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1.
Benef Microbes ; 14(4): 385-400, 2023 Sep 20.
Article in English | MEDLINE | ID: mdl-38661390

ABSTRACT

Honey bee colonies form a complex superorganism, with individual and social immune defences that control overall colony health. Sometimes these defences are not enough to overcome infections by parasites and pathogens. For that reason, several studies have been conducted to evaluate different strategies to improve honey bee health. A novel alternative that is being studied is the use of beneficial microbes. In a previous study, we isolated and characterised bacterial strains from the native gut microbiota of honey bees. Four Apilactobacillus kunkeei strains were mixed and administered in laboratory models to evaluate their potential beneficial effect on larvae and adult bees. This beneficial microbe mixture was safe; it did not affect the expression of immune-related genes, and it was able to decrease the mortality caused by Paenibacillus larvae infection in larvae and reduced the Nosema ceranae spore number in infected adult honey bees. In the present study, we aimed to delve into the impact of the administration of this beneficial microbe mixture on honey bee colonies, under field conditions. The mixture was administered in sugar syrup using lyophilised bacterial cells or fresh cultures, by aspersion or sprayed and feeder, once a week for three consecutive weeks, in autumn or spring 2015, 2017 and 2019. Colony strength parameters were estimated before the administration, and one and three months later. Simultaneously different samples were collected to evaluate the infection levels of parasites and pathogens. The results showed that administering the beneficial microbe mixture decreased or stabilised the infection by N. ceranae or Varroa destructor in some trials but not in others. However, it failed to improve the colony's strength parameters or honey production. Therefore, field studies can be a game-changer when beneficial microbes for honey bees are tested, and meticulous studies should be performed to test their effectiveness.


Subject(s)
Larva , Nosema , Bees/microbiology , Animals , Nosema/physiology , Larva/microbiology , Gastrointestinal Microbiome , Probiotics/pharmacology , Probiotics/administration & dosage , Honey , Paenibacillus larvae
2.
Actas urol. esp ; 40(8): 499-506, oct. 2016. tab, graf
Article in Spanish | IBECS | ID: ibc-156170

ABSTRACT

Objetivos: El objetivo de este estudio fue estimar la supervivencia global y desagregada en función de la escala de Gleason, de la edad y del tipo de extensión de una cohorte de pacientes con diagnóstico de cáncer de próstata avanzado de acuerdo a la práctica clínica habitual. Material y método: Se utilizó un diseño de estudio observacional y retrospectivo. En cada paciente se registraron variables clínicas como el tipo de extensión (metastásico o localmente avanzado), escala de Gleason, edad, fecha de diagnóstico, fecha del último contacto con el sistema de salud y el estado vital en el último contacto. Se utilizaron técnicas estadísticas de supervivencia de tipo univariante y multivariante. Los métodos de supervivencia paramétrica permitieron calcular la supervivencia media mediante extrapolación. Se analizaron 219 pacientes atendidos en el sistema de salud público entre 2008 y 2011. El análisis mostró diferencias estadísticamente significativas en la supervivencia en función de la escala de Gleason, la edad y el estadio. La supervivencia mayor se dio en el subgrupo menor de 75 años, con extensión local y categoría de bajo riesgo en la escala de Gleason (19,41 años) y la menor en el subgrupo opuesto, que fue de 0,97 años. La supervivencia de los otros subgrupos se movió entre esos valores extremos. Conclusión: La principal aportación de este trabajo consiste en calcular por primera vez la supervivencia media del CPA en España en relación con las variables de nuestra población de estudio. Esta información permite al clínico predecir la esperanza de vida de cada paciente según sus factores pronósticos


