Revisão da leucina aminopeptidase no trabalho de parto pré-termo e na pré-eclampsia / Revision of the leucine aminopeptidase in preterm labor and in preeclampsia

Realizou-se um levantamento bibliográfico, com artigos de revisão, analisando e discutindo os trabalhos publicados sobre os efeitos da leucina aminopeptidase e aminopeptidase A no trabalho de parto pré-termo e na pré-eclampsia. A proposta deste trabalho sobre o tema é que grande parte das questões de saúde materna parece pueril, principalmente quanto ao atendimento voltado para os cuidados maternos, no qual, a cada 20minutos, morre uma mulher em decorrência de parto, no mundo todo. Por isso, tais doenças poderão receber mais atenção do que outras. Esse fato fez com que houvesse certa preocupação com o índice de natalidade e morbidade materna, bem como morbidade e mortalidade perinatal. Portanto, abordou-se sobre sua biologia geral, fisiologia de reprodução, síntese de evolução genética, habitação, alimentação, manejo, dor e eutanásia, técnicas de riscos desenvolvidos na experimentação animal, estudos de experimentos farmacológicos e toxicológicos observados dentro dos artigos de revisão. Embora tendências atuais preconizem a utilização de métodos alternativos (estudo in vitro), os modelos animais, como as ratas, apresentam como principal vantagem o fornecimento de informações sobre o organismo como um todo, fato que não é obtido com outros métodos, o que ainda possibilita o seu emprego em pesquisas científicas.

We have carried out a literature review, with review articles, analyzing and discussing the works that have already been published on the effects of leucine aminopeptidade and aminopeptidase A in pre-term labor and preeclampsia. The proposal of this work on the subject is that most of the issues of maternal health seems childish, especially for service oriented maternity care, where, every 20 minutes, a woman dies due to childbirth, worldwide. Therefore, such diseases may receive more attention than others. This led to worry about the birth rate and maternal morbidity and perinatal morbidity and mortality. Therefore, it was addressed their general biology, physiology of reproduction, synthesis of evolution, genetics, housing, feeding, pain and euthanasia techniques developed for animal experimentation risks, studies of pharmacological and toxicological experiments observed within the review articles. Although current trends have preconized the use of alternative methods (in vitro study), animal models, such as rats, have as main advantage the provision of information on the organism as a whole, a fact that is not achieved with other methods, which also allows its use in scientific research.

First Isolation of Streptococcus gallolyticus subsp. pasteurianus from a Korean Patient with Severe Septic Shock / 대한임상미생물학회지

A 60-year-old man presented with a 1-day history of fever, vomiting, and diarrhea. He was diagnosed with severe septic shock on the basis of a body temperature of 38.9degrees C, heart rate of 92/min, respiratory rate of 25/min, WBC count of 22,970/microL, C-reactive protein (CRP) level of 136 mg/L, blood urea nitrogen (BUN) of 34.0 mg/dL, and creatinine of 2.98 mg/dL. On blood culture, Gram-positive cocci were detected in all 6 bottles. Small grayish non-hemolytic colonies were found on blood agar plates after incubation at 37degrees C for 2 days. The isolates were negative for catalase and L-pyrrolidonyl-beta-naphthylamide hydrolysis, and positive for bile-esculin and leucine aminopeptidase activity. The strain was identified as Streptococcus gallolyticus subsp. pasteurianus using Vitek 2 GP II systems. We performed 16S rRNA gene sequencing and detected 100% identity with S. gallolyticus subsp. pasteurianus strain CIP 107122T (1,345/1,345-bp). The patient recovered after receiving ampicillin-sulbactam. This is the first report of phenotypic and genetic identification of S. gallolyticus subsp. pasteurianus causing severe septic shock in a Korean patient.

