Phages of Pseudomonas aeruginosa: response to environmental factors and in vitro ability to inhibit bacterial growth and biofilm formation.

AIMS: To examine effects of various environmental factors on adsorption and inactivation of Pseudomonas aeruginosa-specific phages: δ (family Podoviridae), J-1, σ-1 and 001A (family Siphoviridae) and their ability to inhibit bacterial growth and biofilm formation. METHODS AND RESULTS: The phages examined in the study were clonally different, as revealed by RFLP. The temperature in the range 7-44°C had no influence on the adsorption of Podoviridae, but did affect Siphoviridae adsorption, particularly 001A. All phages were significantly stable at pH 5-9, and phages δ and 001A even at pH 3. Most of the examined carbohydrates and exopolysaccharides of the original host efficiently inactivated phage δ, while phages σ-1 and J-1 were inactivated considerably only by the amino acid alanine. Silver nitrate efficiently inactivated all the phages, while Siphoviridae were more resistant to povidone-iodine. Serum of nonimmunized rats had no influence on phage inactivation and adsorption. Only phage δ showed ability to effectively inhibit in vitro bacterial growth and biofilm formation. CONCLUSIONS: The examined environmental parameters can significantly influence the adsorption and viability of Ps. aeruginosa-specific phages. The phage δ is a good candidate for biocontrol of Ps. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides important data on Ps. aeruginosa-specific phage adsorption, inactivation and in vitro lytic efficacy.

Relationship between the prenatal exposure to low-level of mercury and the size of a newborn's cerebellum.

Exposure to methylmercury at any stage of central nervous system development could induce alterations and result in severe congenital abnormalities. Total mercury level in maternal hair during pregnancy correlates well with blood levels of methylmercury and with total mercury levels in fetal brain. A prospective study has been conducted and a total of 137 childbearing women living at the coastal region with term, normal pregnancies were included and their newborns evaluated by ultrasonography. Mothers and their newborns are divided in two groups according to their hair mercury levels; examined group with high body levels of mercury (≥ 1 µg/g) and control group with low body levels of mercury (<1 µg/g). Neurosonographic examination was conducted to all newborns. Two dimensions of cerebellum in the sagital-medial plane have been measured: maximum height and width starting from the roof of the fourth chamber. Majority of mothers had hair mercury levels lower than 1 µg/g (N = 107). Mean value was 0.88 µg/g (SD 1.24), ranging from 0.02 to 8.71 µg/g. There was no significant difference between the two groups when it comes to the width of cerebellum (Mann-Whitney test: Z = 1471; p = 0.141). However, comparison related to the length of cerebellum shows statistically significant smaller cerebellum in newborns whose mother had hair mercury levels higher than 1 µg/g (Mann-Whitney test: Z = 2329; p = 0.019). Our results lead to a conclusion that prenatal exposure to, what we consider to be, low-levels of methylmercury does influence fetal brain development detected as decreased size of newborn's cerebellum. From a clinical point of view, a question related to the influence of prenatal low-level methylmercury exposure on fetal neurodevelopment remains open. Our further objectives are to direct the research towards performing detailed neuropshychological tests on children at the age of 18 months. Such tests could indicate the presence of subtle neurological or neuropsychological deficits.

Kinetics of degradation of hydrocarbons in the contaminated soil layer.

This work presents kinetic parameters of the removal of hydrocarbons in the course of bioremediation of the soil contaminated with crude and oil derivatives, with special emphasis paid to polycyclic aromatic hydrocarbons. Bioremediation was carried out on a laboratory scale, with continuous and discontinuous recirculation of water through the soil column inoculated with the adapted bacterial population. Biodegradation of hydrocarbons can be described by the following equation--ln C = ln C0- kt--which is most often used in the literature to describe degradation of hydrocarbons, and by the equation--ln C = ln C0- kt0.5--which is determined on the basis of experimental data (except for pyrene and chrysene, for which a linear equation was used). A comparison of the two models showed a significant difference in the calculated remediation times, indicating that it is not realistic to expect a simple kinetic model to provide precise and accurate descriptions of concentrations during different seasons and in different environments. The results indicated that once contaminated with hydrocarbons, soil can be slowly brought to the previous state provided that no new contamination has occurred.

Neonatal outcome in discordant eutrophic twins: twin growth.

