Characterization of a fission yeast gene, gpa2, that encodes a G alpha subunit involved in the monitoring of nutrition. The Schizosaccharomyces pombe gpa2 gene was cloned by hybridization with a cDNA for Dictyostelium discoideum G alpha 1. It encodes a homolog of G-protein alpha-subunits with 354 amino acids and a predicted molecular mass of 40,522. Disruption of gpa2 slows cell growth but is not lethal. Cells defective in gpa2 mate and sporulate readily in the presence of plentiful nutrition, bypassing the requirement of nitrogen starvation for the initiation of sexual development. These phenotypes mimic those of cells defective in cyr1 encoding adenylyl cyclase. The level of cAMP in gpa2 null mutants is only one-third of the wild-type level. Mutations in gpa2 that are likely to inhibit the GTPase activity of the gene product cause a slight increase in intracellular cAMP levels and result in leaky sterility. The cAMP level reaches 20 times as high as the wild-type level if a cell carries both this type of gpa2 mutation and a null mutation in pde1 encoding phosphodiesterase. Cells defective in gpa2 fail to produce cAMP in response to glucose stimulation. These results suggest that Gpa2 is involved in the determination of the cAMP level according to nutritional conditions, most likely as a positive regulator of adenylyl cyclase.


[Quantitative characteristics of neuronal composition of the ventral part of the cat's medial geniculate body]. Calculation of numerical density of neurons in ventral part of cat's medial geniculate body (vMGB) was made. It was shown that 1 mm3 of vMGB tissue contains 29,460 neurons. After 6 months from unilateral removal of the auditory cortex the quantity of large (supposedly thalamocortical) neurons in ipsilateral vMGB reduced on average by 78.1%, but of small ones--only by 10.7%.


SSL1, a suppressor of a HIS4 5'-UTR stem-loop mutation, is essential for translation initiation and affects UV resistance in yeast. The SSL1 locus was identified as a trans-acting suppressor that restores HIS4 expression despite a stem-loop structure in the 5'-UTR. SSL1 encodes an essential protein of 52 kD with features characteristic of a protein with multiple zinc fingers. The mechanism of SSL1 suppression is not related to altering his4 transcription or removing the stem-loop sequence from the 5'-UTR; rather, 3- to 5-fold increases in His4 translational expression are observed indicating a post-transcriptional mechanism for SSL1 suppression. SSL1 suppressor mutants that are conditional for growth have altered polysome profiles at the restrictive temperature, and their cell-free extracts are thermolabile in their ability to translate exogenously added mRNA. In addition, the mechanism of suppression appears to be specific for stem-loop structures placed near the 5' end of the message as opposed to a stem-loop located at a downstream position in the 5'-UTR. These observations suggest a role for this protein in promoting translation initiation presumably at the level of ribosomal binding to mRNA. Surprisingly, SSL1 suppressor mutations that are shown to confer an in vivo and in vitro defect in translation initiation also rendered yeast hypersensitive to UV irradiation. This latter phenotype was observed previously with a mutation in the SSL2 suppressor gene, which encodes the yeast homolog of the human gene ERCC-3, for which a defective form causes xeroderma pigmentosum. In light of the related effects of mutations in the SSL1 and SSL2 genes, the encoded proteins may functionally interact both to promote DNA repair and perform an essential function during translation initiation.


The basic region of AP-1 specifies glucocorticoid receptor activity at a composite response element. Unrelated factors collaborate at composite response elements to confer novel patterns of transcriptional regulation. For example, AP-1 and glucocorticoid receptor bind and mutually affect their activities at a 25-bp composite element, plfG. We found that different members of the AP-1 factor family that behave similarly in the absence of receptor are strikingly distinct in its presence: They specify opposite (enhancement vs. repression) regulatory actions by the receptor. Four amino acids within the AP-1 DNA-binding domain were identified as crucial determinants of receptor regulatory activity at plfG. We conclude that interactions of factors from separate transcription factor families at composite response elements provide a mechanism by which a single factor can regulate both positively or negatively, and a potential resolution of the apparent functional redundancy within regulatory factor families.


Specificities of protein-protein and protein-DNA interaction of GABP alpha and two newly defined ets-related proteins. The ets-related protein GABP alpha interacts with the four ankyrin-type (ANK) repeats of GABP beta to form a high-affinity DNA-binding complex that recognizes a site important for herpes simplex virus type I immediate early gene activation. To investigate the selectivity and specificity of the GABP complex, we have isolated two new ETS family members, termed ER81 and ER71. ER81 and GABP alpha were present in most tissues of adult mice, whereas ER71 was restricted to testis. We have compared the DNA-binding specificities of these proteins by binding site selection. GABP alpha, ER71, and ER81 recognized the common pentanucleotide DNA sequence 5'-CGGAA/T-3'. Although subtle differences were observed for nucleotide preferences flanking this pentanucleotide core, the overall similarity of the selected sequences was most striking. Given the observation that GABP alpha interaction with GABP beta requires its intact ETS domain, we further compared the ability of GABP beta to interact with other ETS proteins. GABP beta did not augment the DNA-binding activity of the highly similar ETS domains of ER81, ER71, or Ets-1. Moreover, probing of total tissue extracts with radiolabeled GABP beta demonstrated its exceedingly stringent specificity for GABP alpha. Given that the DNA-binding specificities of these ETS proteins are similar and that the protein-protein interactions between GABP beta and GABP alpha are highly specific, we conclude that the protein interactions determine the target site selection by GABP alpha.


An embryonically expressed gene is a target for c-Myc regulation via the c-Myc-binding sequence. We have used a subtraction/coexpression strategy involving two different tumors derived from c-myc-bearing transgenic mice to identify a gene that is a target for c-Myc regulation. The gene, expressed in certain embryonic and adult tissues and in several (but not all) c-myc-based tumors, bears a functional c-Myc-binding sequence located 3' to its transcription start site. This sequence is required for the binding of a nuclear protein complex which, by antibody analysis, includes c-Myc. This site is also required for expression of a reporter gene in chimeric constructs transfected into c-myc-overexpressing cells and, conversely, requires c-myc cotransfection for its enhanced expression in COS cells. Furthermore, transfection of c-myc blocks the normal down-regulation of this gene, which occurs in embryonic stem cells as they undergo differentiation. This target gene encodes an anonymous cDNA (ECA39) found previously to be amplified in a teratocarcinoma cell line.


Carboxy-terminal elements of c-Myb negatively regulate transcriptional activation in cis and in trans. The c-Myb protein plays a key role in normal hematopoiesis, and truncation results in its activation to a transforming protein. Truncation of the c-Myb carboxyl terminus also greatly increases its transcriptional activating activity. The role of specific carboxy-terminal domains in negative regulation was investigated using Myb and Myb fusions with GAL4, LexA, or VP16. Negative regulatory activity of the carboxyl terminus in cis resides in at least two regions. A sequence in one of these regions can also inhibit transcriptional activation by Myb, Myb-VP16, or LexA-Myb proteins in trans. Regulation in trans, or suppression, is independent of c-Myb DNA binding and, therefore, likely involves protein-protein interaction. Suppression does not require the presence of a predicted heptad leucine repeat structure on either molecule. The target of suppression is a sequence that contains part of the minimal Myb transcriptional activation domain. This sequence can confer suppressibility on fusion proteins containing heterologous DNA-binding or transcriptional activation domains.


Properties and localization of DNA methyltransferase in preimplantation mouse embryos: implications for genomic imprinting. Preimplantation mouse embryos contain very high levels of DNA methyltransferase activity. We show here that the form of DNA methyltransferase (DNA MTase) in early embryos differs from the form found in other cells and tissues by a slightly higher mobility on gel electrophoresis. Levels of DNA MTase were found to be very high throughout preimplantation development even though levels of 5-methylcytosine (m5C) in nuclear DNA are known to undergo a substantial decline in the same period. Confocal laser scanning microscopy of mouse embryos stained with DNA MTase-specific antibodies showed striking developmentally regulated changes in the distribution of DNA MTase. From the oocyte stage to the four-cell-stage, most DNA MTase was concentrated in peripheral cytoplasm, and nuclei did not contain detectable DNA MTase. In four- and eight-cell embryos, DNA MTase was seen in cytoplasmic granules; and in eight-cell embryos, DNA MTase was also present in large amounts in nuclei. Nuclei of blastocysts stained only faintly, whereas the cytoplasmic granules remained prominent. Paradoxically, DNA MTase was found to be at its highest levels in nuclei at a developmental stage where levels of m5C in DNA are decreasing most rapidly. Changes in methylation patterns in preimplantation embryos are therefore proposed to be under the control of unidentified regulatory factors rather than DNA MTase itself; these regulatory factors could be members of the group that contains the products of the Ssm-1 and Imp-1 genes, which are involved in the regulation of genomic imprinting.


Site-specific cross-linking of mammalian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing. We have used a site-specific cross-linking strategy to identify RNA and protein factors that interact with the 5' splice site region during mammalian pre-mRNA splicing. Two different pre-mRNA substrates were synthesized with a single 32P-labeled 4-thiouridine residue 2 nucleotides upstream of the 5' splice site. Selective photoactivation of the 4-thiouridine residue after incubation of either substrate under splicing conditions in HeLa nuclear extract resulted in cross-links to the U5 snRNA and the U5 snRNP protein p220. These ATP-dependent interactions occur before the first step of splicing. The U5 snRNA cross-links map to a phylogenetically invariant 9-nucleotide loop sequence and do not require Watson-Crick complementarity to the 5' exon. Cross-links of this position in the pre-mRNA to U1, but not to U2, U4, or U6 snRNAs, were also observed. The kinetics of U1 and U5 cross-link formation are similar, both peaking well before reaction intermediates appear.


