Multi-effect of the water-soluble Moringa oleifera lectin against Serratia marcescens and Bacillus sp.: antibacterial, antibiofilm and anti-adhesive properties.

AIMS: To evaluate the antibiofilm potential of water-soluble Moringa oleifera seed lectin (WSMoL) on Serratia marcescens and Bacillus sp. METHODS AND RESULTS: WSMoL inhibited biofilm formation by S. marcescens at concentrations lower than 2·6 µg ml-1 and impaired bacterial growth at higher concentrations, avoiding biofilm formation. For Bacillus sp., the lectin inhibited bacterial growth at all concentrations. The antibiofilm action of WSMoL is associated with damage to bacterial cells. WSMoL did not disrupt preformed S. marcescens biofilms but was able to damage cells inside them. On the other hand, the lectin reduced the number of cells in Bacillus sp. biofilm treated with it. WSMoL was able to control biofilm formation when immobilized on glass surface (116 µg cm-2 ), damaging S. marcescens cells and avoiding adherence of Bacillus sp. cells on glass. The Bacillus sp. isolate is member of Bacillus subtilis species complex and closely related to species of the conspecific 'amyloliquefaciens' group. CONCLUSION: WSMoL prevented biofilm development by S. marcescens and Bacillus sp. and the antibiofilm effect is also observed when the lectin is immobilized on glass. SIGNIFICANCE AND IMPACT OF THE STUDY: Taking together, our results provide support to the potential use of WSMoL for controlling biofilm formation by bacteria.

Antibiofilm activity of Cobetia marina filtrate upon Staphylococcus epidermidis catheter-related isolates.

We report the antibiofilm activity by the sponge-associated bacterium Cobetia marina upon Staphylococcus epidermidis clinical isolates obtained from central venous catheters. Antibiofilm activity/antimicrobial susceptibility correlation might predict the action of the metabolite(s) upon Staphylococcus epidermidis in the clinic, making it a possible adjuvant in therapies against biofilm-associated infections.

Hydrolyzable tannins from Poincianella (Caesalpinia) microphylla fruits: Metabolite profiling and anti-Trichomonas vaginalis activity.

Poincianella (Caesalpinia) microphylla fractions were analyzed by LC-DAD-MS and by statistical tools to determine the possible bioactive compounds against T. vaginalis. Tannins enriched-fractions toxicity (M5 and M10) were analyzed in in vitro against human red cells and in in vivo model of Galleria mellonella. Twenty-six compounds were detected from the P. microphylla fractions. The main compounds identified were hydrolyzable tannins (gallotanins and ellagitannis), such as O-digalloyl hexoside, O-digalloyl HHDP -hexoside, tri-O-galloyl HHDP-hexoside, O-galloyl HHDP-DHHDP-hexoside and their isomers. In addition, procyanidin dimer, epicatechin, ellagic acid and O-(digalloyl) quinic acid were also identified. Based on univariate statistical analyses, stronger correlations with the anti-T. vaginalis properties were observed for the compounds 7 (O-galloyl HHDP-DHHDP-hexoside - Geraniin isomer), 3 (O-digalloyl HHDP -hexoside - mallotinic acid), 2 (O-digalloyl) quinic acid), 1 (O-digalloyl hexoside) and 9 (unknown). Tannins enriched-fractions (M5 and M10) presented anti-Trichomonas activity (IC50 70.41 µg/mL and 142.1 µg/mL, respectively) and no toxicity in the in vivo model of G. mellonella. This innovative approach allowed us to identify likely bioactive compounds in the extracts, although the mechanism(s) underlying anti-trichomonal activity encompass a complex trait.

Myricetin protects Galleria mellonella against Staphylococcus aureus infection and inhibits multiple virulence factors.

