Search

International guidelines for the use of Rhesus D immunoglobulin: a comparative study and the contribution of fetal Rhesus genotyping The introduction of immunoglobulin D (Anti-D, Rhogamâ) has been a major advancement in the prevention of Rhesus D alloimmunisation. However, the required dose and timing of administration remain unclear. Fetal Rhesus genotyping, to administer Anti-D more selectively, is not yet available as part of standard care for Rhesus negative pregnant women in Belgium. We performed a comparative literature search, using the most recent international guidelines regarding the use of Anti-D and fetal Rhesus genotyping. There is international consensus that Anti-D should be administered prophylactically between 28-34 weeks of gestation and especially postpartum. The recommended dose and number of administrations vary strongly in different countries. There is no international consensus regarding the necessity to administer Anti-D nor the dose of Anti-D to be administered in the event of possible sensitisation during pregnancy. The results of our guideline study suggest that several possible causes of sensitisation require a lower dose of Anti-D than the currently available dose in Belgium (1500 IU). One could consider introducing a lower dose in Belgium in order to use Anti-D more efficiently. Nonetheless, further research concerning the ideal dose per indication and the ideal timing of administration is indicated. Fetal Rhesus D genotyping has a sensitivity of 99.9% and specificity of 99.2% when performed after 11 weeks of gestation. Given the high sensitivity and specificity of fetal Rhesus genotyping, a standard implementation of this test in Belgium certainly seems worth considering.

Noninvasive prenatal testing in multiple pregnancies: retrospective analysis of the fetal fraction distribution This article describes the distribution of the fetal fraction (FF) in noninvasive prenatal testing (NIPT) in multiple pregnancies. A retrospective study of 514 NIPT experiments was performed in 484 women with an (originally) multiple pregnancy from January 1st, 2017 to December 31st, 2019 at the Center for medical genetics in Ghent (Belgium). The FF distribution in vanishing twin, twin and triplet pregnancies was significantly different with a mean of 14%, 18.1% and 23.7% respectively. Test failure due to a low FF was higher in dichorionic-diamniotic (DCDA) twins (9.5%) compared to monochorionic-diamniotic (MCDA) twins (0%). The mean ratio FF based on the Y-chromosome (FFY)/total fetal fraction in male-male pregnancies was 0.96, in male-female pregnancies 0.43 and in female-female pregnancies 0.0. In vanishing twin pregnancies, the duration of persistence of cell-free fetal DNA (cffDNA) from the demised cotwin was up to 7 weeks. Test failure due to a low FF seems to appear more frequently in DCDA twins compared to MCDA twins, which could be explained by the higher FF threshold in DCDA twins (8%) in comparison to MCDA twins (4%). In mixed pregnancies, on average each fetus seems to contribute equally to the FF, although this does not seem to apply to all mixed pregnancies, giving rise to the possibility of an unequal contribution and a higher risk of a false negative result.

Flavobacterium columnare , a member of the phylum Bacteroidetes , causes columnaris disease in wild and aquaculture-reared freshwater fish. The mechanisms responsible for columnaris disease are not known. Many members of the phylum Bacteroidetes use type IX secretion systems (T9SSs) to secrete enzymes, adhesins, and proteins involved in gliding motility. The F. columnare genome has all of the genes needed to encode a T9SS. gldN , which encodes a core component of the T9SS, was deleted in wild-type strains of F. columnare . The F. columnare Δ gldN mutants were deficient in the secretion of several extracellular proteins and lacked gliding motility. The Δ gldN mutants exhibited reduced virulence in zebrafish, channel catfish, and rainbow trout, and complementation restored virulence. PorV is required for the secretion of a subset of proteins targeted to the T9SS. An F. columnare Δ porV mutant retained gliding motility but exhibited reduced virulence. Cell-free spent media from exponentially growing cultures of wild-type and complemented strains caused rapid mortality, but spent media from Δ gldN and Δ porV mutants did not, suggesting that soluble toxins are secreted by the T9SS. IMPORTANCE Columnaris disease, caused by F. columnare , is a major problem for freshwater aquaculture. Little is known regarding the virulence factors produced by F. columnare , and control measures are limited. Analysis of targeted gene deletion mutants revealed the importance of the type IX protein secretion system (T9SS) and of secreted toxins in F. columnare virulence. T9SSs are common in members of the phylum Bacteroidetes and likely contribute to the virulence of other animal and human pathogens.

