Correction of experimentally produced vesicoureteric reflux in the piglet by intravesical injection of Teflon.

Vesicoureteric reflux was produced in eight piglets by opening their bladders and slitting the anterior intravesical wall of the ureter. Cystography confirmed the presence of bilateral reflux in six piglets and unilateral reflux in two. Six to eight weeks later the bladder was again opened and Teflon paste injected in the space behind the intravesical ureter, thereby creating a support for the submucosal ureter. Cystography four to six weeks after injection of Teflon paste showed absence of reflux in all animals. Intravenous pyelography showed obstruction at the vesicoureteric junction in only one of the 14 treated ureters and this was later confirmed at necropsy. Animals were followed up from one to six months and then were killed. Gross examination of the vesicoureteric region showed a well circumscribed subureteric Teflon mass of firm consistency, retaining its shape and position at the site of the injection. Histological examination showed encapsulation of the implant by a thin layer of fibrous tissue and a foreign body granulomatous reaction with histiocytes and giant cells within the implant. Hence it is technically feasible to correct experimentally produced vesicoureteric reflux in the piglet by intravesical injection of Teflon paste--a relatively inert material. It may now be possible to treat vesicoureteric reflux in man by endoscopic injection of Teflon behind the intravesical ureter.     

An Inexpensive Vibrating Microtome for Sectioning Fixed Tissue

A tissue sectioner which uses a vibrating razor blade and a simple mechanism for the elevation of the tissue can be constructed for less than fifty dollars. The razor blade is fixed to the vibrator of a hair clipper and a nut and bolt serve as the tissue advance mechanism. A metal disc attached to the nut is used for the stage. The tissue advance mechanism is placed inside a machined teflon cylinder which provides a smooth surface upon which the razor assembly is moved. Fixed tissue may be sectioned uniformly at a thickness of 50 μm or more. This device has the advantages of portability, rapidity of sectioning, and inexpensive construction.     

The use of heat detection aids in estrus synchronization programs

The use of tailhead paste and chin-ball markers for estrus detection in beef cows following estrus synchronization was evaluated. The estrous cycles of 85 cows were synchronized with a prostaglandin analogue in a two-injection program. At the time of the second injection, each cow's tailhead was painted with an estrus detection paste and the cows were placed in pastures with four intact bulls with chin-ball markers. Blood samples for plasma progesterone assays were obtained at the time of each injection and daily for five days following the second injection. Based on progesterone values, tailhead paste was incorrect 27% of the time in indicating estrus and chin-ball markers were incorrect 24% of the time. Detecting particular cows in estrus in groups of synchronized cows is difficult even with estrus detection aids.     

Plant DNases are potent therapeutic agents against Echis carinatus venom-induced tissue necrosis in mice

Echis carinatus envenomation leads to severe tissue necrosis at the bitten site by releasing DNA from immune cells that blocks the blood flow. An earlier report has shown that exogenous DNase 1 offers protection against such severe local tissue necrosis. Tricosanthus tricuspidata is a medicinal plant and the paste prepared from its leaves has been used extensively for the treatment of snakebite-induced tissue necrosis. Most studies including reports from our laboratory focused on plant secondary metabolite as therapeutic molecules against snakebite envenomation. However, the involvement of hydrolytic enzymes including DNase in treating snake venom-induced tissue necrosis has not been addressed. Several folk medicinal plants used against snakebite treatment showed the presence of DNase activity and found to be rich in T. tricuspidata. Further, purified T. tricuspidata DNase showed a single sharp peak in reversed-phase high-performance liquid chromatography (RP-HPLC) with an apparent molecular mass of 17 kDa. T. tricuspidata DNase exhibited potent DNA degrading activity performed using agarose gel electrophoresis, spectrophotometric assay, and DNA zymography. In addition, purified DNase from T. tricuspidata was able to neutralize E. carinatus venom-induced mouse tail tissue necrosis and normalized elevated serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels 30 minutes post venom injection. T. tricuspidata DNase was also able to reverse E. carinatus venom-induced histopathological changes and collagen depletion in mice tail tissue. All these observed pharmacological actions of T. tricuspidata DNase were inhibited by sodium fluoride (NaF). This study provides scientific validation of the traditional use of T. tricuspidata leaf paste in the healing of snakebite-induced tissue necrosis and might be exploited to treat snake venom-induced local toxicity.     

