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PubMed1988

Use of leeches in plastic and reconstructive surgery: a review.

Leeches possess properties that make them uniquely able to assist with venous compromised tissue. Their saliva contains an anticoagulant and a histamine-like vasodilator that promote local bleeding, a local anesthetic, and hyaluronidase that promotes the local spread of the other leech salivary secretions into the wound/bite. In addition, active pharyngeal peristalsis further promotes the egress of venous blood. Resurgence in the use of leeches has been stimulated by Upton in the United States and Mahaffey in Europe. Currently, leeches are used at many microsurgical centers to provide critical venous outflow for compromised tissue replantations and transfers that might otherwise be unsalvageable. As the use of leeches becomes more widespread, knowledge of leech biology and physiology is important. This review reports on Hirudo medicinalis, the species used most often medically in Europe and the United States.

Animals↗
PubMed1994

[True and artificial leeches and their use in otology. Images from the history of otorhinolaryngology, presented with instruments from the Ingolstadt German Medical History Museum collection].

Based on the concepts of humoral pathology in former centuries bloodletting was used as remedy to great extent, systematically by puncturing a vein or locally by scarification and cupping glasses. In the 18th and 19th century leeches were introduced and applied in huge numbers to cure all conceivable affections including diseases of the ear. J. M. G. Itard in Paris in 1821 for treating various types of hearing disorders recommended to place leeches at the anus or in the nose to imitate bleeding of haemorrhoids or nose-bleeds, which were considered natural resources of the body to cope with disorders of the humours. H. Schwartze in Halle in 1885 still followed this suggestion. W. R. Wilde in Dublin in 1842 and A. F. von Tröltsch in Würzburg 1867 advocated local blood-letting by leeches around the external ear. The surgeon Ch. L. St. Heurteloup in Paris 1840 invented an artificial leech, which was soon widely used in otology and ophthalmology. After the first World War the application of leeches became obsolete. The humoral pathology had found new approaches: immunology including stimulation of the inherent bodily means of defence, and rheology, particularly haemodilution. The historical development and the technique of applying live and artificial leeches in otology are described in detail.

Animals↗
PubMed1988

Attachment to an endogenous laminin-like protein initiates sprouting by leech neurons.

Leech neurons in culture sprout rapidly when attached to extracts from connective tissue surrounding the nervous system. Laminin-like molecules that promote sprouting have now been isolated from this extracellular matrix. Two mAbs have been prepared that react on immunoblots with a approximately equal to 220- and a approximately equal to 340-kD polypeptide, respectively. These antibodies have been used to purify molecules with cross-shaped structures in the electron microscope. The molecules, of approximately equal to 10(3) kD on nonreducing SDS gels, have subunits of approximately equal to 340, 220, and 160-180 kD. Attachment to the laminin-like molecules was sufficient to initiate sprouting by single isolated leech neurons in defined medium. This demonstrates directly a function for a laminin-related invertebrate protein. The mAbs directed against the approximately equal to 220-kD chains of the laminin-like leech molecule labeled basement membrane extracellular matrix in leech ganglia and nerves. A polyclonal antiserum against the approximately equal to 220-kD polypeptide inhibited neurite outgrowth. Vertebrate laminin did not mediate the sprouting of leech neurons; similarly, the leech molecule was an inert substrate for vertebrate neurons. Although some traits of structure, function, and distribution are conserved between vertebrate laminin and the invertebrate molecule, our results suggest that the functional domains differ.

Animals↗
PubMed1991

Extracellular matrix molecules in development and regeneration of the leech CNS.