Objectives: The aim of this study was to determine the overall and disaggregated survival based on the Gleason score, age and extent of a patient cohort diagnosed with advanced prostate cancer according to standard clinical practice. Material and method: We used an observational and retrospective design for the study. For each patient, we recorded clinical variables such as the extent (metastatic or locally advanced), Gleason score, age, date of diagnosis, date of last contact with the health system and the vital status during the last contact. We used univariate and multivariate statistical techniques of survival. The parametric survival methods enabled us to calculate the mean survival using extrapolation. We analysed 219 patients treated in the public health system between 2008 and 2011. The analysis showed statistically significant differences in survival depending on Gleason score, age and stage. The longest survival was in the subgroup younger than 75 years, with a local extent and a low-risk category on the Gleason scale (19.41 years), and the shortest survival (0.97 years) was in the 75 years or older group. The survival of the other subgroups ranged between these outliers. Conclusion: The main contribution of this study is that it is the first to calculate the mean survival of advanced prostate cancer in Spain in terms of the variables of our study population. This information helps clinicians predict the life expectancy of each patient according to their prognostic factors


Subject(s)
Humans , Male , Aged , Survivorship/physiology , Prostatic Neoplasms/mortality , Prostatic Neoplasms/pathology , Neoplasm Staging/methods , Life Expectancy/trends , Survival Rate , Age Factors , Retrospective Studies , Prognosis , Observational Study , Spain/epidemiology
3.
Actas Urol Esp ; 40(8): 499-506, 2016 Oct.
Article in English, Spanish | MEDLINE | ID: mdl-27174571

ABSTRACT

OBJECTIVES: The aim of this study was to determine the overall and disaggregated survival based on the Gleason score, age and extent of a patient cohort diagnosed with advanced prostate cancer according to standard clinical practice. MATERIAL AND METHOD: We used an observational and retrospective design for the study. For each patient, we recorded clinical variables such as the extent (metastatic or locally advanced), Gleason score, age, date of diagnosis, date of last contact with the health system and the vital status during the last contact. We used univariate and multivariate statistical techniques of survival. The parametric survival methods enabled us to calculate the mean survival using extrapolation. We analysed 219 patients treated in the public health system between 2008 and 2011. The analysis showed statistically significant differences in survival depending on Gleason score, age and stage. The longest survival was in the subgroup younger than 75 years, with a local extent and a low-risk category on the Gleason scale (19.41 years), and the shortest survival (0.97 years) was in the 75 years or older group. The survival of the other subgroups ranged between these outliers. CONCLUSION: The main contribution of this study is that it is the first to calculate the mean survival of advanced prostate cancer in Spain in terms of the variables of our study population. This information helps clinicians predict the life expectancy of each patient according to their prognostic factors.


Subject(s)
Prostatic Neoplasms/mortality , Prostatic Neoplasms/pathology , Age Factors , Aged , Humans , Male , Neoplasm Grading , Neoplasm Staging , Retrospective Studies , Spain/epidemiology , Survival Rate
4.
Actas Urol Esp ; 40(1): 3-10, 2016.
Article in English, Spanish | MEDLINE | ID: mdl-26115777

ABSTRACT

OBJECTIVES: To present a National Registry of patients with prostate cancer as monitored through active surveillance, with the intention of testing the hypothesis that cancer-specific mortality in very low-risk and low-risk patients is less than 5% at 15 years. MATERIAL AND METHODS: A multicentre observational study (AEU-PIEM/2014/0001) sponsored by the Spanish Association of Urology was conducted using their platform for multicentre studies. The clinical-pathological inclusion criteria were as follows: cT1a-cT3a, PSA ≤ 20 ng/ml, initial minimum biopsy of 10 cores, number of affected cores ≤ 3, 1st Gleason score of 3 and 2nd Gleason score ≤ 4 and a known prostate volume (in cc). A unified follow-up was not established for all recruiting centres; however, a survey was conducted that reflects the follow-up characteristics based on a number of tangible parameters that allow for their comparison. With the same philosophy of flexibility, the use of certain biomarkers and multiparametric MRI was not considered necessary for inclusion. RESULTS: We describe the Registry's characteristics and possibilities, as well as the preliminary results from the 324 patients included in its first 5 months of operation in the 15 recruiting centres. We also report the clinical-pathological variables, biomarkers, radiodiagnosis technique and quality-of-life questionnaires considered for the database, as well as the possibilities for indefinite follow-up, remaining open to any active treatment recognized in clinical guidelines. CONCLUSIONS: The AEU-PIEM/2014/0001 represents an extremely useful tool for all Spanish urologists for multicentre clinical research. The registry will undoubtedly enable the dissemination of active surveillance of our patients in a more coordinated manner, thus maintaining the advantages of optimised opportunistic screening for prostate cancer without resulting in overtreatment.