OBJECTIVE: To compare maternal characteristics and neonatal outcome in discordant twin gestations (DT) and concordant twin gestation (CT). METHOD: Maternal and neonatal data base of live twins >25 weeks' gestation (N=351 pairs) were reviewed for antepartum complications, labor beginning, mode of delivery, neonatal complications, malformations and perinatal mortality. The chi-squared analysis and Student t-tests were used to analyze the differences between discordant and concordant premature and term twin pairs, and appropriate for gestational age (AGA) twins, separately. RESULTS: DT occurred in 15.1% of all twin pregnancies. In preterm and term DT there were significantly more elective cesareans. Growth discordance among preterm and term eutrophic twins was not connected with increased neonatal death or other complications, except higher incidence of early neurological signs in term DT. CONCLUSION: We strongly believed that prematurity and not discordant growth of eutrophic twins has important influence on neonatal outcome.

Cyclic variation of integrated backscatter: dependence of time delay on the echocardiographic view used and the myocardial segment analyzed.

To determine the influence of myocardial anisotropy in ultrasonic tissue characterization, we measured the time delay (and magnitude) of the cyclic variation of myocardial integrated backscatter from specific segments visualized in the 4 standard transthoracic echocardiographic views. The cyclic variation data in 10 myocardial regions were obtained from analyses of 2-dimensional integrated backscatter images from 23 healthy subjects. Resultant values (mean +/- SD) for the time delay were as follows: parasternal long-axis view: 1.08 +/- 0.17 (septum) and 1.00 +/- 0.14 (posterior wall); parasternal short-axis view: 1.03 +/- 0.16 (anterior septum), 1.03 +/- 0.14 (posterior wall), 2.22 +/- 0.71 (lateral wall), and 1.65 +/- 0.66 (posterior septum); apical 4-chamber view: 1.08 +/- 0.31 (septum) and 2.20 +/- 0.79 (lateral wall); and apical 2-chamber view: 1.68 +/- 0.62 (inferior wall) and 2.04 +/- 0.72 (anterior wall). Hence, results of this study indicate that myocardial ultrasonic characterization that uses the cyclic variation is influenced by the echocardiographic view and the specific segment of the left ventricle.

Fetal tachydysrhythmia associated with nonimmune hydrops.

A case of fetal tachydysrhythmia with intermittent episodes of bradycardia associated with severe nonimmune hydrops was detected at 36 weeks of pregnancy. Paroxysms of supraventricular tachycardia were documented postnatally. Induction of labor and postnatal administration of digoxin showed to be a rational therapeutic approach in this case. In spite of some unfavorable clinical predictors the treatment resulted in a benign course of the disease.

Dependence of "apparent" magnitude on the time delay of cyclic variation of myocardial backscatter.

The goal of this study was to determine if the "apparent" magnitude of the cyclic variation, defined as the difference between the values of integrated backscatter at end-diastole and end-systole, was dependent on the corresponding time delay. We measured the cyclic variation in four myocardial segments of the parasternal short-axis view in 23 healthy subjects. The "apparent" magnitude, actual magnitude, and time delay were compared for each segment. Measured time delays were: 2.22+/-0.71 (lateral wall); 1.65+/-0.66 (inferior septum); and approximately 1.0 for the anterior septum and posterior wall. Segments exhibiting large time delays (> 1.0) resulted in a reversal in sign of the "apparent" magnitude of cyclic variation in one instance, and underestimated the true magnitude in both cases. Thus, estimates of the "apparent" magnitude of the cyclic variation are dependent on the associated time delay, whereas a properly defined magnitude is not.

Chromosomal anomalies in abnormal human pregnancies.

OBJECTIVE: The aim of this study was to describe the cytogenetic observations on abnormal human pregnancies (anembryonic pregnancy, early fetal loss, and hydatidiform moles), and to detect the most frequent or typical chromosomal aberration for anembryonic pregnancy and early fetal loss. STUDY DESIGN: Abnormal pregnancies were divided into three clinical and morphological groups: (a) anembryonic pregnancy; (b) early fetal loss, and (c) hydatidiform mole. Of the 119 karyotyped tissue samples, 42 (35%) were from anembryonic pregnancies, 64 (54%) from early fetal losses, and 13 (11%) were from hydatidiform moles (6 complete and 7 partial moles). Long-term cultures of chorionic villi and GTG-banding techniques were used for chromosome analysis. RESULTS AND CONCLUSION: The overall frequency of chromosome anomalies among the 119 karyotyped spontaneous abortions was found to be 37.8%. Trisomy (double trisomy included) accounted for 35.6% of all aberrations, followed by polyploidy (33.3%), mosaicism (11.1%), structural abnormalities (4.4%), and monosomy X (2.2%). Although the difference was not statistically significant, single trisomy was the predominant chromosome abnormality found in anembryonic pregnancies (64.3 %) while in cases of early fetal loss, trisomy (double trisomy included) (38.9%) and triploidy (27.8%) were quite frequently present. The frequency of triploidy among all chromosomal abnormalities was 28.9%, and 53.8% of them were found in partial hydatidiform mole. The rest of them were almost exclusively found in early fetal losses. Complete hydatidiform moles (androgenetic in origin) were present in 13.3% of all aberrations, of which 83.3% had a 46,XX, and the rest of them had a 46,XY karyotype.

