Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “LEECHES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 991 records · Page 55Linked to original sources
PubMed2001

Use of a mechanical leech in a reverse-flow radial forearm flap: case report.

The reverse-flow radial forearm flap provides excellent coverage for distal upper-limb defects. It is simply raised and does not require microsurgical skills. However, since its vascular pedicle is reversed, its venous outflow can be significantly diminished because of the venous valves. The authors present the case of a 16-year-old patient with a sagittal amputation of the radial aspect of the right thumb, who manifested at the time of surgery marked venous engorgement of a reverse-flow radial forearm flap. This was successfully relieved by the placement of a mechanical leech consisting of a Silastic rubber catheter--of the kind used to gain central vascular access in newborns--introduced in the lumen of the reversed vein at the extremity of the flap. This permitted intermittent evacuation of blood from the flap postoperatively, contributing to the success of this procedure. The technique used is detailed and pertinent literature is reviewed.

Adolescent↗
PubMed1986

Segmental differentiation in the leech nervous system: specific phenotypic changes associated with ectopic targets.

The issue of whether interactions between central neurons and the targets they innervate are involved in establishing segment-specific phenotypic differences in the central nervous system was evaluated by studying the effect of naturally occurring ectopic targets on individual identified neurons of the leech Hirudo medicinalis. The specimens studied were found to have additional sex organs located in segments adjacent to those containing the normal structures. We report our observations on neuronal morphology, the presence or absence of specific neurons, and total ganglionic cell number and discuss some of the possible ways in which the observed changes might have arisen.

Animals↗
PubMed1990

Cell position and developmental fate in leech embryogenesis.

The o and p blast cell bandlets of the leech Theromyzon rude, which normally produce two different sets of identifiable cells designated the "O" and "P" fates, respectively, form an equivalence group: in embryos experimentally deprived of their p bandlet, the blast cells of the adjacent o bandlet may "transfate" and take on the P fate. Loss of the p bandlet is not, however, a sufficient condition for transfating of the o bandlet. Rather, loss of the p bandlet allows the o bandlet to shift into ectopic positions, and it is the ultimate position of the o bandlet that mandates which fate--O or P--the blast cells will take on. Therefore, the choice of the pluripotent o blast cells to follow either the O or P developmental pathway depends on their perception of positional cues provided by cells outside the equivalence group rather than on a direct interaction with p blast cell equivalence group members.

Animals↗
PubMed1994

Cable properties of a straight neurite of a leech neuron probed by a voltage-sensitive dye.

We measured a time-resolved map of electrical activity in a thin straight neurite (1.5 microns thick, 500 microns long) at a resolution of 8 microns and 0.4 ms. The neurite was obtained by guided outgrowth of an identified neuron of the leech on lanes of extracellular matrix protein. The electrical signals were detected by a fluorescent voltage-sensitive dye. We observed the voltage that was caused by an action potential elicited at the soma and by a Gaussian hyperpolarization induced at the soma, respectively. We compared the data with numerical solutions of the cable equation using the Hodgkin-Huxley parametrization. We could attribute the experimental results of depolarization and of hyperpolarization to the propagation of an action potential along an "active" cable and to the spread along a "passive" cable, respectively, if we assigned rather high specific resistances to the cytoplasm (RI = 250 omega.cm) and to the membrane (RM = 22 k omega.cm2). This assignment explained the slow velocity of 150 microns/ms of a pulse by active propagation and the limited range of 200 microns of a pulse by passive spread.

Action Potentials↗
PubMed1996

Cloning and expression of sialidase L, a NeuAcalpha2-->3Gal-specific sialidase from the leech, Macrobdella decora.

