Biomedical subjects

J Malecha

Publications and source records attributed to J Malecha.

At least 19 recordsLinked to original sources

PubMed2004

Effects of a surgical treatment of congenital cervicovaginal agenesia.

OBJECTIVE: Congenital agenesis of the uterine cervix and vagina is one of the rarest congenital defects. The objective of this manuscript is to present our experience and effects of a surgical treatment in five girls with inborn agenesis of the uterine cervix and vagina, who were operated in our clinic. METHOD: The vagina is reconstructed in four stages: (1) formation of vaginal recess; (2) phantomization of vagina; (3) joining of the reconstructed vagina with the uterus; (4) revision and correction of the junction. GnRH analogs were administered to avoid menstrual blood flow into the peritoneal cavity during phantomization. RESULTS: Patency of the uterovaginal junction was maintained with a #6 intubation tube. Normal menstruation was restored in all patients. Atresion of the canal at the uterovaginal junction was disclosed after 3-8 months in three of the patients. Patency was restored with surgical hysteroscope and the canal was mechanically distended. Clinical and ultrasonographic followup in the patients will continue.

Adolescent↗

PubMed2003

GnRH analogues as an adjuvant therapy for ovarian cancer patients.

OBJECTIVES: Lowering gonadotropin levels with gonadotropin-releasing hormone (GnRH) analogues in patients with ovarian cancer remains open to debate. The aim of this study was to assess the results of treatment in stage III and stage IV ovarian cancer patients who had surgery supplemented with chemotherapy, radiotherapy, and GnRH analogues. Gonadotropin levels were monitored during treatment. METHODS: The study group comprised 69 patients aged 27-70 years, stratified according to the type of treatment. The overall disease-free, 5-year survival rates and the frequency of remissions were analyzed. Hormonal tests [follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] were performed in 58 patients. Associations were checked between gonadotropin levels, clinical findings, and survival. The results were statistically compared. RESULTS: Statistically significant differences were noted when chemotherapy was supplemented with GnRH analogues and/or radiotherapy. Administration of GnRH analogues resulted in significantly lower levels of LH than of FSH. Levels of FSH were significantly lower in patients surviving at least 5 years or in complete remission at the time of this study. CONCLUSIONS: Combined therapy can produce favorable results in late-stage ovarian cancer, and GnRH analogues have an important role in treatment strategy.

Adult↗

PubMed2001

Cloning and expression analysis of a cDNA that encodes a leech hemerythrin.

We report the cDNA sequence of a leech hemerythrin. A cDNA was isolated from a Theromyzon tessulatum cDNA library and encodes a 120 amino acid protein of about 14 kDa. The predicted protein contains the hemerythrin signature sequence and the iron ligand residues previously identified in crystal structures of hemerythrin and myohemerythrin. The protein displayed the highest identity to myohemerythrin, a non-heme iron-binding protein described in sipunculids. Expression analysis indicated that the mRNA is widely expressed in leech and is stage specific in appearance, being absent after the two first blood meals, appearing after the last blood meal during the period preceding oogenesis and disappearing after egg laying.

Amino Acid Sequence↗

PubMed2000

Therostasin, a novel clotting factor Xa inhibitor from the rhynchobdellid leech, Theromyzon tessulatum.

Therostasin is a potent naturally occurring tight-binding inhibitor of mammalian Factor Xa (K(i), 34 pm), isolated from the rhynchobdellid leech Theromyzon tessulatum. Therostasin is a cysteine-rich protein (8991 Da) consisting of 82 amino acid residues with 16 cysteine residues. Its amino acid sequence has been determined by a combination of techniques, including Edman degradation, enzymatic cleavage, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) on the native and s-beta-pyridylethylated compound. Sequence analysis reveals that it shares no significant homology with other Factor Xa inhibitors except for the putative reactive site. Moreover, it contains a signature pattern for proteins of the endothelin family, potent vasoconstrictors isolated in mammal and snake venom. Therostasin cDNA (825 bp) codes for a polypeptide of 82 amino acid residues preceded by 19 residues, representing a signal peptide sequence. As for the other known inhibitors of Factor Xa, therostasin is expressed and stored in the cells of the leech salivary glands.

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↗

PubMed1999

[Ovarian cancer in girls].

DESIGN: The aim of our study was a clinical analysys of diagnosis, operative treatment and monitoring of epithelial ovarian cancer in girls. MATERIALS AND METHODS: Authors described 11 epithelial malignant tumors of ovaries in girls aged 12-16. Clinical aspects, pathology stage, grade treatment, monitoring and survival were evaluated. Three of the patients had ovarian cancer. A 19-year old patient with stage Ia of epithelial ovarian cancer was managed conservatively. Four years later she give birth. For 10 years no malignant of has been observed. Seven patients with ovarian borderline epithelial tumors were treated. The tumors in 5 patients were of serous and in 2 of mucinous type. CONCLUSIONS: We suggested that patients of stage I who wish to preserve childbearing function may be treated with unilateral salpingo-ovariectomy and chemotherapy.