Subject(s)
Prostatic Neoplasms/therapy , Registries , Watchful Waiting , Adult , Aged , Humans , Male , Middle Aged , Prospective Studies , Prostatic Neoplasms/mortality , Retrospective Studies , Societies, Medical , Spain , Survival Rate , Time Factors , Urology
5.
Rev. neurol. (Ed. impr.) ; 49(7): 354-358, 1 oct., 2009. tab, ilus
Article in Spanish | IBECS | ID: ibc-94835

ABSTRACT

Introducción. La craniectomía descompresiva aumenta la supervivencia en los infartos malignos de la arteria cerebral media (ACM). Se analizan los signos radiológicos y clínicos que predicen la evolución maligna del infarto de la ACM,y factores asociados a un peor pronóstico. Pacientes y métodos. Se estudian 30 pacientes divididos en tres grupos: pacientes operados, y pacientes no operados con ingreso en cuidados intensivos o en planta de neurología. La técnica quirúrgica consistióen la creación de una ventana ósea de al menos 10 cm de diámetro y apertura dural. Para la valoración inicial del paciente se utilizó la escala de Glasgow y la escala de ictus del National Institute of Health, y para el seguimiento, la escala modificadade Rankin, el índice de Barthel y la Glasgow Outcome Scale a los seis meses. Resultados. Los pacientes más jóvenes tienen un mejor pronóstico funcional que los mayores de 60 años. La desviación de la línea media mayor de 10 mm se asocia con un peor pronóstico, al igual que volúmenes de tejido infartado mayores de 350 cm3. Menor puntuación en la escala de Glasgow al ingreso se asocia a peor pronóstico vital y a mayor número de secuelas, así como su disminución durante el ingreso. Conclusiones. La edad condiciona la presencia de secuelas en estos pacientes. La presencia de signos clínicos de herniación (anisocoria, menor puntuación inicial o descenso importante en la escala de Glasgow) y radiológicos (desplazamiento de la línea media, volumen infartado) implica un peor pronóstico. La cirugía precoz en aquellos pacientes en que estuviera indicada reduce el número de secuelas y aumenta la supervivencia (AU)


Introduction. Decompressive craniectomy increases the survival rate in cases of malignant middle cerebral artery (MCA) stroke. The imaging and clinical signs that predict a malignant progression of stroke of the MCA are analysed, together with factors associated with a poorer prognosis. Patients and methods. The study involved 30 patients, who were divided into three groups: patients who had undergone surgery, and patients who had not undergone surgery but were admitted to intensive care or to neurology wards. The surgical procedure consisted in creating a bone window with a diameter of at least 10 cm and a dural opening. The initial evaluation of the patient was performed using the Glasgow scale and the National Institute of Health stroke scale; follow-up was carried out using the modified Rankin scale, the Barthel index and the Glasgow Outcome Scale at six months. Results. Younger patients have a better functional prognosis than those over 60 years of age. A deviation of more than 10 mm from the mean line is associated with a poorer prognosis, as are volumes of infarcted tissue above 350 cm3. Lower scores on the Glasgow scale on admission are associated with a poorer prognosis for survival and a higher number of sequelae, as well as their reduction during hospitalisation. Conclusions. Age conditions the presence of sequelae in these patients. The presence of clinical signs of herniation (anisocoria, lower initial score or important drop on the Glasgow scale) and imaging signs (displacement of the mean line, volume of infarcted tissue) imply a poorer prognosis. Early surgery in those patients in whom it is indicated reduces the number of sequelae and increases the rate of survival (AU)


Subject(s)
Humans , Infarction, Middle Cerebral Artery/surgery , Decompressive Craniectomy , Statistics on Sequelae and Disability , Risk Factors , Anisocoria/complications , Glasgow Outcome Scale
6.
Rev Neurol ; 49(7): 354-8, 2009.
Article in Spanish | MEDLINE | ID: mdl-19774529

