Coexistence of antithrombin deficiency and suspected inferior vena cava atresia in an adolescent and his mother - case report and clinical implications.

BACKGROUND: Antithrombin deficiency (ATD) is an autosomal dominant thrombophilia presenting with varying phenotypes. In pediatric patients with ATD, thrombosis typically develops during the neonatal period or adolescence. However, to date there are no consistent recommendations on the therapeutic management of children with ATD. Inferior vena cava atresia (IVCA) belongs to a range of congenital or acquired vena cava malformations and is described as an independent risk factor for thrombosis. The present case report explores two cases of combined ATD and IVCA in an adolescent and his mother. CASE PRESENTATION: A 14-year-old male presented with extensive deep venous thromboses (DVTs) of both lower extremities as well as an IVCA. The patient had previously been diagnosed with an asymptomatic ATD without therapeutic consequences at that time. His mother was suffering from an ATD and had herself just been diagnosed with IVCA, too. The DVTs in the adolescent were treated by systemic anticoagulation and catheter-directed local thrombolysis causing favourable results. Yet, despite adequate oral anticoagulation the DVTs in both lower extremities reoccurred within 1 week after the patient was discharged from hospital. This time, thrombolysis could not be fully achieved. Surprisingly, probing and stenting of the IVCA was achieved, indicating an acquired IVCA which could have occurred after undetected thrombosis in early childhood. Genetic analyses showed the same mutation causing ATD in both son and mother: heterozygote missense mutation c.248 T > C, p.(Leu83Pro), within the heparin binding domain of antithrombin. This mutation was never reported in mutation databases before. CONCLUSIONS: To our knowledge this is the first case report discussing combined ATD and IVCA in two family members. Since ATDs present with clinical heterogeneity, taking a thorough family history is crucial for the anticipation of possible complications in affected children and decisions on targeted diagnostics and therapeutic interventions. Affected families must be educated on risk factors and clinical signs of thrombosis and need an immediate diagnostic workup in case of clinical symptoms. IVCA in patients with ATD could occur due to thrombotic occlusion at a very early age. Therefore, in case of family members with IVCA and ATD ultrasound screening in newborns should be considered.

Development of a novel fluorine-18 labeled deuterated fluororasagiline ([(18)F]fluororasagiline-D2) radioligand for PET studies of monoamino oxidase B (MAO-B).

The objective of this study was to synthesize and evaluate a novel fluorine-18 labeled deuterium substituted analogue of rasagiline (9, [(18)F]fluororasagiline-D2) as a potential PET radioligand for studies of monoamine oxidase B (MAO-B). The precursor compound (6) and reference standard (7) were synthesized in multi-step syntheses. Radiolabeling of 9 was accomplished by a two-step synthesis, compromising a nucleophilic substitution followed by hydrolysis of the sulfamidate group. The incorporation radiochemical yield from fluorine-18 fluoride was higher than 30%, the radiochemical purity was >99% and the specific radioactivity was >160GBq/µmol at the time of administration. In vitro compound 7 inhibited the MAO-B activity with an IC50 of 173.0±13.6nM. The MAO-A activity was inhibited with an IC50 of 9.9±1.1µM. The fluorine-18 version 9 was characterized in the cynomolgus monkey brain where a high brain uptake was found (275% SUV at 4min). There was a higher uptake in the striatum and thalamus compared to the cortex and cerebellum. A pronounced blocking effect (50% decrease) was observed in the specific brain regions after administration of l-deprenyl (0.5mg/kg) 30min prior to the administration of 9. Radiometabolite studies demonstrated 40% of unchanged radioligand at 90min post injection. An efficient radiolabeling of 9 was successfully established and in the monkey brain 9 binds to MAO-B rich regions and its binding is blocked by the selective MAO-B compound l-deprenyl. The radioligand 9 is a potential candidate for human PET studies.

Synthesis and biological evaluation of novel propargyl amines as potential fluorine-18 labeled radioligands for detection of MAO-B activity.