As neurons grow to their targets their processes elongate, branch and form specialized endings into which are inserted appropriate ion channels. Our aim has been to analyse the role of the extracellular matrix molecules laminin and tenascin in inducing growth and in determining the form and physiological properties of growing neurites. A preparation in which development and regeneration can be followed at the cellular and molecular level in the animal and in tissue culture is the central nervous system (CNS) of the leech. In leech extracellular matrix (ECM) both laminin and tenascin are present; the molecules are structurally similar but not identical to their vertebrate counterparts. Tenascin extracted from leech ECM shows a typical hexabrachial structure whereas laminin shows a typical cruciform structure in rotary shadowed preparations. Leech laminin purified by means of a monoclonal antibody is a molecule of about 1000 kDa, with a polypeptide composition of 340, 200, 180 and 160 kDa. Substrates that contain tenascin or laminin produce rapid and reliable outgrowth of neurites by identified cells. A remarkable finding is that the outgrowth pattern produced by an individual neuron depends in part on its identity, in part on the substrate upon which it is placed. For example, a Retzius cell grows in a quite different configuration and far more rapidly on laminin substrate than does another type of neuron containing the same transmitter (serotonin); and the pattern of outgrowth of the Retzius cell is different on laminin and on the plant lectin Con A (concanavalin A). Thus Con A induces the growth of processes that are shorter, thicker, more curved and contain fewer calcium channels than those grown on laminin. To determine whether laminin can also influence neurite outgrowth in the animal, immunocytological techniques have been used to follow its distribution in the extracellular matrix of normal, developing and regenerating leech CNS. In adult leeches neuronal processes in the CNS are not in contact with laminin which is confined to the surrounding extracellular matrix. In embryos however, laminin staining appears between ganglionic primordia along the pathways that neurons will follow. Similarly, after injury to the adult CNS, laminin accumulates at the very sites at which sprouting and regeneration begin. How the laminin becomes redistributed to appear in the region of injury has not yet been established. Together these findings suggest a key role for laminin and for other extracellular matrix molecules.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Saxitoxin differentiates between two types of Na+-dependent potentials in the Retzius cell of hirudinid leeches.

The effects of tetrodotoxin (TTX) and saxitoxin (STX) on the action potentials recorded in Ca2+-free solution in the absence of Ca2+ and K+ currents were investigated in the Retzius cell of three hirudinid leech species (Hirudo medicinalis, Macrobdella decora and Poecilobdella granulosa) and in the glossiphoniid leech Haementeria ghilianii. In the four leech species, stimulation of the Retzius cell in the presence of 25 mmol l-1 tetraethylammonium chloride (TEA), 3 mmol l-1 4-aminopyridine (4-AP) and 2mmol l-1 Mn2+ evoked prolonged action potentials consisting of an initial fast-rising spike followed by a plateau lasting several hundreds of milliseconds. The amplitude and duration of both components of action potentials recorded under these conditions were dependent on [Na+]o. In the hirudinid leeches the initial spike was unaffected by TTX and STX whereas the plateau was selectively blocked by micromolar concentration of STX. In Haementeria both the initial spike and the subsequent plateau were sensitive to nanomolar concentrations of STX and TTX with an estimated ED50 of approximately 20 nmol l-1 for inhibition of Vmax of the fast spike. The results suggest that there are two types of ionic currents mediating the two distinct components of Na+-dependent action potentials in the Retzius cell: (1) a fast-inactivating one, presumably underlying the normal spike which is TTX- and STX-resistant in hirudinid leeches but sensitive to both agents in Haementeria and (2) a low-threshold, prolonged current which underlies the plateau recorded from these cells in the absence of Ca2+ and K+ currents and which is selectively blocked by STX in hirudinid leeches but sensitive to both STX and TTX in Haementeria. It is likely that the variable kinetic and pharmacological properties that characterize the various Na+ potentials in these identified homologous neurones may be of functional significance and result from differences in the molecular structure of their Na+ channels.

4-Aminopyridine↗
PubMed1987

Tinkering with successful synapse regeneration in the leech: adding insult to injury.

In the leech, synapse regeneration in adults and synapse formation during embryonic development can be studied in single, identifiable cells that make precise connections with their targets. Certain cellular components, such as synaptic targets and glia, were selectively destroyed to study how the regenerating axons locate their targets, what triggers axons to start growing and what stops them. The results showed that glia and targets play only a limited role in synapse regeneration and in axon degeneration. For example, contact with the synaptic target may inhibit sprouting and availability of targets may promote it. Comparative studies on axon growth and synapse formation by interneurones in embryos showed that regeneration does not simply recapitulate embryonic development. There are clearly separate constraints on the two processes. Axon survival is a different problem. Although isolated axon segments can survive for up to a year in the leech, temperature is a major factor in survival. Axon segments in a tropical leech that regenerates synapses well at 31 degrees C degenerated within 2-3 weeks at this elevated temperature, even when regeneration was prevented. In similar leeches at room temperature (22 degrees C), segments survived for months. Overall, results in the leech support the idea that degeneration as well as regeneration share fundamental mechanisms with other invertebrates and the vertebrates, including mammals. Perhaps long-lived axon segments and other features of the leech that speed or encourage functional regeneration can now be made to operate in repair of the mammalian nervous system.

