Hyperproduction of the sigma subunit of RNA polymerase in a mutant of Escherichia coli. A mutant of Escherichia coli K12 is described in which sigma and alpha subunits of the DNA-dependent RNA polymerase (EC 2.7.7.6) are produced at the rates much higher than in the normal strain. The rate of synthesis for sigma subunit was found to be at least 10-times higher, though the rapid degradation of sigma polypeptides accompanied with the accelerated synthesis precludes accurate estimation of the extent of hyperproduction. The alpha subunit synthesis was about 5-times higher in this mutant than in the control, and excess alpha polypeptides produced were as stable as the bulk of protein under the conditions employed. Genetic analyses of the mutant by conjugation and by transduction with phage P1 revealed that at least three distinct but closely linked mutations are responsible for hyperproduction of the sigma subunit; one (sig-1) is located very close to rif, and the others (sig-2 and sig-3) at the argH-bfe and metB regions, respectively. The results further indicate that the accelerated synthesis of alpha subunit is due to a mutation also located at the metB region. The present finding suggests that the synthesis of sigma subunit is subject to a complex control that can be affected by a number of cellular processes. The possible involvement of the core polymerase in determining the rate of synthesis of sigma subunit is discussed.


The effect of lexA and recF mutations on post-replication repair and DNA synthesis in Escherichia coli K-12. We have examined lexA1 uvrA6 and recF143 uvrBdelta derivatives of Escherichia coli K-12 for post-replication repair and DNA synthesis after UV irradiation. Compared to corresponding lex+ rec+ strains, we found that the lexA and recF cells were defective in (1) converting short DNA segments synthesized after irradiation to DNA of normal size; (2) synthesizing high molecular weight DNA after irradiation; (3) transferring pyrimidine dimers from irradiated DNA into unirradiated daughter strands. Our results support the hypothesis that after UV irradiation the formation of large DNA molecules in excision-deficient cells of E.coli depends directly or indirectly upon joining short DNA segments into longer strands, concomitant with the transfer of DNA from irradiated tamplates into unirradiated daughter strands. This process appears to require the activity of lexA and recF genes.


Localized mutagenesis of the aroE-strA section of the Escherichia coli chromosome coding for ribosomal proteins. In order to obtain E. coli strains altered in ribosomal proteins the following isolation technique was used: Phage P1 grown in a streptomycin resistant E. coli strain, was mutagenized by hydroxylamine or nitrous acid, and was used to transduce into a strain auxotrophic for aroE. Transductants with streptomycin resistance and aroE prototrophy were selected and tested for their growth at various temperatures (20 degrees, 30 degrees and 42 degrees) and their response to different antibiotics. Ribosomes from seventeen transductants with an altered response to temperature or antibiotics were isolated. They were tested for alterations in their ribosomal subunit profiles by sucrose centrifugation and for altered ribosomal proteins by two dimensional gel electrophoresis. Two strains showed accumulation of 50S ribosomal precursors and three strains had an altered 50S protein L18. This protein belongs to the 5S RNA-protein complex having GTPase and ATPase activity.


Studies on bacteriophage T7 DNA synthesis in vitro. I. Resolution of the T7 replication system into its components. A soluble extract prepared from T7-infected E. coli is able to initiate DNA synthesis on an exogenous T7 DNA template. We have developed a fractionation procedure to resolve and identify the proteins required for T7 DNA synthesis. By this method we have purified the following T7 replication-related proteins (each greater than 50% pure as judged by sodium dodecyl sulfate gel electrophoresis): T7 DNA-binding protein (27,000 daltons), T7 RNA polymerase (105,000 daltons), T7 DNA polymerase (gene 5-protein, 85,000 daltons, plus host-factor), T7 DNA ligase (40,000 daltons), and T7 DNA-priming protein (65,000 daltons). The T7 DNA-priming protein, synthesized between 7.5 and 15 min following infection, was not detectable if the infecting phage carried an amber mutation in gene 4. Using an in vitro complementation assay which specifically measures the stimulation of DNA synthesis in an extract prepared from T7 gene 4-mutant infected cells, we have purified the DNA-priming protein about 2,000-fold. The purified priming protein preparations are essentially free of endonuclease, exonuclease, DNA ligase and DNA polymerase activity, but they do contain measurable DNA-dependent RNA synthetic acitvity. The enzyme is rapidly inactivated by heating to 46 degrees C and by treatment with N-ethylmalemide. In the presence of T7 DNA-binding protein and all four ribonucleoside triphosphates, the DNA-priming protein enables T7 DNA polymerase to initiate DNA synthesis on intact duplex T7 DNA. Closer studies of its enzymatic function as well as of the possible roles of the other proteins in the T7 replication system will be presented in the accompanying paper.


Increased spontaneous reversion of certain frameshift mutations in DNA polymerase I deficient strains of Escherichia coli. A tenfold increase in the spontaneous reversion frequency of two of six lacZ frameshift mutations tested was observed in strains containing the following DNA polymerase I mutations--polA1, polA5, polA6, polAex1, res-3 and resA1. Reconstruction experiments indicated that this increase was not the result of a selective effect. Only a fourfold increase in frameshift mutations was found in strains containing a polA107 mutation. Both the polAex1 and polA107 mutations result in defective 5' to 3' exonuclease activity and do not affect polymerizing activity, but have different effects on frameshift mutation. A polA mutation on the chromosome induced frameshift mutations in a gene on an F episome. None of three auxotrophic mutations studied showed high frequency reversion in the presence of the polA1 or polA6 mutations.


Compatibility properties of P1 and PhiAMP prophages. Although phages P1 and PhiAmp are heteroimmune (Chesney and Scott, 1975 and Yarmolinsky, unpublished), their plasmid prophages are incompatible. Thus, the immunity and compatibility systems are two distinct regulators of phage replication. The two prophages, and plasmid P15B (Ikeda, Inzuka and Tomizawa, 1970) constitute a new compatibility group, designated Y.


[BCG-septicemia and mucocutaneous candidiasis due to familial combined immunodeficiency (authors transl)]. The history of a patient with mucocutaneous candidiasis and BCG-septicemia is reported. Combined immunodeficiency has been diagnosed as the underlying cause. A sister of the patient died at seven months of age under similar circumstances. The importance of the immunological diagnosis in severe bacterial and mycotic infection during early infancy has been emphasized. In such cases vaccinations with life vaccine are strongly contraindicated.


[Light and electron microscopic study of melanosis coli]. The authors describe the results of light- and electronmicroscopic investigation of the colonic mucosa in melanosis, occurring at a 35 years old male patient. In the tunica propria mucosae pigment storage macrophages described histologically can be recognized as well by the aid of electron microscope. In contrast to the light-microscopic picture, electronmicroscopically separation of the epithelial cells of the mucosa, accumulation of foreign material in the enlarged intercellular spaces, accumulation of granular-fibrillar matrix in the tunica propria and presence of plasma cells with increased secretory activity can be revealed. It is assumed, that separation of epithelial cells of the mucosa goes with the disturbance of the absorptive function. It can not be excluded, that electronmicroscopically revealed lesions of the coolnic musoca aggravate the severe obstipation. For this reason early diagnosis of the melanosis coli and ceasing the medication with athranol-glycosida-containing laxatives are very important.


Effects of sodium arsenite on the survival of UV-irradiated Escherichia coli: inhibition of a recA-dependent function. Epidemiological studies and clinical observation suggesting potential hazards of arsenic compounds in increasing the incidence of cancer have been in complete contradiction with experimental findings in animals. Because of the predominance of skin cancers in the epidemiological reports, we decided to investigate the possibility that arsenic compounds might interfere with DNA repair. Using Escherichia coli as a test system, we show that this is indeed the case. Sodium arsenite, at concentrations of 0.1 mM and higher, decreases the survival of ultraviolet-irradiated E. coli WP2, a strain which possesses the full complement of repair genes. The effect of the arsenite increases with increasing ultraviolet dose. Similar results were obtained with the excision repair deficient strains WWP2 (uvrA) and WP6 (polA). Sodium arsenite had no effect on the survival of a recA mutant, WP10. Survival of ultraviolet-irradiated WP5 (exrA) was enhanced by sodium ardenite, the effect being greatest at low ultraviolet doses. It is postulated that arsenite inhibits a recA-dependent step in DNA repair. To account for the increased survival of the exrA mutant, we suggest that in the absence of the exr+ gene, the arsenite-sensitive recA-dependent function is deleterious. The ability of arsenite to inhibit DNA repair may account for the clinical and epidemiological reports linking arsenicals with an increased incidence of cancer.


The effect of streptomycin resistance, caffeine and acriflavine on ultraviolet light-induced reversion to tryptophan independence in strains of Escherichia coli B/r. When the excision proficient strain E. coli WP2 Hcr+ trp- was grown to stationary phase by glucose starvation in M-9 minimal medium before UV -irradiation, the ability of nutrient broth enrichment of minimal medium to enhance trp- leads to Trp+ reversion was greatly reduced. Less than 50% of the Trp+ revertants were found to be ochre suppressors. However, in the WWP2 Hcr- strain, 75-86% of the tested revertants were ochre suppressors. This indicates that, under the cultural conditions employed, many potential suppressor mutations were removed by excision repair in the presence of broth enrichment. Broth enhancement of reversion also occurred in the Hcr- strain, which indicates that a less error-prone mode of recombination repair functions under minimal growth conditions. An Hcr+ strr derivative of WP2 Hcr+ was more resistant than its strs parent to the lethal effect of UV light and showed a lower UV-induced Trp+ reversion frequency. The percentage of Trp+ revertants that were due to ochre suppressors was markedly reduced in the strr strain. The Hcr- strr strain also had a lower UV-induced Trp+ reversion frequency than its strs parent. The excision repair inhibitor caffeine had little effect at lower UV doses on increasing Trp+ reversion in both Hcr+ strains. Acriflavine, however, was effective at lower UV doses in enhancing reversiin of the Hcr+ strains and the degree of enhancement increased with the dose. Acriflavine appeared to specifically enhance the number of ochre suppressing Trp+ revertants. In both Hcr- strains (strs and strr) caffeine (500 mug/ml) had no effect on survival but reduced the UV-induced Trp+ reversion frequency acting as an antimutagen. In contrast, acriflavine (2 mug/ml) decreased survival and increased the Trp+ reversion frequency of the Hcr- strains. The data on spontaneous Trp+ reversion frequencies show that the Hcr+ strs strain had a higher spontaneous reversion frequency than the Hcr- strs strain on all plating media. Further, caffeine was shown to reduce spontaneous Trp+ reversion in both Hcr+ and Hcr- strains while acriflavine increased the spontaneous reversion frequencies of both strains.