The aim of this project was to synthesize and evaluate three novel fluorine-18 labeled derivatives of propargyl amine as potential PET radioligands to visualize monoamine oxidase B (MAO-B) activity. The three fluorinated derivatives of propargyl amine ((S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)-pent-4-en-2-amine (5), (S)-N-(1-fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (10) and (S)-1-fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (15)) were synthesized in multi-step organic syntheses. IC(50) values for inhibition were determined for compounds 5, 10 and 15 in order to determine their specificity for binding to MAO-B. Compound 5 inhibited MAO-B with an IC(50) of 664 ± 48.08 nM. No further investigation was carried out with this compound. Compound 10 inhibited MAO-B with an IC(50) of 208.5 ± 13.44 nM and compound 15 featured an IC(50) of 131.5 ± 0.71 nM for its MAO-B inhibitory activity. None of the compounds inhibited MAO-A activity (IC(50) > 2 µM). The fluorine-18 labeled analogues of the two higher binding affinity compounds (10 and 15) (S)-N-(1-[(18)F]fluoro-3-(furan-2-yl)propan-2-yl)-N-methylprop-2-yn-1-amine (16) and (S)-1-[(18)F]fluoro-N,4-dimethyl-N-(prop-2-ynyl)pentan-2-amine (18) were both prepared from the corresponding precursors 9A, 9B and 14A, 14B by a one-step fluorine-18 nucleophilic substitution reaction. Autoradiography experiments on human postmortem brain tissue sections were performed with 16 and 18. Only compound 18 demonstrated a high selectivity for MAO-B over MAO-A and was, therefore, chosen for further examination by PET in a cynomolgus monkey. The initial uptake of 18 in the monkey brain was 250% SUV at 4 min post injection. The highest uptake of radioactivity was observed in the striatum and thalamus, regions with high MAO-B activity, whereas lower levels of radioactivity were detected in the cortex and cerebellum. The percentage of unchanged radioligand 18 was 30% in plasma at 90min post injection. In conclusion, compound 18 is a selective inhibitor of MAO-B in vitro and demonstrated a MAO-B specific binding pattern in vivo by PET in monkey. It can, therefore, be considered as a candidate for further investigation in human by PET.

Synthesis and evaluation of [¹8F]fluororasagiline, a novel positron emission tomography (PET) radioligand for monoamine oxidase B (MAO-B).

The aim of this study was to synthesize and evaluate a novel fluorine-18 labeled analogue of rasagiline (6) as a PET radioligand for monoamine oxidase B (MAO-B). The corresponding non-radioactive fluorine-19 ligand, (1S,2S)-2-fluoro-N-(prop-2-yn-1-yl)indan-1-amine (4), was characterized in in vitro assays. The precursor compound (3aS,8aR)-3-(prop-2-yn-1-yl)-3,3a,8,8a-tetrahydroindeno[1,2-d][1,2,3]oxathiazole 2,2-dioxide (3) and reference standard 4 were synthesized in multi-step syntheses. Recombinant human MAO-B and MAO-A enzyme preparations were used in order to determine IC(50) values for compound 4 by use of an enzymatic assay employing kynuramine as substrate. Radiolabeling was accomplished by a two-step synthesis, compromising a nucleophilic substitution followed by hydrolysis of the sulphamidate group. Human whole hemisphere autoradiography (ARG) was performed with [(18)F]fluororasagiline. Blocking experiments with pirlindole (MAO-A), L-deprenyl and rasagiline (MAO-B) were conducted to demonstrate the specificity of the binding. A positron emission tomography (PET) study was carried out in a cynomolgus monkey where time activity curves for whole brain and regions with high and low MAO-B activity were recorded. Radiometabolites were measured in monkey plasma using gradient HPLC. Compound 4 inhibited MAO-B with an IC(50) of 27 nM and MAO-A with an IC(50) of 2.3 µM. Radiolabeling of precursor 3 and subsequent hydrolysis of the protecting group towards (1S,2S)-2-[(18)F]fluoro-N-(prop-2-yn-1-yl)indan-1-amine (6) was successfully accomplished with an radiochemical yield of 40-70%, a radiochemical purity higher than 99% and a specific radioactivity higher than 200GBq/µmol. ARG demonstrated selective binding for [(18)F]fluororasagiline (6) to MAO-B containing brain regions, for example, striatum. The initial uptake in the monkey brain was 250% SUV at 4 min post injection. The highest amounts of radioactivity were observed in the striatum and thalamus as expected whereas in the cortex and cerebellum lower levels were observed. Metabolite studies demonstrated 30% unchanged radioligand at 90 min post injection. Our investigations demonstrated that the new ligand [(18)F]fluororasagiline (6) binds specifically to MAO-B in vitro and has a MAO-B specific binding pattern in vivo. Thus, it could serve as a novel potential candidate for human PET studies.