两种排龈技术的临床效果比较

目的 比较排龈线与排龈膏两种排龈技术的临床使用效果,以确定排龈膏是否比排龈线更易于被患者和临床医师所接受和推广。方法 作者在对151例门诊患者198颗牙,行冠修复时,于取模过程中分别应用排龈线和排龈膏两种排龈技术,分别记录操作时间、观察印模效果及患者舒适度等。结果 两种排龈技术在印模效果上都取得良好效果,在操作的时间和产生的疼痛程度上有着明显的差异。结论 两种排龈技术均可减少牙体预备时对牙龈组织的损伤及出血,帮助取得清晰的印模,但排龈膏便捷的操作方式及无痛操作理念更易被患者及临床医师所接受。     

Characterisation of a thermophilic L-glutamate dehydrogenase biosensor for amperometric determination of L-glutamate by flow injection analysis

Carbon paste wax electrodes incorporating thermophilic L-glutamate dehydrogenase, NADP and a polymeric toluidine blue O (poly-TBO) mediator have been characterised for the amperometric determination of L-glutamate at 313-318 K in a flow injection analysis (FIA) system. The biosensors exhibit good sensitivity, mechanical stability and reproducibilty, unlike carbon paste- or carbon wax-based electrodes under the same conditions. The carbon paste wax electrode responds linearly to L-glutamate up to 40 mM, the detection limit is 0.3 mM and the RSD (n = 10) for 5 mM L-glutamate was 7.6%. The response to some potential interferents has been quantified. Addition of finely ground hexaammineruthenium (III) trichloride ([Ru(NH3)6]Cl3) to the carbon paste wax electrodes decreases the FIA peak width and increases the peak current. The metal complex appears to accelerate the rate of oxidation of NAD(P)H by poly-TBO.     

Effect of Antisense Suppression of Endopolygalacturonase Activity on Polyuronide Molecular Weight in Ripening Tomato Fruit and in Fruit Homogenates

Fruit of tomato (Lycopersicon esculentum Mill.) in which endopolygalacturonase (PG) activity had been suppressed to <1% of wild-type levels were slightly firmer than nontransgenic controls later in ripening. Enzymically inactive cell walls were prepared from these ripening fruit using Tris-buffered phenol. When extracted with chelator followed by Na2CO3, the amounts of pectin solubilized from cell walls of nontransgenic control or from transgenic antisense PG fruit were similar. Size-exclusion chromatography analysis showed that, relative to controls, in antisense PG fruit polyuronide depolymerization was delayed in the chelator-soluble fraction throughout ripening and reduced in the Na2CO3-soluble fraction at the overripe stage. Reduced pectin depolymerization rather than altered extractability thus may have contributed to enhanced fruit firmness. Substantially larger effects of suppressed PG activity were detected in tomato fruit homogenates processed to paste. In control paste the majority of the polyuronide was readily soluble in water and was very highly depolymerized. In antisense PG paste the proportion of polyuronide solubilized by water was reduced, and polyuronides retained a high degree of polymerization. The suppression of fruit PG activity thus has a small effect on polyuronide depolymerization in the fruit but a much larger effect in paste derived from these fruit. This indicates that in the cell wall PG-mediated degradation of polyuronide is normally restricted but that in tissue homogenates or in isolated cell walls this restriction is removed and extensive pectin disassembly results unless PG is inactivated.     

Treatment of vesicoureteric reflux by endoscopic injection of Teflon.

Thirteen girls with grade III-V vesicoureteric reflux were treated by endoscopic injection of Teflon paste behind the intravesical ureter. Fourteen of the 18 treated ureters showed complete absence of reflux after one injection of Teflon. Three ureters required a second injection of Teflon for successful treatment of the reflux. One ureter with grade IV reflux was converted to grade II reflux. Properly carried out, this procedure corrects reflux. It takes less than 15 minutes, may be done as a day procedure, and avoids open surgery. There have been no complications.     

