have chronic obstructive pulmonary disease, 14 showed plasma ACTH elevation. However, 3 of these patients with the highest concentrations subsequently manifested carcinoma or carcinoma in situ.


Primary meningococcal pneumonia.

Three cases of pneumonia caused by Neisseria meningitidis group Y are reported. From the results of these cases, the following conclusions were made. N. meningitidis probably can cause serious infection without preceding blood stream invasion. Primary meningococcal pneumonia is not rare; it has no distinctive clinical presentation; and it may not be recognized by routine expectorated sputum cultures. In addition, it may be associated with recent influenzal and adenoviral infections. Lastly, meningococci of the serogroup Y are capable of causing serious disease. Antimicrobial susceptibility studies showed that all three group Y isolates were sensitive to sulfadiazine and rifampin as well as to penicillin, ampicillin, erythromycin, and chloramphenicol.


[Chromatography of penicillins on a thin layer of sorbents. The utilization of "Silufol" plates].

Carbon metabolism of P. chrysogenum under conditions of periodical addition of the nutrients was studied. It was found that a proper rate of the carbon source addition to the culture was of significant importance for intensive biosynthesis. The use of carbon for the energetic and constructive needs was not the same at different fermentation periods.


[Use of coulometric titration for elucidating the mechanism of the oxidation of 6-APA alkaline breakdown products by halogens].

Penaldinic acid and penicillamine were formed on alkali decomposition (1 N NaOH) of 6-APA for 20 minutes at room temperature, penicillamine being completely oxidized to disulphide by the air oxygen. Coulometric titration of the alkali decomposition products showed that generated chlorine in 0.5 N HCl solution or bromine in a week acid solution of KBr oxidized them with participation of 7 electrones. Generated iodine did not practically oxidize the 6-APA decomposition products during the coulometric titration.


Immunoglobulin deposits in lepromatous leprosy skin. Presence of deposits in apparently uninvolved skin and occurrence of serum antiepithelial antibodies.

Immunoglobulin deposits were detected in ten of 13 biopsy specimens from apparently uninvolved skin of patients with lepromatous leprosy. There were deposits of IgM at the dermoepidermal junction in the skin of five patients, and deposits of IgM along the dermal collagen and elastic fibers in the skin of the other five. The deposits were eluted with acid buffers and high molarity salt solution. Circulating IgG antibodies to intercellular substance of epithelial cells, similar to those present in pemphigus vulgaris, were found in 25% of patients with lepromatous leprosy who were studied. These antibodies appeared to be different from the skin-bound immunoglobulin deposits.


Subclinical neuropathy at "safe" levels of lead exposure.

Electrophysiological methods revealed subclinical neuropathy in 26 workers, exposed from 1 to 17 years to lead and whose blood lead (PbB) values had never exceeded 70mu/100ml, as ascertained by checking the monitor reports of the factory and by careful exposure history. The PbB determinations had been tested repeatedly and had been found valid. The main findings were slowing of the maximal motor conduction velocities of the median and ulnar nerves and particularly the conduction velocity of the slower fibers of the ulnar nerve. Electromyographical abnormalities comprised fibrillations, diminution of the number of motor units on maximal contraction, and an abnormally long duration of the units. Earlier similar measurements from heavily exposed workers had been even more abnormal. Thus, a dose-response relationship exists on a group basis. Since the regular monitoring of PbBs in most workers during their entire period of exposure excludes the possibility of a body burden out of proportion to the PbB slight neurological damage is produced at exposures hitherto regarded as quite safe.


Synovial sarcoma.

Of 34 cases with synovial sarcoma, the five-year survival rate was 36%. A high local recurrence rate results when local excision is performed. Wide excision which may necessitate amputation is the treatment of choice. Prophylactic and even therapeutic node dissections are ineffective in increasing survival because of the disease. Evaluation of radiation therapy was impossible, although some patients obtained significant paliation. Adriamycin appears to have a tumoricidal effect and provided clinically significant responses in several patients.