Global methylation in 'Valencia' orange seedlings associated with rootstocks and Huanglongbing.

Citrus farming is one of the main activities that contributed to the Brazilian trade balance, with citrus seedling being the most important input in the formation of orchards to guarantee high productivity and fruit quality, which fundamentally depends on the chosen genetics. The present study aimed to analyze the existence of epigenetic variability in 'Valencia' orange plants on rootstocks, associated or not with HLB, through the quantification of the global methylation of its genome, in order to support works on genetic improvement and crop production. For this purpose, this work was carried out in greenhouse in a completely randomized experimental design, with 5 treatments and 6 replicates per treatment, each seedling being considered a replicate, namely: T1 = "Valencia" orange grafted onto "Rangpur" lemon, inoculated with HLB; T2 = "Valencia" orange grafted onto "Swingle" citrumelo, inoculated with HLB; T3 = "Valencia" orange grafted onto "Rangpur" lemon, without HLB inoculation ; T4 = "Valencia" orange grafted onto "Swingle" citrumelo, without HLB inoculation ; T5 = "Valencia" orange in free standing. The DNA was extracted from leaves and the ELISA test (Enzyme-Linked Immunosorbent Assay) was carried out, based on the use of receptors sensitive to 5-mC., to measure the relative quantification of global methylation between genomic orange DNAs . Since the control treatment (T5) consists of "Valencia" orange in free standing, it could be inferred that both the normal grafting technique in the seedling formation process and the inoculation of buds infected with HLB are external factors capable of changing the methylation pattern in the evaluated plants, including the DNA demethylation process, causing an adaptive response in association with the expression of genes previously silenced by genome methylation.

Effect of loading on the development of nerve fibers around oral implants in the dog mandible.

Occlusal forces cause stress which morphologically affects the supporting tissues of implants. The aim of this study was to examine the effects of occlusal forces on the distribution of neurofilament protein (NFP)-positive nerve fibers in the tissue of peri-implant bone. The bilateral 2nd, 3rd and 4th mandibular premolars and the 1st molars were extracted from three mongrel dogs. After 4 months of healing, 4 screw-type implants were inserted in the oral cavity. Three months after insertion, the implants on the molar site were loaded by occlusal forces, while those on the premolar site were unloaded. After a further 3 months, the dogs were sacrificed, and specimens were prepared for immunohistochemical NFP-positive staining by the labeled-streptavidin-biotin method. Many NFP-positive nerve fibers were found in the tissues of the loaded site when compared with the unloaded site. These fibers were localized in both the bone marrow space and in the peri-implant fibrous tissue. They had two types of nerve endings: simple free nerve endings, and nerve endings with tree-like ramifications. The present results suggest that loading by occlusal force causes an increase in the number of NFP-positive nerve fibers, many of which have free nerve endings in the peri-implant tissue. The possible role of these NFP-positive nerve fibers is discussed.

Noninvasive measurement of cerebral blood flow with (99m)Tc-hexamethylpropyleneamine oxime single-photon emission computed tomography and 1-point venous blood sampling.

BACKGROUND AND PURPOSE: The arterial and venous blood concentration of technetium 99m-labeled hexamethylpropyleneamine oxime ((99m)Tc-HMPAO) reaches an equilibration more rapidly than other CBF tracers. We hypothesized that (99m)Tc radioactivity of a venous sample at equilibrium, which is similar to that of an arterial sample, would allow estimation of the integrated input function for the clinical measurement of CBF by use of single-photon emission CT. METHODS: In 53 patients with stable cerebrovascular disease, the radioactivity of a venous sample 5 minutes after injection of (99m)Tc-HMPAO was correlated with 5-minute arterial blood radioactivity and the first 5 minutes of the integrated arterial curves of the lipophilic tracer. The measured CBF values were compared with those of xenon 133. RESULTS: The radioactivity of 5-minute venous blood was almost equivalent to that of 5-minute arterial blood (r(2) = 0.987; y = 0.993x + 1.63; P : <0.0001). The correlation between the venous blood radioactivity and the integrated arterial lipophilic fraction was excellent (r(2) = 0.935, P : <0.0001). A strong correlation was obtained between (99m)Tc-HMPAO and (133)Xe CBF values (r(2) = 0.825, P : <0.0001). CBF values were reproducible (coefficient of variation, 8.6%). CONCLUSIONS: This approach is fast, simple, and an alternative to continuous blood sampling in clinical quantitative (99m)Tc-HMPAO CBF studies.

