Developmental patterns in human blood-brain barrier and blood-cerebrospinal fluid barrier ABC drug transporter expression.

When drugs exert their effects in the brain, linear extrapolation of doses from adults could be harmful for children as the blood-brain barrier (BBB) and blood-CSF barrier (BCSFB) function is still immature. More specifically, age-related variation in membrane transporters may impact brain disposition. As human data on brain transporter expression is scarce, age dependent [gestational age (GA), postnatal age (PNA), and postmenstrual age (PMA)] variation in immunohistochemical localization and staining intensity of the ABC transporters P-glycoprotein (Pgp), breast cancer resistance protein (BCRP), and multidrug resistance-associated proteins 1, 2, 4, and 5 (MRP1/2/4/5) was investigated. Post mortem brain cortical and ventricular tissue was derived from 23 fetuses (GA range 12.9-39 weeks), 17 neonates (GA range 24.6-41.3 weeks, PNA range 0.004-3.5 weeks), 8 children (PNA range 0.1-3 years), and 4 adults who died from a wide variety of underlying conditions. In brain cortical BBB, immunostaining increased with age for Pgp and BCRP, while in contrast, MRP1 and MRP2 staining intensity appeared higher in fetuses, neonates, and children, as compared to adults. BCSFB was positively stained for Pgp, MRP1, and MRP2 and appeared stable across age, while BCRP was not detected. MRP4 and MRP5 were not detected in BBB or BCSFB. In conclusion, human BBB and BCSFB ABC membrane transporters show brain location and transporter-specific maturation.

Quality of life and costs of patients prior to colorectal surgery.

Objective: To assess the quality of life and societal costs of patients prior to colorectal surgery in the Netherlands.Methods: This study is embedded in a previous randomized controlled trial (SANICS II). The quality of life was measured using EQ-5D-5L questionnaires. The iMTA medical consumption questionnaire (iMCQ) and the iMTA productivity costs questionnaire (iPCQ) were used to identify and measure healthcare and productivity costs. Subgroup analyses were performed based on age and gender.Results: A total of 178 patients were included in the cost analysis and a total of 161 patients in the quality of life analysis. The three-month mean societal cost per patient amounted to €3,211 of which €1,459 was due to productivity losses. The mean utility was 0.88 per patient. Gender was an important predictor in quality of life with men scoring significantly higher than women (0.92 versus 0.82) at p < 0.0001.Conclusion: Colorectal cancer represents a high economic burden in the Netherlands. Further research with repeated cost and quality of life measurements would be needed to explore the change over time and the effects of surgery.

The contribution of mast cells to postoperative ileus in experimental and clinical studies.

The persistent phase of postoperative ileus (POI) is mediated by inflammatory activation of the resident myeloid immune cell population in the gut wall, likely elicited by neurogenic activation. Mast cells are thought to play a critical role in this inflammatory response and involvement of mast cells in POI has been investigated and described thoroughly in experimental studies. Intestinal manipulation (IM) leads to degranulation of mast cells, resulting in an increase in mast cell proteases in peritoneal fluid and gut tissue. The inflammatory infiltrate formed in the intestinal wall thereby impairs gastrointestinal motility. In the clinical study by Berdun et al., the experimentally known association between mast cell degranulation and delayed motility is shown in a clinical setting. These findings are important and open up therapeutic opportunities to reduce or prevent POI. In this mini-review, the role of mast cells in POI is discussed. Furthermore, an update is given on the involvement of the inflammatory response in POI and potential therapeutic strategies.

A DFT based equilibrium study on the hydrolysis and the dehydration reactions of MgCl2 hydrates.

Magnesium chloride hydrates are characterized as promising energy storage materials in the built-environment. During the dehydration of these materials, there are chances for the release of harmful HCl gas, which can potentially damage the material as well as the equipment. Hydrolysis reactions in magnesium chloride hydrates are subject of study for industrial applications. However, the information about the possibility of hydrolysis reaction, and its preference over dehydration in energy storage systems is still ambiguous at the operating conditions in a seasonal heat storage system. A density functional theory level study is performed to determine molecular structures, charges, and harmonic frequencies in order to identify the formation of HCl at the operating temperatures in an energy storage system. The preference of hydrolysis over dehydration is quantified by applying thermodynamic equilibrium principles by calculating Gibbs free energies of the hydrated magnesium chloride molecules. The molecular structures of the hydrates (n = 0, 1, 2, 4, and 6) of MgCl2 are investigated to understand the stability and symmetry of these molecules. The structures are found to be noncomplex with almost no meta-stable isomers, which may be related to the faster kinetics observed in the hydration of chlorides compared to sulfates. Also, the frequency spectra of these molecules are calculated, which in turn are used to calculate the changes in Gibbs free energy of dehydration and hydrolysis reactions. From these calculations, it is found that the probability for hydrolysis to occur is larger for lower hydrates. Hydrolysis occurring from the hexa-, tetra-, and di-hydrate is only possible when the temperature is increased too fast to a very high value. In the case of the mono-hydrate, hydrolysis may become favorable at high water vapor pressure and at low HCl pressure.

