Paternal age and first trimester placental size and growth: The Rotterdam Periconceptional Cohort.

INTRODUCTION: Despite a noticeable trend of delayed fatherhood, less is known about the impact of paternal age on the paternally programmed placenta. We hypothesize that paternal aging affects seminal quality and as such induces ageing-related epigenetic alterations that influence placental growth. Our main aim is to investigate associations between paternal age and first trimester (vascular) placental growth trajectories. METHODS: Pregnant women were enrolled before 10 weeks of gestation in the Rotterdam Periconceptional Cohort (Predict study). Placental volumes (PV) and utero-placental vascular volumes (uPVV) were measured at 7, 9, and 11 weeks gestation. Associations between paternal age and PV and uPVV were investigated using linear mixed models and the maximum likelihood ratio test to test non-linear relationships. We adjusted for gestational age, fetal sex, parental smoking and maternal age, BMI, education and parity, and stratified for conception mode. RESULTS: From 808 pregnancies we obtained 1313 PV and from 183 pregnancies 345 uPVV measurements. We show no associations between paternal age and PV (p = 0.934) and uPVV (p = 0.489) in our total population or in pregnancies conceived naturally (PV p = 0.166; uPVV p = 0.446) and after IVF/ICSI (PV p = 0.909; uPVV p = 0.749). For example, PV was 0.9% smaller (95% CI -5.7%-7.1%) in fathers aged 40 compared to 30 years old at 9 weeks gestation in the total study population. DISCUSSION: We are not demonstrating a significant impact of paternal age on first trimester placental growth in a tertiary care population. Given the trend of increasing paternal age, our study should be repeated in the general population.

Treatment of acute myocardial infarction in the sub-arctic region of Norway. Do we offer an equal quality of care?

Patients, relatives, healthcare workers and administrators are concerned about the quality of care offered. We aimed to explore the treatment of acute myocatrdial infarction (AMI) in Northern Norway, compare it with the national figures, and document whether there is an equal quality of care or not. The retrospective study included data on patients' treatment for AMI. The following sources were employed. The Norwegian Patient Registry, National Quality of Care Database, Norwegian Myocardial Infarction Registry and data from the National Air Ambulance Services of Norway. The period 2012-2014/15 was studied and the variables were: incidence of AMI, gender and age adjusted rates of AMI and revascularization (PCI, CABG) based on patient's place of living (according to hospital catchment area) and 30-day survival rate. The annual incidence of AMI was 9% higher in the northern region. Significant incidence variations (2.7-5.9 AMI/1000 inhabitants) between the hospitals' catchment areas were revealed. The 30-day survival rate varied between 85.1-92.1% between hospitals. The variation in revascularization/AMI rate was 0.72-1.54. Air amublance services' availability varied through the day. In conclusion, significant variations in the AMI rate and an unequal service within the region was revealed.

Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma.

BACKGROUND: MicroRNAs (miRNAs) regulate expression of many cancer-related genes through posttranscriptional repression of their mRNAs. In this study we investigate the proto-oncogene MYCN as a target for miRNA regulation. METHODS: A luciferase reporter assay was used to investigate software-predicted miRNA target sites in the 3'-untranslated region (3'UTR) of MYCN. The miRNAs were overexpressed in cell lines by transfection of miRNA mimics or miRNA-expressing plasmids. Mutation of the target sites was used to validate MYCN 3'UTR as a direct target of several miRNAs. To measure miRNA-mediated suppression of endogenous N-myc protein, inhibition of proliferation and inhibition of clonogenic growth, miRNAs were overexpressed in a MYCN-amplified neuroblastoma cell line. RESULTS: The results from this study show that MYCN is targeted by several miRNAs. In addition to the previously shown mir-34a/c, we experimentally validate mir-449, mir-19a/b, mir-29a/b/c, mir-101 and let-7e/mir-202 as direct MYCN-targeting miRNAs. These miRNAs were able to suppress endogenous N-myc protein in a MYCN-amplified neuroblastoma cell line. The let-7e and mir-202 were strong negative regulators of MYCN expression. The mir-101 and the let-7 family miRNAs let-7e and mir-202 inhibited proliferation and clonogenic growth when overexpressed in Kelly cells. CONCLUSION: The tumour-suppressor miRNAs let-7 and mir-101 target MYCN and inhibit proliferation and clonogenic growth of MYCN-amplified neuroblastoma cells.

The medicinal chemistry of short lactoferricin-based antibacterial peptides.