A kinetic analysis of spontaneous rho- mutations in yeast. Spontaneous mutation to the petite state at the level of the individual cell was studied in a haploid strain of yeast by the technique of pedigree analysis. Results indicated that (1) the mutability of rho+ cells within a population in log phase is variable; (2) rho+ mitotic buds are, on the average, about 50% more mutable than the rho+ cells from which they arose; (3) the mutability of a rho+ cell tends to decrease as it produces consecutive buds: (4) the probability that a mother cell will become rho- at or immediately subsequent to cell division is, on the average, one third the probability that its bud will be rho-; (5) most, if not all spontaneous rho- mutant cells contain mitochondrial DNA as judged from suppressiveness measurements. The data indicate that the spontaneous production of a mutant cell is a multi-step process. Neither a replicative advantage of defective mitochondrial DNA nor the existence of a "master" mitochondrial genome provides a satisfactory explanation of the process. Either selective dispensation of defective mitochondria to the bud at cytokinesis or normal retention by the mother cell of factors influencing the amplification or rate of induction of defective mitochondrial DNA could be involved.


Repair of UV-induced DNA damage and survival in yeast. I. Dimer excision. The amount of pyrimidine dimer UV photoproduct lost from the DNA of irradiated yeast cells during dark incubation has been measured in various conditions. It was found that no dimers were lost when cells were incubated in saline. When the cells were incubated, with aeration, in a full growth medium, dimers were lost, most excision being complete within 4 h. Not all dimers were lost and the number lost was a function of UV dose. Maximum loss, amounting to 50 000 dimers per genome was observed after 4000 or 6000 erg/mm2 of UV. At higher doses, the number excised declined. Making the assumptions that dimers are the principal lethal product of UV, that a single dimer remaining in its genome is enough to prevent a cell from multiplying and that excision is the principal dark-repair process in yeast, these data were incorporated into the repair term of an expression relating survival to repair8 and it was found that the survival of yeast at doses up to 2000 erg/mm2 of UV could be quite accurately predicted. This is the first time it has been possible to account for survival in terms of measured repair. It is suggested that the divergence of the predicted and observed curves at higher doses is due to other processes known to exist in yeast.


Ethidium bromide mutagenesis in yeast: protection by anaerobiosis. The mutagenesis by ethidium bromide, an intercalating dye, which induces the mutation from wild type (rho+) to the cytoplasmic respiratory deficient petite (rho-) in Saccharomyces cerevisiae, was studied under aerobic and anaerobic conditions. During growth of anaerobic cells at pH 6.5, ethidium bromide at a concentration of 2 mug/ml is unable to induce rho- mutants whereas under aerobic conditions the entire population is converted into rho- cells within 1 generation at the same drug concentration. With ethidium bromide 10 mug/ml 98% of the anaerobic cells are transformed into rho- in 5.5 h (more than 2 generations). In non-growing conditions, ethidium bromide 10 mug/ml has no effect in anaerobic cells. 3 h adapted cells used as control, are converted into rho- in 8 h. Increasing the ethidium bromide concentration to 20 mug/ml resulted in the appearance of some rho- mutants in the anaerobic population but marked at the same time the onset of a detectable toxic effect of the drug.


Treatment of Parkinson's disease with aporphines. Possible role of growth hormone. To avoid the main drawbacks of prolonged treatment with levodopa (involuntary movements and the "on-off" phenomenon), we administered apomorphine by mouth to 14 patients with Parkinson's disease. This treatment caused azotemia, which we circumvented by switching to N-propylnoraporpine, whose nephrotoxic dose (80 mg six times per day) was larger than its therapeutic dose (10 to 15 mg six times per day). Slowly increasing doses induced significant improvement (P less than 0.005) in all 24 patients studied, transitory mental aberrations in seven, and release of growth hormone in three patients tested. In patients previously on prolonged levodopa administration, the dyskinesia and "on-off" phenomenon were almost identical with N-propylnoraporphine, but both drawbacks were reduced or abolished in six patients by coadministration of alpha-methyldopa hydrazine plus levodopa. This coadministration seemed to abolish tachyphylaxis. We conclude that N-propylnoraporphine is very useful in the treatment of Parkinson's disease.


Detection of a liver-membrane autoantibody in HBsAg-negative chronic active hepatitis. Investigation of humoral immunity against hepatocellular membrane antigens in patients with chronic active hepatitis and other liver diseases showed two different immunofluorescence patterns of IgG on hepatocyte membranes. A linear pattern was seen in HBsAg-negative hepatitis, but HBsAg-positive cases and some of protracted, acute hepatitis B had a granular pattern. In patients with IgG bound to hepatocytes, continuing necrosis of parenchymal liver cells was seen. Conversely, hepatocytes without bound IgG were found in cases of chronic active hepatitis in remission, acute viral hepatitis without HBsAg and chronic persistent hepatitis, in "healthy" HBsAg-carriers and in patients with fatty liver or alcoholic cirrhosis. A liver-membrane autoantibody in serum, proved by fixation on membranes of isolated rabbit hepatocytes, could be demonstrated only in HBsAg-negative chronic active hepatitis with elevated IgG-concentrations. The results support the existence of different pathogenetic types of chronic active hepatitis, a so-called autoimmune type and a hepatitis virus-B-induced type.


Secretory component deficiency. A disorder of the IgA immune system. We studied a 15-year-old boy with chronic intestinal candidiasis who had normal serum IgA levels without IgA in his secretions. There was an elevated number of peripheral blood lymphocytes bearing surface IgA. In addition, the lymphocytes cultured in vitro with pokeweed mitogen produced IgA as well as other immunoglubulins. Despite this evidence of normal IgA synthetic capacity, the patient had greatly diminished levels of IgA in the saliva and jejunal fluid, and, as estimated by 14C-L-leucine incorporation, could not synthesize IgA locally at intestinal-mucosal sites. Finally, the patient had no detectable free secretory component in saliva or jejunal fluid in contrast to normal persons and to patients with IgA deficiency. The basis of this disorder is probably a defect in the homing of IgA precursor cells to secretory sites or in the selective proliferation/differentiation of IgA cells at such sites.


Etiologic heterogeneity of neural-tube defects. We classified 106 stillborn and live-born infants with anencephaly, meningomyelocele, meningocele and encephalocele according to the recognized causes of these malformations. Six different causes were identified, including both genetic and nongenetic disorders; 12 per cent had nongenetic disorders, a chromosome abnormality, or an encephalocele as part of the autosomal recessive Meckel syndrome. Therefore, for this 12 per cent genetic counseling normally provided for isolated anencephaly, meningomyelocele or encephalocele would have been incorrect. If all infants were considered together regardless of cause, the precurrence and recurrence rates of similar malformations in the sibs were 5.2 and 1.7 per cent respectively. However, if infants with other disorders, especially the Meckel syndrome, were excluded, the precurrence and recurrence rates for isolated anencephaly, meningomyelocele and encephalocele among white infants were only 1.7 per cent and 0 per cent. These rates are much lower than the risk of 5 per cent currently being used in genetic counseling in the United States.


In vitro study of bovine oligodendroglia. Oligodendroglia were prepared by 'Ficoll' density gradient centrifugation from the centrum ovale of fetal and adult bovine brains. When cultivated in Rose Chambers, and provided an air bubble was included in the chamber during the cultivation, processes developed on cells around the circumference of the bubble. A sizeable air phase seems to be important for process formation in isolated bovine glial preparations. Various culture systems, media and additions to the cultures were examined for their effect on the behavior of the cultures. Fibroblast overgrowth occurred in oligodendroglial cultures from fetal brains in media supplemented with fetal bovine serum (FBS) but not in medium 199 supplemented with 2.5% FBS.


[Some observations on prognosis in congenital hydrocephalus (second report) -with reference to the physical handicap in survival cases (author's transl)]. The prognosis of surgically treated congenital hydrocephalus has lately proven to give a remarkably increased expectation of life for patient. However, numerous reports hve shown that the prognosis of congenital hydrocephalus have not always shown favourable turns from the viewpoints of physical and mental disability. This problem has been considered many times over the past years. But, an important problem has been left unsolved. The children of social and physical disability are most serious problem, so we attempt to obain a new interpretation about the influence factors upon the prognosis. Present report consists of follow-up studies of long surviving patients who have studied the fatty acid metabolism in the cerebrospinal fluid during postoperatively, the influeence on prognosis of complicated central nervous system malformation and discussed motor and mental disability in survival cases. Since lipids are essential factors in composing the brain tissues, quantitative composition of fatty acid inC.S.F. was determined by the method of gaschromatography using the arachidic acid as an indicator...


An ultrastructural and radioimmunoassay study of anterior pituitary somatotrophs following pituitary portal vessel infusion of growth hormone releasing factor. The hypophysial portal vessels and anterior pituitary glands of adult male rats were surgically exposed, cannulated and infused for 1 min with saline, growth hormone-releasing factor (GH-RF), and dbcAMP. After cessation of infusion, anterior pituitary glands were collected at 1, 5, 15, 30 or 60 min for electron microscopic and ultrastructural cytochemical examination. Before and after cannulation of a portal vessel a 1-ml sample of blood was collected at 1, 5, 15, 30 or 60 min from the femoral vein for RIA of growth hormone. When viewed ultrastructurally, the initial response following the infusion of GH-RF into a portal vessel was one of granule release. Emiocytic activity was observed at all time intervals studied. This response was followed 30 min later by evidence of increased protein synthesis. Significant increases in plasma GH levels were present at 1, 5, and 15 min following infusion of GH-RF but not at 30 or 60 min. Preliminary analysis of the RIA data suggests that dbcAMP was significantly more potent than GH-RF in elevating radioimmunoassayable plasma GH levels. The results suggest that similar mechanisms of synthesis and release were involved.


Effects of coincubation of the pituitary and hypothalamus of intact and castrate male rats and the influence of LH-RH on pituitary 5 alpha-reductase activity1,2. Pituitary 5 alpha-reductase activity in intact male rats increases after the pituitary is incubated with the hypothalamus. Incubating the pituitary of castrate rats with the hypothalamus of intact rats relatively inhibits pituitary 5alpha-reductase activity. Coincubation of the pituitary and hypothalamus of castrate rats, or the pituitary of intact with the hypothalamus of castrate males, does not elicit changes in pituitary 5alpha-reductase activity. Different amounts of LH-RH in the incubation medium can modify 5alpha-reductase activity, i.e., activate it in the intact pituitary and inhibit it in the castrate pituitary. Hypothalamus from intact rats, which according to SHIN et al. [1974] is 'rich' in LH-RH, induces changes in pituitary 5alpha-reductase activity. The LH-RH-'poor' hypothalamus of castrate rats does not cause changes in this enzyme activity. The results suggest that there is a very close relationship between LH-RH and 5alpha-reductase activity in the pituitary.