ABSTRACT

INTRODUCTION: Decompressive craniectomy increases the survival rate in cases of malignant middle cerebral artery (MCA) stroke. The imaging and clinical signs that predict a malignant progression of stroke of the MCA are analysed, together with factors associated with a poorer prognosis. PATIENTS AND METHODS: The study involved 30 patients, who were divided into three groups: patients who had undergone surgery, and patients who had not undergone surgery but were admitted to intensive care or to neurology wards. The surgical procedure consisted in creating a bone window with a diameter of at least 10 cm and a dural opening. The initial evaluation of the patient was performed using the Glasgow scale and the National Institute of Health stroke scale; follow-up was carried out using the modified Rankin scale, the Barthel index and the Glasgow Outcome Scale at six months. RESULTS: Younger patients have a better functional prognosis than those over 60 years of age. A deviation of more than 10 mm from the mean line is associated with a poorer prognosis, as are volumes of infarcted tissue above 350 cm3. Lower scores on the Glasgow scale on admission are associated with a poorer prognosis for survival and a higher number of sequelae, as well as their reduction during hospitalisation. CONCLUSIONS: Age conditions the presence of sequelae in these patients. The presence of clinical signs of herniation (anisocoria, lower initial score or important drop on the Glasgow scale) and imaging signs (displacement of the mean line, volume of infarcted tissue) imply a poorer prognosis. Early surgery in those patients in whom it is indicated reduces the number of sequelae and increases the rate of survival.


Subject(s)
Decompressive Craniectomy , Infarction, Middle Cerebral Artery/surgery , Adult , Age Factors , Aged , Aged, 80 and over , Decompressive Craniectomy/methods , Decompressive Craniectomy/statistics & numerical data , Female , Glasgow Outcome Scale , Humans , Infarction, Middle Cerebral Artery/complications , Infarction, Middle Cerebral Artery/diagnosis , Infarction, Middle Cerebral Artery/pathology , Male , Middle Aged , Prognosis , Survival Rate , Treatment Outcome
9.
Actas Dermosifiliogr ; 97(9): 581-2, 2006 Nov.
Article in Spanish | MEDLINE | ID: mdl-17173762

ABSTRACT

Pleomorphic fibroma of the skin was described by Kamino et al 1 in 1989. It is a benign fibroblastic proliferation with cytologic atypia. This article describes a case of a myxoid variant of pleomorphic fibroma of the skin.


Subject(s)
Fibroma/pathology , Skin Neoplasms/pathology , Fibroma/surgery , Hallux , Humans , Immunohistochemistry , Male , Middle Aged , Skin Neoplasms/surgery
10.
Actas dermo-sifiliogr. (Ed. impr.) ; 97(9): 581-582, nov. 2006. ilus
Article in Es | IBECS | ID: ibc-049188

ABSTRACT

El fibroma pleomórfico de la piel fue descrito por Kamino et al en 1989. Es una proliferación fibroblástica benigna en la que existe atipia citológica. Este artículo describe un caso de una variante mixoide del fibroma cutáneo pleomórfico


Pleomorphic fibroma of the skin was described by Kamino et al 1 in 1989. It is a benign fibroblastic proliferation with cytologic atypia. This article describes a case of a myxoid variant of pleomorphic fibroma of the skin


Subject(s)
Middle Aged , Male , Humans , Leiomyosarcoma/diagnosis , Leiomyosarcoma/therapy , Fibroma/complications , Fibroma/diagnosis , Immunohistochemistry/methods , Diagnosis, Differential , Histiocytoma, Benign Fibrous/complications , Histiocytoma, Benign Fibrous/diagnosis , Histiocytoma, Benign Fibrous/surgery , Liposarcoma/complications , Liposarcoma/diagnosis , Carcinoma/complications , Carcinoma/diagnosis
11.
J Chem Phys ; 120(18): 8815-23, 2004 May 08.
Article in English | MEDLINE | ID: mdl-15267813