This review discusses antibacterial peptides from the perspective of development into clinically useful chemotherapeutic drugs using short lactoferricin based peptides as examples. The review shows how important features for antibacterial activity can be identified and explored using the molecular properties of a range of natural and non-natural amino acids. The results have been further refined quantitatively using a "soft-modelling" approach where important structural parameters that influence the antibacterial activity of 15-residue model peptides were identified. The review describes how this knowledge is utilised to generate pharmacophores for antibacterial efficacy. These pharmacophores turn out to be surprisingly small and relatively consistent between typical Gram-negative and Gram-positive bacteria leading to the discovery of a novel class of short synthetic cationic antimicrobial peptides. These compounds are found to have high antibacterial activity against several bacterial strains that are resistant to commercial antibiotics, and are promising as future clinical candidates for treatment of infections caused by several clinically relevant pathogens.

Facile degradative lactonization of Gln-Arg and Gln-Phe hydroxyethylene dipeptide derivatives.

We have found that hydroxyrthylene (HE) dipeptide analogs of Gln-Arg and Gln-Phe are usually susceptible to acid catalyzed lactonization. The synthesis of substrate-based transition state analog inhibitors of botulinum neurotoxin metalloprotease inhibitors that contain the Gln-Arg or the Gln-Phe HE units is complicated by this facile degradative lactonization.

Screening of first degree relatives of patients operated for colorectal cancer: evaluation of fecal calprotectin vs. hemoccult II.

UNLABELLED: Fecal calprotectin (CPT) is elevated in the majority of patients with known colorectal cancer (CRC), but the specificity is not clarified. AIM: To evaluate if a CPT test (PhiCal ELISA) was more sensitive than Hemoccult II test in detecting colorectal neoplasia, and to obtain reference values in subjects with normal colonoscopy. To evaluate a possible relation between number and extent of dysplasia of adenomas in first degree relatives of patients with CRC and the stage of the carcinoma in the index casus. Further to study the prevalence of CRC and adenomas in the first degree relatives of patients operated for CRC. METHOD: In a multicenter study, 253 first degree relatives of patients with CRC, aged 50-75 years (mean age 60 years) underwent colonoscopy after having delivered stool samples and three Hemoccult II slides. RESULTS: In 237 first degree relatives from 148 patients with CRC, polyps were found in 118 (50%). Seventy three (31%) had adenomas and 17 had adenomas > or =10 mm. Five had asymptomatic cancers. The specificity of fecal CPT for adenomas at cut off levels 15 mg/l. The sensitivity of Hemoccult II for adenomas was 8%, and 4/5 of patients with carcinoma had negative Hemoccult II. The specificity for adenomas was 95%. CONCLUSION: Fecal CPT test was more sensitive than Hemoccult II in detecting colorectal neoplasia but the specificity was lower. In a high risk group like first degree relatives of patients with CRC, there are good reasons to consider fecal CPT as a first test in selecting patients for endoscopy.

Bulky aromatic amino acids increase the antibacterial activity of 15-residue bovine lactoferricin derivatives.

A model peptide, FKCRRWQWRMKKLGA, residues 17-31 of bovine lactoferricin, has been subjected to structure-antibacterial activity relationship studies. The two Trp residues are very important for antibacterial activity, and analogue studies have demonstrated the significance of the size, shape and aromatic character of the side chains. In the current study we have replaced Trp residues in the model peptide with bulky aromatic amino acids to elucidate further the importance of size and shape. The counterproductive Cys residue in position 3 was also replaced by these aromatic amino acids. The largest aromatic amino acids employed resulted in the most active peptides. The peptides containing these hydrophobic residues were generally more active against Staphylococcus aureus than against Escherichia coli, indicating that the bacterial specificity as well as the antibacterial efficiency can be altered by employing large hydrophobic aromatic amino acid residues.

The role of tryptophan in the antibacterial activity of a 15-residue bovine lactoferricin peptide.

Bovine lactoferricin is a 25-residue antibacterial peptide isolated after gastric cleavage of the iron transporting protein lactoferrin. A 15-residue fragment, FKCRRWQWRMKKLGA of this peptide sustains most of the antibacterial activity. In this truncated sequence, the two Trp residues are found to be essential for antibacterial activity. The anchoring properties of Trp, as have been observed in membrane proteins, are believed to be important for the interaction of Trp containing antibacterial peptides with bacterial cell membranes. We have investigated the molecular properties which make Trp important for the antibacterial activity of the 15-residue peptide by replacing Trp with natural and unnatural aromatic amino acids. This series of peptides was tested for antibacterial activity against Echerichia coli and Staphylococcus aureus. We found that neither the hydrogen bonding ability nor the amphipathicity of the indole system are essential properties for the effect of Trp on the antibacterial activity of the peptides. Replacement of Trp with residues containing aromatic hydrocarbon side chains gave the most active peptides. We propose that aromatic hydrocarbon residues are able to position themselves deeper into the bacterial cell membrane, making the peptide more efficient in disrupting the bacterial cell membrane. From our results the size, shape and aromatic character of Trp seem to be the most important features for the activity of this class of Trp containing antibacterial peptides.