The Drosophila RNA-binding protein RBP1 is localized to transcriptionally active sites of chromosomes and shows a functional similarity to human splicing factor ASF/SF2. An RNA-binding protein gene (rbp1) from Drosophila melanogaster, encoding an RNA recognition motif and an Arg-Ser rich (RS) domain, has been characterized. The predicted amino acid sequence of rbp1 is similar to those of the human splicing factor ASF/SF2, the Drosophila nuclear phosphoprotein SRp55, and the Drosophila puff-associated protein B52. Northern and immunohistochemical analyses showed that rbp1 is expressed at all stages in all tissues and that the RBP1 protein is localized to the nucleus. Consistent with a role in mRNA metabolism, indirect immunofluorescence reveals that the RBP1 protein colocalizes with RNA polymerase II on larval salivary gland polytene chromosomes. RBP1 protein made in Escherichia coli was tested for splicing activity using human cell extracts in which ASF has been shown previously both to activate splicing and to affect the choice of splice sites in alternatively spliced pre-mRNAs. In these assays, RBP1 protein, like ASF, is capable of both activating splicing and switching splice site selection. However, in each case, clear differences in the behavior of the two proteins were detected, suggesting that they have related but not identical functions. The general nuclear expression pattern, colocalization on chromosomes with RNA polymerase II, the similarity to ASF/SF2, SRp55, and B52, along with the effect on alternative splicing shown in vitro, suggest that rbp1 is involved in the processing of precursor mRNAs.


Spatial regulation of proneural gene activity: auto- and cross-activation of achaete is antagonized by extramacrochaetae. The spatially restricted activities of achaete (ac) and scute (sc) are thought to define proneural clusters of potential sensory organ precursor cells in the imaginal discs of Drosophila. These genes encode transcriptional regulators of the basic helix-loop-helix (bHLH) class. We have found that direct, positive transcriptional autoregulation by the ac protein and cross-regulation by sc are essential for high-level expression of the ac promoter in the proneural cluster pattern and that autoactivation is important for the bristle-promoting function of the ac gene. These auto- and cross-regulatory activities are antagonized in a dose-dependent manner by the inhibitory HLH protein encoded by the extramacrochaetae (emc) gene. We have found that emc is expressed in the wing imaginal disc in a pattern strongly complementary to that of the proneural clusters. Our results indicate that emc plays an essential early role in defining territories of bristle-forming potential.


Gene regulation in two dimensions: the proneural achaete and scute genes are controlled by combinations of axis-patterning genes through a common intergenic control region. The mechanisms that generate precise patterns of discrete cell types within developing fields are not well understood. One model for analyzing how cells interpret positional information in two dimensions is the regulation of proneural cluster formation within insect segments. Two adjacent proneural regulatory genes, achaete and scute, are expressed coincidently in cell clusters at reproducible anteroposterior (AP) and dorsoventral (DV) coordinates within the Drosophila embryo from which single neuroblasts later arise. Here, we show that the AP and DV position of these clusters is regulated through a common cis-acting region between the genes under the initial control of the products of the pair-rule and DV polarity genes and is later maintained by selected segment polarity genes. The combination of proneural gene activation/repression in AP stripes and repression within specific DV domains positions each cluster of achaete/scute expressing cells within segments; interactions between these cells then determine individual cell fates.


Two mammalian helix-loop-helix factors structurally related to Drosophila hairy and Enhancer of split. We report the molecular characterization of two novel rat helix-loop-helix (HLH) proteins, designated HES-1 and HES-3, that show structural homology to the Drosophila hairy and Enhancer of split [E(spl)] proteins, both of which are required for normal neurogenesis. HES-1 mRNA, expressed in various tissues of both embryos and adults, is present at a high level in the epithelial cells, including the embryonal neuroepithelial cells, as well as in the mesoderm-derived tissues such as the embryonal muscle. In contrast, HES-3 mRNA is produced exclusively in cerebellar Purkinje cells. HES-1 represses transcription by binding to the N box, which is a recognition sequence of E(spl) proteins. Interestingly, neither HES-1 nor HES-3 alone interacts efficiently with the E box, but each protein decreases the transcription induced by E-box-binding HLH activators such as E47. Furthermore, HES-1 also inhibits the functions of MyoD and MASH1 and effectively diminishes the myogenic conversion of C3H10T1/2 cells induced by MyoD. These results suggest that HES-1 may play an important role in mammalian development by negatively acting on the two different sequences while HES-3 acts as a repressor in a specific type of neurons.


The Drosophila hedgehog gene is expressed specifically in posterior compartment cells and is a target of engrailed regulation. cDNAs were isolated that represent transcripts of the Drosophila segment polarity gene, hedgehog (hh). Sequence analysis reveals a motif characteristic of a transmembrane domain, suggesting that the hh protein is membrane-associated. hh expression in epidermal cells is confined to the posterior compartments and coincides precisely with that of engrailed (en). Despite the similar patterns of expression in the cellular blastoderm, hh expression is independent of en, but hh expression becomes sensitive to and dependent on en during the extended germ band stage. The ectopic expression of hh that is normally induced in patched (ptc) mutant embryos does not appear in ptc en double mutants. We discuss these findings in terms of the relationship between en and hh, and the role of the hh function.


A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. A starvation-inducible DNA-binding protein was discovered as a result of the analysis of proteins synthesized in 3-day-old cultures of Escherichia coli. This 19-kD protein, designated Dps, is abundant in starved cells. In vitro, Dps forms extremely stable complexes with DNA, without apparent sequence specificity. When complexed with Dps, DNA is rendered DNase resistant. Mutant cells lacking Dps show dramatic changes in the pattern of proteins synthesized during starvation. The mutants also fail to develop starvation-induced resistance to hydrogen peroxide, an agent that can cause oxidative damage to DNA in vivo. These results have prompted us to postulate that Dps plays an important role both in gene expression and DNA protection during stationary phase. The existence of similar proteins, heretofore with no known function, in bacterial species distantly related to Escherichia coli suggests that Dps may define a novel class of widely conserved DNA-binding proteins.


[Specificity of thyroid follicular cells]. Thyroid follicular cells, as polarised endocrine cells in fish, amphibia, reptile, birds and mammalia, release Tg molecules, mainly via apical cytoplasm. These products known as colloid, the amount, contents, staining affinity, density and absorptive capacity, differs clearly even in follicules of the same gland. The pathways of secretion by "a regulated type of exocytoses" are clear signs of cell memory acquired during evolution and genetical program. However, the character of thyroid follicles appearance, as well as structural and ultrastructural organization of follicular cells, proliferative rate, nature of their internal membranes and the other organelles, might be altered by numerous endogenous and/or exogenous factors. Our earlier data, obtained examining thyroid follicular cells in the experiments as are: experimental goitre and iodine prophylaxis, enzootic goitre in the animals, after internal or X-rays irradiation, tissue culture and/or transplantation, might be summarized as follow. Thyroid follicular cells are polarized and their synthetic capacity and pathways of Tg molecules secretion are closely related with proliferative rates of granular and agranular ER, nature of their cavities and vesicles as well as localization of both ER and Golgi complexes. The main pathway of Tg and the other substances are released via apical cells by exocytosis. Their activity, storage capacity, the amount and properties of colloid are dependent on the balanced neuroendocrine cells activities. The character of colloid absorption, expressed in the number and size of microvillis and protrusion, as well as cytoskeleton, colloid droplets and the appearance of other apical cytoplasmic organelles, could be considered as a signs of stimulative hormonal or/and neuronal influences. The fate of reabsorbed Tg molecules, as dependent on the lysosomal activities, and release of thyroid hormones, is closely related on ultrastructural follicular cells properties. However, many endogenous and/or exogenous factors might affect both synthetic and secretory activities by stimulation, or retardation and even inhibition modifying genetic program and cell memory. As a results Tg molecule synthesis and maturation, as well as both Tg and thyroid hormones release, are released directly into blood circulation, as are usually a cases under conditions of thyroid hormones deficiency.


[The role of enkephalinase (neutral endopeptidase) in neurogenic inflammation of the respiratory tract]. In addition to the cholinergic and adrenergic nervous systems, a new noncholinergic and nonadrenergic nervous system has recently been described, involving the afferent sensory nerves in the airways. Many irritants (dusts, chemicals) stimulate these sensory nerves to release neuropeptides. Among these neuropeptides, the "tachykinins" exist in sensory nerves of airways (substance P, neurokinin A). These tachykinins have the ability to affect multiple cells in the airways and to provoke many responses including smooth muscle contraction, mucus secretion, plasma extravasation and neutrophil adhesion. This series of effects is termed "neurogenic inflammation". Using the respiratory tract as experimental model, it has been shown that: a) substance P (SP) is widely distributed in afferent fibers in the vagus, b) SP-immunoreactivity has been demonstrated in the epithelium, in airway smooth muscle, near blood vessels and submucosal glands, c) substance P and other tachykinins are released from sensory nerve terminals during stimulation electrically and by capsaicin, d) local administration of substance P mimics the effect of sensory nerve stimulation, e) smooth muscle contraction, gland secretion and plasma leakage, normally induced by nerve stimulation or noxious stimulus, are absent in tissues pretreated with the substance P depleting agent capsaicin or with tachykinin antagonists. These findings indicate that peptidergic nerve fibers are involved in the local regulation of tone of smooth muscle, regulation of blood flow, vascular permeability, and mucus secretion. We released that degradative mechanisms could play an important role in modulating tachykinin effects, just as acetylcholinesterase modulates effects of acetylcholine released from nerve terminals. We discovered that a membrane-bound enzyme called enkephalinase (also called neutral endopeptidase, EC 3, 4, 24, 11), located on specific cells that contain tachykinin receptors, modulate the action of tachykinins by cleaving and thus inactivating them. Our studies demonstrate that viral infection or cigarette smoke potentiate various effects of tachykinins by decreasing tissue enkephalinase activity. Thus, down-regulation of enkephalinase activity in specific tissues can modify the extent of neurogenic inflammation, and this modification could be important in the pathogenesis of diseases in airways and other tissues that contain tachykinins.


