Anti-Mullerian-Hormone during pregnancy and peripartum using the new Beckman Coulter AMH Gen II Assay.

BACKGROUND: AMH levels determined by the conventional AMH assay declined during pregnancy and postpartum. A new Beckman Coulter AMH Gen II assay removes the potentially assay-interfering complement which is activated in pregnancy. The aim of this study was to evaluate if the decline of AMH levels in the serum of pregnant women during the course of pregnancy and peripartum was assay-dependent and thus artificial. METHODS: In this cross-sectional study prepartal blood samples were collected from 62 patients (median age 30.6 years [interquartile range: 25.6 - 34.5]) in the third trimester of pregnancy and again 1-4 days after delivery between 2011 and 2012. In another cohort of 11 patients (median age 34.1 years [interquartile range: 32.6 - 37.8]) blood samples were taken in different trimesters of pregnancy between 1995 and 2001. The conventional and the modified AMH assay were performed in the same patient serum samples. We used the conventional and the modified AMH-Gen-II ELISA (Beckman Coulter, Immunotech, Webster, USA) for the assessment of AMH levels. The Wilcoxon signed rank test was used for determining differences between AMH levels pre- and postpartum. The method of Bland and Altman was applied for analyzing the agreement of both methods for determining AMH levels. RESULTS: AMH values peripartum were lower than those expected in fertile non-pregnant women of comparable age. An overall mean difference of 0.44 ng/ml was observed between the conventional and the modified assay. Measurements with the modified assay showed a significant decline of postpartal levels compared with prepartal levels which is consistent with values obtained using the conventional assay (both p < 0.00001). Compared to the longitudinal measurements of AMH levels determined using the conventional assay, AMH levels obtained using the modified assay suggest a steeper decline of values during the course of pregnancy. CONCLUSION: By comparing the conventional assay for AMH determination with the modified assay the present study confirmed that AMH levels decline during the course of pregnancy and early after delivery.

Unlocking the Potential of Substrate Quality for the Enhanced Antibacterial Activity of Black Soldier Fly against Pathogens.

Globally, antibiotics are facing fierce resistance from multidrug-resistant bacterial strains. There is an urgent need for eco-friendly alternatives. Though insects are important targets for antimicrobial peptides, it has received limited research attention. This study investigated the impact of waste substrates on the production of antibacterial agents in black soldier fly (Hermetia illucens L.) larvae (HIL) and their implications in the suppression of pathogens [Bacillus subtilis (ATCC 6051), Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853), and Escherichia coli (ATCC 25922)]. The 20% acetic acid (AcOH) extract from market waste had the highest antibacterial activity with an inhibition zone of 17.00 mm, followed by potato waste (15.02 mm) against S. aureus. Hexane extract from HIL raised on market waste also showed a significant inhibitory zone (13.06 mm) against B. subtilis. .Minimum inhibitory concentration (MIC) values recorded were 25 mg/mL against all test pathogens. The fastest time-kill of 20% AcOH extract was 4 h againstB. subtilis, E. coli, ,andP. aeruginosa. Lauric acid was also identified as the dominant component of the various hexane extracts with concentrations of 602.76 and 318.17 µg/g in HIL reared on potato and market waste, respectively. Energy from the market waste substrate correlated significantly (r = 0.97) with antibacterial activities. This study highlights the key role of substrate quality and extraction methods for enhancing the production of antibacterial agents in HIL, thus providing new insights into the development of potential drugs to overcome the alarming concerns of antimicrobial resistance.

Soluble endoglin versus sFlt-1/PlGF ratio: detection of preeclampsia, HELLP syndrome, and FGR in a high-risk cohort.