[Progression of silent cerebral infarction in Japanese: a hospital-based study].

Little is known about the risk factors for progression of silent cerebral infarction (SCI). We evaluated the 5-year change in high signal intensity on T2-weighted MR images of the brain in 50 asymptomatic patients with SCI. SCI progression was evaluated by the sum of the high intensity score (HIS) and the number of new lesions found on reexamination. Progression of SCI was seen in 19 patients (38%) on the 5-year follow-up examination. The baseline HIS in the whole brain was 8.02 +/- 6.93 and the follow-up HIS was 8.7 +/- 7.16 (p < 0.001). SCI progression was frequently seen in the corona radiata (p < 0.02 or < 0.05) or posterior periventricular white matter (p < 0.05). Patients with poor control of hypertension (> 160/95 mmHg) have a higher prevalence of SCI progression (9/12 (75%) vs. 10/38 (26.3%); OR = 8.4, p < 0.005) than the patients with effective control of BP (< or = 160/95 mmHg). The progression group showed a significantly higher baseline-HIS in the whole brain than the nonprogressing group (9.7 +/- 5.6 vs. 7 +/- 7.5; p < 0.05). Neither age percentages of alcohol drinkers, male sex, current smokers, hypertensives, hyperuricemia, hypercholesterolemia, diabetes, obesity, left ventricular hypertrophy, ischemic heart disease, nor differences in the controls of diabetes and hypercholesterolemia during the study were different between the progressing and the nonprogressing groups. Our data showed that poor control of high blood pressure and the degree of T2 high intensity in the baseline were strong predictors of SCI progression.

Indium-111 platelet scintigraphy in a patient with chronic disseminated intravascular coagulation.

The authors describe a relation between the in vivo localized consumption of In-111 platelets and results of laboratory tests in a 82-year-old man with an aortic graft prosthesis who had chronic disseminated intravascular coagulation. In-111 platelets accumulated markedly over the abdominal aortic graft with a shortened platelet survival time and reflected changes in hematologic studies, which showed low platelet counts and a high concentration of fibrinogen degradation products in the plasma. If a strong clinical likelihood of local disseminated intravascular coagulation exists, then In-111 platelets may be valuable in localizing the focus of platelet consumption.

Solid-phase synthesis of substituted guanidines using a novel acid labile linker.

A novel acid labile linker for solid-phase synthesis of substituted guanidines has been developed. Its synthetic utility is exemplified by high-yielding pyrazole displacement with structurally and electronically diverse sets of aliphatic and aromatic amines. The final cleavage is achieved by treatment with 95:5 trifluoroacetic acid/water for 1 h. The corresponding guanidines were obtained in high purity (80-95%) and good isolated yields (50-95%). The scope and limitations of this linker were further demonstrated by the solid-phase synthesis of an 880-member library of individual trisubstituted arylguanidines employing pyrazole displacement with a set of 11 anilines and two subsequent Mitsunobu N-alkylations with sets of 10 and 8 alcohols, respectively.

Light and scanning electron microscopic study on the lingual papillae in the newborn sea otter Enhydra lutris.

The tongue of Enhydra lutris (sea otter), died from brain contusion at the first day of its age, was investigated by light and scanning electron microscopy in this study. Three types of papillae (filiform, fungiform and circumvallate papillae) were present on this tongue. In our evaluation, the filiform papillae were divided into four shapes which were conical, club-shaped, large horny and triangular papillae. The different types of filiform papillae were located in different area of this tongue. The fungiform papillae were divided into two shapes, which were hemispherical or club-shaped, and were distributed among the filiform papillae on the dorsal and the margin of the tongue. These papillae had many taste buds in the epithelium of the dorsal aspect of them. The circumvallate papillae were present in a V-shaped row at the posterior part of the lingual body, and many taste buds lay in the epithelium of both the lateral walls facing the trench and dorsal aspect of the papillary body, and then the ducts of the serous glands opened into the floor of the trenches. The lingual mucosa of the interpapillae area at the pharyngeal part of the tongue contained taste buds and the ducts of the mixed glands opened there. Based on our examination, the lingual papillae of this newborn sea otter had many features which were described to be peculiar to young stage of mammals.

