Autogenous dental transplants: description of a clinical case.

The dental transplant was already in use more than 2,000 years ago, reaching a peak in the Middle Ages, but it was only after the 1950s that this subject appeared in the scientific literature. This article describes the case of a young woman whose mandibular left second and right first molars had to be extracted and were replaced using the maxillary third molars. The positive clinical and radiographic results over a 5-year period encourage the use of this technique.

Purification of pig heart benzylamine oxidase.

A benzylamine oxidase (E.C. 1.4.3.6) has been purified from pig heart. Western blot analysis showed that the enzyme cross-reacts with a polyclonal antibody raised against homogeneous, crystalline pig plasma benzylamine oxidase (BAO). A subunit molecular mass of 97 kDa obtained by SDS electrophoresis is identical to the plasma enzyme. The purification procedure consisted of sequential DEAE-cellulose, DEAE-Sephadex, Con A-Sepharose, Sephadex G 200 and hydroxyapatite columns. The specific activity of the purified enzyme was 0.037 µmol min-1mg-1 at 37ûC and the Km for benzylamine was estimated to be 29 µM. The enzyme was inhibited by carbonyl reagents such as semicarbazide and á-aminoguanidine. Phenylhydrazine reacts mole to mole with the enzyme giving a peak at 425 nm. The copper content was 2 g-atom/mole of enzyme.

Semicarbazide-sensitive amine oxidase in pig heart.

A semicarbazide-sensitive amine oxidase (SSAO) (E.C.1.4.3.6) has been purified from pig heart. Western blot analysis showed that the enzyme reacts with a polyclonal antibody raised against homogeneous crystalline pig plasma benzylamine oxidase (BAO). A subunit molecular mass of 97 KDa obtained by SDS electrophoresis is identical to the plasma enzyme. The purification procedure consisted of sequential DEAE cellulose, octyl-Sepharose, Con A-Sepharose and hydroxyapatite columns. Two peaks of activity were obtained on octyl-Sepharose which were found to be kinetically and immunologically indistinguishable. The specific activity of the purified enzyme was 0.045 mumol/min/mg of protein at 37 degrees C and the Km for benzylamine was estimated to be 63 microM. The enzyme was inhibited by carbonyl reagents such as semicarbazide but was insensitive to the effect of pargyline.

Evidence of semicarbazide-sensitive amine oxidase activity in guinea pig tissues.

Lung, heart tissues and blood plasma of guinea pigs were investigated to see if tissue-bound semicarbazide-sensitive amine oxidase activities with a high affinity for benzylamine (Bz.SSAO) were present in this species as well as in others. This paper shows that these enzymic activities are present in guinea pig lung and heart where they are mainly localized in the cytosol and in microsomal fraction. These activities have a high affinity for benzylamine and appear unable to oxidize histamine at an appreciable rate in agreement with the observation that the purified Bz.SSAO of guinea pig skin shows weak histaminase activity. These guinea pig Bz.SSAO activities show some homology with the pure pig plasma benzylamine oxidase. They crossreact with the antibodies raised in the rabbit against the pig plasma enzyme. Benzylamine oxidase activity was also found in guinea pig blood plasma.

Semicarbazide-sensitive amine oxidase activity of guinea pig dorsal skin.

A semicarbazide-sensitive amine oxidase activity with a high affinity for benzylamine (Bz.SSAO) (E.C. 1.4.3.6) is present in guinea pig dorsal skin. This enzymic activity oxidized benzylamine, histamine, 1,4-methylhistamine and acetylputrescine and was inhibited by semicarbazide and by B24 (3,5-diethoxy-4-aminomethylpyridine), a selective inhibitor of Bz.SSAO enzymes. It cross reacted with the antibodies raised against pure pig plasma benzylamine oxidase. Immunohistochemistry showed that it was localized in fibroblasts. Bz.SSAO activity of guinea pig dorsal skin increased during the process of skin healing. A treatment of the wounds with 3 micrograms of b-FGF significantly accelerated the process of skin healing and the increase of Bz.SSAO activity.

Skin wound healing: some biochemical parameters in guinea-pig.

Hairs were removed from the dorsal skin of guinea-pigs and 5-6 wounds (7 x 7 mm) were surgically induced by totally removing the epidermal and part of the dermal surface. They were then allowed to heal. The newly formed wound tissues were dissected at different times during the process and analysed by biochemical and histological methods. Hydroxyproline, proteins, DNA and semicarbazide-sensitive amine oxidase (SSAO) were measured, as were [14C]leucine and [3H]thymidine incorporation in some samples. The peroxidase-like activity of plasma albumin and the histology of wounds stained with haematoxylin-eosin were also studied. It was shown that SSAO enzymes, which are present in normal guinea-pig skin and have a high affinity for benzylamine are localized in fibroblasts. During skin healing in the newly formed tissue there was an increase in protein content which reached a maximum after 4-6 days; DNA content also increased. The rate of incorporation of [3H]thymidine and [14C]leucine paralleled DNA and protein content, respectively. The content of hydroxyproline had greatly decreased with respect to that in normal skin after 2-10 days. SSAO activity increased much less than DNA after 4 days whereas after 10-11 days it increased more than DNA, thus indicating that at this time it was probably produced by fibroblasts. No significant increase in the peroxidase-like activity of albumin was observed 4, 8 or 11 days after surgery. Treatment of the animals with methylprednisolone acetate (20 mg kg-1, i.m.) two days before surgery decreased the rate of skin healing but did not alter the level of albumin peroxidase activity of the plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