The angiogenic factors sFlt-1 and PlGF play an established role in the detection of preeclampsia (PE). Recent data suggest that sEng might contribute to the pathogenesis of PE. However, only a few studies so far have addressed its role.This monocentric cross-sectional study of high-risk pregnancies aims to compare the levels of sFlt-1/PlGF ratio and sEng depending on different placental-related adverse pregnancy outcomes. The statistical analysis takes into account Pearson's correlation coefficient between angiogenic factors, the area under the curve estimates (AUCs) for detection, and adjusted odds ratios (aOR) with 95% confidence intervals (95%-CIs). The analysis included 206 patients: 60 controls, 90 PE (59 EOPE, 35 LOPE), 94 FGR, and 35 HELLP cases. Some outcomes overlapped because FGR commonly complicated PE and HELLP syndrome. Serum levels of sFlt-1/PlGF and sEng correlated with each other. Higher levels were observed in HELLP syndrome and EOPE cases. AUCs for sFlt-1/PlGF ratio and sEng were, respectively, 0.915 (95%-Cl 0.87-0.96) and 0.872 (95%-Cl 0.81-0.93) in PE, 0.895 (95%-Cl 0.83-0.96) and 0.878 (95%-Cl 0.81-0.95) in HELLP syndrome, 0.891 (95%-Cl 0.84-0.94), and 0.856 (95%-Cl 0.79-0.92) in FGR.aORsfor sFlt-1/PlGF ratio and sEng were, respectively: 2.69 (95%-Cl 1.86-3.9) and 2.33 (95%-Cl 1.59-3.48) in PE, 2.38 (95%-Cl 1.64-3.44) and 2.28 (95%-Cl 1.55-3.4) in FGR, and 2.10 (95%-Cl 1.45-3.05) and 1.88 (95%-Cl 1.31-2.69) in HELLP syndrome. In addition, the aORs between sFlt-1/PlGF and sEng were very similar but higher for PE and FGR than HELLP syndrome.In conclusion,sEng performed similarly to sFlt-1/PlGF to detect placental dysfunctions.

Electron impact ionization of furanose alcohols.

Electron impact ionization of the gas phase 3-furanol, tetrahydro (3-hydroxytetrahydrofuran, 3HTHF) and 2-furanmethanol, tetrahydro (alpha-tetrahydrofurfuryl alcohol, THFA) molecules has been studied both experimentally and theoretically. The electron induced positive ion formation has been investigated experimentally using a crossed electron/neutral beams technique in combination with a quadrupole mass spectrometry. The mass spectra of both molecules have been determined at the incident electron energy of 70 eV. The ionization efficiency curves for each parent cation and a number of fragment cations have been measured near the threshold, and the corresponding appearance energies have been derived using an iterative fitting procedure based on the Wannier threshold law, taking into account the incident electron energy resolution. The appearance energies of the parent cations were experimentally determined to be (9.620+/-0.058) eV for (C(4)H(8)O(2)(+)/3HTHF) and (9.43+/-0.12) eV for (C(5)H(10)O(2)(+)/THFA), which are in a good agreement with G3MP2 calculated results: 9.480 and 9.419 eV, respectively. The most abundant cations in the mass spectra were determined to be 57 amu for 3HTHF and 71 amu for THFA, with the corresponding experimentally determined appearance energies of (10.22+/-0.10) eV and (9.574+/-0.062) eV, respectively. With the help of the energies calculated at B3LYP and G3MP2 levels of theory, the possible fragmentation patterns were discussed.

Ionization of doped helium nanodroplets: complexes of C60 with water clusters.

Water clusters are known to undergo an autoprotonation reaction upon ionization by photons or electron impact, resulting in the formation of (H(2)O)(n)H(3)O(+). Ejection of OH cannot be quenched by near-threshold ionization; it is only partly quenched when clusters are complexed with inert gas atoms. Mass spectra recorded by electron ionization of water-doped helium droplets show that the helium matrix also fails to quench OH loss. The situation changes drastically when helium droplets are codoped with C(60). Charged C(60)-water complexes are predominantly unprotonated; C(60)(H(2)O)(4)(+) and (C(60))(2)(H(2)O)(4)(+) appear with enhanced abundance. Another intense ion series is due to C(60)(H(2)O)(n)OH(+); dehydrogenation is proposed to be initiated by charge transfer between the primary He(+) ion and C(60). The resulting electronically excited C(60)(+*) leads to the formation of a doubly charged C(60)-water complex either via emission of an Auger electron from C(60)(+*), or internal Penning ionization of the attached water complex, followed by charge separation within {C(60)(H(2)O)(n)}(2+). This mechanism would also explain previous observations of dehydrogenation reactions in doped helium droplets. Mass-analyzed ion kinetic energy scans reveal spontaneous (unimolecular) dissociation of C(60)(H(2)O)(n)(+). In addition to the loss of single water molecules, a prominent reaction channel yields bare C(60)(+) for sizes n=3, 4, or 6. Ab initio Hartree-Fock calculations for C(60)-water complexes reveal negligible charge transfer within neutral complexes. Cationic complexes are well described as water clusters weakly bound to C(60)(+). For n=3, 4, or 6, fissionlike desorption of the entire water complex from C(60)(H(2)O)(n)(+) energetically competes with the evaporation of a single water molecule.