[A device for rapid elevation of plasma methotrexate (MTX) concentration and its maintenance in high-dose MTX therapy].

For 5 osteosarcomas and one synovial sarcoma, 24 courses of high-dose methotrexate (MTX) therapy were performed. At MTX doses of 300 mg/kg with several infusion patterns, the plasma MTX concentrations were measured by fluorescence polarization immunoassay. In the various types of infusions, five hours oblique and five hours bolus infusions were well maintained at 1,000 micromol/l for several hours. In particular, five hours bolus infusion of MTX needed only 20 minutes to reach 822 micromol/l and maintained 1,000 micromol/l during almost the entire infusion period. Optimization of chemotherapy for sarcomas by MTX requires individual adaptation of the infusion pattern.

Preparation and properties of A-B-A type block copolymer membranes consisting of poly(N-hydroxypropyl-L-glutamine) as the A component and polyisoprene as the B component.

A-B-A type block copolymer (GIG(P)) membranes consisting of poly(N-hydroxypropyl-L-glutamine) (PHPG) as the A component and polyisoprene (PI) as the B component were prepared by carrying out aminoalcoholysis reaction with 3'-amino-1-propanol and a crosslinking reaction with 1,8-octamethylenediamine (OMDA) on membranes of the starting block copolymer (GIG) membranes consisting of poly(gamma-benzyl L-glutamate) (PBLG) and PI. It was shown that the effective crosslink density was proportional to the molar % of OMDA in the reaction mixture. The relation between their bulk structure and membrane properties was investigated, such as the swelling ratio q in a pseudo-extracellular fluid (PECF), tensile properties, and enzymatic degradation behaviour of the membranes in PECF. The tensile properties of the hydrophilic membranes were highly dependent on q in PECF, and on the hydrophobic portions in molecular chains, whose behaviour was typical of an elastomer. Biodegradation of samples in vitro by papain indicated that the rate of degradation was also highly dependent on q of membranes in PECF.

Synthesis and structure-activity relationships of 3-(2-imidazolyl)thiomethyl cephalosporins.

The synthesis and structure-activity relationships of a series of 3-(2-imidazolyl)thiomethyl cephalosporins are described. Among the compounds, 7 beta-[2-(2-amino-1,3-thiazol-4-yl)-(Z)-2-methoxyiminoacetamido]-3- [(4,5-dicarboxyimidazol-2-yl)thiomethyl]-3-cephem-4-carboxylic acid (1) exhibited potent activity against both Gram-positive and Gram-negative bacteria including Pseudomonas aeruginosa. We also estimated lipophilicity of the compounds from the chromatographic log k' value of reversed-phase HPLC. The relationship between lipophilicity and biological activity showed that compound 1 had the most suitable lipophilicity.

Characterization of serine proteinases isolated from rat submaxillary gland: with special reference to the degradation of rat kininogens by these enzymes.

From the homogenate of rat submaxillary gland, two kinds of serine proteinases, named tentatively proteinases A and B, were isolated and their chemical properties and activities toward rat kininogens were examined, in comparison with those of submaxillary kallikrein. Proteinase A with Mr of 28,200 rapidly cleaved high-molecular-weight (HMW) kininogen into a protein of 67 kDa, which retained thiol-proteinase inhibitory activity, but had lost the correcting activity of HMW kininogen on the prolonged clotting time of Fitzgerald trait plasma. It liberated bradykinin from HMW kininogen but did not liberate kinin from T-kininogen and did not degrade T-kininogen. On the other hand, proteinase B with Mr of 30,400 showed a very weak activity for the liberation of kinin from T-kininogen and the cleavage of T-kininogen at pH 8.0. However, the enzyme extensively degraded T-kininogen at pH 4.5. Proteinase B also degraded HMW kininogen at pH 4.5 and pH 8.0, but liberated bradykinin only at pH 8.0. Thiol-proteinase inhibitory activities of HMW kininogen and T-kininogen were inactivated after the incubation with proteinase B at pH 4.5 but not at pH 8.0, while the correcting activity of HMW kininogen on the Fitzgerald trait plasma was inactivated at pH 4.5 and 8.0. The NH2-terminal amino acid sequences of proteinases A and B were different from each other, and distinguishable with those of serine proteinases in rat submaxillary gland so far reported. These results provide evidence that in addition to the known kallikrein, there exist at least two kinds of serine proteinases in rat submaxillary gland, both of which liberate bradykinin from rat HMW kininogen at pH 8.0 and modulate the functional activities of HMW kininogen and T-kininogen, degrading these proteins at pH 8.0 or 4.5.