Animals↗
PubMed2001

Regulation of Na(+) transport across leech skin by peptide hormones and neurotransmitters.

An increase in intracellular cyclic AMP concentration stimulates transepithelial Na(+) transport across the skin of the leech Hirudo medicinalis, but it is unclear how cytosolic cyclic AMP levels are elevated in vivo. In search of this external stimulus, we performed Ussing chamber experiments to test several peptide hormones and neurotransmitters for their effect on Na(+) transport across leech dorsal integument. Although all the peptide hormones under investigation significantly affected ion transport across leech integument, none of them mimicked the effect of an experimental rise in intracellular cyclic AMP level. The invertebrate peptides conopressin and angiotensin II amide inhibited short-circuit-current- (I(sc)) and amiloride-sensitive Na(+) transport (I(amil)), although to slightly different degrees. The vertebrate peptide hormones 8-arginine-vasopressin and 8-lysine-vasopressin both produced an inhibition of I(amil) comparable with that caused by angiotensin II amide. However, 8-lysine-vasopressin reduced I(sc), whereas 8-arginine-vasopressin induced a moderate increase in I(sc). The neurotransmitter dopamine, which occurs in the leech central nervous system in relatively large amounts, and its precursor l-dopamine both induced large decreases in I(sc) and I(amil). However, the reactions evoked by the catecholamines showed no pronounced similarity to the effects of intracellular cyclic AMP. Two other neurotransmitters known to occur in leeches, serotonin (5-hydroxytryptamine) and gamma-n-aminobutyric acid (GABA), had no influence on transepithelial ion transport in leech skin.

Amiloride↗
PubMed1999

Cellular expression of a leech netrin suggests roles in the formation of longitudinal nerve tracts and in regional innervation of peripheral targets.

Netrins are secreted, diffusible proteins that direct axonal growth. To study the functions of netrins in the relatively simple and easily accessible nervous system of the leech Hirudo medicinalis, we have cloned a leech netrin and have characterized its expression during embryogenesis. By probing a leech cDNA library at low stringency with chick netrin probes, we have identified a complete cDNA clone that bears significant sequence similarity to netrins of other species. In situ hybridization and dye filling of individual neurons show that this leech netrin is expressed by several identifiable central neurons in every segmental ganglionic primordium during early stages of embryogenesis. Some of these neurons, including the bipolar cells which are thought to be involved in setting up longitudinal tracts, express this gene only transiently during embryogenesis, while others continue to express it in the adult. In addition, leech netrin is expressed by ventral but not dorsal longitudinal muscle cells in each segment before central neurons project their axons to the periphery. These highly specific expression patterns are consistent with the hypothesis that leech netrin plays a role in forming the major interganglionic neuronal tracts and in defining ventral versus dorsal domains of peripheral innervation.

Amino Acid Sequence↗
PubMed1989

[Epistaxis caused by leeches].

Among the conditions which can cause epistaxis are intranasal or nasopharyngeal foreign bodies. A living foreign body, or parasite, in the nose or nasopharynx is rare in western countries, but in other parts of the world is a fairly common cause of epistaxis. In the past 5 years 17 cases of epistaxis due to infestation with leeches were treated in our 2 departments. In 14 patients, 9-27 years old, the complaint was of recurrent episodes of mild epistaxis and blood-spitting several days before admission. 3 patients were admitted hemorrhaging profusely from the nose and mouth. Examination showed a green-brown mass protruding from the nasopharynx behind the soft palate, which is every case was a blood-engorged leech. Treatment consisted of removing the leech by applying a forceps to the middle of the leech's body and giving a quick pull. Bleeding ceased immediately after removal of the leech. All the patients had drunk water from brooks, streams or fresh water lakes. Therefore, in areas in which leeches are present only boiled or filtered water should be drunk.

