[Clinical results of cefuroxime (CXM) therapy in pediatric infections (author's transl)].

Cefuroxime, a new cephalosporin C antibiotic, was administered to 15 children with respiratory tract infection, urinary tract infection, or subcutaneous tumour. The following results were obtained. 1) CXM 30 approximately 100 mg/kg/day were used in treatment of respiratory tract infection. Eight of the eleven patients treated responded to the therapy. 2) CXM 45 approximately 75 mg/kg/day were given to 3 patients with urinary tract infection. Excellent results were obtained in all these cases. 3) One patient with subcutaneous tumour responded to CXM therapy. 4) Clinical isolates from the foci involved, i.e., Staphylococcus aureus (4 strains), Group A Streptococcus hemolyticus (1 strain), Streptococcus pneumoniae (1 strain), Haemophilus influenzae (1 strain), and Escherichia coli (3 strains) were all eliminated by CXM therapy except 2 unassessable strains. 5) No noteworthy side effect was noted.


[Maintenance of the circulation and the restoration of heart function in the donor body by means of direct mechanical heart massage for subsequent transplantation (an experimental study)].

The possibility of maintenance of circulation and subsequent restoration of the function of the heart in the organism of the donor, after 10-minute arrest of circulation, for transplantation of the heart was investigated. Study of the hemodynamic, biochemical, metabolic, and structural changes in the myocardium after 2-hour mechanical massage without and with pharmacological protection of the myocardium (glucose--insulin--potassium, hydrocortisone, dipyridamole), without and with restoration of cardiac activity in the organism of the donor and the recipient after transplantation of the second heart by the method suggested by Moory et al. in artificially created left ventricular insufficiency showed that direct mechanical massage of the heart may be used for its short-term preservation in the donor organism.


[Kidney in chronic arterial hypertension: "resetting" and the role of the renal medulla in its development].

In the system of long-term maintenance of the arterial pressure level by means of the renal excretion mechanism described by Guyton et al., the renal medulla with its counterflow-concentrating multiplier and powerful source of natural natriuretics, the prostaglandins, plays the role of a "control device", a regulator setting the level of arterial pressure required for fulfilling the complete volume of renal excretion of salts and water. Increase in systemic arterial pressure causes the development of a complex of structural-functional changes in the kidney, the common purpose of which consists in preventing the loss of salts and water by the organism by shifting parameters of the excretory renal function in such a manner that the normal (balanced with the supply of salts and water to the organism) volume of excretion may occur with higher values (kidney "resetting"). The central role in accomplishing this "resetting" is played by the medullary excretory mechanism which is closely connected, due to the absence of autoregulated blood flow in the medulla, with systemic arterial pressure. When the kidney reaches the "resetting" this mechanism provides for a sufficiently full volume of excretion only when the arterial pressure increases, maintaining its fixed (higher) level by means of the feedback mechanism. In hypertension caused by overdosage of corticosteroid hormones or excess of salt in the diet the increase of pressure develops as a compensation for the occurring deficiency of the medullary excretory mechanism.


Reactivity of gomerular afferent and efferent arterioles in renal hypertension.

Renal tissue from neonatal hamsters was grafted into the cheek pouch of adult hamsters. Renal hypertension was induced in 28 hamsters, and sham operation was performed in 27. When blood pressure became elevated in hypertensive hamsters (10 to 12 days), the renal microcirculation of both groups was evaluated by direct microscopy. Wall and luminal diameter and vascular responses to norepinephrine (NE) and angiotensin II (AII) were determined on glomerular afferent and efferent arterioles and on the corresponding cheek pouch arterioles. In the hypertensive hamsters, the luminal diameter of the afferent arteriole increased, as did the wall/lumen ratio of the efferent arteriole. Although all vessels responded to direct application of NE and AII in both groups, the response of the afferent to AII and the response of the efferent to both NE and AII was greater in the hypertensive hamsters as compared to the sham-operated hamsters. These results indicate that during the development of renal hypertension structural alterations of glomerular vessels and the selective vascular responses to vasoactive agents would lead to an increased glomerular capillary pressure.


Crescentic glomerulonephritis without immune deposits: clinicopathologic features.

Of 46 patients with acute crescentic glomerulonephritis involving 20 to 90% of glomeruli, 16 had no definable systemic disease and no significant glomerular immune deposits by immunofluorescent or electron microscopy. Anti-GBM antibody and circulating immune complexes were further excluded by radioimmunoassay and Raji cell assay in all patients tested. Clinical features included a 10:6 male:female ratio, mean age of 58 years (range, 13-77), disease duration of less than 3 months, rapidly deteriorating renal function, and frequent pulmonary manifestations. Nine patients had oliguria, serum creatinine concentrations over 6 mg/100 ml, and required dialysis, but three of these patients subsequently recovered renal function. These three patients and seven patients with creatinine concentrations of less than 6 mg/100 ml have not progressed to chronic renal failure. In this series, idiopathic acute crescentic glomerulonephritis without immune deposits was more common than was immune complex or anti-GBM nephritis. The clinical, laboratory, and pathologic characteristics of these patients were similar to those reported in anti-GBM and immune-complex-induced glomerulonephritis. These observations expand the spectrum of rapidly progressive crescentic glomerulonephritis. They suggest that glomerular immune deposits may be less important than other factors in determining the extent of renal injury and subsequent clinical course in crescentic glomerulonephritis.


Effects of dopamine on canine intrarenal blood flow distribution during hemorrhage.

Renal blood flow distribution was measured in control dogs, dogs given dopamine, hemorrhaged dogs, and dogs hemorrhaged plus infused with dopamine with a modification of 85Kr washout. Kidneys injected with 85Kr through a renal arterial cannula were removed at several specific intervals after injection, rapidly frozen, and sectioned transversely so that pieces of tissue could be isolated and counted for radioactivity. In the normotensive animals, dopamine appeared to produce a mild vasodilatory effect in the subcortical outer medulla (flow increased 50%). Hemorrhage reduced renal regional flow throughout the kidneys. Subcortical outer medullary flow, however, appeared to be proportionately better maintained than were the more peripheral renal regions, so that all regions had similar flows. Hemorrhaged animals receiving dopamine infusion had statistically significantly higher cortical blood flows than did the animals simply hemorrhaged. From this study, it is impossible to determine if the cortical vasodilation during hemorrhage was a direct or indirect effect on the renal vasculature; however, improved perfusion of the renal cortex during hypotension may partially explain the improved renal function reportedly produced by dopamine infusion in patients in shock.


Recurrence of dense deposits in transplanted kidneys: I. Sequential survey of the lesions.

Serial specimens from transplanted kidneys were obtained in 11 patients with dense deposit disease (DDD). The recurrence of DDD was obvious in 9 patients and appeared very early after grafting. Three different types of evolution of the lesions were observed. In 4 patients no modification of the lesions occurred with time, and in 2 of them the dense alteration alone persisted without any other glomerular changes. In 3 patients a progression of the lesions was observed, whereas a regression occurred in 1 other patient. From these observations the following sequence of the morphologic changes can be proposed: the dense alteration appears first and constitutes the specific marker of that disease; the C3 deposition in the kidney occurs later following the appearance of the dense lesion.


Recurrence of dense deposits in transplanted kidney: II. Serum complement and nephritic factor profiles.

Dense deposit disease of the kidney is a rare form of chronic glomerulonephritis frequently associated with serum complement abnormalities (low C3 levels) and a circulating C3 convertase activator of the alternative pathway, the C3 nephritic factor (NF). Eleven patients with end-stage dense deposit disease underwent kidney transplantation. Of the 11, 7 had pretransplant low C3 and NF. In the posttransplant period, persisting low C3 levels were associated with persisting NF, although not quantitatively so. The original glomerular lesion recurred in the graft within 6 months in 9 of 11. Of these 9, 2 had no complement abnormalities either prior to or after transplantation. Pretransplant complement abnormalities were rapidly corrected in 4 of 7 patients whether or not recurrence of the original lesion occurred. Thus, serum complement profiles before and after transplantation are neither predictive nor indicative of recurrence.


Experimental glomerulonephritis induced by in situ formation of immune complexes in glomerular capillary wall.

An experimental model of glomerulonephritis was produced by the in situ formation of immune complexes directly in the glomerular capillary wall. Perfusing the lectin concanavalin A (Con A) into the left renal arteries of rats led to its binding diffusely to the glycoproteins of the glomerular capillary wall of only that kidney in each animal. The subsequent reaction with anti-Con A antibody (either administered systemically or actively induced) resulted in an exudative and proliferative glomerulonephritis confined to the Con A perfused kidney. Immunofluorescence disclosed the diffuse deposition of immunoglobulin, Con A, and C3 in the perfused, but not the unperfused kidney. The quantitative relationship between antigen and antibody binding and histologic outcome was determined. Since lectins have been found in mammalian tissues, as well as in infectious agents that are pathogenic in man, a series of events conceptually similar to this in situ model may occur in some cases of glomerulonephritis in man.


Fractional excretion of sodium after renal transplantation.

After renal transplantation low urinary sodium concentration (UNa) has been used to diagnose acute rejection (AR), for the early phase of AR is often associated with reduced renal perfusion. Early postoperative graft failure without low UNa favors the diagnosis of ischemic tubular damage (ATN). As fractional excretion of filtered sodium (FENa) better reflects glomerulotubular balance in renal sodium handling, FENa was analyzed during the first 2 weeks in 118 renal allografts. From data on 41 transplants with good early renal function (GEF), a temporal profile of FENa was obtained and used to evaluate the behavior of FENa by means of standardized FENa (Z score). Individual subjects followed their own profile with a small deviation (delta Z less than 1.4 for 2 days). In 31 instances, acute rejection was diagnosed. In 14 with AR, the Z score deviated little; 2 responded to methylprednisolone given intravenously. In 17 with AR, the Z score fell significantly (delta Z greater than 1.5 for 2 days), an average of 2.6 days before the first rise in serum creatinine concentration; 15 responded to treatment. The difference between these two groups was significant (P less than 0.001). This functional heterogeneity and different responses to treatment may indicate different immunologic mechanisms which damage different target cells in the graft in AR. In 46 patients with acute tubular necrosis after cadaver kidney transplantation FENa was significantly higher than it was in the GEF group as early as the first posttransplantation day and approached normal as the renal function recovered. This behavior of FENa was clearly different from that in AR.


[A panorama of medical history of acute iron poisoning (author's transl)].