[Comparative studies on multiple forms of serum cholinesterase in various species].

Multiple forms of serum cholinesterase (ChE) were compared in 8 species by electrophoretic technique and the following characteristics were noted. The first moving fraction markedly hydrolyzed butyrylthiocholine and the activity was not inhibited by 10(-5)M eserine in the serum of some rabbits tested. Electrophoretic patterns of the ChE were obtained by use of two thiocholines as substrate, and the number of fractions against acetylthiocholine were more than against butyrylthiocholine in dogs, miniature pigs, rabbits, and hamsters. The activities of ChE fractions of dogs (C3), miniature pigs (C1, C2), rabbits (C1), and hamsters (C3) were inhibited by 6.1 X 10(-2)M caffein but not by 10(-4)M ethopropazine, which suggests that the fractions are all true-ChE.

A kinetic study on the effects of anticoagulants on the interaction of fibrinogen and thrombin.

The inhibitory effect of heparin and antithrombin III (AT) on the interaction of fibrinogen and thrombin was investigated in preference to studies on heparinizing devices. The turbidity was measured kinetically as a measure of the concentration of fibrin polymer formed in the system. It was found that AT did not act on fibrinogen but, rather, on thrombin, and the main role of heparin is to accelerate the AT-thrombin reaction. On the other hand, dextran sulfate (DSc) did not accelerate the AT-thrombin reaction. When heparin and AT were incubated with thrombin, inhibition did not depend on the mixing order but on the incubation time. Thus, a ternary complex of heparin, AT, and thrombin was supposed to form for the inhibition. The reaction of heparin with fibrinogen and thrombin in the presence of AT was well-explained by assuming a Freundlich-type adsorption of heparin analogous with the reaction of heparin with fibrinogen.

[A statistical approach to pruritus senilis].

The purpose of this study is to analyse the cause of itching and explore the relationships between pruritus senilis and various stimuli. Sample of 132 men and 238 women with ages ranging from 60 to 96 years were used in our study. Pruritus senilis is not significantly related to cosmetics and soap. Nevertheless, statistically significant correlations were found between pruritus senilis and physical stimuli (e. g., warmth, coldness, dryness). These observations could suggest that pruritus senilis is caused by changes of temperature.

[A statistical approach to Yatabe-Guilford personality inventory and manifest anxiety scale of the elderly].

In nursing, if we know an elderly person's mental condition, and that which does not make him/her too tired, this method can be very useful. Using this method we predicted the MAS score from scores of Y-G scales. We obtained the MAS score results by the following formula: MAS score not equal to 0.31 X Lack of objectivity +0.41 X Inferiority feelings -0.15 X Ascendance +0.34 X Y-G Nervousness +0.30 X Y-G Depression +9.54.

Synthesis and antibacterial activity of 6- and 7-[2-(5-carboxyimidazole-4-carboxamido)phenylacetamido]-penicillins and cephalosporins.

The synthesis and antibacterial activity of a series of beta-lactam antibiotics having a 5-carboxyimidazole-4-carbonyl group attached through an amide linkage to ampicillin, cephaloglycin or their analogs are described. Some compounds of this series were found to possess high activity against Pseudomonas and other Gram-negative bacteria.

Optimal control of a semibatch fermentation.

This work is concerned with the optimization study of the semibatch fermentation by which an amino acid is produced. The particular fermentation studied is the synthesis of lysine by the auxotrophic mutant. Applying Green's theorem to the maximization problem was proposed, and it succeeded in determining the feed rate of the substrate that maximized the production rate of the desired product.