Many clinicians, even those without graphological knowledge, are fascinated by the uniqueness of handwriting. No wonder, since the development of one’s handwriting is the result of a unique, complex developmental process. Fascination in handwriting led to the intriguing study of graphology as a means of understanding human behaviour. There are different schools of graphology, one being the ‘trait school’ which interprets handwriting according to single signs and the other being the holistic approach, which advocates the analysis of handwriting as a Gestalt. The trait-method does not do justice to the multi-faceted nature of personality, but the holistic approach seems to be a powerful expressive projective technique. The reliability of the holistic method was put to the test by comparing clinical findings of a patient to a ‘blind’ handwriting analysis of the same patient done independently by a professional graphologist. Graphological inferences made from the five (5) unique composite profiles, show strong similarities with diagnoses on Axis I and Axis II (DSM-IV-TRTM) (APA 2000) of the same patients, which confirms the value of graphology in psychological assessment.

An acardiac twin is probably the most severe malformation found in humans and is found in 1:100 of all monozygotic twin pregnancies and in about 1:30 of all monozygotic triplets. There are several types of acardiac fetuses; in fact, two major theories exist on their origin. Arterio-arterial anastomoses between the umbilical cord of the normal and the acardiac twin are responsible for the clinical presentation of a Twin Reversed Arterial Perfusion or TRAP sequence. Possible complications for the healthy twin are circulatory overload, which can lead to congestive heart failure, polyhydramnios (and, thus, preterm labor), and eventually intra-uterine demise. Furthermore, perinatal complications can occur due to the acardiac mass itself and hypoxia and intra-uterine growth retardation of the normal twin. Several risk factors to estimate the severity of the anomaly and the consequences for the healthy ‘pump’ twin have been identified. Using several ultrasonographic features, a distinction between TRAP sequence and twin-to-twin transfusion syndrome and other anomalies must be made. Treatment of the anomaly can either be conservative (reducing the polyhydramnios) or invasive. The latter consists of umbilical cord occlusion or interstitial vessel obliteration of the acardiac twin. Literature review has pointed out the benefits of the second option.

A selective reversed-phase liquid chromatography/mass spectrometry (LC/MS(n)) method is described for the identification of azithromycin impurities and related substances in commercial azithromycin samples. Mass spectral data are acquired on an LCQ ion trap mass spectrometer equipped with an atmospheric pressure chemical ionization interface operated in positive ion mode. The LCQ provides on-line LC/MS(n) capability, making it ideally suited for identification purposes. In comparison with UV detection, this hyphenated technique provides as its main advantage efficient identification of novel substances without time-consuming isolation and purification procedures. Using this technique, six novel related substances detected in commercial azithromycin samples have been studied.

An optimized capillary electrophoresis method for the analysis of doxycycline is described. The influence of methanol as organic modifier, buffer pH, buffer concentration, capillary length, column temperature, Triton X-100 and methyl-beta-cyclodextrin was systematically investigated. A central composite design was performed in order to optimize the method. The optimal separation conditions were: capillary, uncoated fused-silica [40 cm (32 cm effective length) x 50 microm I.D.]; background electrolyte, a solution of 145 mM sodium carbonate and 1 mM EDTA brought to pH 10.3-methanol (89:11, v/v); temperature, 15 degrees C; voltage, 12 kV. The method showed good selectivity, repeatability, linearity and sensitivity. Six commercial samples were quantitatively analyzed. The results were compared with those established by the liquid chromatography method from the European Pharmacopoeia.