The paper describes briefly the fatal course taken by some of the 720 acute cases of poisoning with iron compounds as documented in literature between 1826 and the present time. The various concomitant conditions and the genesis, as well as the fatal disease pattern, are communicated. The various pathogenetic factors and nosological variants of acute iron poisoning, as well as the epidemiology of juvenile iron poisoning, are discussed. Finally, the paper points to numerous deficiencies in the exact diagnosis of acute cases of iron poisoning.


Immunofluorescence of deparaffinized, trypsin-treated renal tissues. Preservation of antigens as an adjunct to diagnosis of disease.

The present study compares formalin-fixed, deparaffinized and trypsin-treated (DTT) renal tissue sections with frozen sections for detecting immune complex deposition. Both DTT and frozen sections from 52 renal biopsies were stained directly with fluorescein isothiocyanate-conjugated antihuman IgG, IgM, IgA, fibrinogen, and C3. DTT sections alone were stained indirectly for C3 using a double conjugate technique. Antigen presence or absence on DTT section was accurately detected in 90 per cent of biopsies for immunoglobulins and fibrinogen and in 75 per cent for C3 when compared to frozen section. Furthermore, antigen deposition was found in 21 per cent of biopsies only on DTT sections (usually because frozen tissue lacked glomeruli). Frozen sections alone were treated with reagents of the fixation-embedding process and directly stained for IgM and C3 to determine antigen stability. Alkaline or neutral formalin pH helped maximize antigen preservation. DTT sections are a valuable tissue source for adjunctive diagnosis of renal disease.


Hormonal requirements for basement membrane collagen deposition by cultured rat mammary epithelium.

Alveoli and ducts isolated from virgin rat mammary glands synthesize basement membrane collagen (typeIV) in primary culture. Using purified antibodies to type IV collagen, prominent intracellular and extracellular fluorescence is observed in the epithelium. No fluorescence is observed with antibodies to collagen type I and III. From quantitation of the incorporation of [14c]proline-labeled proteins, 1.5 to 2.5 per cent of the newly synthesized proteins are collagen. Type IV collagen from these cultures was biochemically identified on the basis of (1) the high ratio of labeled 3-hydroxyproline to 4-hydroxyproline (1:10), (2) the gel electrophoretic pattern of the collagenase-sensitive proteins precipitated with 1.7 M NaCl, (3)the failure of the collagen to bind to diethylaminoethyl-cellulose, and(4)the immunologic cross-reactivity with mouse tumor type IV is identical with that of type IV collagen from other sources. When the supportive hormones, insulin, prolactin, hydrocortisone, progesterone, and estradiol are removed from the cultures, there is a 90 per cent reduction in the amount of  [3H]proline recovered in collagen synthesis coincides with only a 30 percentdrop in the growht rate and a 20 per cent drop in total protein synthesis of the sells over the 24-hour period without hormones. Pulse-chase experimout hormones. Pulse-chase experiments revealed an enhanced turnover of collagen following hormone withdrawal. This system may be an in vitro model of collagen turnover in mammary gland in involution.


Behaviour of macrophages to Mycobacterium leprae. A tissue culture study.

Macrophage culture was performed on 45 healthy adults who were initially categorised on the basis of their Lepromin reactivity using Dharmendra antigen. There were 25 individuals in the Lepromin positive group and 20 in the Lepromin negative group. The cultures were challenged with M. leprae and the macrophages studied at varying time intervals of culture. No difference was evident in the behaviour of cultured macrophages to M. leprae in both the groups. The Lepra bacilli were phagocytosed by the macrophages and retained essentially intact till the termination of culture. The Lepromin reactivity of a healthy individual was not reflected on the macrophage function in vitro. The observations suggested against the possibility of an intrinsic genetically determined macrophage defect in dealing with M. leprae.


A study of scrotal biopsy in subsided cases of lepromatous leprosy.

Scrotal biopsies were obtained from 38 cases of lepromatous leprosy who were clinically subsided and had negative skin smears. Twenty six (68.4%) of these cases revealed bacilli in the dartos muscle. None except one showed a specific lesion in the dartos. Bacilli obtained from 2 out of 7 cases multiplied in the mouse foot-pad. Bacilli were found to be persisting in the dartos muscle despite prolonged treatment before as well as after clinical subsidence of the disease. The persisting bacilli in this situation could be a cause of relapses.


Echolalia: issues and clinical applications.

Echolalic behaviors have been reported within the context of various pathologies but have remained poorly defined. Consequently, it is not easy to determine whether and to what extent normal repetition can be separated from pathological echoing. Hence, it is unclear whether the occurrence of echolalic behavior may be useful for differential diagnostic purposes. Also, much room is left for controversies about the clinical management of echolalic behavior. This article reviews the various conditions associated with echolalia and the role of repetitions in normal language behavior. Suggestions are made in terms of the various dimensions along which echolalic behavior should be assessed, as well as of the desirability of particular intervention techniques.


The speech-language pathologist's role in understanding the genetics of Van der Woude syndrome.

A family report of three generations of Van der Woude syndrome (cleft lip or palate and lip pits) is presented to show that the speech-language pathologist may play an important role in identifying patients with this defect. Understanding the genetic basis of this syndrome and documentation of family history through pedigree analysis can be instrumental in identifying other family members potentially affected with speech disorders. Because the diagnosis of Van der Woude syndrome was made in one member of the family in the pedigree, three other family members who had never received previous diagnosis or treatment of their speech disorders were identified. Other family members were referred for genetic counseling. The concepts of autosomal dominant transmission, variable expression, incomplete penetrance, and the pedigree technique for recording family history are also presented.


Structure of functional A salina -- E coli hybrid ribosome by electron microscopy.

Small 40S Artemia salina and large 50S Escherichia coli ribosomal subunits can be assembled into 73S hybrid monosomes active in model assays for protein synthesis. The reciprocal combination -- small 30S E coli and large 60S A salina -- fails to form hybrids. The 73S hybrid particles strongly resemble homologous 70S E coli and 80S A salina monosomes. The morphologic differences between the corresponding eukaryotic and prokaryotic ribosomal particles, established by electron microscopy, do not significantly affect the assembly and mutual orientation of 40S A slina and 50S E coli subunits in the heterologous monosome. The fact that the structure of the interface, the supposed site of protein synthesis, is preserved in the active hybrid implies that retention or loss of biologic activity of hybrid ribosomes is determined by the extent of conformational changes in the interface.


Public health education and the control of hypertension in the United States.

In recent years, national and regional health education programs have sought to increase public awareness concerning the risks of asymptomatic hypertensive disease. Such programs have fostered community screening and encouraged long-term care. As a result of these intensive educational efforts, larger numbers of persons with hypertension are now aware of blood pressure elevations and the pool of treated hypertensive patients has grown in number. Recently observed declines in mortality due to cardiovascular disorders in the United States may relate to efforts committed to the control of hypertension. Although a wide range of alternative explanations for these major declines are possible, changes in risk factors and, more pertinently, effective large-scale management of hypertension are plausible explanations for these salutary secular trends.


Impact of alcohol, drug abuse and mental health treatment on medical care utilization. A review of the research literature.

This monograph reviews and assesses twenty-five studies that examined the question of whether treatment for mental illness, alcohol abuse or drug abuse reduces subsequent medical care utilization. In general, the studies found that such a reduction did take place. Twelve of thirteen studies found reductions of 5 to 85 per cent in medical care utilization subsequent to a mental health intervention. The median reduction was 20 per cent. The thirteenth study found that mental health services provided in a new neighborhood health center in a medically underserved neighborhood were followed by a 72 per cent increase in medical care encounters. The remaining twelve studies found reductions of 26 to 69 per cent in either medical care utilization or surrogate measures of such utilization subsequent to treatment for alcohol abuse. The median reduction was 40 per cent. The drug abuse literature in this area is sparse and primarily indirect. Although many of the studies suggested that alcohol, drug abuse or mental health (ADM) treatment was a cause of the subsequent reduction in medical care utilization, such causality was not definitively established, due to frequent methodological limitations, such as inadequate comparison groups, short time spans, small samples and lack of trend analysis. In addition, the studies focused primarily on outpatient psychotherapy in organized health care settings, particularly health maintenance organizations (HMOs), and on alcoholism treatment provided through employee-based programs and HMOs. Only very limited policy implications on such topics as health insurance and the linkage of health and ADM services can be drawn from the current body of literature. There is a need for additional research that is broader and more rigorous. To encourage such research, methodological recommendations for future studies are presented.


[Automated instrument for determining visual acuity].

The developed APZ instrument, due to a high degree of its automation, allows examination of visual acuity within minimum time spent by the physician. This procedure can be made in conditions of individual visit and mass-screening of patients. Measurement's limits age from 0.1 to 2.2. The visual acuity check-up can be performed automatically on a present program, or remotely by a physician. The APZ-I will find its application among instrumentation of ophthalmological cabinets in polyclinics, hospitals, medical centres, in special cabinets, etc.


Hypoparathyroidism.

Recent advances in our understanding of the physiologic actions of PTH and vitamin D have clarified certain aspects of the pathogenesis, classification, and management of hypoparathyroidism. Central to pathogenesis and categorization is the recognition that hypoparathyroidism may result from PTH deficiency, ineffectiveness, or resistance, with a resultant inability to stimulate adenylate cyclase in target tissues. This aberration in adenylate cyclase activity impairs certain physiologic responses such as renal phosphate excretion and renal calcium reabsorption that are required for proper calcium homeostasis. Also critical is the subnormal production of 1 alpha,25-dihydroxycholecalciferol (1,25-DHCC). Although the precise mechanism for the deficiency of 1,25-DHCC remains unclear, one may hypothesize that in hormone-deficient or hormone-ineffective hypoparathyroidism, decreased synthesis results from the absence of the two recognized stimuli for 1 alpha-hydroxylase--bioactive PTH and hypophosphatemia. Provision of either one of these stimuli would then be expected to restore 1,25-DHCC to normal levels, which could explain the calcemic response to PTH in these patients. There is some evidence that the synthesis of 1,25-DHCC may be "primarily" affected in PTH-resistant hypoparathyroidism, and thus may be unresponsive to any of the known stimuli. It remains conceivable, however, that during normocalcemic phases, such patients may improve their renal cyclic AMP and phosphaturic responses to PTH, with associated improvement in 1,25-DHCC synthesis. Certain acquired forms of PTH resistance such as hypomagnesemia and end-stage renal disease may also be associated with defective 1-hydroxylation. Whether occurring primarily or as a secondary process, the subnormal production of 1,25-DHCC may influence calcium and skeletal metabolism directly or by modifying response to PTH. The availability of 1,25-DHCC provides an effective and physiologically meaningful mode of therapy for most cases of hypoparathyroidism.