Effect of propranolol on essential tremor. The investigators tested the effect of 120 mg propranolol daily on 21 patients with essential tremor using a double-blind cross-over method and electrical recording of tremor amplitude and frequency. The patients varied in age between 15 and 60 years and had a mean tremor frequency of 10 cps. Propranolol had no effect on the tremor frequency but reduced the amplutide in 15 of the patients. Propranolol was most effective in older patients and in those with slow tremor frequencies.


Masculinizing ovarian tumors.

Masculinizing ovarian tumors. In a study of 194 masculinizing ovarian tumors from the Ovarian Tumor Registry, the criteria for reclassifying these tumors on a histologic basis are discussed. It is proposed that the terms "gynandroblastoma" and "arrhenoblastoma" be discarded in favor of "gonadal stromal," and that the general classification of "lipoid tumor" can be avoided if careful search is made for specific histologic structures. Finally, it is suggested that the malignancy rate for gonadal stromal tumors, Sertoli-Leydig tumors, hilus cell tumors and adrenal rest tumors is closer to 5% rather than 25-30%.


Placenta percreta.

Placenta percreta. A patient with an unsuspected placenta percreta presented with intraperitoneal bleeding at 33 weeks' gestation. Because of the clinical picture and a lecithin/sphingomyelin ratio of 1:1, this patient was treated conservatively until 38 weeks' gestation when an L/S ratio test indicated fetal maturity. The pregnancy was successfully terminated by cesarean hysterectomy.


Pregnancy in a patient with paroxysmal nocturnal hemoglobinuria. A case of successful gestation, delivery, and postpartum recovery in a woman with paroxysmal nocturnal hemoglobinuria (PNH) is presented. The rarity of pregnancy in patients with PNH and the nature and high rate of complications in such cases are discussed. Finally, suggestions are made for both antepartum and postpartum management of these unusual and complicated pregnancies.


Pregnancy associated with hypergonadotropic hypogonadism. A case of a 30-year-old female with secondary amenorrhea and relative hypergonadotropic hypogonadism is presented. The patient demonstrated persistently elevated levels of gonadotropins in spite of clinical and laboratory evidence of estrogen production. Laparoscopic directed biopsy revealed a total absence of ova, but in spite of this finding, conception ensued. Variations in ovarian sensitivity throughout the female reproductive period and in certain pathologic states are discussed, as well as the apparent limitation of single laparoscopic directed biopsies for confirming the diagnosis of premature menopause. Finally, the theoretical use of exogenous estrogen to induce ovulation in patients with the ovarian insensitivity syndrome is discussed.


Antenatal infection: adequate protection against hyaline membrane disease? It has been argued that fetal and placental infections decrease the incidence of hyaline membrane disease (HMD). However, others contend that this is not so. We performed a rigidly controlled clinicopathologic investigation of one group of infants with evidence of severe antenatal infection compared with another group free of infection. This study shows that placental infection correlated positively with neonatal sepsis and that in this series of patients neither infection nor prolonged rupture of membranes is associated with a decrease of HMD. Our data do not support the proposal that antenatal infections protect the neonate against later development of HMD.


Topical 5-fluorouracil in the treatment of carcinoma in situ of the vulva. The clinical and pathologic response of carcinoma in situ of the vulva in a 42-year-old woman treated with topical 5-fluorouracil is reported. Except for one persistent area of involvement in the anal region which required local excision, the result of treatment was excellent, with no evidence of recurrence after 2 years' followup. This important adjunct in the treatment of intra-epithelial carcinoma deserves long-range investigation.


Pleuroperitoneal endometriosis.

Pleuroperitoneal endometriosis. A nearly unique case is presented of pleural and peritoneal endometriosis with bloody pleural effusion and 4700 cc of bloody ascites. Theories of pathogenesis, differential diagnosis, and treatment are discussed. This represents another of the protean manifestations of endometriosis to complement the many others described in the literature.


Serum FSH and synthetic LHRG response in pregnant women at term and in the newborn. Serum immunoreactive FSH was undetectable in a) pregnant women past 38 weeks of gestation, b) newborn infants, and c) anencephalic infants. The intravenous administration of 100 mug of synthetic LHRH elicited no FSH release in each instance. These results seem to indicate that the absence of FSH in serum in pregnant women in the last trimester, as well as in the newborn, is due to the suppressive effect on the anterior pituitary of the increased amount of circulating sex steroids.


Clinical evaluation of two weekly dose schedules of 'IC-140'. 4-N, N-bis (2-Chloroethyl) amino phenyl N (P-carboxyphenyl) carbamate, in the treatmentment of solid tumors 1,2. 53 patients with intractable malignant diseases that were treated with a new alkylating agent, 4-N,N-bis (2-chloroethyl) amino phenyl N (P-carboxyphenyl) carbanate, also knows as 'IC-140' were evaluated. The records of an additional 41 patients entered on this study could not be assessed from the standpoint of toxicity. At the dose level of 150 mg/m2/week, severe leukopenia (less than 2,000) and thrombocytopenia ( less than 75,000) were encountered in 23 of 34 patients. On the other hand, at the 100 mg/m2/week dose level, the severe toxicity was reduced to 8 out of 19 patients. Tumor response was evaluated in 43  patients. The overall response was 23% (29% at the 150 mg/m2, 13% at the 100 mg/m2) and the duration of the response varied from 3 to 32 weeks with a mean duration of 13 weeks. Responses were noted in patients with ovarian, renal, lung, hepatic, breast carcinomas and lymphosarcoma.


The trophoblast theory of cancer (John Beard, 1857-1924) revisited. Beard's theory can be restated in a modified form in modern terms in the following way. Cancer represents primarily trophoblastic tissue derived either from an aberrant germ cell or from a somatic cell whose normally repressed 'asexual generation' genes are abnormally reactivated ('derepressed'). The variety of tumors, other than teratomas, may be due to a parallel chance derepression of some genes of somatic ('sexual gneration') characters. This would be a defensive reaction against intramural parasitization by trophoblast and would result in the differentiation and hyperplasia of normally present more primative somatic cells.


Comparison of tissue reaction to nylon and prolene sutures in rabbit iris and cornea. The tissue reaction in rabbit iris and cornea to prolene suture is equivalent to the tissue reaction to nylon suture. The prolene suture retains its tensile strength indefinitely and this may make it particularly useful in situations where a permanent suture is desired, such as in refractive keratoplasty, scleral buckle, or to suture an intraocular lens to the iris. The prolene suture also has a greater elasticity and this property may be advantageous in some clinical applications. We feel that a clinical study on humans is now warranted and we have started such a study.


The evaluation of 7-0 Polyglactin 910 suture in cataract surgery. The new synthetic absorbable suture, Polyglactin 910, represents a significant step toward absorbable suture perfection. In 218 cataract surgical procedures, the performance of Polyglactin 910 was compared to that of Chromic Catgut and Chromic Collagen. The results obtained in this series of cases is the basis for this article. When compared to Chromic Catgut and Chromic Collagen, Polyglactin 910 consistently provided greater tensile strength, improved handling, significantly decreased tissue reaction, more batch-to-batch uniformity, superior wound tensile strength retention, and a predictable absorption rate that is virtually completed in 35 days. Both nonprotein and nonantigenic, Polyglactin 910 evoked less tissue reaction and proved more versatile and reliable than either of the other sutures.


Immunization of mice and guinea-pigs against Salmonella dublin infection with live and inactivated vaccine. The immunogenicity of a number of avirulent rough Salmonella dublin mutants was compared in mice and guinea-pigs. Live vaccine prepared from Strain HB 1/17 at doses of 5 X 10(7) per mouse usually gave an immunity of between 70 and 80% but in certain experiments the results were more variable and always poorer. This strain gave a cross protection of 28,5% to S. typhimurium in mice. In guinea-pigs it evoked an average protection of approximately 46% to homologous challenge and approximately 26% to challenge with S. tryphimurium. Strain 5765 protected up to 80% of mice against S. dublin infection and was generally superior to Strain HB 1/17 in this respect. It was, however, less effective in protecting mice against S. tryphimurium (20%). In guinea-pigs it was also less effective than Strain HB 1/17, giving 34% protection against homologous and 20% against heterologous challenge. Other strains also produced immunity in mice but they were not studied in detail. Formalin-inactivated alum-precipitated vaccine prepared from avirulent smooth strain and containing 0,5% packed cells proved to be extremely effective in protecting mice against S. dublin infection. It produced an average immunity of 75% and was often 100% effective. It also protected 60% of mice against challenge with S. tryphimurium. In guinea-pigs it was, however, totally ineffective against challenge with both S. dublin and S. tryphimurium.


Infected total hip replacement after dental procedures. Three cases are reported in which there was a worrisome association between dental work and an infected total hip replacement. The patients had long asymptomatic intervals subsequent to implantation of prosthetic hip joints. After dental procedures, infections became apparent in these hips. Such infections carry an enormous and crippling morbidity. The potential complications of transient bacteremia in the patient with a cardiac valvular prosthesis are appreciated and the importance of prophylactic antibodies for dental work in such patients is well known. Although we emphasize that there is no proof that the infections in our patients were metastatic from the mouth, the sequence of events is suggestive. We recommend prophylactic antibiotics for dental work in the patient with a total hip replacement.


A fractured mandible, from initial operation to removal of tantalum mesh. Report of a case. Report is made of a case of fracture of the angle of the mandible. Treatment was attempted with the Sampson pericortical bone clamp, but was unsuccessful. Routine use of intraosseous wire led to a localized osteomyelitis, without union of the fracture. Treatment then was made with a particulate marrow graft contained within a tantalum mesh screen. The screen was removed 30 months postoperatively.