[Effects of various quantities of dietary iodine on total serum T3 and T4 in gilts during various phases of the reproductive cycle]. The effect of different dietary iodine levels (1 p.p.m.-control, 2 p.p.m. and 3 p.p.m.) on the serum levels of thyroid hormones (T3 and T4) has been studied in gilts at various stages of reproductive cycle, as well as on the their reproductive performances. The gilts were the crossbred of Swedish Landrace x Big Yorkshire x German Landrace, selected for reproduction. The serum concentrations of T3 and T4 in four month old gilts (at the beginning of experiment) were normal: 1.48 +/- 0.28 and 46.57 +/- 11.37, respectively. Approximately the same levels of serum thyroid hormones were found in all 3 experimental groups of the gilts at the first oestrus (about 6.5 months) when they were artificially inseminated, at the end of the first third of the pregnancy, at the end of lactation and on the day of first postlactational oestrus. Significantly lower concentrations of serum T4 were only at 1-3 days before parturition, 6-12 hours after parturition and during lactation in all groups of gilts. There were no significant differences in T3 and T4 concentrations as well as in reproductive performances between gilts on the different iodine diet. The authors concluded that 1 p.p.m. of dietary iodine, which is regularly applied in this pig farm, is sufficient to provide normal levels thyroid hormones and normal reproduction. There is not economical and physiological reasons to increase this level of iodine to 2 and 3 p.p.m., since the obtained results showed that these changes don't demonstrated neither positive or negative effects.


[Selenium toxicity in domestic animals]. The earliest written report of selenium poisoning is thought to be the description by Marco Polo of a necrotic hoof disease of horses that occurred in China in 13. century. However recognition of Se as toxic principle come in the early 1930s. Severity of Se poisoning depends on chemical forms of the element, species of animals and routes of administration. The soluble Se salts (Na2SeO3 and Na2SeO4) appear to be among the more toxic compounds; the Se inherent in grains and selenoamino acids (selenomethionine and selenocystine) appear to have relative moderate toxicity; the poorly soluble forms (e.g., elemental Se, Na2Se, SeS2 and diphenyl selenide) are among the least toxic of the Se compounds. In general, toxicity of Se compounds are substantially less when they are administered orally than when they are given parenterally. Rosenfeld and Beath described three clinical types of Se intoxication: acute selenosis, subacute selenosis (i.e., blind staggers type), and chronic selenosis (i.e., alkali disease type). Acute poisoning occurs when high Se content plants are consumed in large quantities within short period. Accidental acute poisoning occurs as consequence of errors in formulation of a Se supplemented diet. The most characteristic sign of acute selenosis is garlic breath due to the pulmonary excretion of volatile Se metabolites. Other signs include lethargy, excessive salivation, vomiting, dyspnea, muscle tremors and respiratory distress. Pathological findings are: congestion of the liver and kidney, fatty degeneration and focal necrosis of the liver, endocarditis and myocarditis. Subacute selenosis ("blind staggers") occurs as a consequence of exposure to large doses of Se over a longer period of time and manifests with neurological signs (e.g., blindness, ataxia, disorientation) and respiratory distress. This form of selenosis is most frequently observed in grazing animals that have consumed Se-accumulated plants. Chronic selenosis ("alkali disease") comes about when animals consume moderate levels of Se (more than 5 mg/kg and less than 40 mg/kg) for period of weeks or months. The usual clinical signs of chronic selenosis in horses, cattle and swine are: loss of hair (horses and cattle lose long hair from the mane and tails), emaciation, hoof lesions and lameness. In advanced cases liver cirrhosis, atrophy of the heart and anemia occur. In swine symmetrical poliomyclomalacia of cervical and lumbal/sacral spinal cord segment has been seen. Sheep seen to be more tolerant and get milder form of the disease. They lose appetite and have reduced gain. In growing chicks reduced gain and feed intake, rough feathers, and characteristics of nervousness has been observed. Reduced egg production, embryonic deformations and reduced hatchability has been observed in hens.(ABSTRACT TRUNCATED AT 400 WORDS)


[Biological activity of bound pesticide residues]. Investigations of bound pesticide residues carried out to date involved the detection of residues in commodities, their identification, determination, bioavailability and possible biological effects. The results obtained in this research and the results of other authors who have shown an increasing concern about the levels, bioavailability and biological effects of bound pesticide residues since the late 1980' are presented in this paper. Based on the results obtained, bound pesticide residues were detected in some commodities ranging from trace amounts to over 50% of the dose level applied. The residue content was influenced by the biochemical structure of the commodity, the compound and its application rate, the length of time from the application to residue analysis as well as on some environmental parameters (primarily temperature and humidity). It was also established that the bioavailability to experimental animals may range from medium to significant provoking adverse biological effects. These effects depended on the compound, its application rate and length of administration. Based on the results obtained so far it was established that bound pesticide residues need to be taken into account when determining maximum residue limits (MRL) as well as when assessing the toxicological risks of pesticide application.


[1H, 13C 15N and 31P NMR spectroscopy of plant seeds: possible applications]. We review here the recent achievements in high resolution NMR spectroscopy of four biologically most important nuclei (1H, 13C, 15N, 31P) applied on plant seeds in vivo or on extracts from the same tissue. Most often used nuclei in NMR analysis of seeds were protons (1H). The application of low resolution NMR for nondestructive analysis of moisture and oil in seeds had a long history--this was actually the only true nondestructive chemical analysis of genetic material. The combination of high resolution 1H NMR with magic angle sample spinning (MAS) enabled some authors to obtain a method for in vivo study of fatty acids composition of seed lipids. A promising technique for in vivo studies of biochemical composition and physiological processes in seeds is 13C NMR, particularly 1H-13C cross-polarization NMR combined with MAS technique. This enabled analysis of lipids in intact seeds, but also obtaining of high resolution spectra from the solid components of the seed matrix (starch, sugars, proteins). An interesting application of 13C NMR was monitoring the biochemical events following seed germination in vivo. Low natural ability of the NMR accessible 15N nucleus restricts the application of 15N NMR for in vivo seed analysis. Nevertheless, some sophisticated experiments combining double cross-polarization 15N/13C with the MAS technique were reviewed. 31P NMR is a promising technique in biological studies. Some recent achievements included detection of energetically important molecules of cells (ADP; ATP, oligosaccharides) as well as compartmentation of the inorganic phosphate (cytoplasmic, vacuolar). There were some indications of the studies made on intact seeds, but most of the 31P NMR work has been done on the extracts from seed material.


[New technology in maize breeding].

[New technology in maize breeding]. Results obtained by several approaches in the application of Biotechnology in maize breeding are reviewed. RFLP technology in the determination of genetic variation; gene transfer by the use of different methods of gene delivery and the determination of gene integration. Three technologies for foreign gene introduction have been applied; injection of plasmid pRT100 neo into archesporial tissue before micro and macro sporogenesis, slightly modified pollen-tube pathway technology and dry seed incubation in plasmid DNA solution. NPTII gene integration was followed by dot-blot and Southern blot analysis of plant DNA of both T1 and T2 plants. Gene expression was analysed by neomycin phosphotransferase activity. Transformed plants contained the selective NPTII gene sequence in an active form. Bacterial gene integration induced several heritable changes of plant phenotype. As an important change, alteration of the flowering time has been used as a criterion for selection and plant propagation to keep transformed progeny. Besides plant genome transformation, endogenous bacteria living in different maize tissue were found. As a perspective approach for biotechnology application in maize breeding biological vaccine construction has been selected. Therefore, antagonistic effect of gram positive bacterial strains to several pathogenic fungi was investigated. Results obtained after in vivo experiments are discussed.


[Chlorophyll synthesis in cotyledons after gamma ray irradiation of black pine seeds]. The radiosensitivity of the greening system of Pinus nigra Arn. cotyledons has been studied in this paper. An exponential relation exists between the effect and dose for chlorophyll synthesis in the dark. Chlorophyll synthesis in the light roughly parallels that of chlorophyll synthesis in the dark. The restoration of chlorophyll was observed both in the light and in the dark. A stimulative effect of low doses of gamma radiation on chlorophyll synthesis was noticed. The radiosensitivity of chlorophyll a and chlorophyll b synthesis varied with the experimental conditions, suggesting that chlorophyll b synthesis might occur independently of chlorophyll a synthesis.


[Membrane binding and soluble lectins in higher organisms]. Lectins defined as proteins other than enzymes and antibodies that possess binding sites for specific carbohydrate sequences, are associated with the cell surface, the cytoplasm, the nucleus, extracellular matrices and body fluids, and are involved in molecular recognition, trafficking of glycoproteins and tissue organisation. Animal lectins are classified into two main groups: the C-type and the S-type lectins, characterised by sequence homology, solubility, sugar specificity, distribution, developmental regulation, calcium-dependency and reducing agents-dependency for their saccharide binding activity. Under SANU Projects supervised by Academician Milovan Jovanović, lectin types C and S were investigated. Special emphasis was placed on asialoglycoprotein receptor of rat hepatocytes, soluble beta-galactoside-binding protein of human placenta and hepatic nuclear lectins. The results of this work are presented and discussed hereinafter.


[Transferrins].

[Transferrins]. The objective of this paper was to summarize the physical and chemical knowledge of the transferrin molecule. The ample data concerning transferrin were presented. Considerable attention was given to the microheterogeneity of the transferrin molecule--the polymorphism, differences of the binding site for iron and possible donation of iron to the cell, number of carbohydrate antennae, degree of sialyzation--in connection to the possible link between microheterogeneity and physiological and pathophysiological status of the organism.


[Regulation of erythropoiesis by erythropoietin]. Regulation of erythropoiesis by erythropoietin, proposed already at the beginning of this century, has been greatly enhanced by cloning and expression of the gene of erythropoietin. Erythropoietin gene functions as a housekeeping gene, so that erythropoietin is constitutively produced and is never absent from the blood. Hypoxia activates erythropoietin gene primarily in the kidney and also to a small extent in the liver. Peritubular interstitial cells in the kidney or the kidney tubular cells produce erythropoietin. The modulation of erythropoietin gene expression in the kidney and the liver differs. Kidney cells produce erythropoietin in all-or-non pattern and as the stimulus becomes more severe more cells produce erythropoietin while in the liver individual cells produce more erythropoietin when the stimulus is more severe. Erythropoietin exerts its effect on erythroid progenitor cells after binding to specific receptors that are expressed on target cells, interacts with those cells in the bone marrow stimulating them into cell cycle, supporting the survival of cycling progenitor cells and permitting their final differentiation into cells synthetizing hemoglobin.