ABSTRACT

The effect of molecular structure on the gamma relaxation dynamics has been studied in a set of aromatic poly(isophthalamide)s. This polymer family differ in the bridge group between phenylene rings [hexafluoroisopropylidene (C(CF(3))(2)) or ether] and also in the presence of t-butyl groups (C(CH(3))(3)) as pendant substituent on the five position of isophthalic ring. The results obtained from wide angle x-ray scattering in the glassy state indicated that both (C(CF(3))(2)) and (C(CH(3))(3)) groups favor the separation between chains, which is reflected on different interchain average distances. Dielectric experiments showed that both bulky groups favor the mobility in the glassy state. Molecular modelling methods were used to know the kind of molecular motions associated to the dielectric relaxation observed below the glass transition temperature.

12.
J Clin Endocrinol Metab ; 86(12): 5934-43, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11739467

ABSTRACT

The role of cGMP in the regulation of human myometrial smooth muscle contractility is at present unclear. cGMP can be synthesized by a cytoplasmic, soluble guanylate cyclase (sGC), which is stimulated by nitric oxide and carbon monoxide, and by particulate membrane-bound GC, which are activated by natriuretic peptides. The aim of this study was to determine whether sGC or pGC are present in nonpregnant and pregnant human myometrium, and whether the activity and expression of these enzymes and the cGMP content change during pregnancy and with labor. Myometrium was obtained from nonpregnant women (n = 12) and pregnant women who were preterm (25-34 wk gestation; n = 12), term (>38 wk) not in labor (n = 14), or term in active labor (n = 12). The cGMP content in myometrium obtained from preterm deliveries was significantly higher than that in tissue obtained from nonpregnant women and decreased at term, especially in laboring groups. Protein and mRNA for sGC, particulate GC-A, GC-B, and the clearance receptor were detected in human myometrium. cGMP in pregnant human myometrium, however, appears to be produced predominantly by sGC and possibly by GC-B, as GC-A was only weakly expressed. sGC activity was greater in myometrium from preterm (nonlabor) deliveries compared those taken at term (in labor), but was down-regulated compared with activity in nonpregnant myometrium. Neither atrial natriuretic peptide nor C-type natriuretic peptide (agonists for GC-A and GC-B, respectively) altered contractility in vitro of myometrium from women at term (not in labor). We conclude that the cGMP/guanylate cyclase system in human myometrium is gestationally regulated and potentially plays an important role in mediating quiescence during early pregnancy. A reduction in cGMP availability may contribute to the switch to contractile activity at term.


Subject(s)
Guanylate Cyclase/metabolism , Myometrium/enzymology , Pregnancy/metabolism , Atrial Natriuretic Factor/pharmacology , Cyclic GMP/metabolism , Down-Regulation , Female , Guanylate Cyclase/genetics , Humans , Immunohistochemistry , Isoenzymes/genetics , Isoenzymes/metabolism , Labor, Obstetric/metabolism , Myometrium/physiology , Natriuretic Peptide, C-Type/pharmacology , RNA, Messenger/metabolism , Receptors, Atrial Natriuretic Factor/genetics , Receptors, Atrial Natriuretic Factor/metabolism , Reference Values , Reverse Transcriptase Polymerase Chain Reaction , Solubility , Uterine Contraction/drug effects
13.
Arch Cardiol Mex ; 71(1): 34-42, 2001.
Article in Spanish | MEDLINE | ID: mdl-11565360