Circulatory reflexes in tetraplegics during artifical ventilation and general anaesthesia. The arterial blood pressure, heart rate and electrocardiograph were recorded, and plasma electrolytes, arterial blood gases and pH, and plasma catecholamines were estimated in seven patients with physiologically complete cervical spinal cord transections who needed intermittent possitive pressure ventilation (I.P.P.V.) or were undergoing urological surgery under general anaesthesia. In the tetraplegics on I.P.P.V., bradycardia, and in two patients even cardiac arrest, occurred during tracheal suction, especially in the presence of hypoxia. In one tetraplegic being anaesthetised, cardiac arrest occurred during endotracheal intubation. This reflex bradycardia and cardiac arrest appeared to be due to a vago-vagal reflex, unopposed by sympathetic activity or by the pulmonary (inflation) vagal reflex. Atropine was effective in preventing this reflex. In the tetraplegics undergoing urological surgery, severe hypertension resulting from visceral stimulation was effectively reduced by halothane. In these patients, control of arterial blood pressure with lower concentrations of halothane may also be achieved with I.P.P.V.


Gonadotropin and testosterone measurements after estrogen administration to adult men, prepubertal and pubertal boys, and men with hypogonadotropism: evidence for maturation of positive feedback in the male. Nineteen male subjects were fiven five daily injections of 17beta-estradiol and circulating levels of estradiol (E2), testerone (T), and gonadotropins were determined by radioimmunoassay before, during, and after the steroid course. Peak levels of E2 attained during the 5 days of treatment ranged from 173-577 pg/ml. Four of seven normal adult men and one castrate man demonstrated suppression of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) with a subsequent rise in LH (positive feedback) while E2 levels remained elevated. A rise in T was associated with the LH increment in the four normal men. Nine pre-, early, or midpubertal boys and two men with hypogonadotropic hypogonadism displayed only gonadotropin suppression after E2 administration. The difference in LH response to estrogen (i.e., positive feedback) between the adult men with normal or elevated gonadotropin levels as compared with the endocrinologically normal boys is significant (P less than 0.01).


A review: relation between invasiveness and the K1 capsular polysaccharide of Escherichia coli. The conclusions from our studies to date may be summarized as follows. (1) Invasive E. coli strains causing neonatal meningitis are encapsulated. At least 80% of those strains inducing mengitis are K1 and approximately 40% of those strains isolated from infants with septicemia but without meningitis are also K1. Invasiveness is best related to the K1 antigen and not to E. coli O and H antigens. (2) The capsular content of CSF strains is not related to their invasiveness. In contrast to observations reporting higher K capsular polysaccharide content and molecular weight of E. coli invading the renal parenychma as compared with those E. coli confined to the bladder or in the stool, there were no differences among DSF K1 strains. Sepculation as to the mechanism of the invasive properties conferred by acidic capsular polysaccharides may be derived from the literature. Unencapsulated or "rough bacteria" are susceptible to the bactericidal action of agammaglobulinemice sera (15, 53). When injected into precolostral (agammaglobulinemic but complement containing), cesarian-delivered, and antigen-deprived piglets, unencapsulated bacteria are rapidly cleared from the circulation. In contrast, smooth bacteria injected into these same animals circulate without detectable splenic or hepatic clearance, multiply, and result in the death of these animals. The mechanism of the resistance of encapsulated bacteria has been postulated to be due to the inaccessibility of the deep somatic antigen structures capable of activating the alternate complement pathway system. Thus, opsoninization and other host complement-dependent activities may of necessity be antibody mediated for encapsulated bacteria. This complement resistance of encapsulated organisms may be quanititative and studies should be done to determine differences among various K1 E. coli strains. (3) K1 strains are widely prevalent among infants, children, and adults and are quickly transmitted to infants. In most cases the source of the infecting strain in diseased infants is the mother. However, transmission from attendants, demonstrable in our studies, is also a possible mechanism. (4) A protective role of serum anticapsular antibodies in animal models has been demonstrated. Our initial observations indicating low serum K1 antibodies in the general population and the finding that K1 antibodies are predominantly IgM in two animal species studied so far suggest that colostral K1 antibodies may be important in conferring immunity to this disease.


Topical application of insulin in the treatment of decubitus ulcers: a pilot study. To test the hypothesis that there will be a significantly greater increase in the rate of healing of decubitus ulcers in subjects who receive topical insulin therapy than in subjects who receive other forms of therapy, a pilot study was conducted, utilizing a two-group before and after design. Decubitus ulcers of six experimental group subjects were treated with a topical application of ten units of regular insulin twice a day for five days; eight control group subjects received one of a variety of topical therapies other than insulin. Data were collected on all subjects for a 15-day period. Although generalizations cannot be based on the results of 14 subjects, data analysis showed that insulin was a safe and effective agent in the healing of small, uncomplicated decubitus ulcers. The results appear to warrant a more extensive study of the subject.


Antiserotonin agents in the treatment of migraine. The authors present the results of own investigations about the use of both pizotifen (SandomigranR) and cyproheptadine (PeritolR). 32 sufferers were treated with pizotifen and showed positive improvement in 68% of the cases. In this group no serious side effects were found. 20 patients before administration of the drug had placebo and this helped to establish the results statistically. 25 sufferers were treated with cyproheptadine and equally positive results in 64% of the cases. In this group were noted infrequent side effects. The results of the two tests were compared with the results of other tests done with other drug. From this it has been established that the antiserotonin drugs are the most effective at present in the treatment of migraine.


Potential biologically active agents. IX. Synthesis of N-piperidino-(morpholino)-methylisatin-3-anils. Sixteen N-piperidino-(morpholino)-methylisatin-3-anils have been synthesised and evaluated for their antimicrobial activity.


[Polyester tendon for atraumatic cervical suture during pregnancy]. There are presented the attempts of operative treatment of so called habitual abortions resulting from uterine cervix insufficiency. Two groups of operations were distinguished, based on the methods of Shirodkar and McDonald. Advantages and disadvantages of these operations, and their modifications used in obstetrical clinical practice are discussed. The importance of the selection of suitable material for the suture was pointed out. The authors developed an atraumatic needle provided with a polyester tendon. There are shown advantageous clinical resutls in 56 patients, out of whom 48 have given birth to viable children. Abortions occurred in 3 cases, premature deliveries in 3 and immature deliveries in 2.


[Study of the antibacterial activity of ftorlon threads]. The study concerned antibacterial properties of the ftorlon threads obtained from acetone or dimethylformamide polymer solution containing antibacterial derivatives of 5-nitrofuran added which were known to be active against the germs most commonly causing the postoperative infection including Staphylococci, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Candida type fungi. Basing of the laboratory test following the method of infected medium and suspension method it was found that the best effects were obtained when using threads produced from the acetone polymer solution added with beta-(5-nitrofuryl-2)-acroleine and those containing furazolidone obtained from acetone or dimethylformamide polymer solution. Antibacterial properties of the ftorlon threads did not change upon autoclaving, gamma irradiation and prolonged storage.


[Method for producing spheroplasts from yeast cells]. A method for producing spheroplasts from yeast cells has been developed. The method involves: 1) prefreezing and thawing of cells in the minimal nutrient medium; 2) treatment with the mixture of 86 mM 2-mercaptoethanol, 5% sodium dodecylsulphate and 6% Triton X-100; 3) enzymic digestion by the digestive juice of grape helix. The formation of spheroplasts was controlled by microscopy and measurement of the optic density of the spheroplast suspension in the incubation medium and in 1% sodium dodecylsulphate. The method has been successfully tested on some bakery yeast: Saccharomyces cerevisiae, Saccharomyces paradoxus, Schizosaccharomyces pombe, Saccharomyces globosus, Saccharomyces carlsbergensis.


[Primary classification of beer infecting yeast by a combined test]. The paper describes the use of crystal-violet and lysine containing media to detect yeast infection in brewing and to classify it. The crystal-violet containing medium helps to separate brewing yeast Saccharomyces cerevisiae and Saccharomyces carlsbergensis from wild yeast of the Saccharomyces genus, whereas the lysine containing medium--from wild yeast of the genera Candida, Debaryomyces, Brethanomyces, Pichia. The test has been used both on museum cultures and on yeast isolated from beer and non-alcoholic beverages.


[Method for determining methionine in plant material]. Methionine was determined in corn, barley, oat, wheat, rye, pea, vetch by the two methods: the microbiological method using the methionine deficient mutant of E. coli and the method using the Hd1200E amino acid analyzer. The results showed a good agreement between the data obtained by the two methods.


[Bactericidal properties of volatile fractions of garlic phytoncids]. Changes in the bactericidal effect of volatile fractions of garlic were examined during storage. B. coli communis, Proteus vulgaris and Staph. pyogenes aureus 209 were used as test-organisms. Bactericidal properties of volatile fractions of garlic phytoncids were most pronounced prior to storage. During storage they reduced. At higher temperature (16 degrees) the decline in the bactericidal properties was much more significant than at low temperature of storage (2 degrees).


[Optimization of cultivation conditions of baker's yeast]. By an integrated factorial experiment optimal proportions of the quantity of inoculation yeast, molasses dosage and degree of their initial water dilution have been established for the periodic yeast cultivation. An equation describing the process has been derived. This equation makes is possible to assess yeast yield at any value of the above parameters within their extreme limits. The yeast cultivation under optimized conditions results in a 25% increase of the economic coefficient coefficient of reproduction as compared with the control, thus approximating the theoretically conceivable value.


Applications of natural-abundance nitrogen-15 nuclear magnetic resonance to large biochemically important molecules. Natural-abundance nitrogen-15 nuclear magnetic resonance spectroscopy of enzymes and other biopolymers is found to be feasible using newly available instrumentation. The long correlation times of such molecules result in short spin-lattice relaxation times, and these in turn allow rapid signal accumulation. The advantages of short T1 values are sometimes offset, however, by unfavorable nuclear Overhauser effects. The dependence of T1 and nuclear Overhauser effects upon correlation time is discussed, and preliminary nitrogen-15 nuclear magnetic resonance results for several biopolymers, including lysozyme, protamines, pepsin, hemoglobin, vitamin B 12, and tRNA, are presented.


How ribosomes select initiator regions in mRNA: base pair formation between the 3' terminus of 16S rRNA and the mRNA during initiation of protein synthesis in Escherichia coli. Initiation complexes formed by E. coli ribosomes in the presence of 32P-labeled A protein initiator region from R17 bacteriophage Rna have been treated with colicin E3 and disassembled by exposure to 1% sodium dodecyl sulfate. Electrophoresis on 9% polyacrylamide gels reveals a dissociable complex containing the 30-nucleotide-long messenger fragment and the 50-nucleotide-long colicin fragment, which arises from the 3' terminus of the 16S RNA. The complex is a pure RNA-RNA hybird; it is apparently maintained by a seven-base complementarity between the two RNA fragments. Detection of this mRNA-rRNA complex strongly supports the hypothesis that during the initiation step of protein biosynthesis the 3' end of 16S RNA base pairs with the polypurine stretch common to initiator regions in E. coli and bacteriophage mRNAs. The implications of our findings with respect to the molecular mechanism of initiation site selection and mRNA binding to ribosomes, the role of rRNA in ribosome function, and species specificity in translation are explored.