Identification and estimation of leucine aminopeptidase activity in the serum and ascitic fluid of patients with schistosomiasis

The activity and electrophoretic mobility or leucine aminopeptidase [LAP] were assessed in the sera of 100 patients with different clinical stages of Schistosoma hacmatobium infestation as well as in 20 sample of ascctic fluids obtained from patients with bilharzial hepatosplenomegaly associated with ascites matched with 20 healthy adult males sample as control. Generally LAP activity was significantly higher in all patients. The activity of serum LAP showed a progressive rise with the advance in the clinical condition. The highest level was encountered in the sera of patients with bladder carcinoma and liver metastasis, leucine aminopeptidase was considerably found in the ascetic fluid. The ratio of the activity or LAP in the ascetic fluid to its activity in the serum of the same patients was 1:4.5 where the enzyme diffuses from the blood to the ascetic fluid. The enzyme detected by electrophoresis, in serum or ascetic fluid LAP migrated as a single component in the alpha1 -globulin faction. The results are discussed and compared with those in the literature

Effect of gallic acid on alkaline phosphatase and peptidase activities in rat intestine.

Gallic acid is a normal constituent of many edible foods, thus directly interacts with epithelial tissue in intestine. In the present study, the effect of gallic acid on intestinal alkaline phosphatase (IAP) and peptidase activities in rat intestine was evaluated. Gallic acid (0.27-0.5 mM) inhibited activities of leucine aminopeptidase (LAP) and -glutamyl transpeptidase (-GTP) by over 90%, compared to controls in rat intestine. In contrast, 0.1-0.6 mM gallic acid either had no effect or stimulated the activity of IAP in rat intestine. The observed inhibition of peptidases by gallic acid was reversible in nature. Kinetic analysis revealed no change in Vmax of LAP (0.42-0.44 units/mg protein) and -GTP (0.22-0.24 units/mg protein), while the values of apparent Km were increased 6-7 fold, exhibiting competitive-type of enzyme inhibition by gallic acid. The values of Ki for LAP and -GTP were 0.037 mM and 0.017 mM, respectively. These observations indicate that gallic acid is a potent inhibitor of brush border peptidases, and thus may interfere in the digestion and absorption of proteins in the intestine.

Morphometric and electrophoretic analysis of 13 populations of Anatolian black pine in Turkey.

The genetic variation in populations of Anatolian black pine (Pinus nigra Arn. subsp. pallasiana (L.) Holmboe.), one of the species covering large areas in Turkey, was investigated. Open pollinated seeds were collected from 13 populations in a natural distribution range. Six characters of seeds (length, width, ratio of length to width, weight/1000 seeds) and seedling characters (cotyledon number and hypocotyls height) and two enzyme systems viz. leucine aminopeptidase (LAP) and glutamate oxaloacetate transaminase, (GOT) were investigated. Significant differences were detected among the populations for the morphological characters. In addition, isozyme patterns of two enzyme systems revealed that LAP has two loci (one with 2 alleles and the other with 3), while GOT has three loci (two with 3 alleles and the third one with 2 alleles). Polymorphic loci were 74% on the average. The mean number of alleles per loci was 1.94 and expected heterozygosity was 19%. The mean total genetic diversity was calculated as 0.203; the mean gene diversity within populations was determined as 0.188, and the average between subpopulations diversity was 0.016. The relative magnitude of genetic differentiation among subpopulations was measured as 0.074 indicating that only 7.4% of the total genetic diversity was there between populations. Average genetic distance was 0.093 according to Gregorius. Nei's genetic distance was 0.022.