Two isoaccepting seryl tRNAs coded by separate mitochondrial genes in yeast.

In S. cerevisiae four isoacceptor mitochondrial tRNAs for serine have been separated by reversed phase chromatography. At least two of these species are products of different genes. In this work the deletion mapping technique has been used to locate two genes for tRNAser. The gene for tRNAser previously localized in the oli I region of the mitochondrial genome has been found to code for tRNAser2, and another gene coding for tRNAser1 has been detected in the region where most of other tRNA genes are found. Results of fine mapping experiments allowed to localize this gene in the proximity of the gene for tRNAarg.


Low ribonuclease activity in cellular lysates of osmotic sensitive Saccharomyces cerevisiae mutants.

Cellular lysates with very low total ribonuclease activities are obtained by lysis of Saccharomyces cerevisiae VY1160 osmotic sensitive mutant cells in 1% sorbitol solution. These lysates could be used for isolation of intact polysomes and messenger RNA molecules, or for studying of specific ribonucleases.


Radiation sensitivity of messenger RNA.

Messenger RNA function is inactivated by irradiation with ultraviolet light. A unit length mRNA (in bases) is 2-3 times more sensitive than a unit length of DNA (in base pairs) with respect to the inactivation of template function. These data stem from four experimental systems all of which do not repair DNA: the translation of E. coli mRNA in rifampicin-treated cells, of T7 mRNA in infected E. coli, of f2 phage RNA in vivo, and of stable mRNA in chromosomeless minicells. The comparison of relative sensitivities to UV is relevant to the technique of UV mapping of transcription units which enjoys increasing popularity in pro- and eukaryotic genetic research.


Regulation of dihydrofolate reductase synthesis in Escherichia coli.

Two clones from the Clarke-Carbon Escherichia coli colony bank were resistant to inhibition by trimethoprim, a potent inhibitor of dihydrofolate reductase. Both clones had elevated levels of dihydrofolate reductase. Furthermore, trimethoprim resistance and elevated enzyme levels were associated with ColE1 plasmids that carried DNA from the trkC ksgA pdxA region of the E. coli chromosome. Plasmid pLC1437a was shown by two criteria to carry the structural gene for dihydrofolate reductase: 1) A partial diploid containing plasmid pLC1437a produced a kinetically-recognizable dihydrofolate reductase that was not present in the parent haploid strain. 2) Plasmid pLC1437a coded for dihydrofolate reductase in vitro. A 1,000 base pair fragment of plasmid pLC1437a containing fol was used as a probe to measure fol mRNA in a mutant strain isolated by Sheldon and Brenner (Molec. gen. Genet. 147, 91-97, 1976). The mutation in this strain, which results in constitutively-high levels of dihydrofolate reductase and in the inability of the strain to grow at 42 degrees C, is cis dominant (Sheldon and Brenner, 1976). The results of kinetic hybridization and pulse-labeling experiments indicated that the regulatory mutant produced elevated levels of dihydrofolate reductase in response to an increased rate of synthesis of fol mRNA.


Construction and expression of a hybrid plasmid containing the Escherichia coli thrA and thrB genes.

In vitro recombination techniques were used to clone the Escherichia coli thrA and thrB structural genes in the plasmid vector pBR322. The chimeric plasmid was analyzed and characterized genetically, by restriction mapping and DNA sequencing. The limited expression of the threonine biosynthetic enzymes in the strain carrying the recombinant plasmid is discussed.


Inheritance of multiple drug resistance in Saccharomyces cerevisiae: linkage to leu1 and analyses of 2 micron DNA in partial revertants.

The inheritance and phenotype of multiple drug resistance in independent multiple drug resistant mutants, two isolated in this laboratory (GR359 and 2-20), and two (DRI 9/T7 and DRI 9/T8) reported by Guerineau et al. (Biochem. Biophys. Res. Commun. 61,462), was investigated. Comparison of resistance to 12 selected drugs showed that the resistance phenotypes of all mutants were similar, although some differences in levels of resistance of each mutant was observed with certain drugs. Mapping of the resistance loci in GR359 and 2-20 revealed tight linkage of both resistance genes to the centromere linked gene leul. 2 micron DNA was analysed by hybridization of 2 micron RNA to EcoRI fragments of a total DNA extract. Eight partial revertants of 2-20, which had been chosen as having a phenotype similar to the 2 micron DNA deficient [cir degrees] isolate DRI 9/T7, revealed the presence of 2 micron DNA. The lack of detectable 2 micron DNA in DRI 9/T7 was confirmed.


IS2-43 and IS2-44: new alleles of the insertion sequence IS2 which have promoter activity.

The sequence of two new IS2 alleles with promoter activity (IS2-43 and IS2-44) is reported. The alleles are identical and are formed by a 17 bp tandem duplication in an AT-rich region of IS2. This created a new RNA polymerase binding site. A mutation was found that increased the frequency of formation of these 17 bp duplications but not of another class of duplications, the "mini-insertions". This suggested that the mechanisms of formation of the two classes of duplications are different.


Characterization of the mutator mutation mut5-1.

The mutator mutation mut5-1 has been characterized with respect to a range of parameters which have been used to describe DNA repair mutants of yeast. No marked effect of the mutation on UV-mutability at lower doses was apparent. Diploids homozygous for the mutation are deficient in UV-induced recombination between the alleles his1-1 and hist1-315, mutation being sufficient to account for all the UV-induced histidine prototrophs. Complementation and mapping studies indicate that mut5-1 is allelic to rad51-1, supporting the conclusion of Hastings et al. (1976) that a mutator may increase spontaneous mutation by modifying repair parameters. Both mut5-1 homozygous and heterozygous diploids give rise to spontaneous or UV-induced segregants which appear to be the products of nondisjunction events. The levels of parameiotic recombination (see Sherman and Roman, 1963; Esposito and Esposito, 1974), sporulation and spore viability observed in mut5-1/mut5-1 diploids indicate that the function encoded by RAD51 is required at 2 times during meiosis. An essential role of the function encoded by RAD51 in mitotic and meiotic recombination is indicated.


Temperature dependent release of beta-beta' subunits of DNA dependent RNA polymerase from the folded chromosome of a dnaAts mutant of Escherichia coli.

DNA-dependent RNA polymerase has been found to be preferentially released at 43 degrees C from the folded nucleoids of an E. coli dnaAts mutant when compared with the same nucleoids at 30 degrees C or with nucleoids of a dnaA+ strain at either 30 degrees or 43 degrees C. The polypeptides released are identical in molecular weight with those of the beta and beta' constituent polypeptides of the core enzyme of a known E. coli RNA polymerase. In addition, these polypeptides are precipitated by specific anti-RNA polymerase rabbit IgG. The implications of the interactions of RNA polymerase with the dnaA gene product are discussed.


Constitutive mutation of cysJIH operon in a cysB deletion strain of Salmonella typhimurium.

In a cysB deletion strain a new mutation, denoted cys-2332 was isolated, which causes the constitutive expression of the cysJIH operon. cys-2332 is closely linked to cysJIH and presumably is located in the initiator region of this operon, rendering its expression independent of the cysB gene product and the internal inducer O-acetyl-L-serine. The presence of sulfite reductase (encoded by cysI and cysJ) activity in a cysB- cys-2332 double mutant indicates that cysG, which is not linked to cysJIH but is required for the synthesis of the sulfite reductase co-factor siroheme, is not controlled by cysB.


Location and characterisation of a new replication origin in the E. coli K12 chromosome.

A segment of DNA located in the region of the E. coli K12 chromosome previously identified by the Rac phenotype can function as a self-replicating plasmid. Evidence is presented that this plasmid, the oriJ plasmid, contains the origin of replication of a defective prophage postulated to be located in this chromosomal region by Low (1973). The plasmid can only be maintained in strains in which this postulated prophage has been deleted. In strains which possess the prophage selection for plasmid maintenance permits the isolation of clones containing new deletions which we postulate are the result of prophage excision.


Physical characterisation of the "Rac prophage" in E. coli K12.

We confirm the hypothesis of Low (1973) that many E. coli K12 strains contain a prophage (the Rac prophage) located a few minutes clockwise of the trp operon on the genetic map. We have used restriction endonucleases and 32P-labelled probes to construct a physical map of this prophage. Some E. coli K12 strains, including AB1157, have lost the entire prophage, apparently by a specific deletion. This is consistent with prophage excision by site-specific recombination. lambda reverse (lambda rev) phages (Zissler et al., 1971) are recombination proficient derivatives of phage lambda in which the phage recombination functions have been replaced by analogous functions (RecE) derived from the host chromosome (Gottesman et al., 1974; Gillen et al., 1977). Our data support the origin of lambda rev plages by recombination between lambda and the Rac prophage following excision of the Rac prophage from the E. coli chromosome. Important experimental data are included in the Figure legends.


Evidence that the ribosomal DNA genes of yeast are not on chromosome I.

Several workers have reported that most of the ribosomal DNA genes (rDNA) of the yeast Saccharomyces cerevisiae are located on chromosome I. More recently, data indicating that the yeast rDNA genes are located on chromosome XII has been presented. In this report, we present additional evidence indicating that most of the yeast rDNA genes are not on chromosome I. Starting from a diploid yeast strain, we isolated ten strains which were monosomic (2n-1) for chromosome I. We found that each of these ten strains contained two copies of the rDNA-containing chromosome. In addition, we show that the earlier evidence indicating that the yeast rDNA genes were on chromosome I cannot be explained by a difference in the yeast strains which were used in the different experiments.


In vitro DNA synthesis in a concentrated yeast lysate.

A system is described in which DNA synthesis can be monitored in a yeast lysate. The observed synthesis has many of the properties of in vivo DNA replication. It is dependent upon replication growing points that were active in vivo. The in vitro synthesis proceeds via low molecular weight intermediates, but these do not mature into larger DNA. There is a specific requirement for rATP. Mitochondrial DNA is also synthesised in this system.


Plasmid pSC101 replication in integratively suppressed cells requires dnaA function.