Promoter-dependent transcription of tRNAITyr genes using DNA fragments produced by restriction enzymes. Two DNA fragments prepared from the transducing bacteriophage strains ø80psuIII+ and ø80hpsuIII+,- by digestion with restriction enzymes contain one tyrosine tRNA gene (suIII+) and two tyrosine tRNA genes (suIII+, su-) in tandem, respectively, a single promoter in both cases, and some additional DNA regions at the two ends of both. Using these fragments, we have studied characteristics of the promoter-dependent transcription of the tyrosine tRNA genes. The promoter-dependent transcripts were shown to correspond to the expected tRNA precursors. Exposure of the transcript from the single gene fragment to an S100 extract from Escherichia coli gave, via intermediates, 4S material which was active in enzymatically accepting tyrosine and contained some modified bases.


Content of elongation factor Tu in Escherichia coli. The content of elongation factor Tu in E. coli B has been determined both by radioimmune assay and by GDP binding. The two assays gave comparable results: cells growing at 2 doublings per hour contained about 8 molecules of Tu per ribosome, whereas those growing at 0.22 doublings per hour contained about 14 molecules per ribosome. These levels resemble those reported for tRNA, in contrast with the 1:1 ratio of factor to ribosomes reported for elongation factors Ts and G.


A method for the isolation of specific tRNA precursors. tRNA affinity chromatography, based on complex formation between tRNAs with complementary anticodons, has been applied to the isolation of specific tRNA precursors. When [32P]RNA, isolated from an Escherichia coli strain containing a thermolabile ribonuclease P, was chromatographed on resin-bound yeast phenylalanine tRNA, precursor tRNAGlu (possessing the complementary anticodon) was specifically retained. Likewise, precursor tRNAPhe was isolated from a column of resin-bound E. coli glutamate tRNA. Both precursor tRNAs isolated were monomeric and may be processed products of an originally larger RNA precursor. Both tRNA precursors contain additional nucleotides beyond the 5'-end of the mature tRNA and have all modified bases found in mature tRNA. The method can be extended to isolate other tRNA precursors by affinity chromatography with different tRNAs. Since the principle of complementary anticodon interaction is not restricted to any particular organism, specific precursor tRNAs from other sources may also be isolated in this way.


Studies on nucleic acid reassociation kinetics: reactivity of single-stranded tails in DNA-DNA renaturation. The reassociation kinetics of Escherichia coli DNA were measured by S1 nuclease resistance and hydroxyapatite binding. While the reaction assayed by hydroxyapatite displays second order kinetics, the S1 nuclease measurements follow a non-second order from, as previously reported by Morrow (Ph.D. Dissertation, Stanford University. 1974). Much of the reaction measured with S1 nuclease occurs between single stranded regions of fragments already bearing duplex structures from previous collisions, and between such regions and totally free single strands. Experimental determinations indicate that the nucleation rate of single stranded regions on fragments also containing duplexes is inhibited by an average factor of 2 to 4.


Histone gene arrangement in the sea urchin, Strongylocentrotus purpuratus. The DNA coding for histones from Strongylocentrotus purpuratus, purified up to 100-fold with the use of Hg+2-CS2-SO4 and actinomycin-CsC1 equilibrium density gradients, has been used to study the clustering of genes coding for different histones and the size of the repeating multigene cluster. When digested with EcoRI restriction endonuclease, the histone DNA is identified in two classes of fragments with molecular weights of 1.15 X 106 and 2.8 X 106, whereas after treatment of the DNA with HindIII restriction endonuclease, histone gene sequences can be identified only in a fragment of 3.95 X 106. Treatment of the DNA with both enzymes simultaneously shows that there is a HindIII site within the smaller EcoRI fragment. Partial digests with HindIII give fragment sizes that appear to be simple multiples of a 3.95 X 106 repeat. Individual histone mRNAs all hybridize to the 3.95 X 106 fragment but only to one or the other EcoRI fragments. The evidence strongly suggests a repeating unit of 3.95 X 106 containing the genes for most, if not all, the histonrs.


Architecture of the Escherichia coli ribosome as determined by immune electron microscopy. Binding sites for antibodies specific to nineteen of the twenty-one ribosomal proteins from the 30S subunit of E. coli ribosomes have been localized on the surface of the 30S ribosomal subunit by immune electron microscopy. The locations of 13 ribosomal proteins from the 50S subunit were similarily assessed. The arrangement of these proteins is illustrated in three-dimensional models of the 30S and 50S ribosomal subunits and of 70S ribosomes. With specific antibodies to six proteins of the 30S subunit we found only one attachment point for each protein. Antibodies against each of nine of the proteins attached at two separate sites that were separated by various distances. Four further proteins were exposed at three or four sites for antibody binding. Altogether eight to ten of the 19 proteins of the 30S subunit have shown antibody attachment sites at remote points on the surface of the ribosome, at distances which are incompatible with globular shapes; these proteins must therefore have elongated or fibrous structures within the ribosome. On the other hand, only two proteins of the 50S subunit, namely L11 and L18, have so far revealed two separated antibody binding sites; proteins L7/L12 occurred, however, at multiple sites.


Separation of lymphocyte chromatin into template-active fractions with specificity for eukaryotic RNA polymerase II or prokaryotic RNA polymerase. When chromatin prepared from WI-L2 lymphocytes by low salt extraction and shearing is centrifuged on a glycerol gradient, one area of the gradient yields chromatin enriched in template activity for Escherichia coli DNA-dependent RNA polymerase (EC 2.7.7.6; nucleosidetriphosphate:RNA nucleotidyltransferase) as compared to Saccharomyces cerevisiae RNA polymerase II (or B). Another area yields chromatin preferred by the eukaryotic enzyme. Kinetic studies indicate that the differences in activity cannot be explained by differences in affinity of the enzymes for the various templates. The DNA isolated from either fraction has a molecular weight of 8.5 X 106. The "yeast active" fraction seems enriched in proteins. Mixing experiments indicate that the yeast enzyme does not alter the template in such a way as to improve it for the bacterial enzyme.


Strand-selective transcription of globin genes in rabbit erythroid cells and chromatin. In order to investigate the symmetry of globin gene transcription, complementary RNA (cRNA) was synthesized using rabbit globin complementary DNA (cDNA) as a template for Escherichia coli DNA-dependent RNA polymerase (RNA nucleotidyltransferase). The cRNA hybridized specifically to its own cDNA template but not to sheep cDNA, rabbit globin mRNA, or poly(dT). Hybridization studies with cRNA demonstrated that RNA sequences transcribed from the DNA strand complementary to the globin gene region (anti-strand) were not present in cellular, total nuclear, or fractionated nuclear RNA from rabbit marrow. Such sequences were detected in RNA transcribed from rabbit marrow chromatin by E. coli or sheep liver RNA polymerases, but amounted to less than 50% of the globin mRNA sequences present in the same transcript. The evidence indicates that globin mRNA transcription is predominantly DNA strand specific.


Hydrogen bonding in yeast phenylalanine transfer RNA. Further analysis of the three-dimensional electron density map of yeast phenylalanine tRNA is presented. Attention is focused on the several types of unique hydrogen bonding that are found in the molecule and a number of sections of the electron density map are presented. These sections are compared with an electron density map of a dinucleoside phosphate. The bases in the helical stem regions are all involved in Watson-Crick hydrogen bonding interactions with the exception of the guanine-uracil base pair. Several additional tertiary hydrogen bonding interactions are described.


Effects of bacteriophage T4-induced modification of Escherichia coli RNA polymerase on gene expression in vitro. After T4 bacteriophage infection of E. coli a complex series of events take place in the bacterium, including gross inhibition of host transcription and discrete changes in the classes of the genes of T4 that are transcribed. Accompanying these changes in the pattern of transcription one finds T4-induced changes in the RNA polymerase (EC 2.7.7.6; nucleosidetriphosphate:RNA nucleotidyltransferase). The effects of modified polymerase on transcription can be advantageously analyzed in a DNA-directed cell-free system for protein synthesis. In this system gene activity is measured indirectly by the amounts and types of proteins sythesized. In the DNA-directed cell-free system this modified polymerase, like normal polymerase, transcribes T4 DNA with a high efficiency but transcribes bacteriophage lambda and host DNA very poorly. Polymerase reconstruction experiments show that modification of the alpha subunit of the RNA polymerase is sufficient for inhibition of host transcription. Host transcription is also inhibited in vitro by T4 DNA. This latter type of inhibition is presumed to involve competition between host DNA and T4 DNA for some factor essential for transcription. The T4-modified polymerase transcribes from T4 DNA many of the same genes as normal unmodified polymerase; it also shows a capability for transcribing certain "non-early" T4 genes which is enhanced in the presence of protein-containing extracts from T4-infected cells.


Regulation of histidine operon does not require hisG enzyme. Mutations are described which delete all or part of the first structural gene (hisG) of the histidine operon of Salmonella typhimurium. Physiological regulation of histidine enzymes occurs normally in strains carrying any deletion that has both endpoints within the hisG gene. Constitutive high operon expression is observed in strains carrying any hisG deletion and an unlinked regulatory mutation, hisT1504. These results strongly indicate that the hisG protein is not an essential component of the mechanism for regulating expression of the histidine operon.


Identification of a gene for the alpha-subunit of RNA polymerase at the str-spc region of the Escherichia coli chromosome. A structural gene for the alpha-subunit of RNA polymerase (nucleosidetriphosphate:RNA nucleotidyltransferase; EC 2.7.7.6) has been identified and mapped between spcA and trkA, near 64 min on the E. coli chromosome. It appears to be coordinately expressed and possibly cotranscribed with the genes for ribosomal proteins S11, S4, and L17.


Electron microscopic observations on the meiotic karyotype of diploid and tetraploid Saccharomyces cerevisiae. Certain strains of Saccharomyces cerevisiae contain visible segments of synaptonemal complex which are apparent components of bivalents in pachytene of meiotic prophase. The synaptonemal complex has the typical width in the frontal plane but is unusually thin in the sagittal plane, thus accounting for its poor visibility. Amorphous densities situated adjacent to the central element occur at intervals suggesting their coincidence with sites of crossing over. Reconstruction of the synaptonemal complex from serial sections has permitted karyotypic analysis. The number of segments of synaptonemal complex and the distribution of their legths is consistent with the genetic map. Two, possibly three, segments enter the nucleolus as if bearing sequences encoding ribosomal RNA. Reconstruction of tetraploid nuclei reveals an approximate doubling of the diploid chromosome number and confirms the pattern of nucleolar entry. Quadrivalent pairing is evident between the pairs of synaptonemal complex segments in the tetraploid nuclei.