Localization of leucine aminopeptidase in the schistosoma mansoni Eggs and in liver tissue from infected mice

Infection with Schistosoma mansoni causes hepatic granuloma formation and fibrosis in response to parasite eggs. The present work localized the leucine aminopeptidase [LAP] in S. mansoni eggs and in liver tissue sections from infected mice. Fresh eggs and livers obtained from infected hamsters were processed and stained with the L-leucine-7-amino-trifluoro-methylcoumarin specific substrate. The L-argnine-7-amino-trifluoro-methylcoumarin and bestatin [leucine aminopeptidase inhibitor] were used to test the LAP substrate specificity and reactivity. The staining pattern for that enzyme in the egg and liver tissue reflects that the leucine aminopeptidase is a major egg constituent distributed in nearly all the egg except the spine. The control substrates confirmed the substrate broad specificity of LAP. It was concluded that the LAP enzyme is a major egg antigen and the target antigen for the antipathology vaccine development studies

Evaluation of leucine aminopeptidase and superoxide dismutase as markers in nasopharyngeal carcinoma

This study was designed to define changes of leucine aminopeptidase [LAP] and total superoxide dismutase [SOD] levels in nasopharyngeal carcinoma [NPC] patients after radiotherapy as a trial to correlate these parameters with disease activity and response to treatment. The study comprised 20 patients with NPC and 20 control subjects of those attending blood bank for blood donation. All patients underwent detailed history taking, full ENT clinical examination, and general examination for draining lymph nodes. Lesions characters and TNM staging were determined. All study participants gave blood samples for estimation of total blood SOD and serum LAP [pre-treatment] and were re-estimated 2 and 4 months after radiotherapy. There was a significant [P<0.05] increase in patients age and number of males [18:2] with significant increased incidence in smokers [16:4]. Eleven patients had stage I, 4 patients had stage II and 5 had stage III lesions. All patients with stage II and III lesions and 7 with stage I were smokers. Pre-treatment levels of LAP showed a significant [P<0.05] increase compared both to control, 2-m and 4-m levels with progressive steady decease with treatment. The levels detected 4-m after treatment showed a non-significant difference compared to control levels. The pretreatment SOD levels showed a significant [P<0.05] increase compared to both control and 4-m levels. Also, the SOD levels detected 2-m after treatment showed a significant [P<0.05] increase compared both to control and 4-m levels. There was a positive significant correlation between pretreatment levels of LAP, [r=0.85 1, P<0.001] and SOD, [r=0.643, P=0.002] and progress of tumor growth manifested as stage. We can conclude that pretreatment determination of total SOD and serum LAP correlate with severity and aggressiveness of NPC and could be used as an aid for staging. Furthermore, serum LAP correlates with response of NPC to radiotherapy and could be used to follow-up patients and as a prognostic marker

Effect of adrenalectomy on rat epididymidis / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the effect of adrenalectomy (ADX) on the epididymidis of Sprague-Dawley rats.</p><p><b>METHODS</b>The histological, biochemical (cholesterol protein, zinc, copper, alkaline and acid phosphatase aryl sulphatase, lactic dehydrogenase and leucine amino peptidase) and hormonal (FSH, LH and testosterone) changes of caput and cauda epididymis in ADX rats were observed.</p><p><b>RESULTS</b>Organ wet weight, histological studies and morphometric measurements indicated a cellular degeneration in caput and cauda epididymis of ADX rats. Serum testosterone level was significantly lower in ADX than in sham-operated rats, while the serum FSH and LH were below the detection limit of 1 mIU/mL. The enzymatic activity was higher in ADX than in sham-operated rats. Epididymal zinc level increased whereas copper level decreased in ADX rats compared to the sham-operated.</p><p><b>CONCLUSION</b>Adrenalectomy leads to degeneration of caput and cauda epididymidis epithelial cells as a result of decreased supply of testosterone.</p>

Ontogenetic patterns and genetic variations in Anopheles (Anopheles) intermedius Chagas, 1908 and Anopheles (Anopheles) mattogrossensis Lutz & Neiva, 1911 (Diptera: Culicidae) in the Brazilian amazon