Uterine activity monitoring during labour--a multi-centre, blinded two-way trial of external tocodynamometry against electrohysterography.

PURPOSE: The aim of this study was to determine the quality of intrapartum uterine activity (UA) monitoring in daily practice during the first and second stages of labour. The total duration of inadequate UA monitoring is quantified in relation to the technique applied, namely, external tocodynamometry (TOCO) or electrohysterography (EHG). MATERIAL AND METHODS: 144 UA recordings, collected from 1st September 2008 until 15th October 2009 from deliveries at the Marien-Hospital Witten, Germany, were analysed by obstetricians based at different centres. The included recordings were from singleton and simultaneously with external TOCO and EHG monitored pregnancies. External TOCO and EHG UA recordings were blinded. RESULTS: The percentages of "adequate" UA recordings in the first and second stages of labour were much higher for the external EHG than the external TOCO mode (p<0.001). All doctors evaluated the UA assessment as "easier" (p <0.001) using the EHG compared with TOCO. CONCLUSION: Intrapartum UA monitoring in -daily practice via the EHG mode provides a more recognisable UA trace than the TOCO.

The microplastic profile of an urban African stream.

Microplastics are small plastic fragments that have been found around the world, however, research into microplastics in Africa and freshwater systems remains insufficient. In this study, the snapshot microplastic profile of an urban stream was assessed in the Braamfontein Spruit, located in Johannesburg the largest city in South Africa. The abundance of microplastics was determined in water, Chironomus sp. larvae and sediment, while in situ parameters were taken to investigate their relationship to the microplastic profile of the different matrices. Microplastics were detected in water (mean of 705 particles m-3), Chironomus sp. larvae (mean of 53.4 particles g-1 wet weight) and sediment (mean of 166.8 particles kg-1 dry weight). The study found evidence of how urban stream characteristics such as a weir, stream depth and velocity could affect the abundance and dispersion of microplastics. The results indicate that areas of increased depth and decreased flow allowed microplastics to settle down to the sediment where benthic macroinvertebrates could ingest these fragments. Large obstructions like a weir also increased microplastic counts in sediment and invertebrates above the obstruction, with a decrease of fragments after the obstruction, however, microplastics in surface water were able to flow over the obstruction and increase in abundance downstream. This study concludes that first order urban streams such as the Braamfontein Spruit may be contributing large numbers of microplastics to higher order streams and large rivers in times of increased flow.

Calcium balance during calcitriol and paricalcitol administration in healthy humans.