ABSTRACT

We reviewed our work and evaluated clinical and angiographic results, as well as the follow up of young adult patients < 40 years that were subject to percutaneous trans-clinical coronary angioplasty (PTCA) with an endovascular prostheses (stent). From January 1995 to December 1999, 896 PCTA's performed using stents in 770 patients. Only one selected group of 30 patients (with 32 procedures, and an average of 1.06 stents per patients. Patients age ranged from 21 to 39 years old with an average of 32.8 + 52, 2, 29 (96%) were male and only one woman (3.3%). Nine patients (30%) had a severe angina, class III Braunwald and 21 (70%) had a previous history of myocardial infarcts. The average percent of arterial obstruction was 90.88 +/- 5.22 and the expulsion fraction. (EF) had a percent of 46.8 +/- 4.3 with ranges of 35 to 60%. Immediate angiographic success was 93.75% in only two patients (6.25%) with a 100% occlusion, and more than twelve weeks. There was no mortality, nor infarct, and no patient was sent to an urgent revascularization surgery. Clinical follow up lasted 50 weeks in all patients with the effort test using the electric Bruce type and for nuclear medicine. Only in 3 patients EF was reported, and tHalium with a slight septal ischemia. Angiographic control was included in 27 (90% patients between the 4th and 6th month. PTA with stent is a successful angiographic and clinical procedure in young adults < 40 years old; it constitutes a complete and efficient therapeutical revascularization procedure, and is an excellent option before undertaking an aortocoronary revascularization surgery.


Subject(s)
Angioplasty, Balloon, Coronary , Coronary Disease/therapy , Stents , Adult , Age Factors , Female , Follow-Up Studies , Humans , Male
14.
Arch Inst Cardiol Mex ; 70(1): 38-45, 2000.
Article in Spanish | MEDLINE | ID: mdl-10855409

ABSTRACT

UNLABELLED: We examined the immediate and short-term outcomes after stenting protected and unprotected left main coronary artery (LMCA) stenoses, in patients with normal ventricular function. Left main coronary artery disease is regarded as an absolute contraindication for coronary angioplasty, because it has been associated with high procedural morbidity and poor mid-term results. Between february 1995 and february 1999, 596 procedures were performed in 468 patients. Ten patients who had disease involving the left main coronary artery were included. They were not candidates for coronary surgery. The post-stent antithrombotic regimens were aspirin and ticlopidine. The procedural success rate was 100% without episodes of subacute thrombosis. Three to six months follow-up angiography was performed in all, restenosis occurred only in two patients, there were two repeat PTCA (20%) and there were no deaths. CONCLUSIONS: Stenting of unprotected and protected left main coronary artery stenoses may be a safe and effective alternative to surgery in carefully selected patients with normal left ventricular function. The results of our study suggests that when patients have prohibitive surgical risks, elective LMCA angioplasty and/or stenting maybe undertaken with a high procedural success rate as an effective alternative to CABG in carefully selected patients. Further studies in larger patient populations are needed to assess late outcome.


Subject(s)
Coronary Disease/surgery , Stents , Aged , Coronary Disease/diagnostic imaging , Follow-Up Studies , Humans , Male , Middle Aged , Radiography , Time Factors
15.
Placenta ; 21(2-3): 142-9, 2000.
Article in English | MEDLINE | ID: mdl-10736236

ABSTRACT

Pregnant women with active systemic lupus erythematosus (SLE) and/or the antiphospholipid syndrome (APS) are prone to recurrent miscarriage, pre-eclampsia, intrauterine growth restriction and premature delivery. Placental dysfunction may account for these complications yet the mechanisms remain uncertain. Amongst these, an inflammatory response in the placental vasculature could play a role, involving recruitment of neutrophils and platelets and the increased endothelial expression of cell adhesion molecules (CAM), central to the recruitment process. The aim of this study was primarily to investigate CAM expression in the fetoplacental vasculature in women with SLE/APS. Circulating maternal concentrations of soluble CAM were also elucidated. There were no differences in CAM immunostaining in placentae from patients with SLE and/or APS compared with controls. In both patients and controls moderate immunostaining for the intercellular adhesion molecule-1 (ICAM-1) was observed in placental vascular endothelium and mild immunostaining was present in the placental villous stroma. Strong immunostaining for platelet endothelial CAM (PECAM) occured in the placental vascular endothelium whereas P-selectin was mildly expressed in the stem vessel endothelium only. Vascular CAM-1 (VCAM-1) and E-selectin were undetectable in either study or control placentae. In contrast, ICAM-1 and VCAM-1 but not E-selectin, as assessed by immunoassay (ELISA), were elevated in maternal serum from SLE/APS patients compared with controls. This study suggests that upregulation of CAM expression and subsequent activation of neutrophil and/or platelet activity within the placental villous tree is unlikely to be a mechanism by which the adverse pregnancy outcome arises in SLE/APS pregnancies. However, maternal endothelial cell activation (ECA) may play a more important role.