[MRI findings in Wernicke encephalopathy]. / MRT-Befunde bei der Wernicke-Enzephalopathie.

Wernicke's encephalopathy (WE) is a consequence of vitamin B1 (thiamine) deficiency and in the majority of cases due to alcoholism. We report here the case of a 26-year-old male alcoholic who had stayed helplessly at home for 4 days until hospital admission. Clinical diagnosis was difficult due to major disturbance of consciousness. MRI showed an increase in signal intensity (T2-, FLAIR-weighted) around the third ventricle, the quadrigeminal bodies, the fornices, the mamillary bodies, the floor of the fourth ventricle and around the aqueduct. These findings were indicative of WE although of unusual extent. In this case MRI correlated well with clinical symptomatology. Therapy with thiamine was started immediately and symptoms as well as MRI findings resolved partially. The presented case illustrates the diagnostic usefulness of MRI in WE especially if the patient is of reduced consciousness and clinical investigation is limited.

Selective activation of human cortical area V5A by a rotating visual stimulus in fMRI; implication of attentional mechanisms.

The human homologue of area V5A of rotation-selective cells in the monkey medial superior temporal area (MST) was identified using functional magnetic resonance imaging (fMRI). It was located within the border region of occipito-temporo-parietal cortex, in four of 10 subjects on both sides, and on the right or left side in three subjects each. The stimulus was a black-and-white sine-modulated windmill presented either stationary or in rotation phases of 1 s duration. Areas V1-V3 did not show up with this paradigm. Focusing attention by mentally counting the number of rotation phases ensured high signal intensity in V5A, whereas moving attention away by counting electric stimuli to the wrist diminished it despite persistent fixation of gaze to the centre of the windmill.

Parallel increase of heterochromatic increment threshold and postadaptation thresholds in Parkinson's disease and in neuroleptic treatment.

Following reports on a predominant loss of blue/yellow contrast sensitivity in Parkinson's disease, we revisited the physiological phenomenon of transient tritanopia. Normative data were collected from 33 healthy individuals using different colour and time combinations. Stimuli of 440 nm wavelength (blue) proved optimal, if flashed for 50 msec within the early phase of a 2 sec pause in the 600 nm adaptation light. These conditions were then applied to 15 patients with Parkinson's disease. We found a parallel increase of increment threshold (P < 0.001) and postadaptation thresholds (P < 0.01), with little change in the extent of transient tritanopia. The same tendency at a lower significance level was found in 15 psychiatric patients under chronic treatment with depot neuroleptics.

Predominant affection of the blue cone pathway in Parkinson's disease.

Luminance contrast sensitivity and colour contrast thresholds were determined in 26 Parkinson patients and 17 normal controls of comparable age. They were psychophysically tested with a colour monitor system. Stimuli consisted of Gaussian enveloped luminance modulated or colour modulated (protan and tritan axis) vertical sine wave gratings with a spatial frequency of 1 cycle/degree. The stimuli subtended 4 degrees in diameter. Thresholds were determined using a two alternative forced choice method. Three different experimental conditions were explored: the detectability of stationary gratings, of moving gratings at velocities of 0, 2.5 and 5.0 cycles/s, and the detectability of horizontal square wave displacement at a frequency of 5 Hz for gratings of specified contrast levels. Intergroup differences were evaluated using two-tailed t tests with Satterthwaite corrections. Consistent and significant differences between normals and patients were found for tritan stimuli in the static and both dynamic conditions, and for luminance contrast stimuli in the displacement condition. Protan stimuli were much less apt to detect differences between the groups. We conclude that the retinal deficit of dopamine in Parkinson's disease is reflected in diminished centre/surround inhibition and that these changes are primarily apparent when vision is tested along the tritan axis, because blue cones are sparsely distributed.

Visual thresholds to low-contrast pattern displacement, color contrast, and luminance contrast stimuli in Parkinson's disease.

We used the computerized Moorfield Vision System to demonstrate specific increases in various perceptual visual thresholds in idiopathic Parkinson's syndrome. Fifteen patients were compared to 13 age-matched normals. Motion detection was impaired maximally (2 p < 0.01 and better in two-tailed t test) at luminance contrasts of 3-7%. Stimulus was an achromatic vertical 4 cycles/degrees sine wave grating subtending 3 degrees x 2 degrees, centered 5 degrees in the nasal field and oscillating at 5 Hz. In addition, stationary color and luminance contrast thresholds were tested with flashed display of 5 degrees x 6 degrees random letters, which were presented for 200 ms (color) and 50 ms (achromatic). Color discrimination was impaired in the tritan axis only (2 p < 0.05 in two-tailed t test). All achromatic stimuli--luminance increments, decrements, and phase reversing stimuli--were equally well seen by patients and controls. We conclude that the dopaminergic deficit of retinal amacrine cells in Parkinson patients can be monitored by combined low-contrast and motion (displacement) stimuli. Future studies will determine if moving colored targets are more effective in discriminating patients from controls than are the achromatic gratings used in this work.