Organic charge transfer complex based printable biosensor

This paper describes the preparation of an organic charge transfer complex (CTC) based printable enzyme electrode. CTC crystals were prepared by mixing TCNQ powder with TTF solution (in acetonitrile). Glucose oxidase (GOD) was adsorbed at the CTC crystal surface in a monolayer. A printable paste was prepared by mixing GOD-adsorbed crystals with a binder and a solvent. This paste was applied to an electrode cavity and vacuum dried. A thin layer of gelatin was cast on the paste filled dried electrode, and cross-linked with glutaraldehyde in the dry condition. The sensors were fixed in a flow injection system, and continuously polarized at 0·15 V and 37°C, and the samples were automatically injected every 30 min. The developed sensors produced a huge response curren with an extended linear range of detection (0–100 mM) and the response was unaffected by the presence of normal oxygen in the buffer solution. The sensor showed excellent stability. The performance of the sensors was significantly influenced by the binder used.     

Recovery of exine from mature pollens and spores

A procedure has been devised for isolation and recovery of exine that is generally applicable to pollens and spores from a range of common species. Particles are suspended in an aqueous solution of 4-O-methylmorphine N-oxide and cyclohexylamine which swells them and loosens the exine layer. Gentle pressure in a teflon/glass tissue grinder releases protoplasts from exine and subsequent treatment with a mixture of cellulase and pectinase destroys attachments between intine and exine layers. The suspension is placed on a NaCl/Percoll step gradient to remove protoplasts and then the exine is cleanly separated from other cellular fragments on a step gradient of CsCl. The entire procedure can be accomplished at room temperature which greatly reduces possibility of chemical or physical modification of the exine. Moreover, the method is readily scaled up to production of exine in gram amounts which allows future study of the structural properties of its major biopolymer, sporopollenin.     

Dephosphorylation-induced structural changes in beta-casein and its amphiphilic fragment in relation to emulsion properties

To promote the understanding of the relationship between emulsifying and molecular properties of proteins/peptides, intact beta-casein (betaCN) and its amphipathic fragment, i.e., betaCN (1-105/107) were dephosphorylated. Dephosphorylation was found not to change significantly their emulsifying properties. Since it is known that the structure of proteins can change upon adsorption onto an interface, the secondary structure of intact beta-casein, its amphipathic fragment, and their dephosphorylated forms, both in solution and after adsorption onto a hydrophobic teflon/water interface, were studied by far-UV circular dichroism spectroscopy. An increased content of secondary structure, especially alpha-helix, was found for all samples after adsorption onto teflon. Dephosphorylation increased the helix-forming propensity, especially for amphipathic fragment of beta-casein. No influence of the secondary structure properties on the emulsion-forming and -stabilizing properties was observed, but a relationship between the maximum surface load and the emulsion-stabilizing properties was found.     

Stable cupola-shaped bilayer lipid membranes with mobile Plateau-Gibbs border: expansion-shrinkage of membrane due to thermal transitions.

Stable bilayer lipid membranes (BLM) with mobile Plateau-Gibbs border (PGB) have been formed. The precondition of the formation was the presence of a lipid coverage on the teflon surface near the hole, where the membrane has been formed. This allowed the movement of the PGB along the teflon surface after transformation of the planar bilayer into a cupola-shaped by bowing of the bilayer due to excess hydrostatic pressure. As a result the giant bilayers were obtained with an area up to two orders larger in magnitude compared with the initial area. Changes in lipid bilayer area depend on the temperature at the phase transition of the lipid. Cooling of the expanded bilayer was followed by a significant shrinkage of the bilayer at temperatures below the main phase transition.     

Dysphonia Due to Unilateral Nerve Paralysis—Treatment by the Intracordial Injection of Synthetics—A Preliminary Report

The injection treatment of unilateral recurrent vocal cord nerve paralysis, successfully performed 40 years ago with paraffin, has recently been reintroduced with non-reactive synthetics. Improvement in the voice far and above that which can be achieved by voice therapy or any practicable surgical procedure has been demonstrated by several investigators. The technique is relatively simple, and there are no significant untoward reactions. Judging from experimental and clinical studies with these inert materials when introduced into other parts of the body, adverse long range effects are not likely to occur in the larynx.Eleven of 12 patients treated by injection of silicone or Teflon paste had substantial improvement in vocal quality and strength. Silicone was absorbed slowly from the injection site and the improvement was not enduring; when Teflon was used, it stayed at the site and the improvement was sustained.     