Staphylococcus aureus is an opportunistic pathogen related to a variety of life-threatening infections but for which antimicrobial resistance is liming the treatment options. We report here that myricetin, but not its glycosylated form, can remarkably decrease the production of several S. aureus virulence factors, including adhesion, biofilm formation, hemolysis and staphyloxanthin production, without interfering with growth. Myricetin affects both surface proteins and secreted proteins which indicate that its action is unrelated to inhibition of the agr quorum sensing system. Analysis of virulence related gene expression and computational simulations of pivotal proteins involved in pathogenesis demonstrate that myricetin downregulates the saeR global regulator and interacts with sortase A and α-hemolysin. Furthermore, Myr confers a significant degree of protection against staphylococcal infection in the Galleria mellonella model. The present findings reveal the potential of Myr as an alternative multi-target antivirulence candidate to control S. aureus pathogenicity.

The parachute-like asymmetric mitral valve and its two papillary muscles.

OBJECTIVES: The morphologic features of parachute-like asymmetric mitral valves are described to discriminate this anomaly from parachute mitral valves. BACKGROUND: Mitral valves with unifocal attachment of chords have been called "parachute valves," independent of the number of papillary muscles. Therefore the anomaly involving two papillary muscles has not received separate attention. METHODS: The gross anatomy of 29 mitral valves with focalized attachment of chords was studied. RESULTS: In 28 of the autopsy specimens asymmetric mitral valves with two papillary muscles were present, and one of the muscles was elongated, located higher in the left ventricle with its tip reaching to the anulus, and attached at both its base and lateral side to the left ventricular wall. The valve leaflets could be directly attached to this abnormal muscle that received few chords or, in three hearts, no chords at all, resulting in an oblique and eccentric orifice. Because of the focalized attachment of chords to one of the two papillary muscles, we call this malformation "parachute-like asymmetric mitral valve," We found only one "true parachute mitral valve," that is, one having a single papillary muscle that received all chords. CONCLUSIONS: The morphologic features of asymmetric mitral valves are essentially different from those of true parachute valves. A distinction between these two anomalies will contribute to recognition by the pediatric cardiologist and surgeon.

[Contribution to the prenatal diagnosis of congenital cardiopathies]. / Contributo para o diagnóstico pré-natal das cardiopatias congénitas.

After concluding a local teaching program to obstetric echographists about prenatal diagnosis of congenital heart disease (CHD), we compared 2 periods of activity in fetal echocardiography, that before, lasting 3 and one half years (G.I) and the other 6 months after (G.II). During the second period 20 newborn consecutively admitted with CHD (G.III) were also studied. Teaching program protocol for the 10 district hospitals visited is described. The results show a rise from 25 to 58 foetus per month observed, comparing the two periods, being 4 and 7% respectively those referred with suspicion of CHD, and 54 and 33% the obstetric accuracy. Total incidence of CHD was 4,2%, 41 in G.I and 6 in G.II. Types of CHD, treatment and evolution are described, being the mortality 68 and 50% respectively. Of the 20 newborn from G.III, 90% had no prenatal diagnosis of CHD, being 30% high risk pregnancies. Mortality in this group was 20%, 50% in the newborn from high risk pregnancies; eight newborns were operated on without mortality, 5 to aortic coarctation and 2 with arterial switch for transposition. It is concluded the necessity of continuing teaching programs so that prenatal diagnosis of CHD may improve and the care of the newborn may so be anticipated and ameliorated.

[The echocardiogram in the first attack of rheumatic fever in childhood]. / O ecocardiograma no primeiro surto de febre reumática na criança.

In order to evaluate the contribution of echocardiography (echo) in the first episode of rheumatic fever, 18 children (mean age: 8.5 +/- 2.9 years) observed between 1987 and 1992 have been retrospectively studied. In 11 of them the echo has been done in the first month after the initial episode. By M-Mode echo the most constant finding was a high left atrium: aorta ratio in all cases with clinical carditis. By 2D-echo, hyperechogenicity of mitral valve leaflets and subvalvular apparatus was a common finding, three patients having mitral valve prolapse, one with dilatation of the mitral ring and the other with leaflets not able to coapt. Doppler-echo displayed in 17 patients mitral valve regurgitation, and in three aortic regurgitation. In three children the diagnosis of carditis was made by echo in the absence of murmur. During follow-up mitral valve regurgitation disappeared in four children. The authors conclude that early echo is very important in all children suspected to have rheumatic fever, specially because mitral valve regurgitation can be demonstrated by color flow mapping even in the absence of cardiac murmur.