Recurrent and founder mutations in the Netherlands: cardiac Troponin I (TNNI3) gene mutations as a cause of severe forms of hypertrophic and restrictive cardiomyopathy.

BACKGROUND: About 2-7% of familial cardiomyopathy cases are caused by a mutation in the gene encoding cardiac troponin I (TNNI3). The related clinical phenotype is usually severe with early onset. Here we report on all currently known mutations in the Dutch population and compared these with those described in literature. METHODS: TheTNNI3 gene was screened for mutations in all coding exons and flanking intronic sequences in a large cohort of cardiomyopathy patients. All Dutch index cases carrying a TNNI3 mutation that are described in this study underwent extensive cardiological evaluation and were listed by their postal codes. RESULTS: In 30 families, 14 different mutations were identified. Three TNNI3 mutations were found relatively frequently in both familial and non-familial cases of hypertrophic cardiomyopathy (HCM) or restrictive cardiomyopathy (RCM). Haplotype analysis showed that p.Arg145Trp and p.Ser166Phe are founder mutations in the Netherlands, while p.Glu209Ala is not. The majority of Dutch TNNI3 mutations were associated with a HCM phenotype. Mean age at diagnosis was 36.5 years. Mutations causing RCM occurred less frequently, but were identified in very young children with a poor prognosis. CONCLUSION: In line with previously published data, we found TNNI3 mutations to be rare and associated with early onset and severe clinical presentation.

Contiguous gene syndrome due to a maternally inherited 8.41 Mb distal deletion of chromosome band Xp22.3 in a boy with short stature, ichthyosis, epilepsy, mental retardation, cerebral cortical heterotopias and Dandy-Walker malformation.

Microdeletions of Xp22.3 are associated with contiguous gene syndromes, the extent and nature of which depend on the genes encompassed by the deletion. Common symptoms include ichthyosis, mental retardation and hypogonadism. We report on a boy with short stature, ichthyosis, severe mental retardation, cortical heterotopias and Dandy-Walker malformation. The latter two abnormalities have so far not been reported in terminal Xp deletions. MLPA showed deletion of SHOX and subsequent analysis using FISH and SNP-arrays revealed that the patient had an 8.41 Mb distal deletion of chromosome region Xp22.31 --> Xpter. This interval contains several genes whose deletion can partly explain our patient's phenotype. His cortical heterotopias and DWM suggest that a gene involved in brain development may be in the deleted interval, but we found no immediately obvious candidates. Interestingly, further analysis of the family revealed that the patient had inherited his deletion from his mother, who has a mos 46,X,del(X)(p22)/45,X/46, XX karyotype.

Lessons from studies on focal segmental glomerulosclerosis: an important role for parietal epithelial cells?

Glomerular diseases are caused by multiple mechanisms. Progressive glomerular injury is characterized by the development of segmental or global glomerulosclerosis independent of the nature of the underlying renal disease. Most studies on glomerular disease focus on the constituents of the filtration barrier (podocytes, glomerular basement membrane (GBM), endothelial cells) or the mesangial cells. Little attention is given to the epithelial cells lining Bowman's capsule, the so called parietal epithelial cells (PECs). This 'lack of attention' is partly explained by the presumed 'passive' function of PECs, which are large, flattened cells that cover Bowman's capsule in a single cell layer and form a barrier between the ultrafiltrate and the periglomerular interstitium, in normal glomerular physiology. A more important reason has been the lack of an established primary role for the parietal epithelium in glomerular diseases. However, in recent years, several studies have demonstrated that PECs are involved in extracapillary proliferation. In addition, PECs can become highly active, proliferating cells, expressing many growth factors, chemokines, cytokines, and their receptors. It was recently demonstrated that PECs also play a part in the development of focal segmental glomerulosclerosis (FSGS). This review summarises current knowledge of the PEC, with emphasis on the role of PECs in the development of FSGS.

Proliferating cells in HIV and pamidronate-associated collapsing focal segmental glomerulosclerosis are parietal epithelial cells.