Changes in the expression of genes were observed during development in populations of Anopheles (Anopheles) intermedius and Anopheles (Anopheles) mattogrossensis. Esterase showed seven zones of activity: EST1 was present in all developmental stages of both species; EST2 was observed only in larvae of A. intermedius and larvae and pupae of A. mattogrossensis, with greater activity in pupae; EST3 and EST5 were present in all development stages, with greater intensity in larvae; EST4 and EST6 showed weak activity in larvae of A. mattogrossensis and was not found in A. intermedius. Leucine aminopeptidase showed four zones of activity, of which LAP1 and LAP2 were found in all stages of A. intermedius, with highest activity in larvae, and in only of A. mattogrossensis. LAP3 was detected in all stages of A. mattogrossensis and in larvae only of A. intermedius. LAP4 was detected only in larvae and pupae of A. mattogrossensis, with greater intensity in pupae. Alpha-Glycerophosphate dehydrogenase showed a single zone of activity, detected in older fourth-instar larvae and becoming more intense from the pupal stage onwards.

Zinc, copper and hydrolytic enzymes in epididymis of hydrocortisone treated rat.

Administration of glucocorticoid (1, 2 and 4 mg) in excess leads to degeneration of epididymides as supported by cellular degeneration, sperm density and morphometric measurements. Zinc level increased statistically after 1, 2 and 4 mg hydrocortisone treatment while copper increased after 1 and 2 mg treatment. Cholesterol, protein and leucine aminopeptidase levels increased and decreased significantly in caput and cauda respectively. Activity of alkaline phosphatase reduced significantly while the treatment of hydrocortisone at different doses elevated acid phosphatase, aryl sulphatase and lactate dehydrogenase activities. Evidently, these changes are as a result of onset of cellular degeneration leading to impairment of metabolic/secretory activity of epididymal cells. The possible involvement of pituitary-testis axis in hydrocortisone induced epididymal degeneration and functional inhibition has been discussed.

Regulatory mechanisms of angiotensin II on the Na+/H+ antiport system in rabbit renal proximal tubule cells. I. Stimulatory effects of ANG II on Na+ uptake

The importance of the kidney in the development of hypertension was first demonstrated by Goldblatt and his colleagues more than fifty years ago. Many hormones and other regulatory factors have been proposed to play a major role in the development of hypertension. Among these factors angiotensin II (ANG II) is closely involved in renal hypertension development since it directly regulates Na+ reabsorption in the renal proximal tubule. Thus the aim of the present study was to examine signaling pathways of low dose of ANG II on the Na+ uptake of primary cultured rabbit renal proximal tubule cells (PTCs) in hormonally defined serum-free medium. The results were as follows: 1) 10-11 M ANG II has a significant stimulatory effect on growth as compared with control. Alkaline phosphatase exhibited significantly increased activity. However, leucine aminopeptidase and gamma-glutamyl transpeptidase activity were not significant as compared with control. In contrast to 10(-11) ANG II stimulated Na+ uptake (108.03 +/- 2.16% of that of control), 10(-9) M ANG II inhibited (92.42+/-2.23% of that of control). The stimulatory effect of ANG II on Na+ uptake was amiloride-sensitive and inhibited by losartan (ANG II receptor subtype 1 antagonist) and not by PD123319 (ANG II receptor subtype 2 antagonist). 2) Pertussis toxin (PTX) alone inhibited Na+ uptake by 85.52+/-3.52% of that of control. In addition, PTX pretreatment prevented the ANG II-induced stimulation of Na+ uptake. 8-Bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP), forskolin, and isobutylmethylxanthine (IBMX) alone inhibited Na+ uptake by 88.79+/-2.56, 80.63+/-4.38, and 84.47+/-4.74% of that of control, respectively, and prevented the ANG II-induced stimulation of Na+ uptake. However, 10(-11) M ANG II did not stimulate cAMP production. 3) The addition of 12-O-tetradecanoylphorbol-13-acetate (TPA, 0.01 ng/ml) to the PTCs produced significant increase in Na+ uptake (114.43+/-4.05% of that of control). When ANG II and TPA were added together to the PTCs, there was no additive effect on Na+ uptake. Staurosporine alone had no effect on Na+ uptake, but led to a complete inhibition of ANG II- or TPA-induced stimulation of Na+ uptake. ANG II treatment resulted in a 111.83 +/- 4.51% increase in total protein kinase C (PKC) activity. In conclusion, the PTX-sensitive PKC pathway is the main signaling cascade involved in the stimulatory effects of ANG II on Na+ uptake in the PTCs.