[Restrictive myocardiopathy in children]. / Miocardiopatia restritiva na criança.

Four children, three males and one female, aged 1.5 months to 11 years, with restrictive cardiomyopathy are described. One had patent ductus arteriosus in association. Hepatomegaly, abnormal second heart sound and signs of pulmonary venous congestion in chest X-ray were the most consistent clinical findings. Diagnosis was based on echocardiographic findings, cardiac catheterization data and results of pathology. The most frequent echocardiographic sign was the enlargement of both atria but with the ventricles within normal size and normal systolic function. Abnormalities in second phase of the ventricular filling were recorded in 2D-echo Doppler and cardiac catheterization, where a dip and plateau morphology of ventricular tracings was recorded. Metabolic studies performed in two children were normal, and any child had hypereosinophilia. Two children died, one was lost for follow-up and another is asymptomatic. Remarks on medical and surgical management are made.

[Fetal cardiology. Bases for the prenatal diagnosis of congenital cardiopathies]. / Cardiologia fetal. Bases para o diagnóstico pré-natal das cardiopatias congénitas.

The bases for prenatal diagnosis of congenital heart disease are presented, in the line of a comprehensive program undertaken by paediatric cardiology and obstetrics. This program is designed to improve the screening of fetal heart diseases by obstetric echographists, particularly those working at peripheral high risk clinics. It includes the visit of paediatric cardiologists to district hospitals for joint meetings with local physicians, going over theoretical aspects, case presentation and four chamber view with outflow tract demonstrations using local ultrasound equipment. Referring protocols, handouts and support media exchange are contemplated. Support texts on the program divulged, on etiology of congenital heart disease, on abnormal fetal echocardiography and on fetal dysrhythmias are presented here in. The need for an organized national effort to increase the prenatal diagnosis of congenital heart disease is emphasized as well as the need to centralize some aspects related to diagnosis and treatment in order to improve the care of newborns with congenital heart disease.

[Prenatal cardiology. Suspicion to confirmation]. / Cardiologia pré-natal. Da suspeita à confirmação.

The mortality rate is high and prognosis is worse among new-borns with prenatal diagnosis of heart malformation, mainly due to factors such as its association with other malformations, and a range of more severe diseases probably resulting from the predominance of the obstetric use of the four chamber view. In this study we retrospectively assessed the range of cardiopathies diagnosed by foetal echocardiography and their evolution, compared with previous years. From January 1994 to December 1995, 1173 foetal echocardiograms were performed at a gestation age of 24 weeks. Sixty-one foetuses (5.2%) had cardiac anomalies, structural in 56 and arrhythmia in 5. The risks and indications were maternal in 37%, foetal in 31%, familial in 17% and environmental in 15%. Three were false negatives (VSD:2; truncus arteriosus: 1). Five died in utero, and 18 were assessed after birth with a mean gestational age of 37 weeks and birth weight of 3 Kg, a caesarean section was performed in 9. All but one were born in central hospitals. Six children were operated on. Two children died, one after surgery. Compared with the four previous years of activity, indication due to foetal risk rose from 6 to 31%, the number of cases diagnosed with heart disease increased from 14 to 30 per year, and the mortality decreased from 59 to 11%. Despite this, we still observe that the vast majority of new-borns who are hospitalised due to a severe heart disease had no prenatal diagnosis, indicating the need to continue our educational policy in this field.

[Fetal arrhythmia. A case load of 4 years and a half]. / Arritmias fetais. Casuística de quatro anos e meio.

In a 4 1/2 year period fetal, echocardiographic studies were performed on 1600 fetuses. In 55 with arrhythmia, 44 had supraventricular ectopic beats, resolved in all, and none had heart disease. Sustained arrhythmias occurred in 11 fetuses. Atrial flutter was present in 3 all with heart disease (Ebstein disease, right atrial tumour and WPW diagnosed after birth). Another 3 fetuses had supraventricular tachycardia (SVT), all with a normal heart. In the bradycardia group, 2 had complete heart block (CHB) associated with AVSD; 2 sinus bradycardia and one had non conducted atrial ectopic beats. Digoxin was the first choice drug for tachyarrhythmia therapy; association with Verapamil, Flecainide, Quinidine and Procainamide was used in 4 of the 6. One fetus with CHB received Orciprenaline with no results. Atrial flutter resolved or improved; in SVT 2 fetuses converted to sinus rhythm and one died in utero. All fetuses with CHB died in cardiac failure. Mortality was 27% (3 cases) in utero and global 36%. In our experience most fetal arrhythmias (90%) were transitory ectopic beats or non lasting bradycardia in normal heart and did not trigger other kinds of arrhythmias. In sustained arrhythmias, heart failure and heart disease had a negative effect on prognosis.