Adolescent↗

PubMed1999

[Laparoscopy in the diagnosis and treatment of pathology of sexual organs in girls].

DESIGN: The aim of our study was a clinical analysis of laparoscopies in girls which were performed in the Clinic of Gynaecological Surgery. MATERIAL AND METHODS: Laparoscopic procedures were carried out between 1996-1998. Patients were divided in to three groups: aged under 15, between 15-19 and over 19. 44% of patients from the youngest group had laparotomy. 43.8% of laparoscopic interventions were in aged group 20-24. CONCLUSION: Laparoscopy is very good diagnostic method in gynaecological diseases. It gave enabled operative treatment in 40-43.8% girls.

Adolescent↗

PubMed1999

[Operative treatment of developmental abnormalities of sexual organs in girls].

DESIGN: The aim of our study was a clinical analysis of operative treatment of developmental abnormalities of sexual organs. MATERIALS AND METHODS: The authors describe developmental abnormalities of sexual organs in girls aged 12-19 79(1.4%) girls were treated from among 5432 surgically treated patients in the course 6 years. Among these abnormalities were the following: agenesia of vagina, agenesia of cervical canal, absence of cervix of uterus, obstructed normal or rudimental vagina, occlusion of hymen. There were complex abnormalities too. Congenital abnormalities were subject to the reconstructive operative treatment. CONCLUSIONS: We recommend that developmental defects should be treated in specialist centres well equipped and experience of in operative procedures and postoperative monitoring.

Adolescent↗

PubMed1996

Purification, sequence analysis, and cellular localization of a prodynorphin-derived peptide related to the alpha-neo-endorphin in the rhynchobdellid leech Theromyzon tessulatum.

Cells immunoreactive to an antiserum specifically directed against vertebrate alpha-Neo-endorphin (alpha-NE) were detected in the internal wall of anterior and posterior suckers of the rhynchobdellid leech Theromyzon tessulatum. These cells have morphological and ultrastructural characteristics close to the "releasing gland cells" of adhesive organs. The epitope recognized by anti-alpha-NE was contained in granules having a diameter of 0.2-0.3 microm. Previous works involving the brain of this leech demonstrate the existence of approximately 14 neurons immunoreactive to the anti-alpha-NE. Following an extensive purification including high pressure gel permeation and reversed-phase high performance liquid chromatography, epitopes contained in both suckers and central nervous system were isolated. Purity of the isolated peptides was controlled by capillary electrophoresis. Their sequences were determined by a combination of automated Edman degradation, electrospray mass spectrometry measurement, and coelution experiments in reversed-phase high performance liquid chromatography with synthetic alpha-NE. The results demonstrate that epitopes recognized by the anti-alpha-NE in the suckers and the central nervous system are identical to vertebrate alpha-NE (YGGFLRKYPK). This finding constitutes the first biochemical characterization of a prodynorphin-derived peptide in invertebrates. Moreover the isolation of this peptide in the annelida establishes the very ancient phylogenetic origin of alpha-NE as well as its conservation in evolution.

Amino Acid Sequence↗

PubMed1996

Structural characterization of a novel neuropeptide from the central nervous system of the leech Erpobdella octoculata. The leech osmoregulator factor.

Purification of a material immunoreactive to an antiserum against the C-terminal part of the oxytocin (Pro-Leu-Gly-amide) and present in the central nervous system of the Pharyngobdellid leech Erpobdella octoculata was performed by reversed-phase high performance liquid chromatography combined with both enzyme-linked immunosorbent and dot immunobinding assays for oxytocin. The amino acid sequence of the purified peptide (Ile-Pro-Glu-Pro-Tyr-Val-Trp-Asp) was established by Edman degradation and confirmed by electrospray mass spectrometry measurement. When injected in leeches, purified or synthetic peptides exert an anti-diuretic effect, the most effective ranged between 10 pmol and 1 nmol. They provoked an uptake of water 1-2 h post-injection. Furthermore, electrophysiological experiments conducted in the leech Hirudo medicinalis revealed an inhibition of the potency of Na+ conductances of leech skin by this peptide. Immunocytochemical studies with an antiserum against synthetic oxytocin-like molecule provided the cytological basis for existence of a neuropeptide, since large amounts of immunoreactive neurons were detected in the central nervous systems of E. octoculata. The purified molecule is both different to peptides of the oxytocin/vasopressin family and is a novel neuropeptide in the animal kingdom. It was named the leech osmoregulator factor (LORF). An identification of the proteins immunoreactive to an antiserum against oxytocin performed at the level of both central nervous systems extracts and in vitro central nervous system-translated RNA products indicated that in the two cases, a single protein was detected. These proteins with a molecular masses of, respectively, approximately 34 kDa (homodimer of 17 kDa) for the central nervous systems extracts and approximately 19 kDa for in vitro central nervous system-translated RNA products were not recognized by the antiserum against MSEL- and VLDV-neurophysin (proteins associated to oxytocin and vasopressin), confirming that LORF did not belong to the oxytocin/vasopressin family.