The influence of omega-3 polyunsaturated fatty acids feeding on composition of fatty acids in fatty tissues and eggs of laying hens.

The paper deals with supplementation of hen diet with oils with increased content of omega-3 polyunsaturated fatty acids (PUFA) and occurrence of fatty acids (FA) in fatty tissues and eggs of laying hens after the respective supplementation. The experiment was carried out on 30 laying hens of ISA BROWN hybrid, divided to three groups (A, B, C). For the period of three weeks the group A was administered flax-seed oil, group B served as a control and group C was fed base rations supplemented with fish oil. In addition to that the diet of groups A and C was supplemented with vitamin E as an antioxidant. Administration of the oils as a source of omega-3 PUFA increased their concentration in both the fatty tissues and egg-yolk which resulted in the change of the ratio of omega-6 and omega-3 PUFA. No influence of flax-seed and fish oil on concentration of alpha-tocopherol in egg-yolk was observed.

[Testicular metastases as primary manifestation of malignant melanoma at known melanocytoma]. / Hoden-Metastasen als Erstmanifestation eines malignen Melanoms bei bekanntem Melanozytom.

BACKGROUND: Secondary tumors of the testes are rare. In about 15 % of the cases they are metastases of a malignant melanoma, there are about 30 reports of such cases in the literature. Most of them describe findings at post-mortem, in only 4 of the previously described cases testicular metastases were the first manifestation of a melanoma. The transformation of a benign, meningeal melanocytoma into a malignant melanoma has only been described once world-wide. The problems in the diagnosis and therapy of this extremely rare tumor are discussed on the basis of a further patient with metastases of the testes as primary manifestation of a malignant melanoma. CASE REPORT: We report on a 74-year-old man who presented with a relatively rapidly developing but less painful swelling of the left testicle. The case history disclosed operations 10 and 3.5 year earlier for an apparently benign melanocytoma at the level of the 11th and 12th thoracic vertebrae, a local recurrence with paraparesis was known at the time of admission. Sonography revealed an inhomogeneous tumor effecting the entire left testicle. Therapy comprised an inguinal, radical orchiectomy. The correct diagnosis was made histologically. Shortly after the operation, a diffuse, subcutaneous and hepatic metastases occurred. CONCLUSION: In the case of a primary manifestation a correct preoperative diagnosis is unusual. Radical orchiectomy is the treatment of choice for a suspected primary testicular tumor. The prognosis is accordingly poor. In elderly patients with unclear testicular tumors metastases from an (occult) tumor disease must be taken into consideration.

Alterations in glutathione peroxidase and superoxide dismutase activities in plasma and saliva in relation to disease activity in patients with Crohn's disease.

Crohn's disease (CD) is a chronic inflammatory condition with uncertain aetiology. Dysfunction of immunoregulatory factors and overproduction of reactive oxygen species (ROS) may contribute to the damage of the gastrointestinal tract. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) are involved in protection of cells from the damaging effects of ROS. The aim of the study was to assess activity of antioxidative stress enzymes, GPx and SOD, in plasma and saliva of patients with active and inactive forms of CD. Forty-seven patients with CD were prospectively enrolled in the study. The control group comprised 25 healthy volunteers. Patients' demographics, clinical features, localization of inflammatory changes, CD history, and treatment were recorded. SOD and GPx were assayed in plasma and saliva samples by ELISA method. CD activity index (CDAI) scores correlated inversely with SOD in plasma (r = - 0.46; P = 0.0012), but not in saliva. No correlations were observed in respect to GPx activities in both plasma and saliva and CDAI. Higher activity of plasma SOD was observed in patients with inactive CD in comparison with active CD (P = 0.004). No significant differences in SOD and GPx activity both in plasma and saliva were found between CD remission group and the control group. We concluded that in active CD the antioxidant defence system was diminished and returned to normal values in remission. Results of SOD and GPx assays in saliva are not conclusive, suggesting that saliva seems to be not an appropriate material for further similar studies.