Subject(s)
Antiphospholipid Syndrome/complications , Antiphospholipid Syndrome/immunology , Cell Adhesion Molecules/metabolism , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/immunology , Placenta/immunology , Pregnancy Complications/immunology , Adult , Case-Control Studies , E-Selectin/metabolism , Female , Humans , Immunohistochemistry , Intercellular Adhesion Molecule-1/metabolism , Pregnancy , Vascular Cell Adhesion Molecule-1/metabolism
16.
J Physiol ; 521 Pt 3: 705-16, 1999 Dec 15.
Article in English | MEDLINE | ID: mdl-10601500

ABSTRACT

1. Endogenous nitric oxide has been proposed to play a role in the control of myometrial contractility in pregnancy. In this study, the expression, localisation and regulation of nitric oxide synthase (NOS) isoforms have been examined in human pregnant myometrium and cultured human myometrial smooth muscle cells, by immunoblotting, immunohistochemistry and reverse transcription-polymerase chain reaction. 2. Immunoblotting of extracts from freshly isolated myometrial tissue, affinity-enriched for NOS proteins by precipitation with ADP-sepharose, revealed expression of endothelial NOS (eNOS or NOS3) in tissues from preterm, term non-labour and active labour at term. Inducible NOS (iNOS or NOS2) and neuronal NOS (nNOS or NOS1) proteins were not detected at any stage of pregnancy. 3. Immunohistochemical detection showed that expression of eNOS protein was restricted to the endothelium of the myometrial vasculature, with no staining detected in myometrial smooth muscle cells. 4. Messenger RNA for all three NOS isoforms was detected, although iNOS and nNOS mRNAs were detectable only with high cycle number, implying a low copy number. 5. NOS isoforms were not detectable in human myometrial smooth muscle cells cultured from term non-labour pregnancies. Cytokine stimulation of cultured myometrial cells did not induce iNOS expression or nitrite accumulation in the culture medium, although both iNOS protein and nitrite release were detected in the human pulmonary epithelial cell line A549. 6. Levels of eNOS protein and of NOS mRNA expression were not correlated with gestational stage, suggesting that endogenously produced NO is not likely to be a modulator of myometrial tone during human pregnancy.


Subject(s)
Myometrium/enzymology , Nitric Oxide Synthase/biosynthesis , Adult , Cells, Cultured , Cytokines/pharmacology , Female , Humans , Immunoblotting , Immunohistochemistry , Isoenzymes/biosynthesis , Labor, Obstetric/physiology , Lipopolysaccharides/pharmacology , Muscle, Smooth/enzymology , Muscle, Smooth/ultrastructure , Myometrium/ultrastructure , Nitrites/metabolism , Pregnancy , RNA, Messenger/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction
17.
Mol Hum Reprod ; 5(9): 880-4, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10460228

ABSTRACT

Human labour is associated with increased prostaglandin synthesis within the uterus. The aim of this study was to examine the expression of the two isoforms of the central prostaglandin synthetic enzyme, cyclo-oxygenase (COX-1 and COX-2) in human myometrium throughout pregnancy and to test the hypothesis that COX in the myometrium may play a role in labour onset. Expression of COX-1 and COX-2 at the mRNA level was analysed using reverse transcriptase-polymerase chain reaction (RT-PCR) and at the protein level using Western blotting. No significant changes of COX-1 RNA or protein expression were observed either with gestational age or labour. COX-2 mRNA and protein expression increased at term with significant up-regulation occurring prior to the onset of labour (P < 0.005). These data would suggest that up-regulation of COX-2, rather than COX-1, mediates increased prostaglandin synthesis in human myometrium at term. The increased COX-2 expression observed preceded labour onset, suggesting that COX-2 has a role in labour onset, rather than its presence merely a consequence of labour.