Maintenance of plasmid pKC17, a derivative of plasmid pSC101, in E. coli requires a functional dnaA gene product. This was demonstrated by segregation experiments using an E. coli dnaA46 mutant, at various temperatures. Growth of this mutant at elevated temperature was allowed by the presence of a P2sig5 prophage. rpoB mutations, which suppress the temperature sensitivity of a dnaA46 mutant permit efficient maintenance of plasmid pKC17 at temperatures up to 40 degrees, conditions which normally inactivate the dnaA46 product.


The flagellar protein of Clostridium tetani.

Clostridium tetani ATCC 19406 was investigated with regard to the flagellar filaments produced by this anaerobic species. Flagellar filaments were removed from the cell bodies by hydrodynamic shear forces and purified by differential centrifugation. Exposure to acid was shown to result in disaggregation of the flagellar filaments into a preparation of flagellar protein containing 3.5% carbohydrate. The protein was judged homogeneous after examination by acrylamide gel electrophoresis in the presence of 4 M urea at several pH levels, and was shown to have a molecular weight of 35,000 daltons. Amino acid analyses indicated the absence of cysteine and tryptophan and a preponderance of acidic residues, epsilon-N-methyllysine was shown to be absent and the N-terminal amino acid was identified as alanine. Analysis of the C-terminal region indicated the sequence -Leu-Leu-Arg. These findings indicated that the obligate anaerobe C. tetani produced flagella filaments similar to previously studied filaments of aerobic and facultatively anaerobic bacteria.


Surface colony growth in a controlled nutrient environment. 3. Dependence of growth constant on nutrient concentration.

The growth rates of yeast surface colonies under constant nutrient conditions were studied as a function of nutrient concentration. This was complementary to the initial experiment to measure the rates under such conditions, and to the investigation of the effects of alcohol. The possibility of fitting an equation of the Michaelis-Menten type to the variation of growth rate with carbon source concentration is considered.


Phenols: a review of their history and development as antimicrobial agents.

Phenols were first isolated in crude form at the end of the eighteenth century. Pure phenol was isolated in 1834 and its structure proved in 1842. In 1860 Küchenmeister first used phenol as a wound dressing in Germany. Lister was to use it in his classical experiments in antiseptic surgery. Substituted phenols, and to some extent phenol itself, are still in use today as antiseptics, disinfectants and preservatives.


In vitro activity of cefamandole, cefoxitin, cefuroxime, and carbenicillin, alone and in combination with aminoglycosides against Serratia marcescens.

Synergistic antibiotic studies were undertaken to compare the effectiveness of two new beta-lactamase resistant cephalosporins, cefamandole, and carbenicillin, with four aminoglycosides against clinical strains of Serratia marcescens. The strains demonstrated various combinations of resistance and/or susceptibility to the antibiotics tested. Tobramycin was the most effective aminoglycoside when used in combination with beta-lactam antibiotics. Carbenicillin and cefamandole demonstrated similar activity with aminoglycosides in synergy experiments. Tobramycin-carbenicillin was found to be the superior pairs as indicated by the total number of strains inhibited. This combination was the only one effective against certain high drug resistant strains and the strain resistant to all four aminoglycosides. Carbenicillin or cefamandole with tobramycin exhibited comparable activity against multiple drug resistant organisms. However, mutants significantly more resistant to cefamandole developed during susceptibility testing. The findings of this study have clinical relevance for treating infections by this formidable pathogen.


Interferon and cell-mediated immunity.

The thesis is introduced that the phenomena of cell-mediated immunity proceed with the necessary involvement of interferon. In view of this thesis various examples of cell-mediated cytotoxicity both antibody-dependent, and antibody-independent are analyzed. The role of interferon in transplantation, allergic diseases and the treatment of cancer is also discussed. Based on this analysis it is concluded that interferon provides for the cytotoxicity of immune and non-immune leukocytes. It is suggested that interferon is responsible for the cytotoxic effect in itself, since it facilitates lethal hybridization. Recognition of the important role of interferon in the mechanism of cell-mediated immunity leads to some practical conclusions: a) The selective inhibition of the activity of type II interferon may be a sufficient measure in cases where now total immunosuppression is applied. b) Periodic treatment of healthy population by lymphokine stimulants may be a method of prevention of cancer. c) Transplantation, allergic diseases, immunotherapy of cancer should be considered in terms of the role of interferon in these processes.


Sudden-death ischemic heart disease and dietary magnesium intake: is the target site coronary vascular smooth muscle?

The etiology of sudden-death ischemic heart disease (SDIHD) remains an enigma. Data will be presented which suggest that SDIHD may be due to hypomagnesemia in and around the coronary arterial and arteriolar vessels. We have found that blood vessels (especially arteries and arterioles) deficient with respect to Mg can undergo constriction and spasm; the greater the reduction in Mg2+, the greater the magnitude of the spontaneous contractile responses. The higher the Ca2+:Mg2+ ratio, the greater are the magnitudes of these contractile responses. A severe deficit in surface membrane Mg2+, in particular, results in intense vasospasm. Using direct in situ high resolution microscopy (3000 x), we have found that a lowering of Mg2+ around perfused arterioles (15--20 microns i.d.) will also result in spontaneous vasoconstriction and, in addition, increased arteriolar resistance, tissue ischemia and reduced venous outflow. We have also found that the constrictor actions of certain circulating vasoconstrictor hormones (i.e., angiotensin, serotonin, acetylcholine) are enhanced when [Mg2+] is lowered below the levels normally found in plasma. Other direct studies, from our laboratory, indicate that [Mg2+]o regulates calcium exchange and content of vascular smooth muscle. In summary, the concept to be presented suggests that a deficiency in dietary Mg2+ is a key factor in the high incidence of mortality noted in SDIHD in nations of the Western world. The hypomagnesemia produces progressive vasoconstriction, vasospasm and ischemia, which, given time, will lead to SDIHD.


The regulation of prostaglandin E1 formation: a candidate for one of the fundamental mechanisms involved in the actions of vitamin C.

Vitamin C stimulates the formation of PGE1 in human platelets. The effect occurs over the physiologically relevant range of concentrations. PGE1 is required for T lymphocyte function and plays a major part in the regulation of immune responses. PGE1 is also important in the regulation of collagen and ground substance metabolism, in cholesterol metabolism and in regulation of responsiveness to insulin. It is proposed that defective formation of PGE1 could account for many of the features of scurvy and for many of the reported therapeutic effects of vitamin C. If correct, vitamin C will be of value only in conjunction with an adequate supply of dihomogammalinolenic acid, the precursor of PGE1. Essential fatty acids, pyridoxine and zinc are all required to achieve this.


Water relations of fetal development.

Normal pregnancy is accompanied by a remarkable increase in total body water, due partly to the growth and development of the conceptus and partly to water retention by the mother. An evaluation of the changes in body water during pregnancy suggests that they could very largely be brought about by the hydrostatic element of osmotic water regulation and are mainly under fetal, rather than maternal control. The results also suggest that pathological disorders of water in pregnancy are essentially secondary, and not primary manifestations of disease.


Allogeneic bone marrow transplantation across the ABO barrier.

Allogeneic bone marrow transplantation in severe aplastic anaemia has been shown to be a worth-while procedure. A case in which a group O patient received a successful marrow transplant from a group B donor is reported. Major ABO incompatibility is not a contraindication to bone marrow transplantation.


Ear-acupuncture relaxation therapy in alcoholics. Report on a follow-up survey.

Ninety-five out of 190 alcoholics treated in 1977 by ear-acupuncture relaxation therapy were selected for a follow-up survey. Sixty-two replied to our letters and were interviewed (by C.F.). Sixteen (25.8%) said that they had maintained total sobriety for over 12 months, while 54 (87%) said that their life-style, drinking pattern and physical and mental health had improved. thus, a significant percentage had been helped to improved health and to a changed way of life.


The investigation of immunological infertility.

Immune reactions to spermatozoa may interfere with fertility in both males and females. These mechanisms may operate in a small, but significant, proportion of infertile couples, particularly when the duration of infertility is greater than three years. Antibodies to sperm may appear both in the blood and in genital tract secretions and their presence should be sought in both sites. The most suitable screening test for circulating anti-sperm antibodies in either partner is a method based on the complement-dependent antibody immobilization of sperm. This test may also be applied to cervical mucus as a routine in females. In males the detection of sperm immobilizing antibodies in blood serves as a reasonable guide to the existence of antibodies in seminal plasma where their presence may be confirmed using a gelatin agglutination test. Further information about the presence of locally secreted antibodies either in cervical mucus or in semen may be gained by using the sperm-cervical mucus contact (SCMC) test in which cross-testing with donor mucus or sperm will identify the site of local antibody production. Results based on immunofluorescence and sperm microagglutination methods are unreliable and lacking in correlation with continuing infertility and the routine use of these tests is not recommended.


Comparison of enteric-coated levodopa with levodopa-carbidopa combination. A double-blind crossover trial.

Twenty-seven patients with idiopathic Parkinson's disease completed a double-blind crossover trial which compared enteric-coated levodopa (Prodopa) with levodopa-carbidopa combination (Sinemet). It was easy to stabilize the patients' condition with either drug, and the dose-sparing effect both of the enteric-coated preparation, and of the levodopa-carbidopa combination was again noted. At the dosages used, the levodopa-carbidopa combination was objectively shown to be more effective in 71% of the patients investigated, although there was no clear personal preference for either preparation when patients compared the two parts of the trial. Both drugs play a valuable role in the treatment of Parkinson's disease.


Comparison of effectiveness of timolol administered once a day and twice a day in the control of blood pressure in essential hypertension.

The effects of a single dose and of two equally divided doses of timolol were compared in a double-blind trial in 15 patients with essential hypertension. The amount of timolol per day remained constant for each patient, but individual patients received different amounts (from 10 mg to 30 mg/day). Timolol combined with a diuretic produced similar lowering of blood pressure in patients who required 10 mg, 20 mg or 30 mg of timolol per day when given either as one dose or in two equal doses. Plasma concentrations of timolol were high enough to be a major contributing factor to the long duration of the hypotensive response.


Imported parasitic infections in Tasmania.

A number of protozoan, trematode, and nematode infections have been imported into Tasmania over an 18-month period. Some of the cases are briefly discussed as they provide features of clinical interest. The increasing problem of imported infections in Australia is emphasized.


Suppurative pyephlebitis and multiple hepatic abscesses with silent colonic diverticulitis.

This report records the first known case in the literature of suppurative pylephlebitis associated with multiple intrahepatic and extrahepatic abscesses and gross hypoalbuminaemia, as a consequence of colonic diverticulitis.


Bovine diaphragmatic hernioplasty using autologous skin grafts.