A capillary zone electrophoresis method for analysis of polymyxin B sulfate is described. In this method, triethanolamine (TEA)-phosphate buffer at pH 2.5 was employed to reduce the adsorption of analyte onto the capillary wall. Methyl-beta-cyclodextrin (M-beta-CD) and 2-propanol (IPA) were found to be necessary for selectivity enhancement. In order to optimize the method and to control its robustness, a central composite design was performed with four parameters, i.e. concentration of M-beta-CD, TEA, IPA and buffer pH. The optimal separation conditions were as follows: capillary, 55 cm (50 microm I.D., 47 cm effective length); 130 mM TEA-phosphate buffer (pH 2.5) containing 5 mM M-beta-CD and 5% IPA; 24 kV (51 microA) applied voltage; column temperature, 20 degrees C. Further, linearity and limits of detection quantification were examined. Three commercial samples were analyzed quantitatively.

The separation of tylosin by micellar electrokinetic capillary chromatography with a mixed micelle system is described. Good selectivity was obtained with sodium phosphate buffer (80 mM, pH 7.5) containing 20 mM sodium cholate and 7 mM cetyltrimethylammonium bromide (CTAB). This method permits tylosin to be separated from its closely related substances within 15 min. The influences of type of buffer, buffer pH, the concentrations of sodium cholate and CTAB were investigated. The robustness of the method was examined for tylosin by means of a full-fraction factorial design. Quantitative results are presented. Using a similar buffer system (80 mM sodium phosphate, pH 6.0, 20 mM sodium cholate and 5 mM CTAB), separation of erythromycin and its main related substances was also obtained. However, detection sensitivity and resolution are not sufficient for analysis of related substances in erythromycin commercial samples.

To determine the effect of mastitis caused by Escherichia coli on expression of CD18 cell surface receptors and to evaluate the involvement and regulation of receptors by lipopolysaccharide (LPS) and cortisol.11 clinically normal lactating Holstein-Friesian cows.Binding of CD18 monoclonal antibodies to neutrophils was studied, using flow cytometry, before and after intramammary inoculation of E. coli organisms. Effect of LPS and cortisol on expression of adhesion receptors was investigated, using a whole-blood model.Expression of CD18 adhesion receptors on bovine neutrophils increased 35% by 12 hours after intramammary inoculation of E. coli. By 24 hours after inoculation, the number of receptors had returned to control values. High cortisol concentrations (100 nmol/L) were seen 12 to 18 hours after inoculation. Addition of LPS to blood induced a 30% increase in the number of CD18 receptors, and maximal number of receptors was expressed at an LPS concentration of 0.1 ng/ml. A decrease in the number of CD18 receptors was induced by incubation with cortisol or dexamethasone before challenge-exposure with LPS.An increase in the number of CD18 receptors on neutrophils is mediated by local production of LPS. Subsequent endogenous release of cortisol may prevent additional increases in the number of receptors.During acute mastitis caused by E. coli, there is an increase in the number of CD18 receptors on circulating neutrophils. Cortisol induces a decrease in the number of CD18 receptors, probably modulating the acute inflammatory response in mammary glands of lactating cows.

The determination of netilmicin sulfate by liquid chromatography using a column packed with poly (styrene-di-vinylbenzene) and pulsed electrochemical detection on a gold electrode is described. The mobile phase consisted of an aqueous solution containing 35 g 1-1 of sodium sulfate, 0.5 g 1-1 of sodium octanesulfonate, 10 ml 1-1 of tetrahydrofuran and 50 ml 1-1 of 0.2 M phosphate buffer (pH 3.0). The total analysis time was not more than 25 min. The effects of the different chromatographic parameters on this selection were also investigated. When a number of commercial samples of netilmicin sulfate was analyzed using this method, eight different components were separated, three of which were of unknown identities.

