On the validity of modeling concepts for the simulation of groundwater flow in lowland peat areas – case study at the Zegveld experimental field

The groundwater flow models currently used in the western part of The Netherlands and in other similar peaty areas are thought to be a too simplified representation of the hydrological reality. One of the reasons is that, due to the schematization of the subsoil, its heterogeneity cannot be represented adequately. Moreover, the applicability of Darcy's law in these types of soils has been questioned, but this law forms the basis of most groundwater flow models. <br><br> With the purpose of assessing the typical heterogeneity of the subsoil and to verify the applicability of Darcy's law, geo-hydrological fieldwork was completed at an experimental field within a research area in the western part of The Netherlands. The assessments were carried out for the so-called Complex Confining Layer (CCL), which is the Holocene peaty to clayey layer overlying Pleistocene sandy deposits. Borehole drilling through the CCL with a hand auger was completed and revealed the typical heterogeneous character of this layer, showing a dominance of muddy, humified peat which is alternated with fresher peat and clay. <br><br> Slug tests were carried out to study the applicability of Darcy's law, given that previous studies suggested its non-validity for humified peat soils due to a variable horizontal hydraulic conductivity <i>K</i><sub>h</sub> with head differences. For higher humification degrees, the experiments indeed suggested a variable <i>K</i><sub>h</sub>, but this appeared to be the result of the inappropriate use of steady-state formulae for transient experiments in peaty environments. The muddy peat sampled has a rather plastic nature, and the high compressibility of this material leads to transient behavior. However, using transient formulae, the slug tests conducted for different initial groundwater heads showed that there was hardly any evidence of a variation of the hydraulic conductivity with the applied head differences. Therefore, Darcy's law can be used for typical peat soils present in The Netherlands. <br><br> The heterogeneity of the subsoil and the apparent applicability of Darcy's law were taken into account for the detailed heterogeneous model that was prepared for the research area. A MODFLOW model consisting of 13 layers in which 4 layers represent the heterogeneous CCL was set up for an average year, assuming steady-state conditions; and for the winter of 2009 to 2010, adopting transient conditions. The transient model was extended to simulate for longer periods with the objective of visualizing the flow paths through the CCL. The results from these models were compared with a 10 layer model, whereby the CCL is represented by a single layer assuming homogeneity. From the comparison of the two model types, the conclusion could be drawn that a single layer schematization of the CCL produces flowpath patterns which are not the same but still quite similar to a 4 layer representation of the CCL. However, the single layer schematization results in a considerable underestimation of the flow velocity, and subsequently a longer travel time, through the CCL. Therefore, a single layer model of the CCL seems quite appropriate to represent the general flow behavior of the shallow groundwater system, but would be inappropriate for transport modeling through the CCL

Block of endplate channels by permeant cations in frog skeletal muscle

Motor endplates of frog semitendinosus muscles were studied under voltage clamp. Current fluctuations by iontophoretic application of acetylcholine were analyzed to give the elementary conductance, γ, and mean open time, τ, of endplate channels. Total replacement of the external Na+ ion by several other metal ions and by many permanent organic cations changed both γ and τ. Except with NH4 + ions, the γ values with foreign test ions were all smaller than expected from the independence relation and their previously measured permeability ratios. The more hydrophobic ions gave the smallest γ values. Foreign permeant cations also depress γ when mixed with Na+ ions. These effects could be interpreted in terms of binding of ions to a saturable site within the endplate channel as they pass through. The site for organic ions would have a hydrophobic component. Similar evidence is given for a metal ion binding site on the cytoplasmic end of the channel accessible to internal ions. Most foreign cations also shortened τ when applied externally. The changes of gating did not seem to be correlated with changes in γ. Thus there is no evidence for control of τ by ions bound within the pore

An educational intervention to improve the quality of care of diabetic patients

Objective. As few studies have addressed intervention for in-hospital care of diabetes mellitus (DM) patients, we set out to investigate whether an educational intervention targeting doctors could improve the quality of care for diabetic patients.Design. An observational interventional study conducted at Pretoria Academic Hospital, a tertiary care hospital.Subjects. Doctors working in the Department of Internal Medicine were the subjects of two interventional sessions on diabetic care, and all diabetic patients admitted to the wards in the above Department were evaluated.Outcome measures. A Diabetes Attitude Scale (DAS-3) and a Diabetes Practice Scale (DPS) were completed by each doctor before and after the interventional educational sessions. Data from diabetic patients in the wards were collected for 5 weeks before and 5 weeks after the interventional training, and these two sets of data were compared to measure the effect of the interventional training.Results. Subscales of the DAS-3 showed an improvement, with a statistically significant improvement in attitude regarding seriousness of DM (P = 0.03), and a trend towards improvement in attitude regarding need for special training and patient autonomy. Most of the items on the DPS improved significantly (P < 0.05).Conclusions. A short educational intervention resulted in an improvement in attitude, knowledge and clinical management of diabetic patients