A 2nd recurrence of diaphragmatic hernia in a buffalo (Bubalus bubalis), which twice previously had been subjected to herniorrhaphy, was successfully treated by hernioplasty, using a fresh autologous skin graft to compensate for the lost tissue.


Human plasma angiotensinogen: a review of purification procedures.

The current status of the purification and characterization of human angiotensinogen is reviewed. One problem encountered in the past has been the copurification of a protein with similar porperties. This protein has tentatively been designated alanine-protein. An efficient separation of angiotensinogen and alanine-protein was obtained on a zinc chelate column. Alanine-protein has been purified and its amino acid and carbohydrate composition determined. The COOH-terminal amino acid and the NH2-terminal amino acid were determined to be serine and alanine, respectively. Alanine-protein exhibited multiple forms on isoelectric focusing.


[Complete heart block and pacemaker therapy in children (author's transl)].

The etiology of congenital and acquired complete heart block is discussed. The prognosis is much better in mostly congenital suprabifurcational blocks, than in the infrabifurcational blocks, which are acquired in most instances. Suprabifurcational blocks are characterized by normal QRS complexes, higher heart rates, and adequate frequency adaptation to exercise. Widened and deformed QRS complexes and fixed heart rates as low as 40--50 beats per minute are typical in infrabifurcational blocks. Whereas children with congenital heart blocks only eventually need pacemakers, the immediate implantation of a pacemaker is mandatory in children with acquired heart blocks after a decrease of the heart rate below 30--40 beats per minute and/or after an Adams-Stokes attack. With demand pacemakers being available, fixed rate systems are obsolete for children. Lithium batteries should be preferred for their smallness and their larger life span. A considerable body of surgical and technical problems due to infection, wire damage, and other complications still awaits solution.


[Middle ear problems associated with cleft palate (author's transl)].

The relation between cleft palate and middle ear disturbances is well known. Often the ENT specialist will see the patient only after the middle ear has been damaged so that it cannot be corrected any more satisfactorily. Therefore early otological diagnosis (including audiometry, impedance measurement, ERA, microscopic examination of the ear drum and essential myringotomy) and early treatment should be performed in the general plan of treatment in cleft patients.


[Group B streptococci: the most common cause of neonatal septicemia (author's transl)].

Between 1965--78 118 newborns with septicemia have been treated in the Children's Hospital of the Free University Berlin. Microorganisms identified were streptococci in 32 cases, 27 of which were group B streptococci (increasing number since 1973). In 1978 group B streptococci were responsible for 44% of all the septicemia cases as well as for 12% of all newborn deaths. The incidence of group B streptococcal septicemia in newborn babies is 1/1000 live births for the Berlin region. 2 patients presented the late-onset type of group B streptococcal neonatal sepsis; both survived having neurological sequelae. 25 newborns belonged to the early-onset group, the mortality rate in this group is 56%. The clinical features, bacteriological findings and risk factors are summarized in table form. There could be an influence related to the maternal blood type. Histological examinations in 5 placentae revealed signs of amniotic infection.


[A specific IgM test for the diagnosis of Epstein-Barr virus infections (author's transl)].

IgM antibodies specific for Epstein-Barr virus (EBV) were measured in 302 patients with high IgG antibody titers to study whether EBV was the cause of disease in children having one or more symptoms of classical infectious mononucleosis. IgM antibodies specific for EBV were found in all patients with the defined clinical picture of infectious mononucleosis. In addition the majority of cases with clinical suspicion of the disease had also specific IgM titers. Besides infectious mononucleosis EBV can also be the cause of other diseases like hepatitis, and lymphadenitis: we found IgM antibodies specific for EBV in 48% of patients with nonbacterial lymphadenitis and in 64% of patients with hepatitis not due to hepatitis A or B virus. In contrast to observations by others we were able to show heterophile antibodies in cases with incomplete features of infectious mononucleosis. IgM antibodies to EBV were found in 4 out of 85 controls only. We conclude that untypical features of infectious mononucleosis can be caused by EBV also. Therefore the determination of specific IgM antibodies to EBV can be helpful in the diagnosis of uncharacteristic EBV infections.


[On baby formular (author's transl)].

1. During the first months of life adapted or partially adapted formula may very well be used to feed the newborn and the young infant if breastmilk is not available. By feeding adapted milk ad libitum, overweight is avoided. This is partially due to the carbohydrate, which exclusively consists of lactose. 2. Many infants are fed "Beikost", or solid food, much too early. The best possible age is during the fourth month of life. The first solid food to be introduced into the diet of an infant should consist of meat and added vegetable. 3. Too much intake of salt and or sucrose should be avoided. 4. Between the fourth to sixth month of life one bottle per day of adapted formula should be exchanged for a "follow-up" formula instead. As follow-up formulas we prefer those without or only with a small content of sucrose.


[Nutrition of the newborn infant (author's transl)].

Feeding problems concerning the newborn infant are of interest as long as pediatrics exist. Controversial opinions were expressed on the beginning, the quantity and the quality of feeding. Pediatricians are in agreement that nutrition has to warrant normal psychosomatic development of the infant. Disturbances of perinatal adaptation should be avoided as far as possible. Food has to contain protein, lactose and fat in a relation which both quantitatively and qualitatively is similar to human breast milk. In the first days of life and in the case of insufficient production of breast milk a formula adapted to breast milk or a 10% solution of glucose should be fed additionally to breast milk. Increased energy demands eventually require the feeding of higher concentrated solutions of oligosaccharides. At the present time it is impossible to comment on the value of the different kinds of feeding of the newborn infant in terms of psychosomatic parameters. Only prospective studies of a duration of more than two decades could given enough information to elucidate the discussed problems.


Physiological and genetic effects of puromycin aminonucleoside on ultraviolet-irradiated Escherichia coli strains.

Puromycin aminonucleoside (PAN) increased significantly the mutation rate of Escherichia coli B/r strains when used in conjunction with certain ultraviolet dosages. PAN (2.5 mM) when added to the post-irradiation medium of hcr+ cells slowed down RNA synthesis to 65%, protein to 76% and DNA to 48% of the control rate. Purine ribosides such as adenosine decreased the inhibitory action of PAN on DNA, RNA and protein synthesis. Quantitatively quite different results were obtained with the hcr- strains. PAN did not increase killing of UV, but decreased the frequency of UV-induced mutations. Antimutagenic purine ribosides decreased the synergistic mutagenic activity of PAN. Increases in DNA synthesis in the presence of antimutagens correspond to reductions in the rate of mutation to streptomycin resistance. The excision of UV-induced pyrimidine dimers was investigated in the presence and absence of PAN. The pattern of repair-inhibition reversion of pre-mutagenic lesions by adenosine suggests that PAN behaves as a feedback inhibitor of purine biosynthesis in UV-irradiated cells. It is probable that this inhibition results in an impairment of repair which produces the increase in mutant numbers.


UV protection and mutagenesis in uvrD, uvrE and recL strains of Escherichia coli carrying the pKM101 plasmid.

The effect of the pKM101 plasmid on UV mutagenesis and survival was examined in DNA-repair-deficient strains of E. coli carrying the uvrD, uvrE and recL mutations. Although enhancement of UV mutagenesis by pKM101 was found in all 3 strains, UV protection was only observed in the uvrD strain. We conclude that the plasmid not only requires lexA+ recA+ functions of the cell, but also those of uvrE+ recL+ for its UV-protective effect.


The intrasanguineous host-mediated assay procedure distribution and retention of yeast in the mouse.

A study of the factors that could affect a method to detect mutations in cells recovered from different organs after intravenous injection in mice, was performed by using the D4 strain of Saccharomyces cerevisiae. The recovery of the yeast cells 5 min to 3 days after injection in the host animal was investigated. The circulation, distribution and localization of the cells were determined, and histopathologic analysis was performed in order to detect possible interactions between the mice and the microorganisms. We found that the yeast cells were trapped primarily in the capillaries of the organs; 3 days after injection no cells were found outside of the tissue-blood vessels. The spontaneous gene-conversion frequency of the yeast cells recovered at different times after injection was increased, but this increase was not time-dependent.


Significance of specific-locus germ-cell mutations in mice.

On the basis of a historical-control incidence of 39 mutants in 688 921 progeny (5.7 mutants/10(5) animals or 0.82 mutations/locus/10(5) gametes) proposals are made for the numbers of test progeny required when screening for possible mutagens using the specific-locus test in mice (7 loci). It is recommended that 25,000 control progeny should be included in each test, to establish homogeneity with the historical controls. This would also be the number of progeny required from treated males, unless significantly positive results had been obtained with smaller numbers. It would appear that the greater sensitivity of post-spermatogonial stages could more than compensate for practical difficulties in sampling these stages, rather than spermatogonia, in screening tests.


Genetic effects of acridine compounds.

Acridines and a very large number of acridine derivatives are used in enormous quantities both in medicine and industry. The mutagenic action of these compounds has been demonstrated in a wide variety of organisms and is known to occur both in the dark as well as in the presence of light (photodynamic action). At the molecular level, acridines have been shown to cause frameshift mutations of both the addition and deletion types, a characteristic which has been of tremendous help in elucidating the nature of the genetic code. These and various other biological effects of acridines, such as inhibition of DNA repair, curing of plasmids and cell-growth inhibition, are examined in this review.


In vivo covalent binding of organic chemicals to DNA as a quantitative indicator in the process of chemical carcinogenesis.

The covalent binding of chemical carcinogens to DNA of mammalian organs is expressed per unit dose, and a 'Covalent-Binding Index', CBI, is defined. CBI for various carcinogens span over 6 orders of magnitude. A similar range is observed for the carcinogenic potency in long-term bioassays on carcinogenicity. For the assessment of a risk from exposure to a carcinogen, the total DNA dmaage can be estimated if the actual dose is also accounted for. A detailed description is given for planning and performing a DNA-binding assay. A complete literature survey on DNA binding in vivo (83 compounds) is given with a calculation of CBI, where possible, 153 compounds are listed where a covalent binding to any biological macromolecule has been shown in vivo or in vitro. Recent, so far unpublished findings with aflatoxin M1, macromolecule-bound aflatoxin B1, diethylstilbestrol, and 1,2-epithiobutyronitrile are included. A comparison of CBI for rat-liver DNA with hepatocarcinogenic potency reveals a surprisingly good quantitative correlation. Refinements for a DNA-binding assay are proposed. Possibilites and limitations in the use of DNA binding in chemical carcinogenesis are discussed extensively.