Saturation and inhibition binding studies, using [3H]CGP-12177 as radioligand, reveal the presence of a single class of beta-adrenoceptors on intact bovine mononuclear leukocytes. Receptor binding is inhibited by beta-adrenergic agonists with a rank order of potency (-)-isoproterenol(-)-epinephrine(-)-norepinephrine, indicating that the receptors are of the beta 2-subtype. By Scatchard analysis, the mean receptor density was 707 +/- 168 receptors/cell and the dissociation constant was 0.89 +/- 0.47 nM. Specific binding was very high and amounted to 80-90%. It is concluded that [3H]CGP-12177 is a promising compound for the reliable determination of the binding characteristics of bovine mononuclear leukocytes.

The semi-micro analysis of the bulk commercial samples of Oxytetracycline, Tetracycline and Chlortetracycline for their water content using Karl Fischer reagent is described. All the samples analysed comply with the limits laid down in European Pharmacopoeia or United States Pharmacopoeia.

Zusammenfassung Effekt einer Ornithininfusion im Dunndarm von Kuhen auf die Milchproduktion und einiger Blutplasmaparameter bei unterschiedlicher Futterungsintensitat Taglich wurde eine Losung mit 36,1 g Ornithin oder die entsprechende Null-Losung kontinuierlich in den Dunndarm von zwei Milchkuhen verabreicht. Die Rationen entsprachen 110% oder 75 % des Energie- und Eiweisbedarfs. Die Milchproduktion und Milchzusammensetzung wurden gemessen und der Blutplasmagehalt an Aminosauren, Glukose, Wachstumhormon (GH) und Insulin analysiert. Das Experiment wurde ein Jahr spater wiederholt. Die Ornithinzugabe hatte keinen Einflus auf die Milchproduktion und die Blutplasmazusammensetzung bei intensiver gefutterten Tieren. Dagegen wurden signifikant hohere Milch-, Laktose- und Fettgehalte gemessen bei nicht so intensiv gefutterten Tieren, wahrend die Eiweisproduktion nicht beeinflust wurde. Der Blutplasmagehalt an essentiellen und nicht essentiellen Aminosauren stieg an, auser bei Histidin und Methionin. Diese beiden Aminosauren konnen als limitierend fur die Milcheiweissynthese angesehen werden. Der Glukoseumsatz wurde ebenfalls durch die Ornithinzufuhr bei Unterfutterung verandert, da der Glukose- und Eiweisspiegel im Blutplasma und die Laktoseproduktion signifikant anstiegen. Die Ornithinzufuhr erhohte ebenfalls die GH-Sekretion bei Unterfutterung. Die vorliegenden Ergebnisse haben gezeigt, das eine Ornithininfusion in den Dunndarm von Kuhen die Milchproduktion stimuliert, wenn die Energie- und Eiweisversorgung unterhalb des Bedarfes liegt. Diese Feststellung wird durch die erhohte GH-Sekretion bestatigt.

A number of different processes clear peptides from the circulation. These will be described and illustrated using examples from a range of peptide hormones and analogues.

Bovine teat beta-adrenergic receptors were investigated with binding studies using 3H-dihydroalprenolol (DHA) as ligand. Inhibition of 3H-DHA-binding by beta 1-(betaxolol, atenolol, practolol) and beta 2-(zinterol, fenoterol, ICI 118551) selective drugs resulted in monophasic displacement curves with Hill coefficients close to 1.0. Computer assisted analysis of these curves did not reveal the presence of more than one class of binding sites. These data suggest that the beta-receptor sites present in bovine teat walls are of the beta 2-subclass.