An amperometric inhibitor biosensor for the determination of reduced glutathione (GSH) without any derivatization in some plants

In this study, an amperometric biosensor based on cucumber tissue homogenate was developed for the determination of glutathione. Cucumber (Cucumis sativus L.) tissue homogenate was used as the biological material. The cucumber tissue homogenate was cross-linked with gelatine using glutaraldehyde and fixed on a pretreated teflon membrane. The principle of the measurements was based on the determination of the decrease in the differentiation of oxygen level which had been caused by the inhibition of ascorbate oxidase in the biological material by glutathione. Determinations were carried out by standard curves which were obtained by the measurement of the decrease in the consumed oxygen level related to glutathione concentration. Optimization and characterization studies of the biosensor were carried out and a linearity in the gamma-L-glutamyl-L-cysteinyl-glycine (GSH) concentration range 0.1-2 microM was obtained when 600 microM ascorbic acid was used as a substrate. The repeatability experiments (n = 7) revealed that for 1.5 microM GSH, the average value (x), standard deviation (S.D.) and variation coefficient (C.V.) were 1.517 microM, 4.72 x 10(-5) 3.11%, respectively. The biosensor useful lifetime was at least 2 months. The results of some plant samples analyzed with the presented biosensor agreed well with the spectrophotometric method (Ellman's reagent) used as a reference.     

Modified carbon paste electrodes for flow injection amperometric determination of isocitrate dehydrogenase activity in serum

A carbon paste electrode modified with the adsorbed products of the electrochemical oxidation of adenosine triphosphate is described. The electrode was applied to the amperometric electrocatalytic detection of the reduced form of both nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate. The catalytic oxidation current shows a linear dependence on the concentration of the reduced form of nicotinamide adenine dinucleotide up to 1x10(-4)M, with a detection limit of 5x10(-9)M. Modified carbon paste electrodes were coated with an electrogenerated film of nonconducting poly(o-phenylenediamine) to obtain a stable amperometric response for at least 150h. In addition to static measurements, determination of both reduced cofactors was carried out in a flow injection analysis system with a thin-layer amperometric detection cell. The electrocatalytic monitoring of reduced nicotinamide adenine dinucleotide phosphate was applied to flow injection measurement of isocitrate dehydrogenase activity in serum. The results were in good agreement with those for the standard spectrophotometric test kit. The proposed method consumed less time and reagents and provided better precision than the standard method.     

Improvement of the Teflonized Slides Used in the Immunofluoresgent Antibody Technique

The indirect fluorescent antibody (IF) test is often performed on glass slides prepared with smears or tissue sections containing an antigen. It is convenient to have several separate and distinct areas of antigen on the same slide, e.g. to compare dilution steps, and to save time when producing the antigen slides. With this end in view a technique for preparing slides with wells in a thin film of teflon-like compound has been elaborated (Goldman 1968). This technique is useful when a great number of slides need to be prepared in advance, e.g. to test a substantial amount of material. It has, however, not been widely used im serology for Babesia, perhaps because any teflon remaining within the wells is stained with the fluorescent dye, thus creating a potentially disturbing background.     

The effect of silica fume on biodegradation of cement paste and its capacity to immobilize strontium during exposure to microbial sulfur oxidation

The disposal of low-level radioactive waste containing isotopes such as strontium by immobilization in cement paste has become common practice. However, the stability of cement paste in the environment may be impaired by sulfuric acid produced by sulfur-oxidizing bacteria. Since biodegradation rates in the environment of most radioactive waste burial sites are too low to be measured, determination of the degradation kinetics of cement paste is a difficult task. This study reports on the development of an accelerated biodegradation system for cement pastes in which the cement paste is exposed to a continuous culture of the sulfur-oxidizing bacterium Halothiobacillus neapolitanus. This system facilitated detection of the biodegradation processes in cement paste after as short a time as 15 days. A comparison of the durability of a cement paste blended with silica fume with that of unblended cement paste showed that the silica fume induced an increase in the leaching of Ca⁺² and Si and enhanced weight loss, indicating rapid deterioration in the structural integrity of the cement paste. The leaching of Sr⁺² from the silica fume amended cement paste was slightly reduced as compared with the non amended cement paste, indicating an increase in immobilization of strontium. Nevertheless, our findings do not support the use of silica fume as a suitable additive for immobilization of low-level radioactive waste.     