A simplified fetal biophysical profile.

OBJECTIVE: To modify the classic fetal biophysical profile (FBP) with the aim of obtaining rapid and accurate information about actual fetal condition in non-compromised fetuses with a subsequent favorable outcome and to be suitable for a number of outclinic patients. METHODS: Four-hundred and ninety-four fetuses from singleton pregnancies in two randomized groups were monitored by the modified FBP (mFBP) and 168 of them after the external vibratory acoustic stimulation (VAS/mFBP). The mFBP was characterized by two main characteristics: non-stress test was excluded and the testing was finished at the moment when all of the three fetal biophysical activities became normal. The external VAS was applied only in cases with no evidence of fetal activity at the start of the FBP. RESULTS: Of the examined fetuses, 326 fetuses in the control group were monitored by the mFBP and there were 316 (96.9%) favorable outcomes and 10 (3.1%) adverse perinatal outcomes. The sensitivity, specificity and positive and negative predictive values of the mFBP score in predicting adverse perinatal outcome were 60, 99, 66.7 and 98.7%, respectively. In the study group of 168 fetuses there were 165 (98.2%) favorable outcomes and three (1.8%) adverse perinatal outcomes. The sensitivity, specificity and positive and negative predictive values of the VAS/mFBP were 66.7, 100, 100 and 99.4%, respectively. The efficiency of the VAS/mFBP in predicting perinatal mortality alone was even higher. After the external VAS and the first 5 min of the modified testing approximately two-fifths (41.8%) of healthy fetuses with a subsequent good outcome exhibited normal in all of the three biophysical activities and approximately two-thirds (65.5%) of them after 10 min. In the VAS/mFBP group of healthy fetuses, during the same time periods, normal breathing movements were observed in 72% and 87% of fetuses, respectively. CONCLUSIONS: According to our results the mFBP and particularly the VAS/mFBP antenatal protocol as a new and rational variant of the FBP could improve fetal assessment allowing in cases of non-compromised fetuses rapid and accurate information about actual fetal well-being. Because of its high accuracy and a reduced testing time the antepartal method with observation of fetal breathing movements after VAS is becoming acceptable as a screening of fetal well-being evaluation in outclinic conditions.

Perforin-expressing lymphocytes in peripheral blood and decidua of human first-trimester pathological pregnancies.

PROBLEM: We have shown previously that the decidua of first-trimester human pregnancy is heavily infiltrated with perforin-positive cells. The aim was to detect expression of perforin in both decidual lymphocytes (DL) and peripheral blood lymphocytes (PBL) in the first trimester of pathological pregnancies: Anembryonic pregnancy and missed abortion. METHOD: Decidual tissue from a normal pregnancy group and from pathological pregnancies was obtained by vaginal curettage. Perforin (an intracellular antigen) and the cell surface antigens CD3, CD4, CD8, CD16, CD56, CD11c, and CD45RA were quantified simultaneously by flow cytometric analysis. RESULTS: In the missed abortion group, we found: 1) a relative decrease in the frequency of both CD4+P+ cells and CD56+P+ cells as well as the mean fluorescence intensity for perforin; 2) a relative increase of CD16+P+ PBL cells; and 3) a relative increase of CD4+ cells in PBL compared with anembryonic pregnancy and normal pregnancy. There was also a significant relative decrease in the proportion of CD4+ and CD8+ cells among perforin-positive PBL in both anembryonic pregnancy and missed abortion. CONCLUSION: Our results show that significant decreases in the prevalence of perforin-positive lymphoid cells, their subpopulations, and mean fluorescence intensity for perforin are associated with pregnancy failure.

Transthoracic echocardiographic detection of coronary atherosclerosis.