Mutagens in coffee and tea.

Coffee prepared in the usual way for drinking contains a substance(s) that is mutagenic to Salmonella typhimurium TA100 without mammalian microsomal enzymes. One cup of coffee (200 ml) contains mutagen(s) inducing 1.4-4.6 X 10(5) revertants under standard conditions. Instant coffee too is mutagenic to TA100 and one cup of instant coffee prepared from 1 g of coffee powder and 200 ml of water induced 5.6-5.8 X 10(4) revertants of TA100. Caffeine-free instant coffee also has similar mutagenicity. Addition of microsomal enzymes abolished the mutagenicity. Black tea, green tea and Japanese roasted tea were also mutagenic to TA100 without S9 mix and one cup of these teas prepared in the ordinary way produced 1.7-3.8 X 10(4) revertants of TA100. Black tea and green tea were also mutagenic to TA98 in the presence of S9 mix after treatment with a glycosidase from Aspergillus niger, hesperidinase. This type of mutagen in one cup of black tea induced 2.4 X 10(5) revertants of TA98.


Mutagens in Japanese pickle identified as flavonoids.

A kind of Japanese pickel was mutagenic to TA100 and TA98 with S9 mix. Its activity was one sixth that of the Chinese pickle from Linhsien County (China), reported previously. Mutagens in this Japanese pickle were isolated; the main component was identified as kaempferol, and a minor component as isorhamnetin. Flavonoids are ubiquitous and the principles identified, kaempferol and isorhamnetin, do not seem to be specific to the pickle vegetable, implying that the mutagenic substances responsible for the high incidence of human cancer in China remain to be identified.


Genetic effect of 3-carbethoxypsoralen, angelicin, psoralen and 8-methoxypsoralen plus 365-nm irradiation in Saccharomyces cerevisiae: induction of reversions, mitotic crossing-over, gene conversion and cytoplasmic "petite" mutations.

The genetic effects of two mono-functional photosensitizing furocoumarins, 3-carbethoxypsoralen (3-CPs) and angelicin, were compared with those of two bi-functional furocoumarins, 8-methoxypsoralen and psoralen in Saccharomyces cerevisiae. A drug concentration of 5 X 10(-5) M plus various doses of 365-nm irradiation at a dose rate of 1.2 kJ m-2 min-1 were used. Per dose of 365-nm irradiation, the frequency of induced nuclear events such as gene mutation and mitotic recombination (conversion and crossing-over) is higher for the bi-functional than for the mono-functional compounds. The higher efficiency of the bi-functional furocoumarins is also evident when the frequency of mutants is expressed as a function of survival. However, the photo-addition of the 4 furocoumarins studied leads to the same response for the induction of recombinational events per viable cell. Amongst genetically altered colonies induced in the diploid strains D5 and D7, the colonies corresponding to the induction of crossing-over are effectively produced by bi-functional furocoumarins, but are rare (D7) or even absent (D5) after treatment with monofunctional furocoumarins. This suggests a certain specificity of genetic alterations produced by the bi-functional agents. 3-CPs is the most effective inducer on the cytoplasmic "petite" mutation in stationary phase cells per unit irradiation dose or per viable cell.


Investigation into the mutagenic activity of azathioprine (Imuran) in different test systems.

The chromosome-damaging effect of azathioprine (Imuran) was investigated in the micronucleus test with mice and rats, in the pertussis-stimulated lymphocyte metaphase test with rabbits, and in children on azathioprine therapy. In the micronucleus test, there was a dose-dependent increase in the number of cells with micronuclei. The dose was given twice in 24 h, i.p., 25, 50 and 100 mg/kg body weight for mice, and 50, 100 and 200 mg/kg for rats. In the lymphocyte test, a dose of 5 and 20 mg azathioprine per kg body weight was given orally to rabbits on three successive days after pertussis injection. Compared with treatment in the control group these treatments induced a significantly increased number of cells with chromosomal abnormalities. These results were confirmed in lymphocyte cultures of children on long-term azathioprine therapy after kidney transplantation, where an increase in the number of structural chromosomal abnormalities was observed.


Mutational studies with diquat and paraquat in vitro.

Diquat and paraquat were assayed in the following tests. (1) Ames test in Salmonella typhimurium (strains TA1535, TA1537, TA1538, TA98 and TA100) with and without rat-liver microsomal fractions. (2) Resistance to 8-azaguanine in Salmonella typhimurium (strain hisG46, TA92 and TA1535. (3) Repair test in Salmonella typhimurium (strains TA1538 and TA1978). (4) Gene mutations in Aspergillus nidulans: 8-AG resistance and methionine suppression (meth A1 locus). (5) Lethal recessive damage in Aspergillus nidulans. (6) Unscheduled DNA synthesis (UDS) in human epithelial-like cells (EUE). Diquat and paraquat were positive in S. typhimurium (in the repair test and the 8-AG resistance system), in A. nidulans (for gene mutations and lethal recessive damage induction) and in EUE cells (UDS induction).


Mutagenicity of nitrosamines formed from nitrosation of spermidine.

5 nitrosamines formed from the nitrosation of spermidine were investigated for mutagenicity using various strains of Salmonella typhimurium in the presence and absence of S9 mix. Using the plate incorporation method, 3-butenyl-(2-propenyl)-N-nitrosamine, 3-hydroxybutyl (2-hydroxypropyl)-N-nitrosamine, 4-hyroxybutyl-(2-hydroxypropyl)-N-nitrosamine, 4 hydroxybutyl-(3-hydroxypropyl)-N-nitrosamine, and in the liquid test 3-hydroxybutyl-(3-hydroxypropyl)-N-nitrosamine were mutagenic in the absence of S9 mix.


Mutagenic and colicine-inducing activity of two antioxidants: pyrogallol and purpurogallin.

The antioxidants pyrogallol and its oxidative derivative, purpurogallin, both induce colicine E2 as well as base substitution and frameshift mutations. Because of the bactericidal effect of purpurogallin, its mutagenicity could be best demonstrated by short-term exposure followed by dilution on the test plates. The colicine-inducing potential of purpurogallin was also observed when tested directly on the plates.


Mutagenicity of pyrrolizidine alkaloids in the Salmonella/mammalian-microsome test.

The mutagenicities of 7 pyrrolizidine alkaloids to Salmonella typhimurium TA100 were demonstrated by a modified Ames's method. The pyrrolizidine alkaloids found to be mutagenic were clivorine, fukinotoxin, heliotrine, lasiocarpine, ligularidine, LX201 and senkirkine. Pre-incubation of these alkaloids with S9 mix and bacteria in a liquid medium was essential for demonstration of their mutagenicities.


In vitro mutagenicity and cell-transformation screening of N-(2,3-epoxy-propyl)-phthalimide.

N-(2,3-Epoxy-propyl)-phthalimide (EPP) was tested for genetic activity in the Salmonella/microsome mutagenicity test. Concentration-dependent mutagenicity was demonstrated in S. typhimurium strains TA1535, TA1537 and TA100 with and without rat S9. It was inactive in strain TA1538, and active without rat S9 in TA98 at the high dose. EPP induced 6-thioguanine-resistant mutants of Chinese hamster ovary cells in the absence of an exogenous activating system. EPP produced dose-dependent enhancement of SA7 virus transformation of primary hamster-embryo cells, and transformed secondary hamster-embryo cells in a non-dose-related fashion. At a dose of 5 g/kg p.o. or i.m., EPP was inactive in the host-mediated assay using C57Bl/6XC3H mice and S. typhimurium strain TA1535. Murine testicular DNA synthesis was not inhibited by oral administration of EPP at 1000 mg/kg.


Cell-mediated immunity in the liver of mice vaccinated against malaria.

Mice can be protected against several species of lethal malaria infection by vaccination, and their recovery correlates well with increased anti-malarial antibody levels, particularly IgG (ref.2). However, there is also a good correlation between protection by vaccines and priming for delayed-type hypersensitivity in the skin, although there is no obvious explanation for this effect. We now report an apparent relationship between protection and a cell-mediated immune response involving the migration of various types of cell capable of killing malaria parasites in vitro to the liver. We suggest that the effect of vaccination is to bring together parasites, specific antibody and nonspecific cytotoxic cells, and that the liver may be a major site for their interaction.


Identification of the protein encoded by the transposable element Tn3 which is required for its transposition.

Protein products have now been identified which account for the entire coding capacity of the transposable element Tn3. Mutations in Tn3 have allowed us to map the genes encoding each of these peptides and to identify their role in transposition. We have found that only a single Tn3-encoded peptide is required for transposition. Expression of this peptide is repressed by the product of a second gene, which is itself autogenously regulated.


Transposition protein of Tn3: identification and characterisation of an essential repressor-controlled gene product.

Fusions that bring lac gene expression under the control of transcriptional and translational signals within the Tn3 element have been used to identify and characterise a Tn3-encoded 'transposase' (Tnp) peptide of MW 100,000 essential for transposition. The gene specifying this product is regulated by the Tn3 repressor protein and is part of a bidirectional genetic unit that includes the repressor gene.


Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9.

The transposable genetic element Tn9 consists of two direct repeats of the insertion sequence IS1 flanking a region of 1,102 base pairs which determines chloramphenicol resistance. Transposition of Tn9 leads to the duplication of a 9-base pair sequence which preexists at the site of insertion. One copy of this sequence is found at each end of the inserted element. The chloramphenicol resistance determined by Tn9, and by various other R plasmids, is due to the synthesis of the enzyme chloramphenicol acetyl transferase (CAT). This enzyme catalyses the formation of acetylated derivatives of chloramphenicol which are inactive as inhibitors of protein synthesis. By using the chain termination technique of DNA sequencing, we have now determined the nucleotide sequence of the 1,102 base pair region between the directly repeated IS1 sequence in the bacterial transposon Tn9 (encoding chloramphenicol resistance). The amino acid sequence of CAT predicted from the nucleotide sequence is identical to that determined by Shaw and coworkers. An analysis of the sequence suggests that the internal 1,102 base pair region is not directly involved in transposition.


Primary structure of a chloramphenicol acetyltransferase specified by R plasmids.