Amino Acid Sequence↗

PubMed1995

Structural characterization of a diuretic peptide from the central nervous system of the leech Erpobdella octoculata. Angiotensin II Amide.

Purification of a material immunoreactive to an antiserum against angiotensin II and present in the central nervous system of the pharyngobdellid leech Erpobdella octoculata was performed by reversed-phase high pressure liquid chromatography combined with both enzyme-linked immunosorbent assay and dot immunobinding assays for angiotensin II. Establishment of the amino acid sequence by Edman degradation, electrospray, and fast atom bombardement mass spectrometry measurements and enzymatic treatment by carboxypeptidase A indicated that this "central" angiotensin II-like material, the first one fully characterized in the animal kingdom, is an angiotensin II amide. This finding constitutes also the first biochemical characterization of a peptide of the angiotensin family in an invertebrate. Synthetic angiotensin II amide exerts, when injected in leeches, a diuretic effect and is, 1 and 2 h postinjection, 100-fold more potent than vertebrate angiotensin II. An identification of the proteins immunoreactive to an antiserum against angiotensin II performed at the level of both central nervous system extracts and in vitro central nervous system-translated RNA products indicated that in the two cases, two proteins were detected. Their molecular masses, which were, respectively, approximately 14 and approximately 18 kDa for the central nervous system extracts and approximately 15 and approximately 19 kDa for in vitro central nervous system-translated RNA products, differ from that of angiotensinogen (approximately 60 kDa), the precursor of vertebrate angiotensin II.

Amino Acid Sequence↗

PubMed1995

Isolation and structural characterization of enkephalins in the brain of the rhynchobdellid leech Theromyzon tessulatum.

This paper reports the purification of four peptides related to enkephalins from the brain of the leech Theromyzon tessulatum. After reverse-phase HPLC purification, the sequence of the enkephalins (YGGFM, YGGFL, FM, FL) was established by a combination of automated Edman degradation, electrospray mass spectrometry measurement, and co-elution experiments in reverse-phase HPLC with synthetic peptides. ELISA titrations performed on each purified peptide indicated that the major amount was borne by the leucine-enkephalin. The ratio of leucine-enkephalin and methionine-enkephalin of 2:1 is in line with previous immunocytochemical data obtained on T. tessulatum brains. The presence of enkephalins in T. tessulatum, an animal belonging to the oldest group of coelomate metazoans (the Annelida) establishes the very ancient phylogenetic origin of opioids and their conservation in the course of evolution.

Amino Acid Sequence↗

PubMed1994

Isolation and structural characterization of a novel peptide related to gamma-melanocyte stimulating hormone from the brain of the leech Theromyzon tessulatum.

This paper reports the purification of a novel pro-opiomelanocortin derivative peptide (a gamma-melanocyte stimulating hormone-like (gamma-MSH-like) molecule) from the brain of the leech Theromyzon tessulatum. After reverse-phase HPLC purification, the sequence of the gamma-MSH-like peptide (YVMGHFRWDKFamide) was established by a combination of automated Edman degradation, electrospray mass spectrometry measurement, enzymatic treatment and co-elution experiments in reverse-phase HPLC with synthetic peptides.

Amino Acid Sequence↗

PubMed1994

FMRFamide-related peptides in the sex segmental ganglia of the Pharyngobdellid leech Erpobdella octoculata. Identification and involvement in the control of hydric balance.

Using enzyme-linked immunosorbent assays, a dot-immunobinding assay and a three-step reverse-phase HPLC separation, four Arg-Phe-amide (RFamide) peptides were purified from sex segmental ganglia extracts of the leech Erpobdella octoculata; FMRFamide, FM(O)RFamide, FLRFamide and GDPFLRFamide. Their amino acid sequences were elucidated by means of a combined approach using antiserum specificity, synthetic-peptide coelution, automated Edman degradation and electrospray mass spectrometry. One of these peptides, GDPFLRFamide, is a novel leech RFamide neuropeptide. Two of the above RFamide peptides are involved in the control of leech hydric balance; one (GDPFLRFamide) is diuretic, the other (FMRFamide) is anti-diuretic. Titration of each purified RFamide peptide indicated a similar amount of each tetrapeptide and of tetrapeptides and heptapeptides. A comparison between RFamide peptides of E. octoculata and molluscs reveals structural similarities supporting the hypothesis for the existence of an ancestral RFamide peptide gene common to leeches and molluscs.