Coronary atherosclerosis is a pathologic process that produces thickening of the walls of the coronary arteries to the point that flow through those vessels may be impaired. This study attempts to use transthoracic echocardiography to detect coronary atherosclerosis. Eighty-nine patients undergoing coronary angiography were examined with a broad-band ultrasonic transducer with a frequency between 3 and 5 MHz. A modified short axis examination was utilized to identify left main and proximal left anterior descending arteries. The examination was recorded digitally and displayed in a 32-cell, quad screen cine loop. Fifty-six of the 89 patients (63%) had obstructive coronary artery disease (CAD) (i.e, at least 1 vessel with 50% obstruction). There were 14 patients with CAD but no vessel had > or = 50% obstruction. Nineteen patients (21%) had angiographically normal arteries. The coronary echograms were judged qualitatively for brightness, uniformity, and persistence (defined as the ability to see segments of the artery walls in more frames than other segments). The length of the coronary artery visualized, the width of the left main coronary artery, and the width of the thickest segment of the coronary artery walls were quantitatively measured. More than 2 cm of the left coronary artery was seen in almost all patients. Segmental changes were noted in 52 of the 56 patients with obstructive CAD, 12 of the 14 patients with nonobstructive CAD, and 3 of the 19 patients with normal arteries. Persistence greatly enhanced the ability to judge the segmental changes. Forty-six patients with obstructive disease had wall thickness > or = 1.5 mm. Only 6 patients with nonobstructive coronary arteries had this wall thickness, and only 1 normal subject had thick walls. The ultrasonic findings were useful in predicting the presence or absence of coronary atherosclerosis to varying degrees of sensitivity and specificity based on the segmental findings and wall thickness measurements. The results of this study indicate that a transthoracic ultrasonic examination of the proximal left coronary artery could be a clinically valuable tool in the qualitative identification of coronary atherosclerosis.

Evaluation using dobutamine stress echocardiography in patients with insulin-dependent diabetes mellitus before kidney and/or pancreas transplantation.

The purpose of this study was to examine the ability of dobutamine stress echocardiography to stratify patients with juvenile onset, insulin-dependent diabetes mellitus who are being considered for kidney and/or pancreas transplantation, into high-or low-risk groups for future cardiac events. Fifty-three such patients underwent dobutamine stress echocardiography before kidney and/or pancreas transplantation. Cardiac events, including cardiac death, nonfatal myocardial infarction, unstable angina, pulmonary edema, and need for coronary revascularization, occurring between the time of the dobutamine stress echocardiogram and the last patient follow-up contact were retrospectively identified. Twenty patients 938%) had an abnormal dobutamine stress echocardiogram. Eleven patients had 15 cardiac events over a mean (+/- SD) follow-up period of 418 +/- 269 days. Event rates were 45% among those with an abnormal, versus 6% among those with a normal dobutamine stress echocardiogram (p = 0.002). The result of the dobutamine stress test independently predicted prognosis in a multivariate analysis (p = 0.003, odds ratio = 12.7). We conclude that dobutamine stress echocardiography accurately stratifies patients with juvenile onset, insulin-dependent diabetes being considered for kidney and/or pancreas transplantation for risk of future cardiac events.

Fetal transverse cerebellar diameter/abdominal circumference ratio in assessing fetal size.

OBJECTIVE: To test the usefulness of the fetal transverse cerebellar diameter/abdominal circumference (TCD/AC) ratio in predicting known small-for-gestational-age (SGA) infants. METHOD: The relationship between fetal TCD and AC throughout the second half of pregnancy was investigated in 635 well-dated, normal pregnancies and examined with regard to gestational age and infant birth weight percentiles. RESULTS: One hundred eighteen (19%) fetuses were excluded due to inadequate visualization of the fetal cerebellum. A strong correlation was noted between gestational age determined by the last menstrual period and both fetal TCD (r2 = 0.91338) and AC (r2 = 0.89361) in fetuses with birth weights between the 10th and 90th percentiles (n = 407; mean 14.4, S.D. 1.2). Although the TCD/AC ratio showed a poor correlation with gestational age (r2 = 0.15788), a slight increase was noted during gestation. A TCD/AC ratio greater than 15.5 was present in 80% of SGA infants when measurements were performed within 1 week of delivery. CONCLUSION: Fetal TCD/AC ratio as a gestational age-independent method could improve diagnostic sensitivity and specificity in the early detection of fetal growth abnormalities.

Fetal biophysical profile and vibratory acoustic stimulation in high-risk pregnancies.