OBJECTIVE: Secondary hyperparathyroidism in hemodialysis patients requires optimal correction of vitamin D deficiency with active vitamin D and analogues. It has been postulated that new vitamin D analogues, i.e. paricalcitol, efficiently suppress parathyroid hormone serum levels (PTH), but do not increase intestinal calcium absorption as much as calcitriol. The effects of calcitriol and paricalcitol on calcium balance can best be characterized under standardized conditions in healthy individuals with normal renal function, because the urinary calcium excretion at steady state corresponds to the net calcium absorption in the gut. METHODS: In a randomized, double-blind, placebo-controlled, 3-way crossover Phase I study in 13 healthy individuals we investigated the changes compared to placebo in PTH and urinary calcium excretion during 6-day treatment periods with paricalcitol (1.5 microg/day) and calcitriol (0.5 microg/day). RESULTS: 24-hour urinary calcium excretion was stable during 6 days of placebo administration. Neither paricalcitol nor calcitriol significantly changed calcium excretion. Urinary creatinine, magnesium and phosphate excretion also remained unchanged over the study periods irrespective of the treatment. However, calcitriol was shown to be effective in reducing iPTH levels during 6 days of treatment (mean reduction 4.03+/-0.69 pmol/l), whereas paricalcitol had no effect. CONCLUSION: Using a dosing ratio of 1:3 for calcitriol:paricalcitol, i.e. the same conversion factor used previously in studies on hemodialysis patients, only calcitriol was able to reduce iPTH levels in healthy individuals. Low-dose calcitriol reduced iPTH levels without raising calcium absorption and without including any hypercalcemia.

Targeted ED-B fibronectin SPECT in vivo imaging in experimental atherosclerosis.

AIM: The extracellular matrix protein ED-B fibronectin (ED-B) is upregulated in inflammatory atherosclerotic lesions. However, functional in vivo imaging of ED-B-containing plaques has not been explored. This study evaluated whether [(99m)Tc]-conjugated AP39 ([(99m)Tc]-AP39), a single-chain antibody specific to ED-B, can be used for in vivo detection of atherosclerotic plaques in Western diet (WD)-fed, apolipoprotein E-deficient (apoE-/-) mice as compared to wildtype (WT) control mice. METHODS: Using SPECT, 12-month-old WD-fed apoE-/- and WT mice were studied 4 hours after injecting [(99m)Tc]-AP39 (148 MBq). Subsequently, mice were sacrificed, thoracic aortas measured in a g-counter, and plaques analyzed using histology, immuno-histochemistry, autoradiography, and morphometry. RESULTS: In vivo [(99m)Tc]-AP39-SPECT imaging of apoE-/- mice demonstrated a significant signal activity in the plaque-ridden thoracic aorta (52.236 ± 40.646 cpm/cm³) that co-localized with the aortic arch and the supra-aortic arteries in MRI scans. Low signal activity (9.468 ± 4.976 cpm/cm³) was observed in WT mice. In apoE-/- mice, the strongest signals were detected in the aortic root, aortic arch and along the abdominal aorta. Autoradiography analysis of aortas from apoE-/- mice confirmed the in vivo observation by demonstrating signal localization in atherosclerotic plaques. The size of autoradiography-positive plaque areas correlated significantly with the size of ED-B-positive (r=0.645, P=0.044) or macrophage-infiltrated (r=0.84, P<0.002) plaques. A significant correlation was found between the sizes of ED-B-positive and macrophage-infiltrated plaque areas (r=0.93, P<0.01). CONCLUSION: [(99m)Tc]-AP39-SPECT in vivo imaging detects inflammatory plaque lesions in WD-fed apoE-/- mice.

Sequence of the gene encoding ribosomal protein L11 from Thermus thermophilus HB8.

The complete nucleotide sequence of the gene encoding ribosomal protein L11 from the extreme thermophilic eubacterium, Thermus thermophilus HB8, was determined. L11 amino acid (aa) sequences from mesophilic and halophilic organisms, as well as from another thermophiles, were compared with the T. thermophilus L11 aa sequence.

Reduced oxidative stress during acellular reperfusion of the rat liver after hypothermic oscillating perfusion.