Malignant gliomas--ploidy and DNA-content before and after therapy.

DNA ploidy and S-phase percentage from nine malignant gliomas (four glioblastomas, four anaplastic astrocytomas grade 3 and one anaplastic oligoastrocytoma grade 3) have been estimated by single cell cytophotometry on biopsy and necropsy specimens. All gliomas from biopsy material showed, with the exception of two diploid tumours, a polyploid-aneuploid DNA-pattern and stem-lines of different ploidy. The most frequent stem-lines were diploid and hyperdiploid. In necropsy material, following treatment i.e. operation and combined drug and radiation therapy, the heterogeneous nature of all malignant gliomas persisted. From seven aneuploid-polyploid gliomas four showed an elevation and three of them a decrease of DI values. Diploid tumors remained diploid. Because of marked heterogeneity of ploidy patterns and the small number of tumors investigated, ploidy changes could not be used for estimation of therapy efficacy and prognosis. Further studies will be necessary to answer this question.

Altered poly(ADP-ribose) metabolism in family members of patients with systemic lupus erythematosus.

OBJECTIVE: To determine whether the previously detected decrease in poly(ADP-ribose) synthesis in systemic lupus erythematosus (SLE) is familial. METHODS: Poly(ADP-ribose) metabolism was studied in the peripheral blood lymphocytes of members of 3 families with lupus and in healthy controls. RESULTS: Synthesis, as determined by the incorporation of 3H NAD into acid precipitable counts was 30% of normal in the patients with SLE (p < 0.001) and 70% of normal in the unaffected family members (p < 0.05). Family members also had increased prevalence of antinuclear antibodies but the presence of these antibodies did not correlate with abnormalities in poly(ADP-ribose) metabolism. CONCLUSION: The healthy biologic relatives of patients with SLE have a decrease in poly(ADP-ribose) synthesis similar to, but less marked than, their affected relatives. Poly(ADP-ribose) polymerase activity appears to be an inherited trait and abnormalities in it may be one of the susceptibility factors for SLE. If this is the case, then further investigation of this gene and its regulation is warranted.

Latency and duration of the muscle silent period following transcranial magnetic stimulation in multiple sclerosis, cerebral ischemia, and other upper motoneuron lesions.

In muscles with sustained voluntary contraction, EMG activity is transiently inhibited after transcranial magnetic stimulation. We recorded this postexcitatory silent period (SP) at 1.5 times individual stimulus threshold level from the first dorsal interosseus muscle in 65 neurologic patients aged 11 to 80 years. When compared with 20 healthy volunteers and a subgroup of patients with peripheral neurologic conditions not affecting the tested pathways, the SP was significantly longer on the paretic side in cerebral ischemia (p < 0.001) and chronic inflammatory CNS diseases, such as multiple sclerosis or neurosarcoidosis (p < 0.01). There was a similar tendency in pyramidal tract lesions due to CNS tumors and spinal cord trauma. In lesser degrees of paresis, SP duration is more sensitive than central motor conduction time (CMCT), but its specificity awaits further evaluation. SP is dependent on the integration of motor excitatory and inhibitory pathways and, possibly, sensorimotor reflex systems. In contrast to SP duration, which proved to be an independent variable giving supplementary information over the usual CMCT measurement, SP onset latency correlates well with CMCT and peripheral nerve conduction slowing, as in polyradiculoneuritis.

Decreased mRNA levels coding for poly(ADP-ribose) polymerase in lymphocytes of patients with SLE.

Poly(ADP-ribose) metabolism is altered in patients with SLE. In order to localize the defect, the levels of poly(ADP-ribose) polymerase-specific mRNA were measured from dot blots of total RNA from peripheral blood lymphocytes. In this preliminary study, eleven patients with SLE and two with antiphospholipid syndrome were compared to three controls. It was found that the mean levels of specific mRNA were ten fold lower in the PBL from SLE patients compared to controls and no overlap of values was seen between the two groups. No such decrease was seen in the PBL from the patients with antiphospholipid syndrome. It is concluded that the defect in poly(ADP-ribose) polymerase metabolism that is seen in SLE patients occurs at the level of transcription or mRNA turnover.