Adolescent↗
PubMed1989

Sustained bleeding after a leech bite in the apparent absence of hirudin.

The bite of the medicinal leech bleeds for many hours. For decades it has been assumed that the remarkably prolonged bleeding time of a leech bite wound is due to hirudin, a specific anti-thrombin secreted by the leech during feeding. By measuring haematological parameters of blood oozing from a leech bite wound on 15 different occasions in 7 human volunteers, we demonstrate that the hirudin-sensitive coagulation parameters, including thrombin-induced platelet aggregation, are prolonged for only 15 min, after which they return to normal. This suggests that excess hirudin secreted by the leech is washed out during this period. However, bleeding from the leech bite wound persists for a mean of 10 h. Platelets in smears of exuding blood show no evidence of spontaneous aggregation, but in vitro platelet aggregation can be induced by exogenous collagen at any time. In view of sustained bleeding in the apparent absence of hirudin, attention is focussed onto an unsuspected factor or factors which may better explain the prolonged bleeding phenomenon.

Animals↗
PubMed1985

[Destabilase: an enzyme of medicinal leech salivary gland secretion hydrolyzes the isopeptide bonds in stabilized fibrin].

The salivary gland secretion of the leech Hirudo medicinalis contains an enzyme termed by us as destabilase, which hydrolyzes the epsilon-(gamma-glutamyl)-lysine bonds as a result of fibrin stabilization by factor XIIIa in the presence of Ca2+. This hydrolysis, apart from the original lysine and glutamine, is characterized by an appearance of lysine and glutamic acid residues. The accumulation of glutamic acid residues leads to spontaneous depolymerization of the destabilized fibrin. As a result, fluid "spots" of destabilized fibrin depolymerization (DFD) begin to appear at the sites of leech secretion application on the surface of stabilized fibrin plates. The DFD activity of the leech salivary gland secretion manifests itself only in case of stabilized fibrin and increases with an increase in the stabilization degree. Treatment of leech secretion with diisopropylfluorophosphate does not affect the enzyme activity, which is completely blocked by monoiodoacetate. The mechanism of action of leech salivary gland secretion and the enzyme isolated from it, i. e., destabilase, was studied, using a synthetic chromogenic substrate - p-nitroanilide-gamma-glutamic acid. The amidolytic activity of leech salivary gland secretion is 2.2 +/- 0.18 nkat/ml, Km(app) for destabilase is 0.6 X 10(-5) M, V = 5.4 X 10(-3) mol/min.

Animals↗
PubMed1982

Characterization of acetylcholinesterase in individual neurons in the leech central nervous system.

Acetylcholinesterase (AChE) activity was measured in cholinergic and non-cholinergic neurons in the central nervous system of the leech. Intracellular AChE was assayed by pretreating intact ganglia with echothiophate to inhibit selectively extracellular enzyme. The concentration of intracellular AChE in cholinergic neurons was 3- to 24-fold higher than that in non-cholinergic cells. The properties of AChE in extracts of leech ganglia were similar to those of "true" acetylcholinesterase, although butyrylthiocholine was almost as good a substrate as acetylthiocholine. There was also cholinesterase activity in leech blood; this enzyme resembled butyrylcholinesterase. Sucrose gradient velocity sedimentation of Triton X-100 extracts of leech ganglia revealed a major peak of AChE activity at 6.5 S and a small peak at 4.3 S. The pattern of activity in the gradient was the same when intact ganglia were pretreated with echothiophate, although the total activity was reduced by 98%. Intact leech ganglia were stained for AChE activity with and without echothiophate pretreatment. In ganglia that had not been exposed to echothiophate, cholinesterase reaction product was deposited primarily on the ganglionic sheath. In pretreated ganglia, on the other hand, cholinesterase activity was concentrated within neuronal cell bodies. Electrophysiological identification and intracellular injection of the fluorescent dye Lucifer Yellow prior to staining were used to confirm that most AChE-positive cells were cholinergic motoneurons. Two previously unidentified neurons staining for AChE were shown to be motoneurons. These results demonstrate that cholinergic motoneurons can be differentiated from other cells in the leech nervous system by their high intracellular concentration of AChE.