BACKGROUND: ATP resynthesis during reperfusion after liver preservation has been shown to be well correlated with the function of transplanted grafts. Nevertheless, the advantages of a cellular energy charge loading during the preservation period are yet not fully understood. This study evaluates the effects of different nucleotide levels at the end of preservation on metabolic changes and oxidative stress during reperfusion. METHODS: Two experimental groups were chosen reflecting different energy charge states after preservation: static cold storage for 10 hr and hypothermic oxygenated oscillating perfusion for 10 hr. In both experimental groups, normothermic ex vivo acellular reperfusion over 40 min was performed. A third group consisted of nonpreserved livers similarly reperfused for 40 min. Superoxide formation was detected by the superoxide dismutase inhibitable reduction of ferricytochrome c added to the normothermic perfusate. RESULTS: Superoxide formation and lipid peroxidation malondialdehyde were significantly lower during reperfusion after the energy charge loading before reperfusion by the hypothermic oscillating perfusion technique. However, oxygen radical formation, liver cell injury (lactate dehydrogenase [LDH] release), and TNFalpha release were significantly higher in energy charge-depleted groups (nonpreserved and cold stored livers). CONCLUSIONS: Hypothermic oscillating oxygenated perfusion led to the elevated energy charge during preservation and led to reduced oxygen radical formation as well as less lipid peroxidation during reperfusion, in contrast to cold stored livers and nonpreserved livers. This suggests a correlation between the energy charge before reperfusion and oxygen radical formation as well as liver injury at reperfusion.

Contrast adaptation in human retina and cortex.

PURPOSE: Although cortical contrast adaptation has been extensively studied with both psychophysical and electrophysiological techniques, little is known about retinal contrast adaptation in humans. METHODS: Retinal and cortical long-term contrast adaptation was assessed with simultaneous measurement of pattern electroretinogram (PERG) and cortical visual evoked potentials (VEPs). This study involved three approaches: sampling of the contrast transfer function from 2.7% to 98% with adaptation to high (98%) and low (7.3%) contrasts, linearity of adaptation effects, and transfer of contrast adaptation between parallel and orthogonal grating orientations. RESULTS: Contrast adaptation affected retinal and cortical recordings quite differently. The VEP showed a sigmoid contrast transfer function, which was shifted toward higher contrasts (by a factor of 1.9), whereas amplitudes at higher test contrasts were enhanced to 127%. The PERG decreased in amplitude to approximately 90%, and the latency was significantly reduced by 4 to 6 msec (P < 0.05). All measured effects were linear with adaptation contrast. Orientation played no role in the PERG results, whereas the VEP was enhanced to 125% when tested parallel and to 150% when tested orthogonal to adaptation. CONCLUSIONS: VEP results confirm and extend previous findings and fit well with single-cell recordings. The PERG findings suggest that retinal contrast adaptation occurs and mainly operates in the temporal domain, comparable to rapid gain-control findings in cats and primates.

Subtractive hybridization of cDNA from small amounts of plant tissue.

A subtractive hybridization method is described that allows the generation of a subtractive gene library from small amounts of plant or other eukaryotic tissues. The method uses paramagnetic oligo-dT beads to capture poly-adenylated mRNA and to synthesize the complementary cDNA on a solid support. The use of magnetic beads facilitates the change of reaction buffers and the removal of primers and minimizes yield losses. Subtracted material obtained from this method can either be cloned directly or used to screen a specific library.

A double-blind comparison of amitriptylinoxide versus doxepine in the treatment of severe depression.

In a double-blind parallel group study the efficacy and safety of amitriptylinoxide were evaluated vs. doxepine in the treatment of in-patients with severe depression. Two groups of 22 patients each received amitriptylinoxide and doxepine respectively at a daily dosage of 120-360 mg for a period of 4 weeks. The total score on the Hamilton Depression Scale (HAMD) was reduced with amitriptylinoxide on an average from 28 +/- 5 before treatment to 12 +/- 8 at the end of treatment, with doxepine from 29 +/- 8 to 13 +/- 11. Of the amitriptylinoxide-treated patients, 12 showed a more than 50% reduction in this score compared with 15 under doxepine. The difference was not statistically significant. Twenty patients in each group experienced adverse drug reactions, the percentage of anticholinergic side effects being equal in the two groups.