Biology of amazonian anopheline: XX. Ontogeny of esterases, leucine aminopeptidase and alpha-glycerophosphate dehydrogenase in Anopheles (Nyssorhynchus) darlingi Root, 1926 (Diptera, Culicidae)

The esterases, leucine aminopeptidase and alpha-glycerophosphate dehydrogenase revealed modifications in gene expressions during the development of Anopheles darlingi. The esterases showed five activity bands, 1 and 2 being more deeply stained during the larval stages than in pupae or adults, esterases 3 and 4 more deeply stained in pupae and adults whereas esterase 5 was present throughout development. Leucine aminopeptidase showed five activity bands: LAP2 and LAP5 were characteristic of larvae, LAP3 was specific for pupae and adults, LAP4 was detected only in pupae, and LAP1 and LAP6 were detected in all stages. Alpha-Glycerophosphate dehydrogenase presented one activity band on starch gel whose intensity increased with development. Two activity bands were detected on polyacrylamide gel (alpha-GPDH1 and alpha-GPDH2) in 4th-instar larvae (old pigmented larvae) and this activity increased with development.

Expression of brush border enzymes in ethanol fed rat intestine.

The effect of feeding ethanol daily for 40 days was studied on various brush border enzymes in rat intestine. Brush border alkaline phosphatase (AP), lactase, gamma-glutamyltranspeptidase (gamma-GTP), p-nitrophenyl (PNP)-beta-D-galactosidase (P < 0.01) and sucrase (P < 0.001) were significantly enhanced while leucine aminopeptidase and PNP-beta-D-glucosidase activities were unaltered in ethanol fed rats compared to the controls. Kinetic studies revealed that an increase in Vmax together with a decrease in affinity in case of gamma-GTP and an increase in Vmax for AP and sucrase were responsible for the observed stimulation of enzyme activities in ethanol administered rats. Significant changes in enzyme activities were observed in different populations of enterocytes along the crypt-villus unit in the ethanol fed animals. These observations suggest that ethanol feeding modifies the brush border enzymes in rat intestine but the underlying mechanisms seem to be distinct in differentiating enterocytes.

Serum leucine amino peptidase levels as a marker in testicular tumours.

Serum levels of leucine aminopeptidase (LAP) were studied in 60 patients with testicular tumours and 26 normal individuals, and were found to be significantly elevated in patients with seminomas, nonseminomas and benign tumours of the testis. After treatment the LAP levels decreased in patients with seminomas, nonseminomas and mixed cell carcinomas indicating response to therapy. The quantitative method used for estimation of LAP, which is simple, quick, accurate and cost-effective, could be used more effectively as a tumour marker in testicular malignancies.

Serum leucine aminopeptidase profile in cancers of gastro-intestinal tract with special reference to hepatic metastasis.

Serum leucine aminopeptidase (LAP) was estimated in 30 cases of gastro-intestinal cancers, and compared with 50 age and sex matched controls. Highly significant increase of serum LAP was seen in 7 patients with hepatic metastasis (p less than 0.001) from adenocarcinoma of stomach, colon and rectum. The enzyme values showed a highly significant increase in carcinoma of colon, when compared with different anatomical sites of the gastro-intestinal tract (p less than 0.001). However, in adenocarcinoma of stomach and rectum, significantly increased level of serum LAP was observed (p less than 0.01) which contrasted sharply with the normal enzyme values in squamous cell carcinoma of oesophagus and anal canal.

Kinetic properties of gamma-glutamyltranspeptidase and leucine-amino-peptidase in developing rat intestine: effect of hormones.