Sialidase L is a NeuAcalpha2-->3Gal linkage-specific sialidase that releases 2,7-anhydro-NeuAc instead of NeuAc from sialoglycoconjugates (Chou, M.-Y., Li, S.-C., Kiso, M., Hasegawa, A., and Li, Y.-T.(1994) J. Biol. Chem. 269, 18821-18826). A 2. 5-kilobase cDNA of sialidase L was cloned by a combination of methods based on polymerase chain reactions. The composite cDNA sequence reveals an open reading frame coding for 762 amino acids, including a putative 28-residue signal peptide at the N terminus that is similar to the signal sequence of the Clostridium septicum sialidase. The result suggests that sialidase L is a secretory enzyme. The coding sequence excluding the putative signal peptide of sialidase L was overexpressed in Escherichia coli. The purified recombinant enzyme was characterized to be as active as the enzyme isolated from the leech. It also possessed the strict NeuAcalpha2-->3Gal linkage specificity and released the unique cleavage product, 2,7-anhydro-NeuAc from sialoglycoconjugates. The deduced amino acid sequence of sialidase L exhibits little similarity with other reported sialidases. However, sialidase L contains a conserved "FRIP region" and four repeating "Asp box" motifs that align well with the corresponding positions of bacterial sialidases. The predicted beta-strand structures near the conserved motifs of sialidase L are similar to those of Salmonella typhimurium sialidase. Several conserved single amino acid residues of bacterial sialidases, including those known to be involved in the active site of Salmonella enzyme, are conserved in the deduced amino acid sequence of sialidase L. This observation suggests that part of the catalytic mechanism of sialidase L may be similar to the ordinary sialidase.

Amino Acid Sequence↗
PubMed1997

The three-dimensional structure of recombinant leech-derived tryptase inhibitor in complex with trypsin. Implications for the structure of human mast cell tryptase and its inhibition.

The x-ray crystal structure of recombinant leech-derived tryptase inhibitor (rLDTI) has been solved to a resolution of 1.9 A in complex with porcine trypsin. The nonclassical Kazal-type inhibitor exhibits the same overall architecture as that observed in solution and in rhodniin. The complex reveals structural aspects of the mast cell proteinase tryptase. The conformation of the binding region of rLDTI suggests that tryptase has a restricted active site cleft. The basic amino terminus of rLDTI, apparently flexible from previous NMR measurements, approaches the 148-loop of trypsin. This loop has an acidic equivalent in tryptase, suggesting that the basic amino terminus could make favorable electrostatic interactions with the tryptase molecule. A series of rLDTI variants constructed to probe this hypothesis confirmed that the amino-terminal Lys-Lys sequence plays a role in inhibition of human lung tryptase but not of trypsin or chymotrypsin. The location of such an acidic surface patch is in accordance with the known low molecular weight inhibitors of tryptase.

Amino Acid Sequence↗
PubMed2000

Theromin, a novel leech thrombin inhibitor.

We purified the most potent thrombin inhibitor described to date from the rhynchobdellid leech Theromyzon tessulatum. Designated theromin, it was purified to apparent homogeneity by gel permeation and anion exchange chromatography followed by two reverse-phase steps of high performance liquid chromatography. The primary sequence of theromin (a homodimer of 67 amino acid residues including 16 cysteine residues) was determined by a combination of reduction and s-beta-pyridylethylation, Edman degradation, trypsin enzymatic digestion, and matrix-assisted laser desorption mass spectrometry measurement. Theromin exhibits no sequence homology with any other thrombin inhibitors. Furthermore, theromin significantly diminishes, in a dose-dependent manner, the level of human granulocyte and monocyte activation induced by lipopolysaccharides. In summary, this potent thrombin inhibitor promises to have high biomedical significance.

Amino Acid Sequence↗
PubMed2002

The unfolding pathway of leech carboxypeptidase inhibitor.