[Susceptibility and the immune response to Trichinella spiralis infection]. The aim of this study was to assess a possible association between parameters of immune response and susceptibility/resistance to T. spiralis infection. The immune response to T. spiralis was monitored in two inbred strains of mice (BALB/c and C57Bl/6), as well as in the most important natural host--swine, by analysing the changes in main lymphocyte populations, the appearance of specific antibodies and worm burden recovery. In the murine model, the lower level of muscle larvae worm burden recovery in mice expressed greater resistance (BALB/c) was associated with manifested clinical signs, earlier development of humoral antibody response and a persistently higher level of CD4/CD8 T cell ratio, compared to T. spiralis-susceptible mice (C57Bl/6). In swine, high susceptibility to infection was also associated with lack of clinical symptoms, a delayed development of an antibody response and persistently low CD4/CD8 T cell ratio.


[Effects of amiodarone on the thyroid gland in euthyroid and hypothyroid animals]. Amiodarone is a benzofurane derivative which contains an appreciable amount of iodine (37%). It is used in cardiology as an antianginal and antiarrhythmic drug. Using guinea-pigs and rats as the animal model systems, the effects of amiodarone on normal and hyperplastic thyroid glands were investigated in eu- and hypothyroid animals. After the amiodarone treatment, considerable differences were observed in the levels of thyroid hormones and in the structure of the gland which proved dependent not only on the length of treatment, but also on animal species and the functional status of the gland before treatment. At extended amiodarone application, the rats with hyperplastic thyroid developed microlesions in epithelium of some follicles.


Socioeconomic factors affecting first marriage and birth rates by sex and age in the total Japanese population. The effects of a wide variety of social, economic and demographic factors on age-specific first marriage and live birth rates in 46 Japanese prefectures were analyzed using stepwise regression analysis for 1970 and again for 1975 after classification of those twenty-two factors by factor analysis. The principal results were as follows: (1) high employment (high income) and social mobility caused by industrialization had a strongly positive influence on the first marriage and birth rates for young females, (2) rural and urban residence factors had positive effects on the marriage and birth rates for young males and females, respectively, (3) old age factor had an inverse effect on the marriage rates for both males and females over a wide range of ages, and (4) young age factor promoted the birth rate for young and middle-aged females. The characteristics of the first marriage and live birth rates in Japan were discussed in the light of these findings.


The effect of modern Balinese Baris dancing exercise (MBBDE) on serum lipid profiles. There is still a lack of information on the effect of regular dancing exercise on lipid profiles. On the other hand, many studies have been carried out on the effect of aerobic exercise on lipid profiles. This study tried to find out the effects of Modern Balinese Baris Dancing Exercise (MBBDE) on serum lipid profiles. Subjects of the study were 30 healthy young male Balinese as an experimental group, and another 30 healthy young Balinese as control group. The MBBDE involved exercise intensity at 70-80% of targeted heart rate, for 50 min period, 3 times per week for 8 weeks. Pre- and post-control group design was applied. Total cholesterol and triglyceride were measured enzymatically. Following MBBDE 3 x 50 min/week for 8 weeks duration, serum level of high density lipoprotein cholesterol (HDL-C) concentration increased significantly from 55.3 +/- 2.32 mg/dl to 63.2 +/- 2.82 mg/dl (p < 0.001). It was also associated with the decrease of total cholesterol concentration from 195.5 +/- 21.10 mg/dl to 161.8 +/- 21.29 mg/dl (p < 0.001); triglyceride concentration from 132.2 +/- 9.65 mg/dl to 110.6 +/- 9.08 mg/dl (p < 0.001); and low density lipoprotein cholesterol (LDL-C) concentration from 113.8 +/- 21.68 mg/dl to 76.9 +/- 20.76 mg/dl (p < 0.001). No significant differences were found in the above parameters in the control group. It is concluded that MBBDE is an aerobic, endurance exercise, and therefore produces beneficial effect on the serum lipid profiles.


Daily energy expenditure in active and inactive persons with spinal cord injury. Daily energy expenditure (DEE) was evaluated seventeen male subjects with spinal cord injury (SSCI), who had active (N = 9) and inactive (N = 8) lifestyles, and in seven normal males. DEE was estimated from the mean 24-hr heart rate and heart rate-energy expenditure relationship determined in an arm cranking exercise. The DEE of SSCI on active days did not differ from those of normal subjects. On inactive days, SSCI had significantly lower DEE than on active days and than normal subjects. In contrast, the mean 24-hr heart rates of SSCI on active days and inactive SSCI were significantly greater than those of normal subjects, suggesting that SSCI, particularly inactive SSCI, exhibited a slight degree of tachycardia when compared to normal subjects. On inactive days, the DEE was fairly independent of the level of spinal cord injury. However, on active days, there was a clear tendency for SSCI with a low lesion level to have a higher DEE. Even if a SSCI with a high lesion level engaged in active sports, his DEE was relatively small. This lower DEE was largely attributable to the smaller functional muscle mass due to the limitation of physical activity.


Analysis of survival rates of sportsmen utilizing Cutler-Ederer method. Few studies have been conducted in Japan on the relationship between sports and the length of human life, i.e. the life-prolonging effect of exercise. In this paper, we conducted such research on 3,113 male graduates of a national university having a faculty of physical education. The subjects were divided according to their academic majors, which included physical education, humanities, and science courses. The following results were obtained: 1) Comparison of ages at death showed that physical education majors were distinctly represented in lower age groups. 2) Comparison of average ages at death by dividing the subjects into 20-year intervals according to their years of birth, revealed that physical education majors do not necessarily live longer and tend to die relatively young. 3) Comparison of average ages at death, including death by war, indicated that physical education majors lived 8.572 and 7.792 years less than humanities and science course majors, respectively. When death by war was excluded, the results were 6.189 and 5.548 years, respectively. Both results were statistically significant. 4) Comparison of cumulative survival rates utilizing Cutler-Ederer method showed that the rates of survival of physical education majors were low.


Investigation and research on classification of productive skills (1)--Actual work and skills in the car manufacturing industry. A survey was conducted at a site for production which is in the process of technological innovation to examine and clarify the trend in current productive skills. In August 1991, a questionnaire survey was given to skilled workers of a car manufacturing company. The number of valid responses was 1,215. The survey included 133 items in the following three areas: (1) nature of workers' productive skills, 40 items; (2) human functions and vocational ability necessary for the work, 60 items; and (3) working conditions, 33 items. The characteristics of the productive skills required in each division were compared and examined based on the final results of the survey. Clear characteristics were found which reflect the actual productive skills introduced into the factories and sections through technological innovation. The nature of the product manufactured at each factory and the actual skills which reflect the work done at the factories are clarified. The survey items used for this research have effectively clarified the characteristic of the productive skills.


Occupational biomechanics of the neck: a review and recommendations. Musculoskeletal disorders of the neck are becoming a major concern in industry. Several studies reported the association of neck pain, discomfort, and symptoms with different occupations in industry. Thus, the main objective of this study is to review and evaluate biomechanical techniques used in analyzing the occupational factors leading to neck pain and disorders. Recommendations for future research are also discussed.


Effects of anticipation, prior-exercise, and breathing frequency to ventilatory response during step exercise: a preliminary study. We examined the effects of anticipation, prior-exercise, and restricted breathing frequency on the ventilatory transient response to bicycle step exercise (75 W, 4 min, 50 rpm), i.e., 1) whether the increase of work rate was anticipated by the subject or not, 2) whether the exercise was preceded by light exercise (25 W), or rest, and 3) whether the exercise entrained the breathing frequency (f: 12.5/min, or 25/min) or not (voluntary). The corresponding step-on exercise was randomly performed at least two to five times by one adult male subject. As a result, a) the initial rapid ventilatory component, phase 1, was not observed when initiated from light exercise, whereas the overshot phase 1 was observed from rest in anticipation and voluntary breathing frequency condition due to the rapid increase of tidal volume; b) compared with the anticipation condition, the phase 1 response of VE in the non-anticipation condition was slower with prior-rest, and not with prior-light exercise; and c) the restriction of the breathing frequency for entraining the exercise rhythm did not affect the initial rapid response, but decreased the fluctuation of VE in the steady state, compared to the condition of voluntary breathing frequency.


A study of perceptual motor load as affected by combined manual and decision task characteristics. This paper presents an experimental study of Perceptual Motor Load (PML) in a Combined Manual and Decision Task (CMDT). The purpose of this study was to compare Michon's scoring method of measuring PML with a new scoring method as affected by a two-stage CMDT under different work-load conditions. The dual task approach, using the simple task of foot tapping as the secondary task, was applied to obtain the quantitative measurement of PML. The experiment consisted of four major conditions, comprised of one of two activations, i.e., either unimanual or bimanual paired with each of the two stimulative repetitions, i.e., either random or sequential. Under each of the four experimental conditions, a central composite design was applied. The design included three levels of information load, three levels of time lag, and three levels of probability distribution of emergency signal occurrences. Twenty male subjects participated in the experiments. No significant difference between the results of both scoring methods was found; however, the new scoring method can contribute a better measure of PML in terms of correlation coefficients between the scores and the performance times.


The effects of various mental tasks on appearance of frontal midline theta activity in EEG. A distinct theta rhythm of EEG in the frontal midline area during performance of mental tasks has been called Fm theta. One of the characteristics of Fm theta is individual differences in its appearance. The effects of various mental tasks and its repetition on appearance of Fm theta were investigated. Adding, correcting wrong words, short-term memory (STM) and counting cubes were imposed 6 times on 7 male students who didn't generate Fm theta at the previous experiment. Counting cubes evoked more Fm theta than the other three mental tasks. In every task the first trial showed little Fm theta, and the appearance time of Fm theta was enhanced by repetition. Type I of Spielberger's State-Trait Anxiety Inventory (STAI-I) demonstrated a higher state anxiety level of the subjects at the first trial. It may be concluded that the amount of Fm theta corresponds to the level in concentration of attention.


Induction of monocytic cell adherence to matrix-bound fibronectin by phorbol ester. The inflammatory response requires the localization of monocytic cells to sites of tissue injury through adherence to extracellular matrix molecules such as fibronectin (Fn), a nonimmune opsonin, which binds to collagen, fibrin, heparin and cell surfaces. Adherence to this molecule of two myeloid cell lines differing in their stage of differentiation, was studied. In the baseline state, U937 monocytic cells bound specifically to matrix-bound Fn, while HL-60 promyelocytic cells bound minimally. Exposure to Phorbol myristate acetate (PMA) dramatically increased binding of both U937 and HL-60 cells to Fn with plateau effects at 10 ng/ml for both cell lines and at 30 and 60 minutes for U937 and HL-60, respectively. Treatment with metabolic inhibitors suggests that PMA stimulation depends at least in part on intact energy metabolism, protein synthesis and cytoskeletal components. This system should help elucidate the early molecular and biochemical events involved in monocyte adherence to the extracellular matrix.