Acetylcholinesterase↗
PubMed1997

Leech immunocytes contain proopiomelanocortin: nitric oxide mediates hemolymph proopiomelanocortin processing.

This report establishes the presence of mammalian-like proopiomelanocotropic hormone (POMC), and six of its peptides, including adrenocorticotropic hormone (ACTH) and melanocyte-stimulating hormone (MSH), in the immune tissues of the leech Theromyzon tessulatum. The 25.4-kDa protein was purified by high pressure gel permeation chromatography, anti-ACTH-affinity column, and reverse-phase HPLC. Its characterization was performed by Edman degradation, enzymatic treatments, and electrospray mass spectrometry. Leech POMC exhibits considerable amino acid sequence similarity to mammalian POMC. Of the six peptides, three showed high sequence similarity to their vertebrate counterparts met-enkephalin, alpha-MSH, and ACTH: 100, 84.6, and 70%, respectively; whereas gamma-MSH, beta-endorphin, and gamma-lipotropin hormone exhibited only 45, 20, and 10% sequence identity, respectively. No dibasic amino acid residues were found at the C terminus of the gamma- and beta-MSH peptides. In contrast, the leech alpha-MSH was flanked at its C-terminal by the Gly-Arg-Lys amidation signal. ACTH and corticotropin-like intermediary pituitary peptide were also C-terminally flanked by dibasic amino acid residues. The coding region of leech POMC was obtained by reverse transcription-PCR using degenerated oligonucleotide primers. Circulating levels of ACTH and MSH were 10 and 1 fmol/microl hemolymph, respectively. Morphine, in a dose-dependent manner, increased the levels of both peptides threefold; this effect was blocked by naloxone treatment. Similar results were found with the anandamide. Leech ACTH was processed to MSH by the enzymes neutral endopeptidase (24.11) and angiotensin-converting enzyme. Leech alpha-MSH had the same activity as authentic alpha-MSH in two bioasssay systems. Taken together, the study demonstrates that POMC is present in invertebrates and its immunoregulatory actions have been conserved during evolution.

Adrenocorticotropic Hormone↗
PubMed1998

Formation and localization of cytoplasmic domains in leech and ascidian zygotes.

Leech and ascidian embryos are well suited for the study of certain developmental processes. Although leeches and ascidians belong to different bilateralia groups (protostomes and deuterostomes, respectively) they share important developmental features and, in particular, the determinate character of their embryogenesis. In both types of embryos this property is related to the presence of specific cytoplasmic domains that are selectively allocated to different blastomeres during cleavage. In this review leech and ascidian eggs and zygotes are compared in terms of the structure of these cytoplasmic domains and of the cellular mechanisms involved in their formation and localization. During meiosis the zygote of leeches and ascidians undergo stereotypic actin-dependent contraction movements related to both the emission of the polar bodies and the formation and relocalization of cytoplasmic domains. After completion of meiosis, during first interphase, monaster microtubules nucleated from the sperm-derived centrosome play a key role in pronuclear migration. In addition, these astral microtubules direct the relocalization of cytoplasmic domains and the translocation and accumulation of organelles in the interior of the zygote. Microtubules and microfilaments, on the other hand, are involved in cortical reorganizations and organelle translocations in both zygote species during interphase and cleavage divisions. In the case of leech zygotes, this process leads to formation of characteristic polar cytoplasmic domains called teloplasms. These domains are selectively inherited by teloblasts, precursor stem cells of ectodermal and mesodermal tissues in the leech embryo. In the ascidian zygote, the cytoplasmic movements observed during interphase and mitosis lead to relocalization of the bulk of a mitochondria-rich domain, called the myoplasm, along with an endoplasmic reticulum-rich domain towards the future posterior pole of the embryo. The myoplasm is inherited by a subset of posterior blastomeres committed to become the primary muscle cells of the ascidian tadpole.

Animals↗
PubMed2005

The Leech method for diagnosing constipation: intra- and interobserver variability and accuracy.