The unfolding and denaturation curves of leech carboxypeptidase inhibitor (LCI) were elucidated using the technique of disulfide scrambling. In the presence of thiol initiator and denaturant, the native LCI denatures by shuffling its native disulfide bonds and transforms into a mixture of scrambled species. 9 of 104 possible scrambled isomers of LCI, amounting to 90% of total denatured LCI, can be distinguished. The denaturation curve that plots the fraction of native LCI converted into scrambled isomers upon increasing concentrations of denaturant shows that the concentration of guanidine thiocyanate and guanidine hydrochloride required to reach 50% of denaturation is 2.4 and 3.6 m, respectively. In contrast, native LCI is resistant to urea denaturation even at high concentration (8 m). The LCI unfolding pathway was defined based on the evolution of the relative concentration of scrambled isoforms of LCI upon denaturation. Two populations of scrambled species suffer variations along the unfolding pathway. One accumulates as intermediates under strong denaturing conditions and corresponds to open or relaxed structures, among which the beads-form isomer is found. The other population shows an inverse correlation between their relative abundances and the denaturing conditions and should have another kind of non-native structure that is more compact than the unfolded state. The rate constants of unfolding of LCI are low when compared with other disulfide-containing proteins. Overall, the results presented in this study show that LCI, a molecule with potential biotechnological applications, has slow kinetics of unfolding and is highly stable.

Amino Acid Sequence↗
PubMed2003

Nested allosteric interactions in extracellular hemoglobin of the leech Macrobdella decora.

Hemoglobin from the leech Macrobdella decora belongs to the class of giant extracellular hexagonal bilayer globin structures found in annelid and vestimentiferan worms. These complexes consist of 144 heme-bearing subunits, exhibit a characteristic quaternary structure (2 x (6 x (3 x 4))), and contain tetramers as basic substructures that express cooperative oxygen binding and thus provide a structural basis for a hierarchy in allosteric interactions. A thorough analysis of the isolated tetramer indicates that it functions as a trimer of cooperatively interacting subunits and a non-cooperative monomer rather than as four interacting subunits. A thermodynamic analysis of the whole molecule favors the application of a nested Monod-Wyman-Changeux model with six cooperatively interacting 12-mer allosteric units. In contrast to the isolated tetramers, all subunits of the tetramers seem to be coupled cooperatively within the oligomerized 144-mer. Thus, besides hemocyanins and GroEL, the hexagonal bilayer hemoglobins represent another class of proteins in which the hierarchical quaternary structure provides the basis for nested interaction in their functional properties.

Animals↗
PubMed2003

Immediate use of medicinal leeches to salvage venous congested reverse pedicled neurocutaneous flaps.

Reverse pedicled neurocutaneous flaps have recently become popular for reconstructing soft tissue defects of the lower extremity. Venous congestion is a relatively common problem in these flaps in diabetic patients and those with electric burns, and this may cause partial or complete loss if capillary perfusion is not re-established urgently. We describe our experience of 13 neurocutaneous flaps, of which five developed venous congestion and were treated successfully with leeches placed immediately.

Adult↗
PubMed2000

Inhibition of induced and spontaneous platelet aggregation by destabilase from medicinal leech.

Destabilase, endo-epsilon-(gamma-Glu)-Lys isopeptidase from the medicinal leech, inhibits arterial thrombus formation in rats. Inhibition of platelet aggregation was supposed to be one of the main mechanisms of this phenomenon. To elucidate this question highly purified destabilase preparations were used. Aggregation was monitored both by a turbidometric method and by a method based on real-time estimation of mean aggregate size. Spontaneous aggregation of human platelets was completely blocked by destabilase. At 5 microM ADP maximal inhibition was 63%. Aggregation induced by PAF (100 nM) and collagen (0.1 mg/ml) was inhibited in the presence of destabilase by 50 and 65%, respectively. This enzyme does not activate adenylate cyclase but inhibits it. We suggest that destabilase interacts with high-affinity binding sites on the platelet plasma membrane, thus providing an anti-aggregating effect. This idea coincides with the data that destabilase primary structure has high homology with some adhesive proteins.

Adenosine Diphosphate↗
PubMed2000

Biochemical evidence of specific trypsin-chymotrypsin inhibitors in the rhynchobdellid leech, Theromyzon tessulatum.