Subject(s)
Isoenzymes/genetics , Isoenzymes/metabolism , Myometrium/enzymology , Pregnancy/metabolism , Prostaglandin-Endoperoxide Synthases/genetics , Prostaglandin-Endoperoxide Synthases/metabolism , Adolescent , Adult , Blotting, Western , Cesarean Section , Cyclooxygenase 1 , Cyclooxygenase 2 , Female , Humans , Labor, Obstetric/physiology , Membrane Proteins , Pregnancy Trimester, Second , Pregnancy Trimester, Third , Reverse Transcriptase Polymerase Chain Reaction
18.
Br J Obstet Gynaecol ; 106(5): 439-45, 1999 May.
Article in English | MEDLINE | ID: mdl-10430194

ABSTRACT

OBJECTIVE: To determine whether corticotrophin releasing hormone plays a role in the regulation of tone in term nonlabouring human myometrium. SETTING: A teaching hospital research laboratory. SAMPLE: Thirty-seven women undergoing elective nonlabour caesarean section under regional anaesthesia. METHODS: Human corticotrophin releasing hormone (1, 10, 100 nmol/L) was added to strips of term, nonlabouring myometrium mounted in an organ bath, and the effect on spontaneous, oxytocin (1 nmol/L) or prostaglandin F2alpha (100 nmol/L) stimulated contractions determined. Cyclic adenosine monophosphate (cAMP) content of the tissue was also determined by enzyme immunoassay. RESULTS: Corticotrophin releasing hormone did not affect myometrial tension development in any of the experimental protocols. cAMP increased transiently after addition of corticotrophin releasing hormone (166.7 +/- 12.7% at 1 minute) but this was not reflected by any change in tension. CONCLUSIONS: This study suggests that despite high maternal plasma concentrations of corticotrophin releasing hormone in pregnancy at term, this peptide is unlikely to play a direct role in the control of myometrial contractility in nonlabouring myometrium.


Subject(s)
Corticotropin-Releasing Hormone/pharmacology , Myometrium/drug effects , Uterine Contraction/drug effects , Cesarean Section , Female , Humans , Oxytocin/pharmacology , Pregnancy
19.
Am J Obstet Gynecol ; 181(1): 180-9, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10411817

ABSTRACT

OBJECTIVE: Placentas from pregnancies complicated by antiphospholipid syndrome often show thromboses and infarction, which may result from aberrations in placental coagulant pathways. We tested the hypothesis that alterations in tissue factor, thrombomodulin, and annexin V expressions contribute to poor pregnancy outcome associated with antiphospholipid syndrome. STUDY DESIGN: Frozen sections from random biopsy samples of the basal plates of placentas from patients with primary antiphospholipid syndrome (n = 9), patients with secondary antiphospholipid syndrome (n = 3), and gestational age-matched control subjects (n = 10) were immunostained for tissue factor, thrombomodulin, and annexin V. Intensity of immunostaining was assessed by means of quantitative image analysis. Annexin V protein expression was evaluated with Western blotting techniques. RESULTS: Tissue factor was expressed in the perivascular cells of the villous vasculature. Thrombomodulin and annexin V immunostaining was localized to the syncytiotrophoblast. There were no differences in the intensity of immunostaining for tissue factor, thrombomodulin, and annexin V between placentas from women with antiphospholipid syndrome and those from control subjects. Western blot analysis of annexin V expression showed no differences between study patients and control subjects. CONCLUSION: Alterations in placental coagulant pathways involving tissue factor, thrombomodulin, and annexin V do not contribute to poor pregnancy outcome associated with antiphospholipid syndrome.


Subject(s)
Annexin A5/metabolism , Antiphospholipid Syndrome/metabolism , Placenta/metabolism , Pregnancy Complications/metabolism , Thrombomodulin/metabolism , Thromboplastin/metabolism , Adult , Antiphospholipid Syndrome/pathology , Blotting, Western , Case-Control Studies , Female , Humans , Immunohistochemistry , Placenta/pathology , Pregnancy , Pregnancy Complications/pathology , Pregnancy Outcome , Prospective Studies
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