BACKGROUND: The data concerning the value of a plain abdominal radiograph in childhood constipation are inconsistent. Recently, positive results have been reported of a new radiographic scoring system, "the Leech method", for assessing faecal loading. OBJECTIVE: To assess intra- and interobserver variability and determine diagnostic accuracy of the Leech method in identifying children with functional constipation (FC). MATERIALS AND METHODS: A total of 89 children (median age 9.8 years) with functional gastrointestinal disorders were included in the study. Based on clinical parameters, 52 fulfilled the criteria for FC, six fulfilled the criteria for functional abdominal pain (FAP), and 31 for functional non-retentive faecal incontinence (FNRFI); the latter two groups provided the controls. To assess intra- and interobserver variability of the Leech method three scorers scored the same abdominal radiograph twice. A Leech score of 9 or more was considered as suggestive of constipation. ROC analysis was used to determine the diagnostic accuracy of the Leech method in separating patients with FC from control patients. RESULTS: Significant intraobserver variability was found between two scorers (P=0.005 and P<0.0001), whereas there was no systematic difference between the two scores of the other scorer (P=0.89). The scores between scorers differed systematically and displayed large variability. The area under the ROC curve was 0.68 (95% CI 0.58-0.80), indicating poor diagnostic accuracy. CONCLUSIONS: The Leech scoring method for assessing faecal loading on a plain abdominal radiograph is of limited value in the diagnosis of FC in children.

Child↗
PubMed1997

Leeches as indicators of dietary mercury exposure in non-piscivorous waterfowl in central Ontario, Canada.

We collected and analysed 113 leeches (Hirudinea) from 17 small lakes in the acid-stressed Muskoka region of central Ontario, Canada to examine the relationship between lake chemistry and mercury (Hg) concentrations in leeches, and thus determine whether leeches and other benthic invertebrates posed a dietary risk of Hg exposure for non-piscivorous waterfowl. Hg concentrations in leeches were generally low and only a few-fold above the detection limit (0.78 ng g(-1) wet weight (ww)). Mean Hg concentration in the bloodsucker Macrobdella decora was 6.94 +/- 0.78 SE ng g(-1) ww (n=49) and was 5.98 +/- 0.46 ng g(-1) ww (n=64) in the scavenger Percymoorensis marmoratis. Leech Hg concentrations were correlated with calcium and dissolved organic carbon concentrations in the water, respectively. These data suggest that leeches are not suitable monitors of Hg (usually as methylmercury) biomagnification in central Ontario lakes, and do not pose a dietary risk to non-piscivorous waterfowl.

Journal Article↗
PubMedDate not supplied

Cannibalism in a population of the medicinal leech (Hirudo medicinalis L.).

Medicinal leeches (Hirudo medicinalis L.) were maintained in large ponds in a commercial leech farm at Biebertal, Germany. The feeding of hungry adult leeches was performed on representative individuals that were placed on cloth soaked with mammalian blood obtained from a local butchery (pig, Sus scrofa). In a second set of experiments, cane toads (Bufo marinus) were used as host organisms. The leeches rapidly attached to the toads, explored the body and sucked blood. After feeding, the fully engorged leeches were placed into the pond or an aquarium. In this artificial habitat, the satiated leeches were attacked by hungry conspecifics, sucked off and killed. This observation demonstrates that H. medicinalis must be classified as a cannibalistic annelid.

Animals↗
PubMed1997

Developmental control of cell division in leech embryos.

During embryogenesis, cell division must be spatially and temporally regulated with respect to other developmental processes. Leech embryos undergo a series of unequal and asynchronous cleavages to produce individually recognizable cells whose lineages, developmental fates and cell cycle properties have been characterized. Thus, leech embryos provide an opportunity to examine the regulation of cell division at the level of individual well-characterized cells within a community of different types of cells. Isolation of leech homologues of some of the highly conserved regulators of the cell division cycle, and characterization of their patterns of maternal and zygotic expression, indicate that the cell divisions of early leech embryos are regulated by cell type-specific mechanisms. These studies with leech embryos contribute to the emerging appreciation of the diverse mechanisms by which animals regulate cell division during early development.

Animals↗