Collapsing focal segmental glomerulosclerosis (cFSGS) is characterized by hyperplasia of glomerular epithelial cells. In a mouse model of FSGS and in a patient with recurrent idiopathic FSGS, we identified the proliferating cells as parietal epithelial cells (PECs). In the present study, we have evaluated the origin of the proliferating cells in cFSGS associated with human immunodeficiency virus (HIV) and pamidronate. We performed a detailed study of glomerular lesions in biopsies of two patients with HIV-associated cFSGS and a nephrectomy specimen of a patient with pamidronate-associated cFSGS. Glomeruli were studied by serial sectioning using light and electron microscopy and immunohistochemistry to determine the epithelial cell phenotype. We used Synaptopodin, vascular endothelial growth factor, and CD10 as podocyte markers, CK8 and PAX2 as PEC markers and Ki-67 as marker of cell proliferation. The newly deposited extracellular matrix was characterized using antiheparan sulfate single-chain antibodies. The proliferating cells were negative for the podocyte markers, but stained positive for the PEC markers and the cell proliferation marker Ki-67. The proliferating PAX-2 and CK8 positive cells that covered the capillary tuft were always in continuity with PAX-2/CK8 positive cells lining Bowman's capsule. The matrix deposited by these proliferating cells stained identically to Bowman's capsule. Our study demonstrates that PECs proliferate in HIV and pamidronate-associated cFSGS. Our data do not support the concept of the proliferating, dedifferentiated podocyte.

Blue five-level frequency-upconversion system in rubidium.

We demonstrate production of continuous coherent blue laser light by using a five-level system in rubidium vapor. Two low-power lasers, at 780 and 776 nm, induce strong atomic coherence in the 5S-5P-5D states. The atoms decay to the 6P excited state, from which stimulated emission produces a coherent blue (420 nm) beam. We have coupled both ground-state hyperfine levels, effecting coherence between four levels. The coherent blue output is enhanced by several mechanisms, including stronger coupling to a larger fraction of the atomic population, operation at a detuning such that the vapor is nominally transparent to the 780 nm pump field, reduced losses owing to optical pumping, and optimal phase matching. We report experimental findings and compare them with results from a semiclassical Maxwell-Bloch model.

Magnetic nanodots from atomic Fe: can it be done?

Laser focusing of Fe atoms offers the possibility of creating separate magnetic structures on a scale of 10 nm with exact periodicity. This can be done by using the parabolic minima of the potential generated by a standing light wave as focusing lenses. To achieve the desired 10-nm resolution, we need to suppress chromatic and spherical aberrations, as well as prevent structure broadening caused by the divergence of the incoming beam. Chromatic aberrations are suppressed by the development of a supersonic Fe beam source with speed ratio S = 11 +/- 1. This beam has an intensity of 3 x 10(15) atoms sr(-1) s(-1). The spherical aberrations of the standing light wave will be suppressed by aperturing with beam masks containing 100-nm slits at 744-nm intervals. The beam divergence can be reduced by application of laser cooling to reduce the transverse velocity. We have constructed a laser system capable of delivering over 500 mW of laser light at 372 nm, the wavelength of the (5)D(4) --> (5)F(5) atomic transition of (56)Fe we intend to use for laser cooling. Application of polarization spectroscopy to a hollow cathode discharge results in a locking system holding the laser continuously within 2 MHz of the desired frequency.

Changes in mRNA expression profile underlie phenotypic adaptations in creatine kinase-deficient muscles.

We have studied the mechanisms that regulate the remodeling of the glycolytic, mitochondrial and structural network of muscles of creatine kinase M (M-CK)/sarcomeric mitochondrial creatine kinase (ScCKmit) knockout mice by comparison of wild-type and mutant mRNA profiles on cDNA arrays. The magnitudes of changes in mRNA levels were most prominent in M-CK/ScCKmit (CK(-/-)) double mutants but did never exceed those of previously observed changes in protein level for any protein examined. In gastrocnemius of CK(-/-) mice we measured a 2.5-fold increase in mRNA level for mitochondrial encoded cytochrome c oxidase (COX)-III which corresponds to the increase in protein content. The level of the nuclear encoded mRNAs for COX-IV, H(+)-ATP synthase-C, adenine nucleotide translocator-1 and insulin-regulatable glucose transporter-4 showed a 1.5-fold increase, also in agreement with protein data. In contrast, no concomitant up-regulation in mRNA and protein content was detected for the mitochondrial inorganic phosphate-carrier, voltage-dependent anion channel and certain glycolytic enzymes. Our results reveal that regulation of transcript level plays an important role, but it is not the only principle involved in the remodeling of mitochondrial and cytosolic design of CK(-/-) muscles.

Nd:YAG laser posterior hyaloidotomy for the treatment of a premacular vitreous floater.