[Fetal echocardiography. The results of a 3-year study]. / Ecocardiografia fetal. Resultado de três anos de estudo.

During the last three years (July 89 to July 92), 875 fetal echocardiographic studies were performed, at a mean gestational age of 26 +/- 6.5 weeks (12 to 40 weeks), with the following main referral indications: family history of congenital heart disease 23%; advanced maternal age 13%; fetal arrhythmia 10%; amniotic fluid abnormalities 8%; fetal malformations 7%; diabetes 6%; four chamber view changes 4%. There were 15 twin pregnancies, two of them thoracopagus. The referral patterns with higher percentage of cardiac pathology were: abnormal four chamber view 56%; fetal chromosomal anomalies 50%; omphalocele 33%; twin pregnancy 33%; IUGR 29%; arrhythmias 20%. The referral patterns with lower percentage of cardiac pathology were: congenital heart disease in the family 0.2%; maternal risk 2%; amniotic fluid volume alterations 2%. The diagnosis of congenital heart disease was made in 37 fetus (4.2%), with the following most frequent: 9 cases with the atrioventricular septal defect, 5 cases with ventricular septal defect, 5 cases with hypoplastic left heart syndrome and 5 cases with tricuspid valve abnormalities. From a total of 78 referred fetal arrhythmias, 25 were confirmed, 20 being ectopic beats without structural heart disease, 3 with tachycardia, and 2 with bradycardia. One newborn with tachycardia and Ebstein's disease died, and the fetus with AV block and left atrial isomerism died in utero. The treatment and follow up of the remaining cases with heart disease are described. The whole mortality in the cases with heart pathology was 65%.(ABSTRACT TRUNCATED AT 250 WORDS)

[Prenatal diagnosis of congenital cardiomyopathies. Current reality in the south of the country]. / Diagnóstico pré-natal das cardiopatias congénitas. Realidade actual no sul do país.

In order to make an actual perspective about prenatal diagnosis of congenital heart disease in the area of influence of our department, a prospective study including 948 fetus and 185 newborn was done, 348 fetus and 20 newborn evaluated during 1993 (group I) and the remaining during 1994 (group II). In both groups indications for fetal echocardiography were mainly maternal (18%) and familiar (14%) factors, but occurrence of CHD were respectively 2% and 0% for them. Fetal factors for echocardiography account for 7%, namely arrhythmias (7%) and obstetric suspicion of CHD (6%), but occurrence of CHD was respectively 13% and 32% for group I and 36% and 48% for group II. In the newborn with serious CHD, risk factors could be identified in 30% in group I and 36% in group II, being respectively 15% and 7% referred for fetal echocardiography. It is concluded that although a rise in the number of fetus evaluated and a better obstetric accuracy have occurred, the rate of prenatal diagnosis of CHD is still very low, pointing to necessity of continuing our actual policy of teaching and spreading this area, specially in the primary health care units.

[Cardiovascular morbidity in children with human immunodeficiency virus infection]. / Morbilidade cardiovascular em crianças com infecção pelo vírus da imunodeficiência humana.