Expression of MHC class I antigen in Morris rat hepatoma and normal liver.

Expression of major histocompatibility complex (MHC) class I antigen in Morris rat hepatoma 7777 and normal rat liver tissues was studied by immunohistochemical techniques, Western blot and radiobinding assay. It was found that hepatoma tissue displayed a pattern of enhanced MHC class I antigen expression compared with normal liver. This phenomenon may represent an uncommon pattern of MHC class I antigen expression of liver tumor. The influence of enhanced MHC class I antigen expression on the development and spread of tumors as well as the possible mechanisms of this enhancement are discussed.

Anticore endotoxin F(ab')2 equine immunoglobulin fragments protect against lethal effects of gram-negative bacterial sepsis.

Gram-negative bacterial sepsis and shock remain a cause of substantial morbidity and mortality in hospitalized patients despite appropriate antimicrobial therapy, fluid resuscitation, and monitoring. We sought to test the ability of equine antibody directed against core endotoxin, a portion of bacterial outer membrane lipopolysaccharide common to many gram-negative microorganisms, to bind to various gram-negative bacteria in vitro, to promote bacterial phagocytosis by leukocytes, and to protect against lethal gram-negative bacteremia in mice. The importance of the IgG Fc leukocyte attachment site was examined by comparing the ability of intact IgG and IgG F(ab')2 fragments to protect against lethality during murine sepsis. A single horse was immunized with Escherichia coli J5, an organism that expresses a portion of core endotoxin extensively on the cell surface. Preimmunization IgG and F(ab')2 possessed no titer as determined by enzyme-linked immunosorbent assay, did not promote in vitro phagocytosis, and did not protect in vivo. Postimmunization IgG and F(ab')2 possessed a significant titer to E. coli J5 whole cell and lipopolysaccharide antigens and provided significant (p less than 0.05) protection in vivo during lethal intravenous sepsis caused by either E. coli J5, E. coli 0111:B4, Klebsiella pneumoniae, or Pseudomonas aeruginosa. Only postimmunization IgG, but not F(ab')2, promoted in vitro phagocytosis of these same organisms. We therefore hypothesized that protection occurred as a result of antitoxin activity rather than opsonization and phagocytosis, as F(ab')2 fragments were as active as the intact molecule. Further studies must be done to determine the role of anticore endotoxin antibody in conjunction with antibiotics so that appropriate clinical studies may be undertaken.

Theoretical study of the reactivity properties of two forms of stobadine.

The cardio- and neuroprotective effect of the pyridoindole stobadine (S) is conditioned mainly by its good radical scavenging properties. It has been showed by EPR experiment, that the ultimate product of the reaction of stobadine with hydroxyl radical is the nitroxyl radical. However, for the unsaturated dehydrostobadine (DHS) the ultimate product was not experimentally determined, although its reactivity with a hydroxyl radical has been detected. Using the quantum chemical method AM1 we calculated the physico-chemical properties of S, DHS and their radicals. For the stobadine alone, the corresponding radical was formed by removing the H* from the NH group of indol, while in the case of DHS we removed the CH3* from the nitrogen in pyridine ring. For S and DHS we calculated the differences in the energies between the parent molecules and the corresponding radicals as well as the spin distributions for the radicals. The results confirmed the differences in the reactivities of S and DHS.

Comparison of effects of D. Penicillamine and dexamethasone on the articular and non articular lesions in immune connective tissue disease induced in rabbits.