Influence of topical exposure to chemical allergens on murine Langerhans cells. Comparison of 2,4-dinitrochlorobenzene with trimellitic anhydride. Chemical allergens differ with respect to the type of hypersensitivity reactions they preferentially elicit. Some chemicals, such as trimellitic anhydride (TMA), have the potential to induce both contact and respiratory hypersensitivity. Other chemicals cause only contact allergy. For example, 2,4-dinitrochlorobenzene (DNCB), a potent contact allergen, appears not to induce respiratory sensitization. In previous studies we have shown that topical exposure of mice to TMA and DNCB, under conditions of equivalent immunogenicity with respect to draining lymph node activation and contact sensitization, caused qualitatively different antibody responses. While the chemicals provoked IgG anti-hapten antibody responses of equivalent magnitude, only TMA induced an IgE response, and DNCB caused a significantly stronger IgG2a response. These data are consistent with the preferential activation by DNCB and TMA of Th1 and Th2 cells respectively. The purpose of the present study was to examine whether the qualitative differences in immune responses stimulated by these chemicals is reflected by variable affects on Langerhans cells (LC) in situ. Mice were exposed to concentrations of DNCB (1%) and TMA (50%) which caused equivalent levels of contact sensitization. Under these conditions topical exposure to DNCB, but not to TMA, or to vehicle alone, resulted in increased expression by LC of Ia antigen. Similar treatment with an irritant concentration (20%) of sodium dodecyl sulphate failed to influence Ia expression by LC. These data indicate that, at concentrations which induce similar levels of skin sensitization, not all contact allergens cause rapid changes in LC Ia expression, and that the qualitative differences in immune responses elicited by chemical allergens DNCB and TMA is associated with variable effects on LC.


Inhibition of the neutrophil oxidative response induced by the oral administration of nimesulide in normal volunteers. The superoxide (O2) production by phagocytes (neutrophils plus monocytes) and the lactoferrin release by neutrophils were measured in normal volunteers before and after the oral administration of the anti-inflammatory drug nimesulide. The chemotactic factor N-formylmethionyl-leucyl-phenylalanine (FMLP) and opsonized zymosan particles (OPZ) were used as activating stimuli. The oral administration of nimesulide lowered the phagocyte ability to generate O2- in response to both FMLP (percent inhibition = 67.62) and OPZ (percent inhibition = 36.75). The lactoferrin release by neutrophils was unaffected, proving that the drug does not affect the exocytosis of specific granules. The results provide direct evidence that the oral administration of nimesulide efficiently reduces the oxidative potential of phagocytes, particularly neutrophils, without interfering with mechanisms related to exocytosis of specific granules and involved in the amplification of the cell responses to inflammatory mediators.


Efficacy of a herbal medicine "sho-saiko-to" on the improvement of impaired cytokine production of peripheral blood mononuclear cells in patients with chronic viral hepatitis. Peripheral blood samples were collected from normal subjects and chronic viral hepatitis patients, and the in vitro capability of the peripheral blood mononuclear cells to produce various cytokine (interleukin-1 beta, interleukin-6, interferon-gamma, and granulocyte-macrophage colony-stimulating factor) were analyzed by adding pokeweed mitogen. We then investigated the effects of a herbal medicine "Sho-saiko-to" on the levels of cytokine production. The production levels of the 4 cytokines were significantly lower in the peripheral blood mononuclear cells of the patients (Patient Group) than in those of normal subjects (Control Group). The addition of Sho-saiko-to to the Patient Group resulted in improved productions of those cytokines, as well as an remarkable improvement of interleukin-1 beta production. The results demonstrated that Sho-saiko-to acts to improve such immunological abnormalities as decreased cytokine productions. Administration of Sho-saiko-to to chronic viral hepatitis patients is also expected to have immunological benefits.


Stress and murine NK cell function: the role of blood loss. It has been previously reported that NK cell function decreases following surgery in mice. To explore the basis of this observation, we compared the relative influence of anesthesia, surgical amputation and bleeding on NK cell cytotoxicity in C57BL/6 mice. After hind limb amputation, including with blood loss, there was a statistically significant decrease in NK cell cytotoxicity and the appearance of splenomegaly on the 4th day postoperative day. The increase in spleen size appeared to be due to either the surgical stress-induced expansion of splenic erythroblasts or erythroblast generation following blood loss. In contrast, if blood loss was minimal there was no suppression of NK cell cytotoxicity following hind limb amputation. Moreover, there was a statistically significant correlation of NK cell activity and the quantitation of total blood loss. Interestingly, the decrement in NK cell activity was not observed if blood transfusion was made, even in the presence of surgical amputation. These observations are important for defining the immune suppression reported following surgery and suggest that in human, chronic blood loss may also be associated with immune suppression.


Effects of cryopreservation on immune responses: VI. An inexpensive method for freezing human peripheral blood mononuclear cells. To determine the true usefulness of the commercially available inexpensive freezing unit (Mr Frosty) for cryopreservation of peripheral blood mononuclear cells (PBMCs), functional assays were carried out on cells that were frozen in this unit. The responses of these cells were compared with those of controlled rate frozen and fresh cells. Cell viability, recovery, proliferative responses to both phytohemagglutinin (PHA) and pokeweed mitogen (PWM), as well as interleukin (IL)-1 secreting activities of cells frozen in this container were closely comparable to controlled rate frozen cells. Although, the mean immunoglobulin (Ig) and IL-2 secreting abilities of these frozen cells were lower than the controlled rate frozen cells, these responses were still comparable to fresh cells in several of the individuals. Based on these results and considering the cost involved in controlled rate freezing method, we recommend this inexpensive unit for freezing PBMCs for subsequent immunological studies.


Age-related changes of the antigen-specific antibody formation in vitro and PHA-induced T-cell proliferation in individuals who met the health criteria of the Senieur protocol. The antigen-specific antibody secretion in vitro after immunisation with the primary T-cell dependent antigen Helix pomatia Haemocyanin (HPH) was investigated in both young and elderly individuals, who all met the health admission criteria for immunogerontological studies as detailed in the SENIEUR protocol. In addition, elderly non-Senieur persons were incorporated in this study. Young and elderly Senieur volunteers were fully comparable in terms of the occurrence of anti-HPH antibody secreting cells after in vitro simulation of peripheral blood mononuclear cells with variable doses of the antigen. In contrast, the non-Senieur elderly showed a lower number of anti-HPH antibody secreting cells in vitro. PHA-conditioned medium did enhance this in vitro response, whereas the addition of IL-2 remained ineffective. The PHA-induced T-cell proliferation was found to be somewhat impaired in elderly Senieur individuals and significantly lower in elderly non-Senieur individuals compared to young healthy persons. Using an immunofluorescence double staining technique after BrdU incorporation, the phenotype of the proliferating cells was determined. Again the total number of proliferating cells was impaired in the non-Senieur elderly. No changes in the relative contribution of CD4+ or CD8+ cells to the number of proliferating cells were found in the different age groups. On the other hand, a significantly lower number of proliferating cells with IL-2 receptor expression were detected in the non-Senieur individuals, which could account for the lack of response to IL-2 in this group. Our study clearly shows that so-called age-associated immune deficiency can be the result of disease and not necessarily of the ageing process itself.


Epidermal growth factor suppresses in vitro senescence in the ability of human umbilical vein endothelial cells to proliferate, but not in the ability to produce prostacyclin. Addition of epidermal growth factor (EGF) to culture medium extended the replicative life span of human umbilical vein endothelial (HUVE) cells in culture. In brief, EGF suppresses the age-related decrease (in vitro senescence) in cell proliferative ability. However, the addition of EGF did not extend the culture period in which prostacyclin (PGI2) is actively produced by the cells. Therefore, EGF does not suppress the age-related decrease (in vitro senescence) in the ability of cultured HUVE cells to produce PGI2. These results suggest that the process of in vitro senescence in the cell proliferative ability is not necessarily correlated with that of in vitro senescence in the ability to produce PGI2.


Influence of ageing on antibody formation in vivo after immunisation with the primary T-cell dependent antigen Helix pomatia haemocyanin. The in vivo antibody response to the primary T-cell dependent antigen Helix pomatia Haemocyanin (HPH) was studied, in order to detect the possible presence of a humoral immune deficiency in ageing. The IgG subclass distribution of the specific antibodies was also determined. In order to define a dose of HPH which could be used to discriminate between the responsiveness of healthy and immunocompromised individuals, we first established a dose-response curve for this antigen in 60 healthy young volunteers. Their responses were compared with the responses of a group of patients suffering from end stage renal failure. The patients who were treated with haemodialysis showed a significantly lower IgM, IgG and IgA anti-HPH antibody response after immunisation with a dose of 30 micrograms HPH, which could be restored by increasing the antigen dose. Patients treated with continuous ambulant peritoneal dialysis and a group of elderly persons, selected according to the Senieur protocol, showed no impairment of antibody formation after immunisation with 30 micrograms HPH, but in the non-Senieur elderly the anti-HPH antibody response was significantly lower. Furthermore, Senieur and non-Senieur elderly persons showed a diminished IgG2 anti-HPH antibody formation, whereas in the elderly non-Senieur individuals and in the patients with renal insufficiency, IgG1 and IgG3 anti-HPH antibodies were also diminished. This study clearly shows that the so-called age-associated immune deficiency can be the result of disease and is not necessarily due to the ageing process itself.