Functional interactions of viral and histocompatibility antigens at tumor cell surfaces. Several lines of evidence are presented to suggest that histocompatibility antigens can be physically associated on the cell surface with viral antigens and possibly other foreign antigens. The lysis of the murine tumor cells EL4 and P388 by syngeneic cytotoxic lymphocytes was inhibited by antisera directed against the H-2 antigens on the tumor cells, consistent with the hypothesis that H-2 antigens are part of the target of the cytotoxic lymphocytes. Moreover, it was found that patching and capping of the H-2 antigens on EL4 cells resulted in the co-patching and co-capping of viral antigens as detected by antisera against Rauscher leukemia virus. Capping of H-2 antigens also resulted in co-capping of determinants detected by an antiserum to the viral protein gp69/71. On the basis of these and other observations, we propose the hypothesis that the H-2 molecules serve as adaptors that combine with viral antigens on the cell surface to form hybrid antigens containing elements of self (H-2) and non-self (virus). The adaptor-antigen complex may then be recognized by a subclass of thymus-derived (T) lymphocytes that possesses a repertoire of receptors directed against hybirds of foreign and H-2 antigens. This raises the possibility that other products of the major histocompatibility complex may have analogous functions.


Immunosurgery of mouse blastocyst.

Immunosurgery of mouse blastocyst. Mouse blastocysts with and without zonae pellucidae are susceptible to complement-dependent antibody cytotoxicity. Exposure of blastocysts to rabbit anti-mouse serum together with complement results in the death of all cells; however, when blastocysts are exposed to antiserum alone and then transferred to guinea pig complement, only the trophoblastic cells are killed. These results suggest that the mouse blastocyst is not permeable for certain antibodies. The inner cell masses can easily be separated from the remnants of trophoblastic cells and are then able to grow and differentiate in vitro. This method of immunosurgery can be used to obtain large quantities of pure inner cell masses in a relatively short period of time.


Alterations in envelope structure of heptose-deficient mutants of Escherichia coli as revealed by freeze-etching. The surface of freeze-etched E. coli strain GR467, a heptose-deficient ("deep rough") mutant derived from CR34, was studied by electron microscopy. The outer membrane of GR467 has an increased ratio of phospholipid to protein, mainly due to a decreased protein content. Freeze-etched CR34 showed structural features indistinguishable for wild-type E. coli, i.e., the primary cleavage occurring in the inner membrane with only minor appearance of cleavage within the outer membrane. In contrast to this, in mutant GR467 most of the freeze-cleavages had taken place along a new plane, presumably in a hydrophobic region of the outer membrane. In this cleavage plane numerous particles were seen. Often the cleavage extended over the entire exposed cell surface; occasionally only a few large plateaus were visible, around which the next deeper cleavage plane, that of the protoplasmic or inner membrane, was discernible. Two spontaneous revertants (R11 and R16) with protein and lipid A levels similar to wild-type cells showed mostly freeze fractures with wild-type characteristics, and only a few cells had retained fracturing properties of GR467. A partial revertant revealed intermediate characteristics. Thus, there appears to be a morphological correlation with the chemical data relating the amount of outer membrane protein with the heptose content of the lipopolysaccharide.


Novel F prime factors able to replicate in Escherichia coli Hfr strains. A novel type of F' plasmid that can replicate extrachromosomally in cells of Hfr strains has been isolated. Genetic analysis of these plasmids indicates that all carry the dnaA-bglA region of the E. coli chromosome as well as ilv+ used as a selective marker. It is suggested that a specific site, designated poh+ (permissive on Hfr), is located in this region, and is essential for these plasmids to replicate in Hfr cells. The most interesting explanation, among others, would be that the poh+ site represents the replication origin of the bacterial chromosome.


Serial propagation of Creutzfeldt-Jakob disease in guinea pigs. The transmission and serial propagation of Creutzfeldt-Jakob disease from man to guinea pigs are reported. The latency, symptomatology, and morphology of the infection during the first four passages are presented. The incubation period between the first and subsequent passages was halved. One hundred percent take, morbidity, and mortality were achieved in all inoculated animals. All guinea pigs developed a subacute spongiform virus encephalopathy with marked neuronal destruction in the cerebral cortex and subcortical grey structures. The neuronal loss resulted in cerebral atrophy and hydrocephalus ex vacuo.


Does somatostatin inhibition of insulin secretion involve two mechanisms of action? Somatostatin, the hypothalamic growth hormone release inhibitory factor (GHRIF), directly inhibits both the first and second phases of insulin secretion. The sensitivities of these two phases of insulin secretion to somatostatin differ remarkably. The first phase of secretion is approximately 25 to 50 times more sensitive to somatostatin inhibition than is the second phase. In addition, somatostatin inhibition of insulin secretion during the second phase is "reversed" by supplemental calcium, whereas the somatostatin effect on the first phase is unaffected by additional calcium. These findings suggest that the cellular events which produce the two phases of insulin secretion are separate processes, and that somatostatin has a dual mechanism of action in inhibiting insulin secretion.


A DNA fragment from the origin of single-strand to double-strand DNA replication of bacteriophage fd. A specific complex is formed between fd DNA, Escherichia coli DNA unwinding protein, and RNA polymerase (EC 2.7.7.6; nucleosidetriphosphate:RNA nucleotidyltransferase) during the first steps in the conversion of the single-stranded viral DNA to the double-stranded replicative form. In this complex a unique DNA fragment of about 120 nucleotides is protected against nuclease digestion. Both the requirements for its isolation and its position on the map of the phage genome indicate that the fragment contains the origin of single-strand to double-strand DNA replication. The isolated DNA fragment possesses double-strand-like characteristics, which protect it from being covered by the DNA unwinding protein and thus indirectly positions the RNA polymerase to the origin of replication.


Mechanism of growth delay induced in Escherichia coli by near ultraviolet radiation. Continuously growing cultures of E. coli B/r were irradiated with a fluence of broad-band near-ultraviolet radiation (315-405 nm) sufficient to cause extensive growth delay and complete cessation of net RNA synthesis. Chloramphenicol treatment was found to stimulate resumption of RNA synthesis, similar to that observed with chloramphenicol treatment after amino-acid starvation. E. coli strains in which amino-acid starvation does not result in cessation of RNA synthesis ("relaxed" or rel- strains) show no cessation of growth and only a slight effect on the rate of growth or of RNA synthesis. These findings show that such near-UV fluences do not inactivate the RNA synthetic machinery but affect the regulation of RNA synthesis, in a manner similat to that produced by amino-acid starvation. Such regulation is believed to be mediated through alterations in concentration of guanosine tetraphosphate (ppGpp), and our estimations of ppGpp after near-UV irradiation are consistent with such an interpretation. These data, combined with earlier published data, strongly suggest that the mechanism of near-UV-induced growth delay in E. coli involves partial inactivation of certain tRNA species, which is interpreted by the cell in a manner similar to that of amino-acid starvation, causing a rise in ppGpp levels, a shut-off of net RNA synthesis, and the induction of a growth delay.


Molecular cloning of DNA from F sex factor of Escherichia coli K-12. We describe the molecular cloning of various DNA segments generated by partial EcoRI endonuclease digestion of the sex factor F. These segments have been analyzed by agarose gel electrophoresis of EcoRI digests and were arranged in a series of overlapping fragments using the EcoRI fragment map of F established by H. Ohtsubo and E. Ohtsubo. The clones isolated demonstrate one or more of the following F-specified functions: inhibition of female-specific phage (T7) multiplication, formation of F pili, surface exclusion, or immunity to lethal zygosis. These properties are discussed in terms of the EcoRI fragments of F that specify them.


Incorporation of fatty acids containing photosensitive groups into phospholipids of Escherichia coli. A series of new fatty acids containing photosensitive groups at different positions on the paraffin chains supported the growth of an auxotroph of E. coli requiring unsaturated fatty acids. The derivatives were 6-, 9-, 11-, and 12-azidostearic acids, 12-azido-oleic acid, 16-azidopalmitelaidic acid, and 12-(4-azido-2-nitrophenoxy)-stearic and -oleic acids. Analyses of the phospholipids from cultures grown in the presence of the first six compounds showed that these derivatives accounted for 16-43% of the total fatty acids. Further analysis of phospholipids from cultures grown with 12-azido-oleic acid, 11-azidostearic acid, or 16-azidopalmitelaidic acid indicated that the azido fatty acids were at the 2-position of the glycerol moieties. The incorporation of these fatty acid derivatives offers a new approach to the study of membrane structure and, in particular, phospholipid-protein interactions by photolysis-induced crosslinking of the fatty acids to the structures in their immediate vicinity.


Chemical and enzymatic synthesis of lactose operator of Escherichia coli and its binding to lactose repressor. The 21-nucleotide-long duplex DNA constituting the lactose operator sequence of E. coli has been synthesized by both chemical and enzymatic methods. The synthetic duplex has the essential feature of the lactose operator as seen by its binding to the lactose repressor. The binding of the synthetic operator fragment to the lactose repressor is specific because it is inhibited by the inducing ligand isopropyl-beta-D-thiogalactoside. Thus, it is now possible to show that a chemically synthesized oligodeoxynucleotide can be specifically recognized by its natural regulatory protein.


Position of aminoacylation of individual Escherichia coli and yeast tRNAs. Transfer RNAs terminating 2'-or 3'-deoxyadenosine were prepared from unfractionated E. coli and yeast (Saccharomyces cerevisiae) tRNAs and purified to remove unmodified tRNAs. The modified tRNA species were assayed for aminoacylation with each of the 20 amino acids to determine the initial position of tRNA aminoacylation. The E. coli and yeast aminoacyl-tRNA synthetases specific for arginine, isoleucine, leucine, methionine, phenylalanine, and valine, as well as the E. coli glutamyl-tRNA synthetase, aminoacylated only those cognate tRNAs terminating in 3'-deoxyadenosine (i.e., those having a 2'-OH group). On the other hand, those E. coli and yeast synthetases specific for alanine, glycine, histidine, lysine, proline, serine, and threonine, as well as the yeast synthetase specific for glutamine, utilized exclusively those tRNAs having an available 3'-OH group on the 3'-terminal nucleoside, while the E. coli and yeast synthetases specific for asparagine, cysteine, and tyrosine, and the yeast aspartyl-tRNA synthetase, utilized both of the modified cognate tRNAs. The only observed difference in specificity between the E. coli and yeast systems was for tRNATrp, which was aminoacylated on the 2'-position in E. coli and the 3'-position in yeast. The results indicate that the initial position of aminoacylation is not uniform for all tRNAs, although for individual tRNAs the specificity has been conserved during the evolution from a prokaryotic to eukaryotic organism.