The effect of D. Penicillamine (DP) at the dose of 50 mg/kg/day, on an immune induced connective tissue disease in rabbit, is compared to that of dexamethasone (Dexa) at the doses of 0.15 and 0.075 mg/kg/day. This model includes polyarthritis and lesions of connective tissue of liver, kidneys and lungs. The result of immunization is initially a non-specific macrophage infiltration and secondarily a specific lymphocyte and plasma-cell infiltration. In short treatment, high dose of Dexa inhibits the non-specific and specific responses while DP modifies only non specific response. In long treatment, Dexa at low dose and DP inhibit the two responses. Data suggest that, in vivo, macrophages is the target cell of DP.

[Antinuclear antibodies: analysis of images and classification of immunofluorescence aspects on nuclei isolated from rat hepatocytes]. / Anticorps anti-noyaux: analyse d'images et classification des aspects d'immunofluorescence, sur noyaux isolés d'hépatocytes de rat.

Image analysis and Fourier transformation of the various nuclear immunofluorescence patterns observed while detecting antinuclear antibodies allow an objective and quantitative definition of the fluorescence. They also point out various IF types hidden by the main pattern, without having to dilute the test serum. They make obvious the difference between speckled and reticular patterns, and reveal the existence of intermediate states. The usual nuclear IF patterns (homogeneous, ring, nucleolar, reticulated, speckled and diffuse) may be grouped, according to their photo emission, into nuclear and subnuclear patterns. The first group includes homogeneous, annular and passive nucleolar IF. The second group is composed of speckled, reticulated, mixed, and active nucleolar IF. Alternatively, these aspects may be grouped into three types: homogeneous nuclear IF (homogeneous and ring), heterogeneous nuclear IF (speckled, reticulated and mixed) and nucleolar IF (active or passive). Diffusion can affect or not these aspects and does not apply to a special type or pattern. Image analysis and the study of the image spatial spectrum lead to automated recognition of the IF types, and later on, to the discrimination of antinuclear antibodies.

Manganese-containing superoxide-dismutase deficiency in polymorphonuclear leukocytes of adults with rheumatoid arthritis.

Superoxide dismutase (SOD) is known to regulate the level of superoxide radicals inside cells. The purpose of this work was to investigate the role of SOD activity in tissue damage produced by superoxide radicals. SOD was measured in polymorphonuclear cells of patients with rheumatoid arthritis and controls. The distinct SOD activities, including manganese-containing and copper-zinc-containing enzymes, were evaluated in cytoplasma and mitochondria of human granulocytes. Except for the comparison between total SOD and cytoplasmic copper-zinc SOD, no correlation was found among the different SOD levels. Moreover, a significant decrease was observed only for cytoplasmic manganese-containing enzyme in granulocytes of adults with rheumatoid arthritis. These data confirm the necessity of evaluation of various SOD classes and suggest the interest of biochemical tests in granulocytes for early diagnosis and better comprehension of tissue damage due to inflammation.

Electro-optic response and switchable Bragg diffraction for liquid crystals in colloid-templated materials.

We report optical switching studies on nematic liquid crystal incorporated into structures based on self-assembled colloids. We compare the electro-optic responses of liquid crystal imbibed into colloid-templated polymers, liquid crystal imbibed in the interstitial space of colloid crystals, and conventional polymer-dispersed liquid crystals. We characterize the Bragg diffraction of our templated liquid-crystal/polymer composites as a function of electric field and measure switching times. The response of liquid crystal in connected networks differs qualitatively from that of liquid crystal in isolated cavities.

Evaluation of a dry, rehydratable film method for rapid enumeration of Staphylococcus aureus.

Results with the new 3M Petrifilm Rapid S. aureus Count (RSA) Plate method were compared with those of the classical Baird-Parker agar (BPA) method for detection and enumeration of Staphylococcus aureus. Studies on 219 bacterial strains demonstrated that the Petrifilm RSA plate is more sensitive than and as specific as the classical BPA method for confirmed identification of S. aureus. Counts of colonies from 71 pure cultures, 61 naturally contaminated food samples, and more than 750 artificially inoculated food samples showed that the Petrifilm RSA method was as effective as the classical BPA method for identification and enumeration of S. aureus. The Petrifilm RSA method gave results in one-third the time required for the classical method.

[The genetic contribution of cows to the prevalence of mastitis in the following generation]. / Genetický podíl krav na prevalenci mastitid v následující generaci.