The presence of two specific trypsin-chymotrypsin inhibitors from head parts of the rhynchobdellid leech Theromyzon tessulatum is reported. Two proteins, anti-trypsin chymotrypsin A (ATCA; 14636.6 +/- 131 Da) and anti-trypsin-chymotrypsin B (ATCB; 14368 +/- 95 Da) were purified by size exclusion and anion-exchange chromatography followed by reversed-phase HPLC. Based on amino-acid composition, N-terminal sequence determination (MELCELGQSCSRD-NPQPSNM), matrix assisted laser desorption-time of flight measurement (MALDI-TOF), trypsin mapping comparison, inhibition constant determination (Ki), and influence on amidolytic activity of different serine proteases, it is demonstrated that ATCA and ATCB are novel and highly potent serine-protease inhibitors of trypsin and chymotrypsin (ATCA: 350fM towards trypsin and chymotrypsin; ATCB: 400 and 75 fM towards trypsin and chymotrypsin, respectively). It is further surmised that ATCA and ATCB are linked, in that ATCB would lead to the formation of ATCA after loss of few amino acid residues.

Amino Acid Sequence↗
PubMed1969

The site of origin of electrical responses in visual cells of the leech, Hirudo medicinalis.

In leech visual cells the presumed light-absorbing structures are microvilli arising from the membrane of what would seem to be a large intracellular vacuole. This vacuole, however, is an extracellular compartment, since it communicates with the intercellular spaces through narrow channels. Therefore, the membrane of the microvilli is-as in other invertebrate visual cells-a part of the cell membrane. Visual responses recorded with an electrode within the vacuole were compared with the intracellular recordings. Following illumination the vacuole becomes negative with respect to the outside fluid, while the cells are depolarized. This finding indicates that inward current penetrates the cell through the microvillar membrane. It is concluded, therefore, that the electrical response (receptor potential) originates as a result of changes in the properties of the light-absorbing membrane.

Animals↗
PubMed1972

The distribution of synapses on a physiologically identified motor neuron in the central nervous system of the leech. An electron microscope study after the injection of the fluorescent dye procion yellow.

The fine structure of a physiologically identified motor neuron in the segmental ganglion of the leech central nervous system and the morphology of synapses on it were studied after injection of the fluorescent dye Procion yellow as a marker. The injected cell and its processes within the neuropil were located in thick or thin sections with fluorescence optics after initial fixation with glutaraldehyde and brief treatment with osmium tetroxide. The same or adjacent thin sections could then be examined in the electron microscope. Comparison with uninjected cells showed that the general features of the injected cell are retained although some organelles are distorted. The main features of the geometry of this neuron are the same from animal to animal: a single large process runs from the soma through the neuropil to bifurcate and enter the contralateral roots. Within the neuropil the main process gives off long branches (up to 150 micro), but these are greatly outnumbered by short branches and spines, one or a few microns in length, which were not appreciated in previous light microscope studies after injection of Procion yellow. Serial thin sections of selected areas along the main process within the neuropil showed that there are synapses on most of the shorter branches and spines; occasional synaptic contacts were also made on the main process itself and on longer branches. At least two morphologically distinct types of synapse could be recognized. A minimum estimate of the total number of synapses on the motor cell is 300, based on their occurrence in reconstructed segments.

Animals↗
PubMed1991

Electrogenic sodium-dependent bicarbonate secretion by glial cells of the leech central nervous system.