A 63-year old man, with a large central vitreous floater, underwent a Nd:YAG laser posterior hyaloidotomy. Although the vitreous floater disappeared from the central optical axis, visual acuity did not improve. Microperimetry performed with the SLO revealed an absolute scotoma, which corresponded well in shape and dimension with the original vitreous floater. This finding suggests that a fragment of the neurosensory retina became detached together with the internal limiting membrane in the process of the vitreous collapse.

[Use of the computer in dentistry: video superimposition and facial reproduction using the expedience of the computer interface]. / L'utilisation de l'ordinateur en odontologie: superposition vidéo et reproduction faciale par le biais d'une interface informatique.

Both video superimposition and computer facial reproduction play a useful role in the process of identification, particularly in the early stages of an investigation or when other more definitive methods may not be available. Video superimposition combined with a computer overlay technique can be used to establish whether a skull outline matches an image of a missing person, being a photograph, slide or videoimage. A computer driven robot can be used to simplify the alignment of the skull with the position of the head on the image. Craniofacial or facial reproduction are used in both 2D or 3D to produce an image which can be used for identification purposes.

Learning phacoemulsification: the surgeon-in-training.

With the increasing trend towards phacoemulsification a perceived increased complication rate during the learning curve gives rise to a dilemma as to the best stage at which a surgeon-in-training can safely learn the technique. We prospectively analysed the complications and visual outcome of the first 160 phacoemulsification procedures performed by three surgeons-in-training. The main outcome measures included posterior capsule tear, vitreous and nuclear loss, surgical re-intervention rate and visual outcome. Posterior capsule tear occurred in 7 eyes (4.4%) and vitreous loss in 6 (3.8%). No nucleus was lost in the vitreous. Surgical re-intervention was required in 1 eye. Best corrected visual acuity was 6/12 or better in 88% of eyes. These results compare favourably with reports of surgeons-in-training learning extracapsular surgery and also with recently reported phacoemulsification series. This study indicates that with careful case selection and supervision phacoemulsification can be a safe procedure.

Learning phaco in a teaching environment.

We prospectively analysed the complications and visual outcome of the first 160 phacoemulsification procedures performed by three surgeons in training under supervision. The main complications were posterior capsular tear (4,35%), vitreous loss (3,3%), nuclear loss (none) and surgical reintervention (one). Postoperative visual acuity was 0,7 or better in 94% of eyes without pre-existing pathology. Only one eye did not have a posterior chamber lens implanted.

ABO bloodgrouping on tooth material.

The ABO bloodgroup was determined on the pulp, dentin and enamel of 35 teeth using the adsorption-elution technique. A bloodstained compress from the extraction wound was used as the reference sample. Twenty teeth were examined within 6 weeks after extraction and fifteen teeth were examined 6-10 months after extraction. It was found, that bloodgrouping on pulp gives fairly good results, whereas the possibilities for correct bloodgrouping seem to be limited for dentin and debatable for enamel. Similar results were found in both groups of teeth.

Definition of subchromosomal intervals around the myotonic dystrophy gene region at 19q.

The localization to 19q of the gene causing myotonic dystrophy (DM) has been defined more precisely by refinement of the physical location of several linked markers. A somatic cell hybrid mapping panel from cells with t(1;19), t(12;19), and t(X;19) translocation products was constructed to define five different intervals across 19q. In addition, we have derived a series of cell hybrids by irradiation of a der(19)-only hybrid to further subdivide the cen-q13.1 region. Using an array of 36 cloned genes, anonymous DNAs, and enzyme markers, we have tested the location of the panel breakpoints and refined the regional assignment of several of these markers. All markers tightly linked to DM are localized mainly within 19q13.2, thus suggesting that the DM gene is also close to this region.

Tight linkage between myotonic dystrophy and apolipoprotein E genes revealed with allele-specific oligonucleotides.

In 16 families with myotonic dystrophy (DM) a novel approach based on use of allele-specific oligonucleotides has been employed to study the linkage relationship between the apolipoprotein E (APOE) gene and DM. Synthetic oligonucleotides, designed to discriminate between APOE alleles epsilon 3 and epsilon 4, enabled us to distinguish heterozygous carriers in a hybridization assay. In a subset of families, the relevant segment of the APOE gene was enzymatically amplified to increase the sensitivity of the method. For DM and APOE, a maximum lod score (zmax of 7.47 was obtained at a recombination frequency (theta) of 0.047 (male theta = female theta). No recombination (maximum lod score of 5.61 at theta = 0.0) was found between APOE and the apolipoprotein CII (APOC2) gene. These results suggest that, in addition to APOC2, APOE is a useful marker for presymptomatic DM diagnosis.