A reagentless amperometric electrode based on carbon paste, chemically modified with D-lactate dehydrogenase, NAD(+), and mediator containing polymer for D-lactic acid analysis. II. On-line monitoring of fermentation process

The production of D-lactic acid by Lactobacillus delbrueckii (ATCC 9649) during fermentation was monitored on-line with a reagentless D-lactate dehydrogenase modified carbon paste electrode in a flow injection system integrated with a filtration sampling device. The time delay between sampling and detection was approximately 6 min. The use of an electropolymerized ortho-phenylenediamine membrane on the elctrode resulted in a very selective sensor response with acceptable stability and sensitivity. The D-lactate concentrations determined on-line agreed well with those determined by a standard method, suggesting that this sensor system is suitable for on-line monitoring of fermentation processes. (c) 1995 John Wiley & Sons, Inc.     

Differential effects of N-methyl-D-aspartate receptor stimulation on growth hormone secretion at specific stages of postnatal development of the male rhesus monkey

The present study attempts to examine the role of N-methyl-D-aspartate (NMDA) receptor in the central regulation of growth hormone (GH) secretion during specific stages of pubertal development of the male rhesus monkey (Macaca mulatta). Infantile (n=4), prepubertal (n=5), peripubertal (n=5) and adult (n=5) intact male rhesus monkeys were given an agonist of NMDA receptor, N-methyl-D,L-aspartate (NMA) (15 mg/kg BW) through a teflon cannula implanted in the saphenous vein. Blood samples were collected 20-60 min before and 40-80 min after the injection of the drug at 10-20 min intervals. NMA was dissolved in normal saline immediately before use and passed through a 0.22 microm filter at the time of injection. All bleedings were carried out under ketamine hydrochloride anesthesia (initial dose 5 mg/kg BW, im followed by 2.5 mg/kg at 30 min intervals). The plasma levels of GH and testosterone (T) were determined by using specific assay systems. The hypothalamic-somatotrope activity under basal conditions was studied by averaging all the GH concentrations obtained before NMA injection, whereas the sensitivity of NMDA receptor to NMA stimulation was determined by comparing basal GH levels immediately before NMA injection at 0 min and GH concentrations obtained 10 min after the injection. The mean basal plasma concentrations of GH in the four groups of animals showed marked age-related differences. The levels of GH were found to be higher in infantile and peripubertal monkeys as compared to those of prepubertal and adult animals. A single iv injection of NMA produced differential effects on GH secretion during specific stages of postnatal development depending upon the level of GH secretion under basal conditions. Whereas NMA had no demonstrable effect on GH secretion in infantile and peripubertal animals in which the basal GH levels were high, it produced pronounced effects on GH secretion in prepubertal and adult monkeys wherein baseline GH concentrations were low. In conclusion, the present study suggests that the glutamatergic component of the control system that governs GH secretion by utilizing NMDA receptor may participate in regulation of age-related changes in the secretion of GH in the male rhesus monkey.     

Determination of aluminum by neutron activation analysis in human lung tissue and in chemicals for dialysis of uremic patients

Aluminum (Al) determination in human lung tissue, in chemicals for dialysis fluids, and in commercially available dialysis solutions is carried out using both Instrumental and Radiochemical Neutron Activation Analysis (INAA and RNAA). For some matrices, pre-and post-irradiation Al separations are tested. Chelex 100 resin and HDEHP, this latter adsorbed both on teflon and polyurethane foam, are employed for the pre-irradiation Al separations. The postirradiation procedures are performed using roughly the same schemes; the conventional Al precipitation as hydroxide is also carried out. Al values found in lungs of workers exposed to Al containing dust appear to be much higher than those found for unexposed people. Chemicals analyzed seem pure enough to give suitable dialysis solutions; for commercially available dialysis fluids, a 10-fold reduction of Al content should be convenient.