The ability to move acid/base equivalents across the membrane of identified glial cells was investigated in isolated segmental ganglia of the leech Hirudo medicinalis. The intracellular pH (pHi) of the glial cells was measured with double-barreled, neutral-ligand, ion-sensitive microelectrodes during step changes of the external pH (pHo 7.4-7.0). The rate of intracellular acidification after the decrease in extracellular pH (pHo) was taken as a measure of the rate of acid/base transport across the glial membrane. Taking into account the total intracellular buffering power, the maximum rate of acid/base flux was 0.4 mM/min in CO2/HCO3-free saline, and 3.92 mM/min in the presence of 5% CO2/10 mM HCO-3, suggesting that the acid/base flux was dependent upon HCO3-. The rate of acid influx/base efflux increased both with the external HCO3- concentration and with increasing pHi (and hence HCO3-i). This suggested that the decrease in pHi was due to HCO3- efflux. The rapid decrease of pHi was accompanied by a HCO3--dependent depolarization of the glial membrane from -74 +/- 5 mV (n = 20) to -54 +/- 7 mV (n = 13). Both this depolarization and the rate of intracellular acidification were greatly reduced by the anion exchange inhibitor 4,4-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS; 0.3-0.5 mM), but were not affected by the removal of external Cl-. Reduction of the external Na+ concentration to one-tenth normal affected the rate of intracellular acidification only in the presence of CO2/HCO3-: the rate increased within the first 3-5 min after lowering external Na+; after longer exposures in low external Na+ the rate decreased, presumably due to depletion of intracellular Na+. Amiloride (1 mM), which inhibits the Na+-H+ exchange in these cells, had no effect on the rate of intracellular acidification. The intracellular Na activity (aNai) of the glial cells was measured to be 5.2 +/- 1.0 mM (n = 8) in CO2/HCO3-free saline; aNai increased to 7.3 +/- 2.2 mM (n = 8) after the addition of 5% CO2/24 mM HCO3-. Upon a change in pHo to 7.0 in the presence of CO2/HCO3-, aNai decreased by an average of 2 +/- 1.1 mM (n = 5); in CO2/HCO3--free saline external acidification produced a transient increase in aNai. It is concluded that, in the presence of CO2/HCO3-, the rate of intracellular acidification in glial cells is dominated by an outwardly directed, electrogenic Na+-HCO3-cotransport. Neurons, which do not possess this cotransporter, acidify at much lower rates under similar conditions.(ABSTRACT TRUNCATED AT 400 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2001

Domain and genomic sequence analysis of bdellin-KL, a leech-derived trypsin-plasmin inhibitor.

Bdellin-KL is a trypsin-plasmin inhibitor from Hirudo nipponia, whose N-terminal sequence was identified as a non-classical Kazal-type. A cDNA clone encoding the inhibitor was isolated by reverse transcription-PCR and 5' rapid amplification of cDNA ends. The cDNA showed an open reading frame of 155 amino acids comprising one signal peptide and two separated domains. The C-terminal domain consists of distinct internal repeats, including HHEE and HHDD. The bdellin-KL sequence, from the constructed genomic library of Korean leech, was determined for the 2109 bases comprising the open reading frame and flanking regions (3' and 5'). The promoter region contains potential regulatory sequence motifs, including TATA, CAAT, and GC boxes. To characterize the properties of each domain, an N-terminal fragment was prepared by limited proteolysis of the intact protein. The inhibitory activity of the region was as potent as that of the intact protein. This suggests that the compact domain plays an important part in the inhibitory action of bdellin-KL. The C-terminal domain was revealed to have binding affinity to ions such as Ca(2+), Zn(2+), Fe(3+), and Fe(2+) without an influence on the inhibitory activity. This study demonstrates that bdellin-KL may be a novel bifunctional protein with two distinct domains.

Animals↗
PubMed1989

High- and low-technology solutions for massive cavernous hemangioma of the face: lasers and leeches to the rescue.

A massive cavernous hemangioma of the face has been resected using a combination of old and new techniques. The ability of the neodymium: YAG laser to cut with excellent hemostasis formed the basis for the resection. Adjuncts to the surgical excision included such high-technology modalities as laser Doppler flowmeters and tissue expanders. Flap viability was salvaged by the lowly but essential common leech. This case is presented to illustrate the fantastic array of modalities available to plastic surgeons for major resections and reconstructions.

Adult↗