Studies on nucleic acid reassociation kinetics: empirical equations describing DNA reassociation. The rate of appearance of duplex DNA renaturation, measured with single strand specific nuclease, deviates significantly from a second order reaction. Measurements reported in paper I of this series indicate an inhibition in the rate of reassociation of single strand tails on partially reassociated molecules by a factor of at least two. Equations are derived that describe the observed form of reassociation kinetics as measured with hydroxyapatite and with single strand specific nuclease. The free parameter that describes the extent of inhibition of nucleation with single strand tails in these equations has been evaluated by least squares methods and agrees with the experimentally measured value.


Visualization of prokaryotic DNA in a regularly condensed chromatin-like fiber. Electron microscopy of disrupted Escherichia coli cells under certain conditions revealed loops of a fiber 120 A in diameter which were attached to the cell envelope and showed a 130 A repeating beaded substructure. These fibers were detected only when the cells were lysed in 0.15 M NaCl solutions directly on the electron microscope supporting films and if the dehydration steps began within 2 min of lysis. Under these conditions examination of cells lysogenic for phage lambda after superinfection with lambda wild type or deletion mutants disclosed short loops of a 120 A diameter fiber free of the cell envelope. Because the contour length of these loops was proportionate to the DNA content of the superinfecting lambda phage, it was concluded that the fibers contained DNA condensed 6.5-fold in blocks of about 250 base pairs.


Comparison of three hemolytic plaque assays using sheep and mouse erythrocytes as antigens in rats. Several hemolytic plaque assays using spleen cells from rats immunized with sheep or mouse erythrocytes were compared. Modifications of the assays were made to define optimal conditions. Mrbc in assays with guinea pig complement yield a much reduced number of plaques than is observed when human complement is used. In contrast, no difference in numbers is noted with Srbc regardless of complement source. The use of Srbc in an assay technique that does not employ agarose is more sensitive than techniques that require agarose; the result is a 50-100% increase in the number of plaques. This increase in plaque number was observed with either guinea pig or human complement.  In contrast, there was considerable clumping of Mrbc in the absence of agarose which tended to obscure distinct plaque formation. Increased sensitivity of plaque development for Srbc in the absence of agarose was due to increased numbers of direct (IgM) plaque detection and not to concomitant detection of cells producing IgG (indirect plaques). Maximal detection of plaques with Mrbc was observed when human complement was used in assays containing agarose. These assays gave a threefold increase in plaque numbers when compared with assays without agarose. It is evident from the above that the most sensitive and reproducible hemolytic plaque assay to be used by an investigator will depend primarily upon the antigen, and on reagents selected for the test.


Evidence for the dark repair of ultraviolet damage in Saccharomyces cerevisiae mitochondrial DNA. Evidence for the dark repair of ultraviolet damage to yeast mitochondrial DNA has been observed. The ultraviolet dose necessary to inflict significant damage to both nuclear and mitochondrial DNA was determined. Cell survival at large doses of ultraviolet light was observed after immediate and delayed plating of yeast onto 1% pyruvate and 1% glucose media. In the highly lethal dose ranges of irradiation an increase in the number of normal colonies appeared after a period of liquid holding and delayed plating. This increase, demonstrated separately on 1% glucose and 1% pyruvate media suggested that the repair of both mitochondrial and nuclear DNA had occurred. After low doses of ultraviolet light an actual decrease in the number of petite survivors was seen after delayed plating, even though the total number of survivors increased. When a known repair inhibitor, caffeine, was added to the liquid holding buffer prior to the delayed plating of yeast, a marked decrease in the number of petites did not occur after delayed plating. Therefore, the decrease in the number of petite survivors after delayed plating following low doses of ultraviolet light is attributed to the repair of yeast mitochondrial DNA.


Effect of short-term castration and starvation upon hypothalamic content of luteinizing hormone-releasing hormone in adult male rats. A sensitive and specific radioimmunoassay for hypothalamic LH-RH has been described. Within 7 days after castration there is a significant decline in hypothalamic content of LH-RH in adult male rats. Total starvation for 7 days does not affect hypothalamic content of LH-RH in either intact or castrated rats.


Origin of antibody-dependent, lymphoid-cell-independent hemolytic plaques in rabbit spleen cell cultures. This report describes a phenomenon that may be a significant source of error in the data acquired from hemolytic plaque assays of lymphocyte cultures. The data show that background antibody in the normal-serum supplement of the culture medium does not actually support initiation of a vigorous primary-type plaque-forming cell (PFC) response to sheep erythrocytes (SE). Instead, the background antibody causes the development of numerous spurious lymphoid-cell-independent plaques (LCIP). In general agreement with the literature, primary-type rabbit spleen cell responses in vitro were low in culture media supplemented with fetal bovine serum, a serum which does not contain background antibodies to SE.


Effects of allogenic and effector cells on the development of antibody-producing cells, in vitro. Effects of allogenic and effector spleen cells on the development of anti-SRBC plaque-forming cells from target spleen cells in vitro were studied. Addition of allogeneic cells resulted in various degrees of suppression of target cell PFC production. This allogeneic suppression could be abolished by pretreating the cells with either uv or X irradiation. Addition of effector cells to target spleen cell cultures markedly suppressed PFC production. Effector cell suppression of PFC production was anti-BAtheta serum-sensitive and radiation-resistant.


Influence of lead and cadmium on the susceptibility of rats to bacterial challenge. Intravenous administration of an acute dose of lead acetate or cadmium acetate enhanced the susceptibility of rats to intravenous challenge with E. coli by approximately 1000-fold. Since equivalent vulnerability of lead- or cadmium-treated rats to killed E. coli was observed, toxicity is probably due to the endotoxin content of the bacteria. This postulate is further supported by the observation that equal doses of the Gram-positive bacteria, Staphylococci epidermidis, failed to elicit lethality in the acute lead-intoxicated rats. The synthetic glucocorticoid, methylprednisolone, prevented lethality induced by the Gram-negative bacteria in lead-treated rats. It did not, however, afford significant protection in cadmium-treated rats in the presence of E. coli. Marked alterations in hepatic morphology were apparent in both lead- and cadmium-treated rats challenged with E. coli.


Effect of acute infection and endotoxemia on zinc absorption in the rat. Intestinal zinc absorption was found to be significantly increased during acute bacterial infection and endotoxemia in the rat. Although serum zinc concentrations were depressed, there was a significant accumulation of 65Zn in the livers of the stressed animals. This study demonstrates that acute inflammation produces a redistribution of zinc within the host, which results in both increased zinc absorption and retention.


Human figure drawings by children with Duchenne's muscular dystrophy. Seventy-two human figure drawings by forty-three patients who had a diagnosis of Duchenne's muscular dystrophy were examined. The study includes a description of these human figure drawings according to eleven emotional indicators and according to directionality quadrants. When the human figure drawings were used as a projective tool, four personality traits of some of the children were identified: physical inadequacy, immaturity, body anxiety, and insecurity. Both the emotional indicators and the quadrant in which the figures appeared were examined in relation to stages of the disease process to see if the human figure drawings of the children might reflect more stress and anxiety at a particular stage of the disease. Suggestions for improvements and recommendations for future study are given.


[On the philosophical genealogy of Freud]. The origins of psycho-analysis, like those of every other medico-psychological study, have their own particular scientific and specific social, historical and philosophical-theoretical presuppositions. Freud's philosophical genealogy is closely linked to classical german philosophy and subsequent philosophical movements. I. Kant, J.-F. Herbart, A. Schopenhauer, F. Nietzsche, E. v. Hartmann, G. T. Fechner, E. Mach, W. Ostwald, L. Feuerbach and others did not only emphasise the significance of drives and the unconscious in human behaviour, they also described many psychological mechanisms from depth psychology, (for example repression, condensation, substitution, sublimation). Some false theoretical trends in psycho-analysis (biologism, psychologism and simplifying psycho-energetics to simplify) can be explained to some extent by the influences mentioned above.


[Carl Hohnbaum, the life and work of a distinguished neurologist, founder of the contemporary Hildburghausen District Hospital]. Carl Hohnbaum was the founder of what is now the Hildburghausen District Mental Hospital. He was born at Coburg in 1780, called to Hildburghausen in 1805 after having been active as a general practitioner for a short time, and appointed chief medical officer of the former Duchy. He devoted much of his time and effort to improving medical care for the inmates, and he thus made a meritorious contribution to the treatment of mental and emotional disorders. Hohnbaum was one of the most productive medical writers of his time. He died in 1855. In an obituary published in the Village Paper he was designated as one of Hildburghausen's "most distinguished and eminent" citizens.


Gilbert's first night anxiety.

Gilbert's first night anxiety. W. S. Gilbert, librettist of the Gilbert and Sullivan operas, suffered from such extreme anxiety at the opening night performance of his plays that he was unable to remain in the theater. This paper explores the dynamics behind Gilbert's first night anxiety, utilizing his literary works as well as biographical and autobiographical sources.


The effects of n-dipropylacetate on the acquisition of conditioned behaviour with negative reinforcement in mice. n-Dipropylacetate (nDPA), at a dose of 100 mg/kg, has a facilitating action on the acquisition of conditioned reactions with negative reinforcement in mice. On the other hand, nDPA reduces the number of conditioned reactions with a dose of 200 mg/kg. These effects of nDPA on conditioned behaviour are correlated with the increase of the level of brain gamma-aminobutyric acid, following administration of nDPA.


The De Carvalho and acid-barium tests in the demonstration of an unsuspected gastro-oesophageal reflux and its related oesophagitis. Importance of the water and acid-barium tests (De Carvalho and Donner tests) was proved in a large series of cases demonstrating an unknown gastro-oesophageal reflux and an associated oesophagitis.