Serum immunoglobulin class and IgG subclass levels and the occurrence of homogeneous immunoglobulins during the course of ageing in humans. Serum levels of IgM and IgA classes and of IgG subclasses were determined and related to the presence of homogeneous immunoglobulin components (H-Ig) in volunteers equally distributed in age groups from 25 to 98 years, who all met the Senieur admission criteria for immunogerontological studies. In addition, sera of non-Senieur volunteers aged 75 years and older were included. Furthermore, the amount of IgD was determined in sera of Senieur individuals equally distributed in age groups from 15 to 98 years. In the Senieur persons, the contribution of the IgG subclasses and the IgM and IgA classes to the pool of serum immunoglobulins remained relatively unchanged during the course of ageing. In comparison with Senieur individuals aged 25-34 years, a slight increase in IgM and IgA levels was observed from the age 35 to 44 onwards and in IgG1 from the age 55 to 64 onwards. The variability of the immunoglobulin concentrations increased during ageing. The most prominent observation was the continuous decline of serum IgD starting in young adults. The non-Senieur persons differed from their Senieur age-matched counterparts mainly by the elevated IgG2 and IgA levels. During the course of ageing, H-Ig mainly of low concentration were detected at an increasing frequency in the Senieur persons and even more frequently in the elderly non-Senieur volunteers. Although in some individuals the elevation of immunoglobulin levels correlated with the appearance of H-Ig within the corresponding isotype, this relationship was not conclusive for all sera investigated. These results suggest that the rise of serum levels of individual immunoglobulin isotypes associated with ageing is usually the consequence of a polyclonal B cell activation. The occurrence of H-Ig and the decline of serum IgD in aged Senieur persons indicate that these are, at least partly, true phenomena of ageing and not always the consequence of disease.


The dynamics of aging and mortality in the United States, 1900-1988. The Strehler-Mildvan modification of the Gompertz relationship between aging and mortality provides a dynamic theoretical model for identifying and separating genetic, environmental and competitive influences upon age-related mortality. The initial method of longitudinal Gompertzian analysis, which utilized linear regression, tended to underestimate both genetic and environmental influences upon age-related mortality. A modified method of longitudinal Gompertzian analysis has been applied to mortality due to stomach cancer, cervical cancer and emphysema. This modified method of longitudinal Gompertzian analysis suggests that the genetic influence upon age-related mortality is essentially the same for both men and women. Moreover, application of this modified method suggests that environmental influences upon age-related mortality in the United States have been declining for the past 20 years for men and for the past 45 years for women.


The effects of exercise mode, swimming vs. running, upon bone growth in the rapidly growing female rat. The purpose of this study was to compare the effects of two programs of endurance training, of equal duration and intensity, on bone development in female rats. Thirty-eight female Wistar rats were randomly assigned to one of three groups: run-trained (RUN), swim-trained (SWIM) or control (CON). The RUN group ran at a speed of 27 m/min up an 8 degrees incline. Swim trained animals swam with 2% of body weight attached to their tails. Training sessions were 2 h/day, 5 days/week and were conducted over a 10-week period. Hindlimb and forelimb muscles were removed upon sacrifice and analyzed for citrate synthase (CS) activity, liver (LG) and muscle (MG) glycogen. The parametrial fat pads were removed, digested with collagenase, and 2-deoxy-D-[3H]glucose uptake measured in isolated cells. Bone weight, length, diameter, ponderal index and bone mineral content (BMC) were measured in the femur and humerus of each animal. The LG, MG, fat cell volume, glucose uptake of the adipocyte and adrenal weight data indicate that the training response was identical. The CS activity of the muscles indicated that mechanical and recruitment patterns of the upper and lower body differ and could be responsible for bone development patterns found in this study. Exercise had a minimal effect on bone growth in the run-trained animals but did stimulate development in the swim-trained animals. The humerus of the SWIM was significantly (P < 0.05) heavier, wider and had a greater BMC when compared with those of the RUN and CON rats. The results of this study indicate that the muscular forces applied by the swim training protocol produced greater bone adaptations than the forces applied by a running protocol of equal duration and intensity.


Oral choline alfoscerate counteracts age-dependent loss of mossy fibres in the rat hippocampus. Mossy fibres represent a major intrahippocampal associative pathway. They consist of axons of granule cells of the dentate gyrus and show an age-dependent loss as do the granule cells of the dentate gyrus. The present study was designed to assess whether long-term treatment of rats with choline alfoscerate in their drinking water would be effective in countering the loss of mossy fibres and of granule cells occurring with aging. Choline alfoscerate is a precursor in the biosynthesis of brain phospholipids and increases the bioavailability of choline in nervous tissue. Male Sprague-Dawley rats of 18 months of age were divided into two groups. One group received a daily dose of 100 mg/kg choline alfoscerate for 6 months; the other group was used as an untreated control. Twelve-month-old untreated animals were used as a reference group. The area occupied by mossy fibres, as well as their density, was significantly reduced in 24-month-old control rats in comparison with 12-month-old rats. The same is true for the density granule cells of the dentate gyrus which was decreased by about 20% in the oldest animals. In choline alfoscerate-treated rats both the area occupied by mossy fibres and their density were significantly higher than in age-matched controls. Moreover, the number of granule neurons of the hippocampus was higher by about 7% in choline alfoscerate-treated than in control 24-month-old rats. The above data suggest that choline alfoscerate treatment counteracts some anatomical changes of the rat hippocampus occurring in old age.


Effect of traditional Chinese herbal medicines on the pharmacokinetics of western drugs in Sprague-Dawley rats of different ages (II): Aminophylline-huan shao tan and aminophylline-pu chung yi chi tang. The effect of Chinese herbal medicines (Huan Shao Tan and Pu Chung Yi Chi Tang) and western drugs (sodium phenobarbital and cimetidine) on the serum concentration and pharmacokinetic parameters of theophylline and cytochrome P-450 of Sprague-Dawley (SD) rats of three different ages were examined. The older rats without pretreatment with Chinese herbal medicines and western drugs exhibited higher serum theophylline concentration and lower pharmacokinetic parameters of theophylline than middle-aged and younger rats (P < 0.05), but there was no difference in cytochrome P-450 activity among the three different ages of rats. All rats when pretreated with sodium phenobarbital showed lower serum theophylline concentration and higher pharmacokinetics parameters of theophylline. Also, the activity of cytochrome P-450 was higher (P < 0.05). When cimetidine was pre-administered in SD rats of three age groups, all rats exhibited lower serum theophylline concentration and higher pharmacokinetics parameters (P < 0.05), but the activity of cytochrome P-450 remained unchanged (P > 0.05). The results were opposite to other studies, probably because the dose and dosing intervals were different. No single effect occurred on the younger and middle-aged rats after pretreatment with Huan Shao Tan and Pu Chung Yi Chi Tang: their serum theophylline concentration, pharmacokinetics parameters and cytochrome P-450 activity were the same as the control group. However, the older rats after pretreatment with Huan Shao Tan or Pu Chung Yi Chi Tang showed lower serum theophylline concentration and higher pharmacokinetics parameters than the younger and middle-aged rats pretreated with similar Chinese herbal medicines. This indicates that Huan Shao Tan and Pu Chung Yi Chi Tang may perhaps improve the elimination of theophylline in older rats. This might be attributed to the increase in hepatic blood flow or in liver volume, since the activity of cytochrome P-450 was not affected by the administration of Chinese herbal medicines.


School and family effects on the ontogeny of children's interests, self-perceptions, and activity choices. In this chapter we have presented two perspectives on the link between social context and the following motivational constructs: self-concept of ability and sense of personal efficacy in specific activity domains; perceptions of the value of skills in various domains; interest in various activities; activity choice; persistence; performance; and general self-esteem. In the first section, we discussed how social-contextual variables in both the family and the home could produce individual differences in the motivational constructs of interest. We presented a general framework for thinking about this issue and summarized our recent empirical work. In the second section, we discussed how systematic changes in the social environments that confront children as they develop could explain age-related changes in the motivational constructs of interest. Again we presented a general framework for thinking about this issue and summarized our empirical work testing the hypotheses generated from this framework. Throughout this section we have argued that optimal development takes place when there is good stage-environment fit between the needs of developing individuals and the opportunities afforded in their social environments. Furthermore, we suggested that the negative changes in motivational variables often associated with early adolescent development result from regressive changes in school and home environments. For example, the transition to junior high school, in particular, often confronts early adolescents with regressive environmental changes such as a decrease in the opportunity to participate in classroom decision making, a decrease in teacher support and teacher efficacy, and an increase in teaching styles and reporting practices likely to induce a focus on relative ability and comparative performance as well as excessive social comparison. Not surprisingly, there is also a decrease in intrinsic motivation and an increase in school misbehavior associated with this transition, and these changes are most apparent among adolescents who report regressive changes in the characteristics of classroom and school social environment. Such motivational changes are not apparent in adolescents who report the more developmentally appropriate shifts in the social context at school. Although our analysis of the family data is not as complete as our analysis of the classroom data, we have found evidence that a similar process may be going on in the family in relation to issues of control and autonomy. Excessive parental control is linked to lower intrinsic school motivation, to more negative change in self-esteem following the junior high school transition, to more school misbehavior, and to relatively greater investment in peer social attachments.(ABSTRACT TRUNCATED AT 400 WORDS)


Motivation for social contact across the life span: a theory of socioemotional selectivity. Older people engage in social interaction less frequently than their younger counterparts. As I mentioned at the start, the change has been interpreted in largely negative terms. Yet when asked about their social relationships, older people describe them as satisfying, supportive, and fulfilling. Marriages are less negative and more positive. Close relationships with siblings are renewed, and relationships with children are better than ever before. Even though older people interact with others less frequently than younger people do, old age is not a time of misery, rigidity, or melancholy. Rather than present a paradox, I argue here that decreasing rates of contact reflect a reorganization of the goal hierarchies that underlie motivation for social contact and lead to greater selectivity in social partners. This reorganization does not occur haphazardly. Self-definition, information seeking, and emotion regulation are ranked differently depending not only on past experiences, but on place in the life cycle and concomitant expectations about the future. I contend that the emphasis on emotion in old age results from a recognition of the finality of life. In most people's lives this does not appear suddenly in old age but occurs gradually across adulthood. At times, however, life events conspire to bring about endings more quickly. Whether as benign as a geographical relocation or as sinister as a fatal disease, endings heighten the salience of surrounding emotions. When each interaction with a grandchild or good-bye kiss to a spouse may be the last, a sense of poignancy may permeate even the most casual everyday experiences. When the regulation of emotion assumes greatest priority among social motives, social partners are carefully chosen. The most likely choices will be long-term friends and loved ones, because they are most likely to provide positive emotional experiences and affirm the self. Information seeking will motivate some social behavior, but for reasons discussed previously, this will also require judicious choices of social partners. Narrowing the range of social partners allows people to conserve physical and cognitive resources, freeing time and energy for selected social relationships. As such, SST is highly consistent with the selective optimization with compensation model of successful aging formulated by P. Baltes and M. Baltes (1990) described above. SST is meant to describe and explain the underlying mechanisms for age-related changes in social behavior. It is not intended to be prescriptive.(ABSTRACT TRUNCATED AT 400 WORDS)