Investigations of mastitis prevalence were performed in 1227 dam-daughter pairs during 1985-1988. From the records of the last 10 years, the findings concerning mammary gland were obtained. Six herds of agricultural co-operatives in the Brno region were investigated. In the early period the Bohemian Pied breed, later the crosses of this breed with Red Holstein and a low genetic proportion of Ayrshire cattle, dominated in the set of cows. Udder health was assessed using the clinical and bacteriological findings in individual udder quarters and the results of the rapid mastitis test NK. Based on this data, the cows were divided into two groups--resistant and susceptible. The lactation order in which the disease appeared for the first time was of great importance. The occurrence of the disease in daughters and udder health in dams were in a significant relation (P less than or equal to 0.01). While nearly 50% of daughters of susceptible dams were affected, it was only 25% in daughters of the resistant group. The assessment of heritability of resistance to mastitis was carried out on the principle of dam-daughter regression. The index of heritability h2 = 0.52 +/- 0.06 was calculated. The results show that the genetic contribution of dams to the susceptibility of daughters to mastitis is significant.

[Acute necrotizing pancreatitis--a constant surgical problem]. / Akutní nekrotizující pankreatitida--trvalý problém chirurga.

The treatment of each patient admitted for the diagnose of acute pancreatitis requires a very individual approach. It is necessary to consider the possibility of severe complications. Currently, there is no specific therapeutic measure to prevent the development of pathogenic processes. In addition to all modes of supportive therapy and surgical sanatation of the local finding, it is necessary to retain the energetic intake by means of parenteral and enteral modes of nutrition provision.

[Correlation of urography, ultrasonography and computer tomography with clinical findings in examinations of the kidneys and retroperitoneum. Possibilities of diagnostic errors]. / Korelácia urografie, ultrasonografie a pocítacovej tomografie s klinickým nálezom pri vysetrení obliciek a retroperitónea. Moznosti diagnostických omylov.

The authors prospectively and retrospectively mutually compared the diagnostic value of urography, ultrasonography (USG), and computer tomography (CT) in four patients, with the diagnosis misstated prior to operation. The most valuable findings were provided by CT. Mere utilization of these methods does not exclude the possibility of diagnostic misstatements. In three patients the ultimate diagnosis was assessed on the basis of operative revision and in one patient, by means of cytologic examination of the renal punctate. Owing to retrospective evaluation of examinations, diagnostically valuable symptoms which could have lead to the correct diagnosis, were distinguished. Hence, examinations must necessarily be evaluated in complex with the general clinical state. (Fig. 6, Ref. 14.).

[Atrial-triggered and ventricular-inhibited ventricular VDD pacing with atrial and ventricular single catheter electrodes--the Phymos MPS pacemaker]. / Sínemi spoustená komorami inhibovaná stimulace komor VDD se sínovou a komorovou elektrodou na monokatetru--Phymos MPS.

Optimal treatment from the haemodynamic aspect in complete atrioventricular block is dual-chamber DDD pacing. It makes possible atrioventricular sequence and adaptability of the frequency of ventricular pacing to the load under the control of the sinoatrial node. DDD pacing calls, however, for an atrial and ventricular electrode on two leads. Italian authors developed in 1984 a pacemaker (PM) PHYMOS (MEDICO ITALIA) which makes possible atrial triggered ventricular VDD pacing by means of a single pass lead with a tripolar electrode. As the first ones in the post-communist countries the authors implanted to six patients a PM PHYMOS MPS. They confirmed that the advantages of the VDD system, as compared with DDD, are the use of a single pass lead. This makes the implantation quicker, simpler and safer and reduces the time of X-ray irradiation and the incidence of electrodes dislocations. Another advantage is the lower incidence of atrial fibrillation and probably also pacemaker mediated tachycardias. The disadvantages of the VDD system are that atrial stimulation is impossible and that the diameter of the single pass lead is somewhat larger. According to data in the literature and the so far small experience of the authors, when the indication is correct--atrioventricular block grade III or II with intact function of the sinoatrial node--the advantages of VDD pacing with a single pass lead, as compared with DDD pacing, predominate over disadvantages.