OBJECTIVES: To determine the influence of the non-stress test (NST) on the efficiency of the fetal biophysical profile (FBP) and to test the clinical usefulness of the FBP and its combination with vibratory acoustic stimulation (VAS) in managing high-risk pregnancies. METHODS: One hundred twenty fetuses of preeclamptic patients were included in a prospective study. Five standard variables of the FBP were observed ultrasonically following NST. In cases of non-reactive NST, external VAS was applied and the FBP score calculated and compared with the FBP score before VAS. RESULTS: Of 120 calculated FBPs, 102 (85%) had normal profile scores before VAS and 104 (86.7%) after VAS. No statistically significant difference was found. The sensitivity, specificity, and positive and negative predictive values of the FBP score in predicting poor perinatal outcome were 94.7%, 94.4%, 75% and 99%, respectively. VAS produced a high conversion (58.8%) of non-reactive NST to reactive fetal heart rate pattern. The false-negative rate of the FBP score was 9.8 per 1000, which did not increase after VAS. CONCLUSIONS: The efficiency of the FBP score was not significantly improved by VAS, although a high conversion of non-reactive to reactive NST was produced. The FBP with its three 'acute biophysical variables' was found to be an accurate method of antepartum assessment even without an NST. The low incidence of perinatal complications among patients with normal FBP scores, permits the conservative management of preterm high-risk pregnancies.

[Oral rehydration in the treatment of acute diarrhea in children]. / Oralna rehidratacija u lecenju dijareje kod dece.

Acute diarrhea is one the major causes of child ailments, regardless of their age and a frequent cause of infancy deaths. The Oral Rehydration is the recommended therapy for all patients suffering from mild and moderate dehydration, whatever the cause or the age of the child. A timely and properly administered oral rehydration effectively supplements the liquids and electrolytes deficiency, compensates for their continued loss and ensures physiological needs. The Oral Rehydration is an effective, simple and safe therapy which mothers can easily implement at home. It substantially reduces the necessity for hospitalizing the children and considerably cuts the treatment cost. Rehydration solutions contain glucose and electrolytes with the concentration of Sodium ringing from 60 to 90 mmol/l.

Decidual-trophoblast interactions: decidual lymphoid cell function in normal, anembryonic, missed abortion and ectopic human pregnancy.

This study was designed to investigate the consequences of decidua-trophoblast interactions on the phenotype, spontaneous and induced proliferation and immunoregulatory potential of decidual leukocytes in normal pregnancies (NP), anembryonic pregnancies (AP), missed abortions (MA) and ectopic pregnancies (EP). Spontaneous proliferation of decidual non-adherent cells (NAD) from pregnancies with viable trophoblast inside the uterus is significantly higher than proliferation of peripheral blood lymphocytes (PBL) from the same groups (P < 0.001 for NP; P < 0.05 for AP). Spontaneous proliferation of decidual NAD cells from NP was higher (P < 0.001) when compared with AP and EP. The induced (PHA and Con A) responses of PBL from women with normal and pathological pregnancies were significantly higher than that of decidual NAD cells (P < 0.001). Higher proliferation of NAD decidual cells was obtained when Con A-stimulated NP were compared with MA and EP (P < 0.01). The interaction of viable trophoblast with intrauterine decidua appears to be a prerequisite for the activation of NAD suppressor cells, since NAD cells from MA produced stimulation instead of suppression, and NAD cells from EP had no suppressive effect. On the contrary, both NAD and adherent (AD) decidual leukocytes from NP and AP produced very strong suppression of PHA or alloantigen-induced PBL proliferation. The contact between trophoblast and AD decidual leukocytes is not necessary for their suppressive function, since even higher suppression is obtained with the cells from ectopic pregnancies.

An immunohistochemical study of leucocytes in human endometrium, first and third trimester basal decidua.

An immunohistochemical quantitative study of leucocyte subpopulations on fresh human endometrium and on biopsy specimens of first and third trimester basal decidua in normal (uncomplicated) pregnancies was performed. The most prominent population in endometrial and decidual stroma of basal decidua are macrophages. B cells as well as gamma/delta T cell receptor positive cells were found occasionally, scattered throughout the endometrial/decidual stroma. CD3+ cells were present in a relatively small number in the endometrium as well as in the first trimester basal decidua, but their number was elevated (doubled) in the third trimester of pregnancy. CD2+ cells showed a slight increase in first trimester basal decidua when compared with both endometrium and third trimester basal decidua. Cells with positive NKH-1 marker (CD56+) showed a significant increase in the first trimester, while in the third trimester their number diminished drastically. CD56:CD3 cell ratio increased to more than five times in first trimester basal decidua, while in the third trimester basal decidua decreased drastically. The mentioned increase of CD56+ cells in the first trimester and that of CD3+ cells at term suggests that these cells could have some specific function(s). However, it still has to be established whether the described quantitative changes of decidual leucocytes in basal decidua during pregnancy are of any importance for the mechanism(s) for the fetal allograft protection.