Total body irradiation in the management of malignant lymphoma. A pilot study for the evaluation of fractionated total body irradiation in the management of disseminated lymphomas is reported. Nine patients were treated in the period February - August, 1973. A telecobalt unit with treatment distance of 328 cm was used. Daily fractions of 15 rad total body dose measured in the midpelvic plane were given for the first 5 days. Thereafter a dose of 10--15 rad per session was given every other day or at longer intervals depending on the leukocyte and thrombocyte count. The total dose for a full course varied between 200 and 265 rad in 5--12 weeks. The results of the study demonstrate the value of total body irradiation in the treatment of patients with stage III lymphosarcoma, and potentially in the management of disseminated mycosis fungoidis.


Immunosuppressive anticancer drugs in man: their oncogenic potential. As therapy of cancer becomes more sucessful, the number of long-term survivors is increasing. In addition, cancer chemotherapeutic drugs are being used as immunosuppressive agents in the chronic treatment of non-neoplastic diseases. The long-range hazards of therapy are a serious consideration in these two groups of patients. This brief review examines the potential oncogenic as well as co-oncogenic hazards of cancer chemotherapy in man. Many of these drugs are oncogenic and/or co-oncogenic in experimental animals.


Granulomatous disease of the intestinal tract (Crohn's disease). In a landmark lecture on this disease, the author presents his experience with some 8,000 cases of regional enteritis and 4,000 of granulomatous colitis. The paper is divided into sections on the history, pathology, pathogenesis, roentgen findings, and course of the disease. The main thrust of the paper is the extensive roentgen analysis of granulomatous disease, with a discussion of differential diagnosis, especially from ulcerative colitis. Through the years, the author has been impressed with certain clinical and roentgen features of this malady (his Ten Principles of Crohn's Disease) which are carefully reviewed.


Computed tomography with the EMI scanner in the diagnosis of primary and metastatic intracranial neoplasms. The findings and diagnostic results in 600 examinations of primary and metastatic intracranial neoplasms performed at one hospital with the EMI scanner are presented and the effectiveness of this method is compared with that of radionuclide imaging, cerebral angiography, and pneumoencephalography. The computed scan proved to be highly reliable in the diagnosis of glioma, cerebral metastases, meningioma, and acoustic neuroma.


Atraumatic evaluation of myocardial revascularization procedures with 43K1. Least-squares regression analysis was computerized to provide functional color images of myocardial perfusion patterns. In 8 of the 9 patients studied, changes in pattern indicated that the revascularization procedure speeded the appearance time and/or net rate of 43K uptake. The functional images correlated well with coronary arteriograms made before and after surgery. This noninvasive isotope technique may prove to be clinically useful in (a) evaluating coronary artery disease, (b) defining the remaining viable myocardium when an aneurysm is present, and (c) determining the effectiveness of the revascularization procedure.


Improved isolation, separation and cytochemistry of living cells. 1. This paper describes improved methods of obtaining, purifying and studying bulk suspensions of isolated living hepatocytes and other cells of adult rats and urodeles. 2. The cells were isolated largely by dissolving the hepatic ground substance through the extracorporeal portal perfusion and further incubation of the excised liver with 0.05% collagenase and 0.1% hyaluronidase. The different kinds of cells were then separated from one another by counter-current centrifugation. The isolated cells were examined by differential interference, phase-contrast, amplitude-contrast, ultraviolet, fluorescence and electron microscopy. Various cytochemical tests were carried out. Whenever possible, for each method of examination, the isolated cells were compared with cells of the same kind which had not undergone isolation. 3. Dye-exclusion, lysochromy, fluorescence and differential interference microscopical analysis indicated viability rates between 75 and 99%. Succinate dehydrogenase activity was preserved at a high level in nearly all isolated cells. In hepatocytes, the essentially extracellular cells. In hepatocytes, the essentially extracellular 'soluble' alkaline phosphatase activity of bile canaliculi was retained. Living hepatocytes were studied by super-modulating methods of microscopy for the first time, with somewhat unexpected findings. It now seems probable that previous methods of tissue preparation produced gross alterations in hepatocyte mitochondria. The assessment of the viability of isolated cells was re-examined. 4. The methods described may permit a more meaningful correlation between biochemical, cytochemical, ultrastructural and biophysical findings than that obtainable by the use of current methods.


Temperature and vision.

Temperature and vision. Published information concerning the influence of temperature in the physiological range (0-40 degrees) on various aspects of the visual process in invertebrates and vertebrates (poikilotherms and homeotherms) is reviewed. Bleaching and regeneration of scotopic and photopic pigments is influenced by temperature; so is the porphyropsin-rhodopsin ratio in fresh-water fishes. Effect on electrophysiological responses such as early receptor potential (ERP), electroretinogram (ERG) and its flicker fusion frequencies (FFF), S-potentials, glial membrane optic nerve and brain responses is presented. Histophysiological responses such as photomechanical or retinomotor responses are influenced by temperature. It also affects behavioural responses such as innate light reflexes, colour change, sensitivity, threshold acuity, FFF, spectral sensitivity and conditioned responses. General aspects such as oxygen consumption of the retina, acid formation and survival times are discussed with reference to their modification by temperature. Theoretical aspects such as the interaction between light and thermal energy in the absorption of a photon by a molecule, and available equations for comparing the effect of temperature on biological processes are also discussed. An attempt is made to draw a coherent picture and to discuss the significance of the thermal influence on vision to the animal. New avenues of research are also indicated. One hundred and eighty-eight (188) references are cited.


Similarity of acetaldehyde and sulfhydryl reagent actions on the perfused guinea pig heart. The effects of acetaldehyde (1 mM) and two known sulfhydryl reagents, diamide (0.25 mM) and cystamine (0.5 mM), on the perfused guinea pig heart were compared and were found to be markedly similar. All compounds increased heart rate and coronary flow. Dichloroisoproterenol blocked the increase in heart rate but not the increase in coronary flow. It is concluded that acetaldehyde may produce its effects by reaction with cellular thiols.


Resting bradycardia of exercise training: a concept based on currently available data. The classic view that the resting bradycardia of exercise training is due to an increase in the tonic discharge of the vagi is no longer tenable; If it were true, full doses of atropine would accelerate the heart to a greater extent in the athletic animal. All investigators agree that the reverse is the case. Some workers report a decrease in cardiac sympathetic influence after training. This would cause a preponderance of vagal influence which may decrease to a lesser extent than the sympathetics or may not change at all. Several studies have demonstrated that training causes a distinct slowing of the intrinsic rate of the pacemaker. It is concluded that two factors operate to lower the resting heart rate after athletic training: 1) slowing of the intrinsic rate of the pacemaker, 2) Increase in cholinergic predominance on the pacemaker frequency as a result of a decrease in adrenergic influence. Vagal influence per se seems either to remain the same or to decrease slightly in the athlete compared to the nonathelete. It appears that slowing of the intrinsic rate plays a more important role, a factor that has been hitherto overlooked. The mechanisms underlying these changes remain obscure.


[Etiology of Paget's disease of bone].

[Etiology of Paget's disease of bone]. The etiology of Paget's disease is just as doubtful in 1975 as it was in 1876 when Sir James Paget described the disease. The authors analyse the etiology on the basis of 100 personal cases and the literature. Although there are undoubtedly familial cases of the disease, investigation of the leucocyte grouping of 46 patients with Paget's disease did not reveal any correlation between occurrence of the disease and the HL-A antigens. Various pathological associations have been reported in the literature and were also found in this series. These associations were at the limits of coincidence (inflammatory rheumatism, diffuse chondrocalcinosis, multiple myeloma...). Metabolic changes (hyperuricaemia, hyperglycaemia, dyslipidaemia) did not appear to be more frequent than in control patients. Involvement of elastic tissue and the presence of pseudocrystalline inclusions in the osteoclasts constitute interesting points for discussion.


[The cardiovascular manifestations of Paget's disease]. After reviewing the old ideas (peripheral syndrome and cardiac insufficiency at high outputs) and more recent ideas (arterial and intracardiac vascular calcification), the authors report their experiences based on the study of the histories of 150 patients suffering from Paget's disease followed for several years. First they emphasize the frequent latency of cardiac manifestations looked for systematically by means of the electrocardiogram particularly during evolutive stages of the disease. Electrocardiographic disturbances were observed 91 times and included 43 cases of conduction disorders and 48 of repolarization disorders; their distribution was studied as a function of various inherent parameters, of the patient (age and sex) and of the disease (its evolutive character and its degree of diffusion). On the basis of the anti-arythmic power of calcitonin found during experiments and obtained excellent results three times. Finally the place of cardiovascular manifestations in the pathogenic hypotheses and nosology of Paget's disease is discussed.


[The neurologic complications of Paget's disease]. There are several neurological complications of Paget's disease. They may concern the medulla, the cauda equina, and also the cerebral trunk, the cranial nerves, and the brain. Complications are rare when the great frequency of Paget's disease is considered. But they should be recognized because they constitute an aspect of the disease the treatment of which is usually well tolerated of even negligible. The signs of involvement of the medulla, of the cauda equina, and those which complicate the basilar impressions usually develop in a progressive and not very spectacular fashion. Nevertheless, they should be recognized as soon as possible so that therapeutic measures may be taken. The results of modern medical treatments, calcitonin, cellulolytic medications, and possibly others are already sufficiently established to justify their use initially. Indications for surgical treatment appear when a check in medical therapy is observed after three months of treatment.


[Radiculomedullary compressions by osteosarcoma and by giant cell tumor of the spine occurring in Paget's disease]. Two complications of Paget's disease are presented : they include an osteogenic sarcoma and a giant-cell tumour (GCT) of the lumbar spine with a radiculo-medullary compression syndrome. The rarity of sarcomatous degeneration of the spine affected by Paget's disease is emphasized and also the unusual character of the spinal GCT which develop in the presence of Paget's disease. As in the present case, it is often difficult to confirm the benign or malignant character of a GCT and the principal criteria of this classification are discussed.


[Neurosensory complications of Paget's disease]. The authors made a bibliographic study and report their conclusions with reference to problems concerning a personal series of 17 patients who had undergone an ocular examination, a cranial radiography studying the joint, a bilateral radiotomographic study of the ossicles and of the petrosal bone, an audiogram, and a bilateral electronystagmogram: 1. the rarity of angioid striae, the existence of which, in the view of the authors, does not allow Paget's disease to be included within the framework of the systemized elastorrhexis, 2. the frequency of ocular vascular lesions, 3. deafness is a constant phenomenon, when the cranial arch is affected usually in combination with other lesions ;  the deafness is sometimes of transmission or sometimes of perception, but it can precede the cranial lesions. The signs of these can be found radiologically in the chain of ossicles and in the cochlea by means of special projections. The part played by basilar pressure in this deafness is negligible. Labyrinth disorders are rarely met.