Naturally occurring isolates of chloramphenicol-resistant bacteria commonly synthesise chloramphenicol acetyltransferase (EC 2.3.28; CAT) in amounts which are sufficient to account for the resistance phenotype and often harbour plasmids which carry the structural gene for CAT. The findings of CAT in such diverse prokaryotes as Proteus mirabilis, Agrobacterium tumefaciens, Streptomyces sp., and a soil Flavobacterium has led to speculation concerning the origin and evolution of the more commonly observed CAT variants specified by plasmids in clinically important bacteria. To provide a more solid basis for studying the evolution and spread of CAT within prokaryotes we chose to determine the complete amino acid sequence of a type I variant of CAT, the variant known to be associated with most F-like plasmids conferring chloramphenicol resistance. The sequence has been determined by combining the results obtained from manual and automated sequential degradation with those obtained by mass spectrometry of peptides generated by enzymatic digestion. The directly determined primary structure is identical with that predicted by the DNA sequence analysis of the chloramphenicol resistance transponson Tn9 known to specify a type I variant of chloramphenicol acetyltransferase.


E. coli ribosomal RNA contains sequences homologous to insertion sequences IS1 and IS2.

The insertion sequence (IS) elements, IS1 and IS2, present in multiple copies in the Escherichia coli chromosome, are transposable genetic elements of known nucleotide sequence. These elements can modulate gene expression, but it is not known whether they normally function in genetic control. To determine whether IS elements could exert control through specific RNA transcripts, we hybridised lambda NNC1857 r14 (carrying IS1) and pBR322 (carrying a portion of IS2) to Northern blots of E. coli RNA. Regions of homology between the IS elements and ribosomal RNA were observed. Computer analysis of reported nucleotide sequences detected large segments of homology between the IS elements and both 23S and 16S rRNA. Additional homologous sequences in phi X174 and a leader region of a ribosomal protein gene cluster were also detected. The homologous sequence between IS2 and 16S rTNA is the same sequence in phi X174 DNA which codes for the ends of the E and D gene and the start of J. The partial IS sequences may represent silent evolutionary remnants or they could modulate the expression of genes carrying these sequences.


Short-term effect of the lesion of the organum vasculosum of the lamina terminalis on hypothalamic LH-RH and serum LH, FSH and prolactin in adult female rats.

The short-term effect of the mechanical lesion of the organum vasculosum of the lamina terminalis (OVLT) was investigated in 4-day cycling female rats. The lesions were performed on the 2nd day of diestrus, and the animals were killed by decapitation 30 h after the lesion. Serum LH, FSH and prolactin and hypothalamic LH-RH content of 3 different parts of the hypothalamus were determined with radioimmunoassay. OVLT lesion caused a significant increase in the LH-RH content of the mid-basal hypothalamus and in serum prolactin levels and a decrease in LH and FSH serum levels. The results support the view that the OVLT may play a role in the control of pituitary gonadotrophic hormone secretion.


Late radiation damage to the central nervous system: a radiobiological interpretation.

The evidence in support of the two opposing views as to the pathogenesis of late radiation damage to the central nervous system after therapeutic doses of radiation are briefly reviewed. Analysis of the radiation dose levels used by various investigators holding opposing views as to pathogenesis of CNS lesions they reported, suggests that both groups of protagonists were in part correct. Both vascular and glial changes are important in the development of late radiation damage to the nervous system; the preponderance of one or the other type of cell damage depends upon the radiation dose. Vascular effects occur at lower dose levels but after a longer latent period than effects mediated through damage to neuroglia. Current radiobiological concepts and their possible importance in establishing the radiosensitivity of the central nervous system are discussed. The origins of both vascular and neuroglial cell mediated changes are discussed in terms of the reproductive loss of radiation damaged cells. This view is supported by some recent experimental findings. The radiobiological basis of some recently investigated treatment modalities aimed at improving the control of intracranial gliomata by radiotherapy are also considered.


[Clinico-immunological study of 6 cases of Behçet's disease].

6 cases of Behçet's disease are reported. Diagnosis was particularly arduous because the typical triad of symptoms (oral aphthae, genital ulcers, uveitis) was masked by secondary disturbances in other organs and systems. Immunology confirmed the presence in these patients of changes in various in vivo and in vitro tests and particularly interesting was the aspecific cutaneous hypersensitivity seen in 5 cases and enhanced lymphocyte blastic transformation. This was seen spontaneously and following PHA. On the basis of the outcome of the studies carried out and the literature data, the possibility that Behçet's disease may contain immunological changes and changes in the mechanisms that regulate quinine, complement and clotting activation is suggested. Therapeutically, confirmation was obtained of the effectiveness, at least temporarily, of corticosteroid treatment and antilymphocyte globulin was experimented for the first time in these patients. This might be used to replace the immune depressant antiblastic substances already successfully employed in patients with Behçet's disease.


Hypothalamic monoamine contents in endotoxin fever of new-born guinea pigs and kittens.

In new-born guinea pigs and kittens Escherichia coli endotoxin did not significantly modify the hypothalamic level of either norepinephrine (NE) or 5-hydroxytryptamine (5-HT). The hypothalamic 5-hydroxyindoleacetic acid (5-HIAA) content was significantly elevated in endotoxin-treated animals, but this elevation was confined to a period following the endotoxin injections by about 60 min in guinea pigs and 75-90 min in kittens.


Simple gas-chromatographic measurement of valproic acid in psychiatric patients: effect on levels of other simultaneously administered anticonvulsants.

A simple, rapid and precise gas-chromatographic procedure which involves one-step extraction and on-column methylation for routine clinical monitoring of plasma valproic acid levels is described. Six other commonly, simultaneously administered anticonvulsants can also be determined by substituting methylene chloride for chloroform for extraction and changing the column temperature. Also, described is the effect of valproic acid on blood levels of other simultaneously administered anticonvulsants.


Fetoscopy in the practice of perinatology and obstetrics.

The clinical indications for fetoscopy in perinatal medicine are encompassing an ever widening circle of inherited diseases and teratologically induced disorders that can now or potentially be diagnosed in utero. Fetoscopy, which has evolved over the past 20 years into a clinically accepted procedure, is not without risk to the pregnant mother and the fetus. When the decision is made to perform the procedure, its merits should outweigh its inherent risks and it should be performed by a perinatologist experienced in the technique. Fetoscopy is appropriate when the fetus is at risk for teratologically induced malformation, inherited blood dyscrasias, sex-linked or autosomal abnormalities with gross malformations, or neural tube disorders that cannot be diagnosed by amniocentesis. Technical improvements should result in increased refinement and wider applicability of the technique in the future.


Psychologic aspects of gynecology and obstetrics.

In the preceding pages, an attempt has been made to emphasize some of the more important psychologic aspects of gynecology and obstetrics. This review should be considered incomplete and the interested reader should refer to the many excellent papers and reviews on each of the topics mentioned and many more not mentioned. If there is any one guiding principle, it is that the obstetrician-gynecologist, more than most specialists, must be able to listen to and to hear the fears and anxieties of his or her patients. The need to include the patient as a partner with a controlling interest in her care is paramount. The gynecologist-obstetrician can truly become the patient's primary physician in the finest sense of the word.


Genetic counseling in primary obstetric care.

Of all health professionals the obstetrician is by far the best-equipped to integrate genetic counseling into health care delivery. As with most aspects of the practice of medicine, genetic information increases daily. The practicing obstetrician, by learning the fundamentals of genetics, can at least delineate at-risk situations. When a new situation arises he can either consult an up-to-date article or book 185--190 or refer the patient to the local genetic counseling clinic for further appraisal. By incorporating prospective counseling and in some cases retrospective counseling into his daily practice, the obstetrician can contribute greatly to the ideals of obstetrics, namely producing healthy children for his patients.


The genetics of intersexuality: clinical and theoretic perspectives.

Recent studies using newly devised serologic techniques have dramatically advanced our understanding of gonadal organogenesis and aberrant sexual differentiation. Thus XY gonadal dysgenesis, XX male syndrome, and XX true hermaphroditism, for example, may be seen as typifying specific errors of regulation, synthesis, or function of a phylogenetically conserved plasma membrane component heretofore recognized solely as the male-specific transplantation antigen of mice. This report seeks to convey the excitement of these recent studies, and to provide the clinician with new and useful insights into the fundamental mechanism of primary sex determination.


[Cultivation of the isolated eye rudiments of birds and mammals].

The cultivated of isolated organs finds ever increasing use for studying urgent problems of normal development and causes of pathogenesis. The system of developing vertebrate eye is one of the models suitable for analysis of a wide range of problems. The organotypical development of isolated eye rudiments with typical morphogenesis and cytodifferentiation of their parts are to be considered as criteria of their complete development. The elaboration of methods of complete cultivation of the early eye rudiments in vitro is one of the urgent tasks of the future, whereas there are methods which allow to grow eye rudiments successfully in vivo.


Eikenella corrodens: a pathogen in head and neck infections.

E. corrodens as a co-existent or primary causal agent in six cases of osteomyelitis or orofacial infections following trauma or elective dental extraction is presented. The importance of this facultative gram-negative bacillus as a head and neck pathogen is discussed.


The bone marrow in polycythemia vera.

The sequential biopsies and the careful clinical and laboratory studies in this large prospective study of PV patients offer a unique opportunity to properly evaluate the diagnostic and prognostic importance of the biopsy and to study the complications of this condition. The results of the study to date confirm and extend previous studies. Because of the long natural history of PV, the results of studies relating to leukemia, other myeloproliferative diseases, myelofibrosis, and effects of therapy are tentative at this time, even though the study is in its eleventh year. Hypercellularity of the marrow, together with hyperplasia and hypertrophy of megakaryocytes, is an almost constant finding in untreated PV. A very few cases (7 of 281) had relatively normal cellularities and normal megakaryocytic concentrations. Whether these findings were the result of sampling errors could not be determined, since only one site was biopsied. In any event, we found no unique clinical or laboratory features to distinguish these patients. At this time, the course of these patients appears to be the same as that of the other patients. Although increases in reticulin were regularly found during the spent phase of polycythemia, the relationship was not a precise one. For example, a moderate to marked increase in reticulin was found in 12 percent of the patients early in the course of the disease and was not predictive that the spent phase with myeloid metaplasia was imminent. In addition, in a given patient with serial biopsies taken over several years, some variability in reticulin was noted among the biopsies. Whether this represented variation in sampling or fluctuation in reticulin content could not be decided at this time. Using the standard criteria for examination of the marrows, we have found it impossible to predict which patients will develop leukemia, since the pretreatment and posttreatment biopsies almost up to the clinical onset cannot be separated from the remainder of the group.


Toxicity of wheat flour proteins and protein-derived peptides for in vitro developing intestine from rat fetus.