UNLABELLED: Abnormalities of cardiovascular structure and function have been described among children with human immunodeficiency virus (HIV). In order to evaluate their occurrence and clinical predictors, 32 children infected with HIV, aged three months to 13 years (mean age = 3.11 +/- 3.51 years) were studied. Perinatal transmission was identified in 90% of the children. Twenty-two patients (pts) (69%) had symptoms, nine being moderately symptomatic and eight severely symptomatic. Fourteen pts had immunological disturbances and eight of them were severely immunosuppressed. Twenty-eight pts (88%) had HIV-1 infection and 6 recent Ebstein-Barr virus coinfection. Nineteen were on zidovudine and 14 on intravenous immunoglobulin treatment. Nineteen cardiovascular abnormalities were found in 15 pts (47%): 11 cases of pulmonary hypertension by echocardiographic criteria (eight of them had interstitial lung infiltrates seen on chest X-ray) and four cases of left ventricular dysfunction requiring anticongestive therapy. Other abnormalities were: patent ductus arteriosus, septal hypertrophy, mitral valve prolapse and pericardial effusion (one case each). Surface ECG displayed right ventricular hypertrophy in four pts, left ventricular hypertrophy in one patient and unspecific ST-T wave changes in two pts. Fourteen pts (44%) had sinus tachycardia with mean heart rate above the 95th percentile on 24-hour Holter monitoring. CONCLUSIONS: Cardiovascular abnormalities are frequent among children with HIV-1 infection in late stages; pulmonary hypertension is the most frequently found cardiovascular anomaly and seems to be related to either chronic or recurrent respiratory disease; cardiological follow-up is recommended for HIV-infected children.

[A resurgence of rheumatic fever. New causes or old attitudes?]. / Ressurgimento da febre reumática. Novas causas ou velhas atitudes?

Bearing in mind that in the last years there has been an increase in rheumatic fever episodes, the authors evaluate the cases recently observed in the department. The data of 3 children born and living in Portugal, the first known outbreak of rheumatic fever observed between June 93 and March 94, were examined. One case presented polyarthritis, another polyarthritis and carditis and the third chorea and carditis. In just one case was the diagnosis of rheumatic fever considered in the beginning, and over-all, failures in the diagnosis and treatment of tonsillitis, and in echocardiographic diagnosis were detected. In view of these examples, the authors conclude that the increasing incidence and morbidity of rheumatic fever is more probably due to forgetfulness of old attitudes than to new causes. Delay in the diagnosis and errors in secondary prophylaxis may influence long term results.

[Acute myocardial infarct in infants]. / Enfarte agudo do miocárdio na criança.

A retrospective study was made of 6 children, with nonsurgical-related acute myocardial infarction (AMI), between January 1987 and December 1994. The ratio for gender was 1 and mean age at AMI was 49 days, 4 cases being associated with congenital heart disease (Fallot's tetralogy, truncus arteriosus and DiGeorge syndrome, one case each, and anomalous origin of left coronary artery, 2 cases). Kawasaki disease and coronary embolisation from thrombosis of the renal vein occurred in the other 2 cases respectively. All developed congestive cardiac failure and cardiomegaly. In the ECG pathologic q waves with more than 35 msec occurred in all, and QT prolongation occurred in 3. Five children (83%) all with AMI in the anterior and lateral wall of the left ventricle died, death being related with cardiac mechanical failure and not with arrhythmias.

Cattle tick-associated bacteria exert anti-biofilm and anti-Tritrichomonas foetus activities.

Research on microbiota in cattle tick and the evaluation of its activity against other microorganisms can contribute to identify new molecules potentially useful to control infections caused by bacteria and protozoa. Biofilms pose increasing problems worldwide, mainly due to their resistance to antimicrobial therapies and host immune response. In this study we investigate the ability Rhipicephalus (Boophilus) microplus-associated bacteria may exhibit to produce anti-biofilm and trichomonicidal compounds. Gut, ovary, salivary glands, and Gené organ were collected from engorged R. microplus female. Homogenates of each tissue were inoculated onto 15 distinct culture media. Anti-biofilm and trichomonicidal activities were analyzed by culturing each bacterium isolated in a liquid medium. Results showed that R. microplus cattle tick microflora varies for different tissues. Bacteria belonging to different genera (Aeromonas, Bacillus, Brevibacillus, Castelaniella, Comamonas, Kocuria, and Microbacterium) were identified. Interestingly, all bacterial species found displayed pronounced activity against Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms, and also against the cattle pathogen Tritrichomonas foetus, confirming the hypothesis that cattle tick could be a source of bacteria active against pathogens. This is the first study showing that bacteria isolated from a tick exert anti-biofilm and trichomonicidal activities.