Neutrophil function as related to neutrophil-endothelial cell interactions. Advances have recently been made in several areas related to neutrophil-mediated tissue damage. Certain chemotactic receptors have been cloned, their relationship to G proteins examined, and their mechanisms of downregulation partly elucidated. Downregulation of chemotactic receptors could be useful in controlling the damage inflicted by neutrophils on nearby tissues. Studies of neutrophil adhesion to endothelium have revealed two classes of adhesion proteins: selectins, and the integrin/integrin receptor system. Selectins are involved in neutrophil margination, while the integrin system participates in the egress of neutrophils into regions of inflammation. The microfilaments are strands of polymerized actin which form a network that pervades the neutrophil cytoplasm. On activation of the neutrophil, changes occur in the state of the microfilament network that appear to act in combination with the integrin/integrin receptor system to cause neutrophils to be retained in capillaries of tissues reperfused after hypoxia. To the extent that reperfusion injury is caused by these retained neutrophils, prevention of their effect may require measures directed at the microfilament network as well as the integrin/integrin receptor system. With regard to mechanisms of neutrophil-mediated tissue damage, recent studies have emphasized the extent to which oxidative systems interact with proteases to augment the damage inflicted by activated neutrophils. The unique efficacy of secretory leukoprotease in preventing neutrophil-mediated proteolysis is discussed.


Tumor necrosis factor in metabolism of disease: hormonal actions versus local tissue effects. Tumor necrosis factor is a cytokine that participates in the mediation of numerous diseases associated with inflammation, cachexia, shock, and tissue injury. Early studies of the biology of TNF delineated its hormonal actions as well as its systemic toxicity. More recent investigations have drawn attention to its paracrine actions that predominate when it is produced locally in the brain or vital organs. For instance, when compartmentalized production of TNF occurs in the central nervous system it directly mediates fever, anorexia, and altered whole-body metabolism. Since these changes are mediated within the neural network they occur independently of simultaneously sampled serum TNF levels. These paracrine actions of TNF have implications for diseases associated with production of TNF in tissues (e.g. HIV cerebritis, multiple sclerosis, cerebral malaria and cancer), because they may differ markedly from the hormone like-actions associated with systemic release. Since TNF may be beneficial in some diseases yet injurious in others, both the hormonal and paracrine actions must be precisely defined in order to formulate novel treatment strategies based on either enhancing its useful effects, or suppressing toxicity.


Can immunocompetent cells and their cytokines play a role in atherogenesis? The human atherosclerotic plaque represents a mixture of inflammation, lipid accumulation and fibrosis. Cholesterol deposits are predominantly found in the core of the plaque, which is surrounded by a fibrotic cap. Much of the cholesterol is intracellular in macrophage-derived foam cells, and other macrophages together with T lymphocytes are intermixed with smooth muscle cells in the fibrous cap. Recent studies have shown that cytokines released by inflammatory cells can regulate cell proliferation, cholesterol metabolism, and differentiation of vascular cells. Interferon-gamma, one of the major secretory products of activated T lymphocytes, inhibits smooth muscle proliferation and reduces expression of alpha-actin. This suggests that T cells, by releasing gamma-interferon, may inhibit the formation of arterial stenosis and spasm. Recent animal experiments support this hypothesis by demonstrating that injections of recombinant gamma-interferon reduce arterial intimal lesions after balloon catheter injury. In similar experiments, it was shown that larger intimal lesions develop in T cell-depleted rats and in athymic nude rats. This shows that T cells inhibit the vascular response to injury and supports the notion that gamma-interferon released by these T cells is an important paracrine regulator of arterial cell proliferation. The cytokines, gamma-interferon and TNF, also downregulate scavenger receptor expression in monocyte-derived macrophages, leading to an inhibition of the transformation of these cells into foam cells. Taken together, these data show that cytokines of the immune system are important regulators of cholesterol deposition and cell proliferation in atherogenesis.


Cytokines as mediators of vascular pathology. Interactions between leukocytes and intrinsic vascular wall cells characterize many inflammatory reactions and contribute importantly to the pathogenesis of many vascular diseases. In view of this intimate involvement of leukocytes in vascular pathology it is important to understand the signals that recruit and activate leukocytes locally in regions of vascular pathology. It is also desirable to delineate the mechanisms by which leukocytes influence the behavior of intrinsic vascular wall cells in ways which may contribute to vascular lesion formation. Mediators elaborated by leukocytes include small molecules including lipid-derived mediators such as prostanoids, leukotrienes, and platelet activating factor. Leukocytes can also produce protein mediators including those currently classified as cytokines. The cytokines, protein mediators involved in inflammation and control of the immune response, derive from all classes of leukocytes studied. Local cytokine networks may orchestrate complex programs of expression of functions of leukocytes and endothelial and smooth muscle cells involved in vascular homeostasis and pathology. Our laboratory has been interested in hyperplastic arterial diseases including atherosclerosis and restenosis following angioplasty treatment of obstructive atherosclerosis. Definitive evidence for roles of cytokines in the pathogenesis of these syndromes are lacking. However, various in vitro and in vivo studies have furnished sufficient information to permit formulation of rather detailed hypotheses or models. In hypercholesterolemic rabbits vascular cell adhesion molecule-l (VCAM-l) may participate in initial monocyte recruitment to prelesional areas of arterial endothelium. Other adhesion molecules including Intercellular adhesion molecule-l (ICAM-l) may also participate in monocyte adhesion to arterial endothelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular mechanism of blood monocyte adhesion to vascular endothelial cells. The blood monocytes adhere to endothelial cells unstimulated and after stimulation by interleukin-1, tumor necrosis factor or other mediators. This process is mediated through specific molecules on both endothelial cells and monocytes. Using specific monoclonal antibodies and molecular cloning several families of molecules involved in leukocyte endothelial cell interaction have been defined. Leukocyte adhesion molecules include the three beta 2 integrins (CD11/CD18 molecules), VLA-4 and the L-Selectin. E-Selectin (ELAM-1), P-Selectin (GMP-140) and receptors of the immunoglobulin superfamily (ICAM-1, ICAM-2 and VCAM-1) are expressed on endothelial cells in basal conditions and after activation. It has been shown that these adhesive molecules are involved in blood monocyte adhesion to endothelial cells. Monocytes from patients with diabetes mellitus had an increased adhesion to endothelial cells in culture. As estimated by flow cytometry CD11b/CD18 expression on diabetic monocytes was increased. Pentoxifylline reduced CD11b/CD18 expression on normal and diabetic monocytes. This effect was associated to a decrease in monocyte adhesion to endothelial cells.


Pentoxifylline in vitro reverses neutrophil chemotactic deficiency induced by interleukin-2 treatment. Patients undergoing immunotherapy with Interleukin-2 experience multiple side effects and are highly susceptible to bacteremia. In a previous study, we confirmed the profound neutrophil chemotactic deficiency induced by Interleukin-2 therapy. Peripheral blood cells exposed to Interleukin-2 in vitro secrete secondary cytokines. The release of tumor necrosis factor into the circulation after Interleukin-2 injection has been proposed as an important mechanism underlying cell function alterations. We tested chemotaxis of neutrophils from normal subjects after incubation with the serum from treated patients. Serums induced a defective chemotaxis of normal neutrophils similar to the one observed with neutrophils from Interleukin-2 treated patients. We have previously demonstrated a dose-dependent reversion of neutrophil chemotaxis after incubation with anti-Tumor Necrosis Factor-alpha antibody. Pentoxifylline is known for counteracting the inflammatory action of tumor necrosis factor. We tested its capability to reverse the chemotactic deficiency of neutrophils induced by treated patient serums. Pentoxifylline was added after incubation of normal cells with patient serum, and the directed chemotaxis was restored. Pentoxifylline may have a significant therapeutic potential for the prevention or treatment of complications related to inappropriately activated neutrophils.


Study of pentoxifylline induced modulation of TNF alpha and interleukin-6 secretion in healthy and septic patients by the use of an ex-vivo model on whole blood. The aim of this work is to reinvestigate the pentoxifylline (PTX) action on TNF alpha and IL-6 production using a whole blood ex-vivo model. 5 healthy controls, 2 septic patients (4 samples) and 3 patients in septic shock (7 samples) have been studied. Our data confirm the inhibitory action of PTX on TNF alpha production in healthy controls. This inhibition is nearly complete for a PTX concentration of 10(-3) M. More surprisingly a suppressive activity of PTX on IL-6 secretion has been found both in controls and septic patients. On the 7 samples of the 3 patients with septic shock no TNF alpha production has been detected. Taken altogether, these observations suggest a potential role for PTX as a modulator of the major cytokines involved in the pathogenesis of septic shock.