A peptic-tryptic-cotazym (PTC) digest of a crude wheat gliadin preparation was obtained under experimental conditions simulating in vivo protein digestion and then fractionated into 10 peaks by ion-exchange chromatography. PTC-gliadin digest and one of its subfractions (coded as fraction 9 according to its elution pattern) were very active in inhibiting in vitro development and morphogenesis of small intestine from 17- and 18-day-old rat fetuses, whereas they were harmless for the culture of jejunum from 21-day-old fetuses. PTC-digest also induced extensive tissue degeneration and necrosis of in vitro cultured small intestinal mucosa from patients with active celiac disease (gluten-induced entheropathy), but did not cause any detectable effect on histologically normal human small intestinal mucosa. Some wheat albumin and gliadin fractions were also tested on in vitro developing small intestine from 17-day-old rat fetus. Among all the tested protein fractions, only one gliadin fraction (coded as alpha 10-gliadin from its gel electrophoretic mobility) exhibited a toxic effect; morphologic alterations induced by alpha 10-gliadin were similar to those induced by PTC-digest and fraction 9.


The working mother in contemporary perspective: a review of the literature.

The purpose of this article is to provide a perspective on the dilemmas working mothers face due to the contradictory messages that emanate from both popular child-rearing books and clinical studies. A review of the working mothers' sections of popular child-rearing books, and an interdisciplinary review of research in the area of maternal employment over the last decade was conducted in order to assess current knowledge in the field. Both popular child-rearing books and books for the working mother tend to reflect the cultural norms of appropriate maternal behavior, but rarely include evidence from empirical research. Findings from a review of literature in medicine, psychology, sociology, and education are discussed in relation to effects of maternal employment on children and families. An understanding of recent research regarding maternal employment will enable the pediatrician to better respond to the needs of the working mothers and their families.


Campylobacter fetus infections in children.

There has been a gradual accumulation of reported Campylobacter fetus human infections since the first description in 1947. The taxonomy of these fastidious and morphologically confusing organisms has been recently revised and, in the past few years, through the use of selective culture medium for stool isolation C fetus has been implicated as a frequent paghogen in children. There are three relatively distinct patterns of human C fetus infection. The first, most frequent, pattern of disease is enteritis which is usually uncomplicated and due to C fetus subsp jejuni. A second form of disease consists of focal infections, often associated with vasculitis and/or chronic bacteremia. These infections, due to C fetus subsp intestinalis, are seen most often in older, debilitated, or chronically ill men. The third pattern, perinatal infections causing abortion, prematurity, and neonatal meningitis, is the least frequent, but these infections are usually fetal to the fetus or infant and are also due to C fetus subsp intestinalis.


Isolated pulmonary leukemic relapse following successful bone marrow transplant in a child with acute lymphoblastic leukemia.

Leukemic relapse and interstitial pneumonitis are common complications for leukemic patients following bone marrow transplantation. We present the case of a successful bone marrow transplantation patient who developed an interstitial infiltrate on chest roentgenogram 212 days post-transplant that was diagnosed by open lung biopsy and found to be a leukemic relapse of the lung parenchyma. No extrapulmonary sites were involved and the infiltrate cleared in three weeks with systemic chemotherapy. Pulmonary function tests continued to demonstrate restrictive disease. The patient remained in remission for nine months following  pulmonary relapse on systemic chemotherapy. This patient illustrates an unusual site of leukemic relapse and the importance of open lung biopsy in the diagnosis of the immunosuppressed patient with a pulmonary infiltrate.


45Ca2+ uptake by dispersed pancreatic islet cells: effect of D-glucose and the calcium probe, chlorotetracycline.

Uptake of 45Ca2+ was studied in dispersed pancreatic islet cells from non-inbred ob/ob-mice. Like whole islets the dispersed cells responded to 20 mM D-glucose with a markedly increased 45Ca2+-labeling of both the lanthanum-nondisplaceable and the lanthanum-displaceable calcium pools. The pronounced effect of D-glucose could not be reproduced with 3-O-methyl-D-glucose, L-glucose, D-mannose, L-leucine, or D-leucine; however, 45Ca2+ uptake was greater in the presence of L-leucine as compared with D-leucine. 45Ca2+ uptake by dispersed cells or whole islets was stimulated severalfold by 100 microM or more chlorotetracycline. At the concentration of only 10 microM, chlorotetracycline had no effect on whole islets and partially inhibited 45Ca2+ uptake by the dispersed cells. The ability of D-glucose to stimulate 45Ca2+ uptake by islets or dispersed cells remained in the presence of 10 microM chlorotetracycline. Islet cell suspensions apparently represent a valid model for studying how Ca2+ interacts with the cells. However, when using chlorotetracycline as fluorescent Ca2+ probe, attention must be paid to it potential ionophoric activity. At only 10 microM, the drug seems to monitor a peripheral pool of Ca2+, some of which may reside in normal transport channels.


Transplantation haemopoiesis. Morphological bone marrow studies after allogeneic marrow transplantation in man for severe aplastic anaemia and acute leukaemia.

Bone marrow (BM) morphology was studied in patients who underwent bone marrow transplantation (BMT) for severe aplastic anaemia (SAA) and acute myeloid leukaemia (AML). Four patients with SAA had marrow transplanted from HLA-identical siblings after conditioning with cyclophosphamide (CY); three cases of AML were treated with allogeneic BMT. The medullary material was generally obtained by aspiration, but treated so as to preserve its architecture; fresh, postvital preparations were particularly useful for this purpose. In SAA, the bone marrow before transplantation showed dense infiltrates composed of macrophages and other inflammatory cells; after BMT, repopulation started at the periphery of the infiltrates, when they were not totally destroyed by CY. Granulocytic repopulation generally preceded erythropoiesis, which always occurred by way of typical erythroblastic reticulocytopoietic islands. Marked but transient dyserythropoiesis was found in two cases; in one of them, it was considered as drug-dependent. Both in SAA and in AML, but more so in the latter, a marked and sometimes imposing macrophagic hyperplasia was found after transplantation. Many macrophages were actively engulfing erythrocytes and nucleated cells. This last type appeared prominent in rejection episodes.


[Thrombopoietin and the control of platelet production (author's transl)].

Under normal conditions the circulating platelet mass is remarkably constant, which suggested early on that a negative feedback mechanism was involved in the control of thrombocytopoiesis. Furthermore, plasma or serum from thrombocytopenic animals or humans has been reported to stimulate platelet production when injected into laboratory animals, thus suggesting that it contained a thrombopoietin(s). During the past decade, the biochemistry and physiology of thrombopoietin have been studied. However, it is not now well established that thrombopoietin is a glycoprotein produced, at least in part, by the kidney. Moreover, particular attention may be directed to its machanism of action.


Homology of the 3' terminal sequences of the 18S rRNA of Bombyx mori and the 16S rRNA of Escherchia coli.

The terminal 220 base pairs (bp) of the gene for 18S rRNA and 18 bp of the adjoining spacer rDNA of the silkworm Bombyx mori have been sequenced. Comparison with the sequence of the 16S rRNA gene of Escherichia coli has shown that a region including 45 bp of the B. mori sequence at the 3' end is remarkably homologous with the 3' terminal E. coli sequence. Other homologies occur in the terminal regions of the 18S and 16S rRNAs, including a perfectly conserved stretch of 13 bp within a longer homology located 150--200 bp from the 3' termini. These homologies are the most extensive so far reported between prokaryotic and eukaryotic genomic DNA.


Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates.

We have used a modification of the Berk-Sharp technique to determine that the 5' termini of the mouse 15 S beta-globin precursor and the mature mRNA have identical map coordinates. The modification involves the use of 5' (or 3') terminally labeled probes; it allows the detection of the precursor in the presence of excess mature mRNA.


Nucleotide sequence determination of a strong promoter of the colicin E 1 plasmid. Analysis of restriction sites protected by RNA polymerase interactions before and after limited transcription.

This paper presents the location and nucleotide sequence of a strong promoter of ColE 1. This promoter is of interest because of its greatly enhanced activity in the supercoiled state of the plasmid DNA (3) and its possible role in the maintenance of the plasmid replicon (4). This strong promoter is located at the restriction endonuclease Hae III f-h site 0.13 map units from the single EcoR 1 site proximal to the origin of DNA replication. The nucleotide sequence of the Hpa II l fragment of ColE 1 which contains this promoter has been determined. Initiation of transcription at this promoter occurred at two positions. Limited transcription by omitting one of the four nucleotide triphosphates allowed transcription to proceed to the fourth (-UTP) and to the twelfth (-CTP) nucleotides respectively. This was used to probe the interaction between RNA polymerase and the ColE 10.13 promoter by means of restriction cutting at the Hae III site at =27 and the Hha I site at +17. RNA polymerase binding alone blocks restriction cutting at the HAE III site but not at the Hha I site. Limited transcrption to the fourth nucleotide resulted in blocking at both sites. Transcription to the twelfth nucleotide resulted in partial cutting at the Hae III site and blocking at the Hha I site.


Studies on transfer ribonucleic acids and related compounds. XXXII. Synthesis of ribonucleotides corresponding to residues 1-5 and 6-10 of tRNAfMet from E. coli and their base conversion analogs.

E. Coli tRNAfMet fragments, C-G-C-G-Gp (bases 1-5), U-G-C-G-Gp (base 1 transition, analog) pG-G-C-G-Gp (base 1 transversion analog) and pG-G-s4U-G-Gp (bases 6-10) were synthesized by triester methods using 2'-O-(o-nitrobenzyl) nucleotides including a 3',5'-bisphosphorylated guanosine derivative. The s4U containing pentanucleotide was derived from the pG-G-C-G-Gp by treatment with liquid hydrogen sulfide.


Aspects of specific protein-DNA interaction; multi-mode binding of the oligopeptide antibiotic netropsin to (A.T)-rich DNA segments.

By means of titration viscometry a number of distinct modes could be resolved for the interaction between the antibiotic netropsin and DNA species of 50, 58, and 69 mole + (A+T) below r = 0.04 netropsin molecules bound per DNA phosphate group. The number of corresponding binding sites increases with a high power of the (A+T) content. The apparent association constants are very high (greater than 10(6) M-1, some perhaps greater than 10(6) M-1) and also rather different for most of the binding sites. It is suggested that some of these interaction modes differ in the number of hydrogen bonds formed between donors of the ligand and acceptors of the binding sites. The interaction modes were characterized quantitatively by their (species-independent) changes of DNA contour length and by the percentage of local DNA stiffening.