Amino Acid Sequence↗

PubMed1993

Biochemical evidence of angiotensin II-like peptides and proteins in the brain of the rhynchobdellid leech Theromyzon tessulatum.

The peptides contained in neurons localized in the brain of the leech Theromyzon tessulatum (Hirudinae, Rhynchobdellida) and showing an immunopositive reaction with an antibody directed against angiotensin II (AII), were purified by reversed-phase HPLC. Three AII-like peptides (P1, P2 and P3) which exhibited the same retention times and chromatographic behaviors as synthetic AVII (fragment 6-8 of AII), AIV (fragment 3-8 of AII) and AII, respectively, were resolved in brain extracts. An identification of the proteins immunoreactive to an anti-AII was performed at the level of both brain extracts and in vitro brain-translated RNA products. The protein detected at the level of the brain extracts (of a molecular mass of approximately 18 kDa) is multipeptidic as it is also recognized by two other antisera, a polyclonal one directed against gamma-MSH and a monoclonal one (Tt159) raised against a leech brain epitope. It could be the pro-AII-like precursor. The protein detected at the level of in vitro brain-translated RNA products (of a molecular mass of approximately 19 kDa) could be the prepro-AII-like precursor.

Angiotensin II↗

PubMed1993

Isolation, structural characterization and biological function of a lysine-conopressin in the central nervous system of the pharyngobdellid leech Erpobdella octoculata.

Several neuropeptides are suspected to act on the control of hydric balance in leeches. One of these peptides, a peptide immunoreactive to an antibody against oxytocin, was previously characterized from the central nervous system of the leech Erpobdella octoculata [Salzet, M., Wattez, C., Verger-Bocquet, M., Beauvillain, J.-C. & Malecha, J. (1993) Brain Res. 601, 173-184]. This paper reports the isolation from the central nervous system of E. octoculata of another peptide of the oxytocin/vasopressin family; a lysine-vasopressin-like molecule. Its purification was performed by reverse-phase high-performance liquid chromatography combined with both dot immunobinding assay and enzyme-linked immunosorbent assay for lysine-vasopressin. The amino acid sequence was established by Edman degradation and confirmed by electrospray-mass-spectrometry measurement. The nonapeptide obtained corresponded to the lysine-conopressin previously isolated from the venom of the mollusc Conus geographus [Cruz, L. L., de Santos, V., Zafaralla, G. C., Ramilo, C. A., Zeikus, R., Gray, W. R. & Olivera, B. M. (1987) J. Biol. Chem. 262, 15821-15824]. In leeches, synthetic lysine-conopressin exerts a diuretic effect which can be compared to that of the arginine-vasopressin-like peptide isolated in the Insect Locusta migratoria [Proux, J., Miller, C. A., Li, J. P., Carney, R. L., Girardie, A., Delaage, M. & Schooley, D. A. (1987) Biochem. Biophys. Res. Commun. 149, 180-186].

Amino Acid Sequence↗

PubMed1993

Oxytocin-like peptide: a novel epitope colocalized with the FMRFamide-like peptide in the supernumerary neurons of the sex segmental ganglia of leeches--morphological and biochemical characterization; putative anti-diuretic function.

A large number of oxytocin (OT)-like neurons were detected in the sex segmental ganglia (SG5, SG6) of three species of leeches belonging to different orders: Theromyzon tessulatum, Hirudo medicinalis and Erpobdella octoculata. In this latter species, an epitope close to the vertebrate OT by its C-terminal part (MSH release inhibiting factor: MIF), localized in granules of a size diameter of ca 120 nm and colocalized with FMRFamide(FMRFa)-like material was demonstrated. With reverse phase-high performance liquid chromatography, evidence was given that the two epitopes (OT and FMRFa) colocalized in the same neurons were biochemically different. A titration of OT per SG indicated that the OT-like amount was considerably higher in sex SG than in non-sex SG (ca. 5 pmol vs. ca. 0.5 pmol). Moreover, at the level of sex SG, this amount was ca. 3-fold higher in immature leeches than in mature specimens. Injections of extracts of SG of E. octoculata and of fragments of OT (Tocinoic acid or MIF) to T. tessulatum, indicated that MIF (the epitope found in the sex SG) and sex SG have the same anti-diuretic effect on the leeches injected. These results pointed to an anti-diuretic role of the leech OT-like substance.

Animals↗

PubMed1993

[Ultrasonography, laparoscopy, neoplastic markers--new possibilities of conservative operative treatment for ovarian neoplasms in girls and young women].

The knowledge of ovarian physiopathology and the employing of modern diagnostic methods (ultrasonography, laparoscopy) allow to avoid to operate patients with functional cysts. Girls and young women should be evaluated individually for ovarian surgical procedures. The operative treatment should be so conservative as possible employing all diagnostic and monitoring methods including specific markers assays.

Adolescent↗