Cellular and molecular aspects of PECAM-1. PECAM-1 is a surface membrane glycoprotein of 130,000 daltons found on platelets, endothelial cells, monocytes, neutrophils, and certain T-cell subsets. Except for bone marrow precursor cells, it does not seem to be found outside the vasculature. Previous studies have shown that PECAM-1 is a member of the immunoglobulin gene superfamily that localizes to the intercellular junction in cells, like endothelial cells, that grow in monolayers. We and other laboratories have been involved in studies designed to define the role of PECAM-1 in various cell types. Mammalian expression vectors have been constructed that express all, or selected domains, of the PECAM-1 protein. These vectors have been used to induce PECAM-1 expression in a variety of both monolayer and suspension cell types. Domain-specific anti-PECAM-1 monoclonal antibodies, synthetic peptides, and heparin were also used to define structurally and functionally important domains of this membrane glycoprotein. Protein synthesis, subcellular localization, and association with the cytoskeletons of various cells expressing PECAM-1 were examined. From these studies we have obtained evidence that: (1) PECAM-1 redistributes to the border of monolayer cells when they contact each other. (2) Border localization may result from homophilic interactions, i.e. PECAM-1 on one cell binds PECAM-1 on the adjacent cell. (3) Suspension cells such as L-cells or neutrophils may also interact via PECAM-1 mediated interactions, but an as yet unidentified ligand probably participates in a heterophilic, rather than homophilic, type of interaction. (4) Intracellular movement of PECAM-1 in both monolayer and suspension cell types requires active participation of the cytoskeleton.(ABSTRACT TRUNCATED AT 250 WORDS)


[A longitudinal radiologic study of the esophagus in an endemic area of Chagas' disease over a 6-year period]. A longitudinal study over six years was undertaken of 494 residents of the municipality of Mambaí, Goiás. Two hundred and twelve (43%) were seropositive in 1975/76 and 199 of 464 of the same patients group, positive in 1980/82 (42.8%). At both examination single radiographs of the oesophagus were obtained immediately after ingestion of 75 ml of barium sulphate solution and a second X-ray taken one minute later. Among the 201 seropositive patients without megaesophagus in the first study 4 (2%) evolved megaesophagus during the six years of observation. During this time, using Rezende's classification, patients with established megaesophagus, changed their group in the following manner. Only one Group I patient changed to group II. Only one group II patient progressed to group IV. Progress of megaesophagus in the affected patients occurred in 2.8% of 212 patients. Also four patients with grade I megaesophagus initially had a normal oesophagogram on the follow up examination. Ten patients had doubtful oesophagogram initially and six on follow up, 75% of these patients were seropositive. This study could indicate that a doubtful oesophagogram is an early sign of megaoesophagus.


[The treatment of chronic osteomyelitis]. From 1986 to 1990, sixty patients were treated for chronic osteomyelitis with surgery and chemotherapy. The clinical and radiologic follow-ups ranged from twelve to sixty months. The surgery was associated with a six-month oral sulfamethoxazole-trimethoprim combination chemotherapy. Clinical cure was achieved when inflammatory signs and symptoms disappeared, as happened to 50 (83.33%) of the single surgery cases, and to 9 (15%) of the multiple surgery cases. For one case (1.67%) osteomyelitis was not cured.


Influence of light conditions on adult mortality and egg-laying of Triatoma brasiliensis Neiva, 1911 (Hemiptera: Reduviidae: Triatominae). We compared, for Triatoma brasiliensis, the egg-laying process and the mortality of adults under conditions of almost permanent darkness and with normal laboratory luminosity. Mortality did not differ between groups. The egg-laying per vial and per female was significantly greater in the group of normal luminosity. We consider that it is not recommendable to keep the adults of this species under complete darkness. Other biological aspects should be analysed in relation to luminosity.


[A retrospective therapeutic study of neurocryptococcosis in 112 AIDS and non-AIDS patients]. A total of 112 AIDS and no AIDS cryptococcosis patients admitted at Emílio Ribas Hospital--São Paulo, Brazil, were treated with amphotericin B (AMB) or amphotericin B and 5 fluorocytosine (5FC). Age, race, predisposing and epidemiological factors, respiratory symptoms were evaluated. Goodman tests applied in three patients groups (I, II and III) with associate or unique therapy revealed: 1. prognostic factors: leukocytes ang glucose showed similar response in groups I, II and III and protein spinal fluid after 1.5g/AMB; 2. India ink tests and Cryptococcus culture were often positive until 1.0g/AMB; 3. significant hypokalemia during monotherapy. Hypo and hyperkalemia had similar data in associate therapy; 4. significant difference in adverse reactions often appeared above 0.7g AMB/250g 5FC; 5. early and late death were common in group III (unique) and group I (no AIDS) and III (2.5 to 4.0g) respectively; 6. similar remission and deaths were verified in AIDS/cryptococcosis. adverse reactions were observed above 0.75g/AMB plus 250g 5FC. Association was important in initial therapy and AMB maintenance permitted late relapses.


Glucantime resistant Leishmania promastigotes are sensitive to pentostam. Growth inhibition in vitro tests were used to study the susceptibility to pentostam of different Leishmania strains involved in cutaneous and mucocutaneous leishmaniasis--one glucantime sensitive strain, three naturally glucantime resistant strains and one glucantime resistant line developed by in vitro drug exposure. Contrasting with the high degree of glucantime resistance, all strains were sensitive to pentostam. These differences suggest that there is some relationship between chemical structure and in vitro activity for these antimonial compounds. These data justify a clinical re-evaluation to compare therapeutic efficacy of glucantime and pentostam in the treatment of leishmaniasis.


[Anti-Leptospira agglutinins in different professional groups in the city of Londrina, Paraná]. Serum samples were obtained from 208 individuals in Londrina, Paraná, Brazil. The serum were analysed for leptospiral agglutinins by agglutination microscopic tests and 28.4% were positive. The highest positivity was found for the serum of garbage collector (46.7%).


Reduction of spleen size in a child with hyperreactive malarious splenomegaly (HMS) treated outside the Brazilian endemic area of malaria with only one course of quinine. We report the clinical picture, treatment and evolution of a child with hyperreactive malarious splenomegaly treated outside the endemic area of malaria. The patient presented gross splenomegaly, proceeded from an area where malaria is endemic, showed increased immunoglobulins levels, high antimalarial antibody titres and hepatic sinusoidal lymphocytosis. The child did not return to an area where malaria is endemic and showed a favorable response to only one course of quinine. The response of this patient to limited antimalarial therapy suggests the importance of reinfection with malaria in the development and maintenance of this syndrome.


[Ataxia telangiectasia: effects of cycloheximide on G2 repair of chromosome damage in lymphocytes cultured in vitro]. The chromosomal sensitivity to the spontaneous and induced DNA damage detected in lymphocytes from patients affected with ataxia telangiectasia (AT) might be related with disturbances in DNA repair mechanisms. We have studied the effect of caffeine, an inhibitor of G2 repair and the enlargement of G2 by 0.5 ug/ml cycloheximide on chromosomal aberration frequencies in AT lymphocytes, both in control and X-ray irradiated conditions. The increase of spontaneous and X-ray induced chromosomal aberrations by caffeine treatments during G2 in AT lymphocytes was higher than in control cells. The number of spontaneous and X-ray induced lesions repaired during G2 in AT cells was higher than in normal cells. The enlargement of G2 duration by 0.5 ug/ml cycloheximide decreased the spontaneous and X-ray induced chromosomal aberrations in AT cells, whereas no such effect was observed in control cells. We postulate that disturbances in the mechanism that control G2 duration might be involved in the high frequency of spontaneous and induced chromosomal aberrations detected in AT lymphocytes.


[Microalbuminuria after unilateral nephrectomy in humans: study of effects of dietary protein]. Microalbuminuria may be due to increased glomerular capillary pressure. In turn, this increased pressure may be associated to augmented protein intake or reduction of renal mass. 85 normal subjects who had a unilateral nephrectomy were studied. Creatinine clearance, microalbuminuria and blood pressure were measured. Hyperfiltration was evaluated by comparison of creatinine clearance before and after nephrectomy. Protein intake was evaluated by a nutritional questionnaire. Hyperfiltration was estimated as 38% and microalbuminuria was not different in patients submitted to nephrectomy (9.8 micrograms/min) compared to controls (9.7). Microalbuminuria post nephrectomy was not correlated to level of hyperfiltration, protein intake, age, blood pressure or time after surgery. These results suggest that the remaining kidney is able to double the excretion of albumin with a likely increase in glomerular filtration and intracapillary pressure.


[Hemoptysis and the adult respiratory distress syndrome as the causes of death in leptospirosis. Changes in the clinical and anatomicopathological patterns]. Human leptospirosis, one of the main urban endemics/epidemics in Brazil, has dramatically grown in the last three decades, especially after floods caused by summer rains. This presentation describes recent changes in the clinical patterns of this pathology in our region, expressed by the emergence of massive haemoptysis and acute respiratory distress syndrome, or both conditions associated. The evident changes in the respiratory structures emerged as a serious life threat and death mechanisms, becoming the main cause of death in leptospirosis among us because of their high incidence. This new face of the disease demands a revision of current concepts about its seriousness and raises speculations about the pathogenesis of such alterations.


[Typhoid fever in school children: by what measures is the modification of the clinical course due to oral vaccination?]. The clinical course of infection by Salmonellae was compared between patients who had been vaccinated against typhoid fever using the Ty21a vaccine and those who had not. Of 2566 bacteriological confirmed cases 84% were infected with S typhi, 14% with S paratyphi B and 2% with S paratyphi A. Among patients with typhoid fever, 34% were treated in hospital, 3.5% had relapses, 5.4% developed complications and 1 patient died (0.05%). Among patients with paratyphoid fever, 18% were treated in hospital, 0.6% had relapses, 1.4% developed complications and there were no deaths. These figures were similar among vaccinated and non-vaccinated cases. A slightly greater proportion of vaccinated cases were treated in hospital (38 vs 30%). Thus, use of oral vaccination against typhoid fever does not alter the clinical course of infection with Salmonellae.


[Alternative treatment for acute asthmatic crisis]. In a double blind study, 20 patients who presented with acute episodes of asthma were randomized to treatment with nebulized salbutamol (5 mg) of subcutaneous salbutamol (0.5 mg). The clinical conditions were not significantly different before or after treatment in both groups. Significant bronchial dilation was obtained to a similar level in both groups. The heart rate increased significantly in the group treated by subcutaneous injection. Other side effects were minor and similar in both groups. Thus nebulized salbutamol may be used to treat acute asthma in selected patients.


[Postoperative radiotherapy in adenomas of the hypophysis]. Postoperative radiotherapy was performed in 58 patients with pituitary adenomas from 1977 to 1987. There were 21 non functioning adenomas and 22 growth hormone producing tumors. Extrasellar extension was found in 59% of patients. Radiotherapy was performed electively in 38 patients and due to tumor recurrence in 20. Megavoltage radiation was used in all patients. A dose of 5000 cGy in 5 weeks was attained in 54 of 58 patients. The recurrence free survival was 81% after a median follow-up of 6.8 years. Hypopituitarism developed in 13 of 58 patients. These findings confirm that external radiotherapy offers a real curative alternative for patients with macroadenomas of the pituitary gland.