Pyrroloquinoline quinone, a method for its isolation and identification by mass spectrometry.

Procedures for the unambiguous detection and for the isolation and mass spectrometric identification of pyrroloquinoline quinone (PQQ) are presented. The procedure involved acid hydrolysis of protein in the presence of phenylhydrazine and successive isolation and identification of the formed adduct using mass spectrometry. In HPLC the phenylhydrazone of PQQ gave many methylated products, of which the predominant compound was the pentamethylated derivative. After reaction of the phenylhydrazone derivative of PQQ (PHPQQ) with ammonia, a product was obtained which did not contain phenylhydrazine and which formed a pentamethylated derivative as the main methylation product. The HPLC profiles of the methylated products of PHPQQ and of its ammonia derivative were very characteristic and could be used for identification in addition to mass spectrometry. However, prolonged treatment of proteins with phenylhydrazine during hydrolysis can result in the formation of a material that resembles PQQ in some aspects of its behaviour. Thus, analysis by MS is essential for unambiguous identification. This analytical procedure was applied to pig plasma benzylamine oxidase, pig aorta lysyl oxidase, pig kidney diamine oxidase and bovine serum albumin with negative results. However, samples of pronase contained variable quantities of non-covalently bound PQQ: this can lead to erroneous identification of PQQ in enzyme after pronase digestion.

The evolution of human periodontal tissues with ageing.

In this research, the structural modifications with ageing of clinically healthy periodontal tissues were analyzed by means of polarization microscopy and morphometrical methods for light microscopy. The new findings may be summarized as follows. The periodontal ligament was found to be widened in the cervical and apical regions. The thickening of cementum with ageing was shown to be accompanied by a modification in the shape of Sharpey's fibres, which in the elderlies were wavy instead of straight as in the control. Lamellar bone, forming an osteone, was found to substitute in part for cementum in one tooth. These results are interpreted as indicating that: (1) late active eruption occurs in man, causing the observed modification in the thickness of periodontal ligament and cementum in the apical region and in the direction of Sharpey's fibres within cementum; (2) cementum may undergo renewal during lifetime and in this case bone may be deposited in contact with dentin.

A method for the isolation and identification of pyridoxal phosphate in proteins.

A method for the isolation and identification of covalently bound pyridoxal phosphate (PLP) contained in some enzymatic proteins is presented. The method involves acid hydrolysis of the protein in the presence of phenylhydrazine, separation of the adduct by elution from Sep-Pak C18 cartridges, isolation by HPLC, and either direct analysis by mass spectrometry with direct electron impact or conversion into trimethylsilyl derivatives followed by gas chromatography-mass spectrometry. Under the prescribed conditions of hydrolysis, PLP forms its phenylhydrazone and is released from the protein and hydrolyzed to the phenylhydrazone of pyridoxal, which shows a typical fragmentation in direct electron impact and in gas chromatography-mass spectrometry after silylation. The yield in phenylhydrazone of pyridoxal is on the order of 50% (+/- 5% SE, n = 15) when PLP is added to 10 mg of protein in amounts ranging from 20 to 40 nmol. Analysis of pig plasma benzylamine oxidase by this procedure confirms the presence of covalently bound pyridoxal phosphate in this enzyme.

Autacoid and beta-adrenergic agonist modulation of N-formylmethionyl-leucyl-phenylalanine evoked lysosomal enzyme release from human neutrophils.

Isoprenaline, histamine and PGE1 inhibit N-formylmethionyl-leucyl-phenylalanine evoked lysosomal enzyme release from human neutrophils. Their effects are dose-dependent and potentiated by 3-isobutyl-1-methylxanthine pretreatment of the cells. The order of activity is PGE1 greater than isoprenaline greater than histamine. The maximum of inhibition afforded by each agonist depends on the amount of the secretory stimulus, since it is higher at lower concentrations of the secretagogue. Isoprenaline effects are competitively antagonized by propranolol and are mimicked by fenoterol and salbutamol. These results suggest that human neutrophil functions are modulated by endogenous control mechanisms, that can also be activated by drugs acting on the same receptors as the endogenous mediators.