Publisher Summary This chapter discusses a study to analyze the secretory activity of the cow's udder after injection of atropine. The effect of atropine on the hourly production and composition of milk was studied in 14 experiments performed on 2 cows. These animals were milked every hour with the aid of intravenous oxytocin injection for a total period of 8 h. Atropine was injected after the 3 rd milking. In 14 control experiments, NaCl 0.9 % was injected instead of atropine. The concentrations of casein and α-1a in fat free milk observed during the last 3 h of the atropine experiments were significantly lower than the corresponding values in the NaCl 0.9% experiments. Milk sodium, chloride, and lactose levels remained constant. As compared with the NaCl experiments, atropine induced significant reductions of the productions of milk (20%), lactose (18%), casein (35%), and α-la (45%) during the last hours of the experiments. It was found that there was a decrease of 20 to 50% of arterial amino acid concentrations, 50 min after the injection of atropine. These effects were accompanied by a decrease of the arterio-venous differences of the amino acids across the udder, amounting to 40–50%.

The synthesis of 6 beta-bromopenicillanic acid labeled with deuterium and tritium in the beta-methyl group is described. The S-sulfoxide of benzyl- or p-methoxybenzyl 6 alpha-bromopenicillanate is refluxed in benzene containing an excess of tert-BuOD, D2O or HTO. After deoxygenation and deprotection of the ester, the labeled 6 alpha-bromopenicillanic acid is epimerized (N,O-bis(trimethylsilyl)acetamide/1,5-diazabicyclo[4.3.0]non-5-ene in CH2Cl2). The two epimers are separated by column chromatography.

beta-Adrenoceptors present in smooth muscles of the cistern wall of teats of lactating cows were identified and characterized by 3H-dihydroalprenolol (DHA) binding. Binding association and dissociation at 37 degrees C were very rapid (t1/2 amounting to 2 min and 1 min 30 sec respectively), stereospecific and saturable. By Scatchard analysis, the mean receptor density was 92 fmol of 3H-dihydroalprenolol bound/mg protein and the dissociation constant was 0.63 nM. The Hill number was 1.01. The order of potency of catecholamines for the binding was (-)-isoproterenol greater than (-)-epinephrine greater than (-)-norepinephrine, indicating a population of mainly beta 2-adrenoceptors. Potency series of 10 antagonists and 15 agonists, determined from binding-inhibition experiments, were composed.

The highly specific alpha 1-adrenoceptor antagonist 3H-prazosin was used to characterize alpha 1-adrenoceptors in smooth muscles of the cistern wall of teats of lactating cows. Binding was rapid, readily reversible, stereospecific and saturable. Scatchard analysis of saturation binding data on bovine teat membranes revealed that 3H-prazosin bound to a single class (Bmax = 128 +/- 13 fmol/mg of membrane protein) of noncooperative sites (Hill coefficient = 0.95 +/- 0.02) with high affinity (KD = 0.47 +/- 0.05 nM). Competition binding studies with 9 antagonists and 8 agonists, indicated that 3H-prazosin labeled the alpha 1-receptor.

3H-Yohimbine and 3H-rauwolscine, both potent and selective alpha 2-adrenergic antagonists, were used to identify alpha 2-adrenoceptors in smooth muscles of the cistern wall of teats of lactating cows. Binding of these radioligands was rapid and readily reversed by 10 microM phentolamine. Saturation experiments in the presence of 3H-yohimbine showed an equilibrium KD value of 6.23 +/- 0.74 nM and a maximum number of sites of 81 +/- 7 fmol/mg of membrane protein. In the presence of 3H-rauwolscine, however, a higher density of alpha 2-receptors (164 +/- 12 fmol/mg protein) with a KD of 6.16 +/- 0.64 nM was found. No cooperative interactions among both binding sites were observed. Both 3H-yohimbine and 3H-rauwolscine binding sites showed alpha 2-adrenergic specificity. On the basis of its higher affinity to the alpha 2-adrenoceptor sites, better ratio of specific to non-specific binding and for other reasons, 3H-rauwolscine appears to be the ligand of choice.