Administration of cortisone and thyroxine produced adult-type increase in the activities of soluble and membrane-bound gamma-glutamyltranspeptidase (gamma-GTP) in suckling rat intestine. Membrane-bound enzyme activity remained unaltered while the soluble enzyme activity was reduced (27%) in insulin-injected pups. Kinetic analysis revealed that the observed changes in the enzyme levels were a consequence of altered Vmax with no change in apparent Km. A 2-fold increase in the Km value was observed in adult gamma-GTP activity compared to that of suckling animals. Membrane-bound and soluble gamma-GTP yielded similar values of the Ea (9.7-13.1 kcal/mole) but exhibited apparent differences in heat stability in the control and hormone-injected groups. Leucine-amino peptidase(LAP) activity was reduced to adult levels in insulin-treated suckling animals. Thyroxine- and cortisone-treatment did not affect soluble activity but significantly (P less than 0.001) augmented the membrane-bound LAP levels. This increase was due to enhanced (54-82%) Vmax with no change in Km. The observed decrease in LAP activity in response to insulin was due to reduced Vmax. There was no change in Ea (8-11.6 kcal/mole) except the value was raised to 19.1 kcal/mole in cortisone-injected pups. Both the soluble and membrane-bound LAP activities were quite resistant to heat inactivation upto 30 min at 60 degrees C except in weanling rats. Thus, the kinetic behaviour of normally developed and precociously induced gamma-GTP and LAP is essentially similar but there are apparent differences in the mode of action of insulin, cortisone and thyroxine in affecting the development of these enzymes.

Serum leucine aminopeptidase estimation: a sensitive prognostic indicator of invasiveness in breast carcinoma.

Serum Leucine Aminopeptidase (LAP) was studied in 25 patients of breast tumors, and compared with 50 age matched controls. Clinical staging was done in 13 cases of breast carcinoma. Serum LAP was increased but not significantly in stage I (P less than 0.1), while in stage II & III, significant increase was observed (P less than 0.05), but a sharp peak was noted in stage IV with metastasis in liver (P less than 0.001). Serum LAP values were correlated with histological sub-types of breast carcinoma. Highest serum LAP values were observed in filtrating duct carcinoma with productive fibrosis (P less than 0.01), followed by medullary carcinoma and the enzyme values were slightly decreased in intraductal carcinomas.

Cyclic changes in salivary lactate dehydrogenase, peroxidase and leucine aminopeptidase during menstrual cycle.

Salivary lactate dehydrogenase (LDH), peroxidase (Px) and leucine aminopeptidase (LAP) activities were scanned in both normally ovulating and anovulating women during entire menstrual cycle. In ovulating women, all the three enzymes exhibited significant increase in the activity on or before the onset of ovulation which was monitored by the shift of the basal body temperature (BBT) as well as the ferning pattern of the cervical mucus. The peak maximum at the midcycle was several times higher than the previous day value in all the six normal women. In anovulatory women, no such remarkable change in the enzyme activities was found throughout the cycle. Salivary LDH and LAP showed peak at the midcycle and at the same time required short time for assay, so the present results are strongly suggestive that the determination of salivary enzyme content may be a convenient method for detecting the day of ovulation.

Significance of serum leucine aminopeptidase in tumours of female genital tract.

Serum leucine aminopeptidase (LAP) was estimated in 57 female patients with tumours of the genital tract and compared with 50 age-matched controls. Serum LAP was increased in leiomyomas (p less than 0.01) but significant rise was seen in leiomyosarcomas and endometrial carcinoma of the uterus (p less than 0.001). No increase in serum LAP level, above mean control value was observed in carcinoma of cervix. LAP levels were also increased in cystadenomas of the ovary (p less than 0.01) and highly significant increase, noted in cystadenocarcinomas and dysgerminoma stage III (p less than 0.001). However, no increase of enzyme activity was observed in poorly differentiated papillary carcinoma of the ovary. The serum enzyme level was higher in cystadenocarcinomas as compared to solid ovarian carcinomas in the same clinical stage.