Hypervascular spinal tumors: influence of the embolization technique on perioperative hemorrhage.

BACKGROUND AND PURPOSE: Corporectomy is an effective treatment for vertebral metastases; however, massive perioperative hemorrhage is often associated with this procedure. We compared preoperative particle, particle-coil, and coil embolizations of hypervascular spinal tumors prior to vertebral body replacement to determine which prevented perioperative hemorrhage most effectively. METHODS: The vertebral tumors of 59 patients were embolized prior to corporectomy. In 26 cases, only coils were used for the proximal occlusion of feeding segmental arteries. Twenty-four patients received a combination of polyvinyl alcohol (PVA) particles and coils, and nine tumors were embolized with particles alone. We compared intraoperative blood loss between the three groups and 10 other patients who did not undergo embolization prior to corporectomy. RESULTS: Estimation of intraoperative hemorrhage showed a median value of 4350 mL in patients without embolization, 2650 mL in cases of coil embolization, 1850 mL in cases of particle-coil embolization, and 1800 mL in cases of particle embolization. The difference between unembolized patients and those who underwent coil embolization was not statistically significant. Particle and particle-coil embolizations showed very similar results, and reduced hemorrhage significantly as compared to unembolized and proximal coil occlusion cases. Residual bleeding came from the venous system and the neighborhood of the embolized region. CONCLUSION: Particle embolization prior to corporectomy can reduce perioperative hemorrhage. The additional benefit of proximal coil occlusion of arterial feeders is questionable.

Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation.

Genetic transformation of Petunia hybrida with a reporter gene and selectable marker gene (35S-bar) was achieved in similar frequencies by pollinating flowers with pollen vacuum-infiltrated with Agrobacterium tumefaciens or applying a drop of Agrobacterium suspension to the stigma immediately prior to pollination. Nine percent of the T1, and 5% of the T2 progeny germinated in nutrient medium with 3 mgl/l BastaR. Polymerase chain reaction assays indicated that of the BastaR-resistant plants, 66% of the T1 plants, and 61% of the T2 plants harboured the GUS gene. Histochemical assays showed that 10% of the putatively transformed T1 plants and 5% of their progeny expressed GUS in leaf tissue, pistils and young anthers. Southern hybridization confirmed genomic integration of the bar gene in one to three places in selected T1 and T2 progeny.

Treatment outcome for a maltreated population: benefits, procedural decisions, and challenges.

OBJECTIVE: To review the benefits, challenges, and procedural decisions to consider when implementing and managing a treatment outcome program for a maltreated population. METHOD: We reviewed the reasons to implement a treatment outcome program, decisions regarding procedures, and challenges likely to be encountered based on literature in the field and the authors' 6 years of experience in developing and maintaining an outcome program at a center specializing in the treatment of maltreated children. RESULTS: The development of an outcome programs requires careful measurement selection, early and ongoing staff-involvement, support from higher management, a well-developed data base and client tracking system, a coordinator and support staff, clinical utility, planning for fiscal impact, and flexibility to contend with challenges. CONCLUSIONS: Based on our experience, the plethora of clinically rich and administratively useful information derived from an outcome program far outweighs the challenges and costs of establishing and maintaining an outcome program.

Comparison of methods for the analysis of 226Ra in water samples.

226Ra is one of the key nuclides among the natural radionuclides from the point of view of radiation protection. For monitoring the radiation exposure risk along the water pathway it has to be measured routinely with low detection limits. Because of the properties of 226Ra and its progenies, a number of quite different methods are possible for this purpose. This paper presents a comparison of routine techniques for the determination of 226Ra in water samples as applied by the authors.

[Contact lenses--possibilities, problems and hazards]. / Kontaktlinsen--Möglichkeiten, Probleme und Gefahren.

Because use of contact lenses has become more common, knowledge of the basics is necessary. The most important kinds and materials of contact lenses, the indications for their application, and especially the important complications are illustrated by some clinical examples. Some simple first-aid measures are presented.