The cell adhesion molecule L1 regulates the expression of choline acetyltransferase and the development of septal cholinergic neurons

Mutations in the L1 gene cause severe brain malformations and mental retardation. We investigated the potential roles of L1 in the regulation of choline acetyltransferase (ChAT) and in the development of septal cholinergic neurons, which are known to project to the hippocampus and play key roles in cognitive functions. Using stereological approaches, we detected significantly fewer ChAT-positive cholinergic neurons in the medial septum and vertical limb of the diagonal band of Broca (MS/VDB) of 2-week-old L1-deficient mice compared to wild-type littermates (1644 ± 137 vs. 2051 ± 165, P = 0.038). ChAT protein levels in the septum were 53% lower in 2-week-old L1-deficient mice compared to wild-type littermates. ChAT activity in the septum was significantly reduced in L1-deficient mice compared to wild-type littermates at 1 (34%) and 2 (40%) weeks of age. In vitro, increasing doses of L1-Fc induced ChAT activity in septal neurons with a significant linear trend (*P = 0.0065). At 4 weeks of age in the septum and at all time points investigated in the caudate-putamen (CPu), the number of ChAT-positive neurons and the levels of ChAT activity were not statistically different between L1-deficient mice and wild-type littermates. The total number of cells positive for the neuronal nuclear antigen (NeuN) in the MS/VDB and CPu was not statistically different in L1-deficient mice compared to wild-type littermates, and comparable expression of the cell cycle marker Ki67 was observed. Our results indicate that L1 is required for the timely maturation of septal cholinergic neurons and that L1 promotes the expression and activity of ChAT in septal neurons

Permeation Mechanisms in the TMEM16B Calcium-Activated Chloride Channels

TMEM16A and TMEM16B encode for Ca2+-activated Cl- channels (CaCC) and are expressed in many cell types and play a relevant role in many physiological processes. Here, I performed a site-directed mutagenesis study to understand the molecular mechanisms of ion permeation of TMEM16B. I mutated two positive charged residues R573 and K540, respectively located at the entrance and inside the putative channel pore and I measured the properties of wild-type and mutant TMEM16B channels expressed in HEK-293 cells using whole-cell and excised inside-out patch clamp experiments. I found evidence that R573 and K540 control the ion permeability of TMEM16B depending both on which side of the membrane the ion substitution occurs and on the level of channel activation. Moreover, these residues contribute to control blockage or activation by permeant anions. Finally, R573 mutation abolishes the anomalous mole fraction effect observed in the presence of a permeable anion and it alters the apparent Ca2+-sensitivity of the channel. These findings indicate that residues facing the putative channel pore are responsible both for controlling the ion selectivity and the gating of the channel, providing an initial understanding of molecular mechanism of ion permeation in TMEM16B

Poisson-Nernst-Planck Systems for Narrow Tubular-like Membrane Channels

We study global dynamics of the Poisson-Nernst-Planck (PNP) system for flows of two types of ions through a narrow tubular-like membrane channel. As the radius of the cross-section of the three-dimensional tubular-like membrane channel approaches zero, a one-dimensional limiting PNP system is derived. This one-dimensional limiting system differs from previous studied one-dimensional PNP systems in that it encodes the defining geometry of the three-dimensional membrane channel. To justify this limiting process, we show that the global attractors of the three-dimensional PNP systems are upper semi-continuous to that of the limiting PNP system. We then examine the dynamics of the one-dimensional limiting PNP system. For large Debye number, the steady-state of the one-dimensional limiting PNP system is completed analyzed using the geometric singular perturbation theory. For a special case, an entropy-type Lyapunov functional is constructed to show the global, asymptotic stability of the steady-state

A Structural Model of the Pore-Forming Region of the Skeletal Muscle Ryanodine Receptor (RyR1)

Ryanodine receptors (RyRs) are ion channels that regulate muscle contraction by releasing calcium ions from intracellular stores into the cytoplasm. Mutations in skeletal muscle RyR (RyR1) give rise to congenital diseases such as central core disease. The absence of high-resolution structures of RyR1 has limited our understanding of channel function and disease mechanisms at the molecular level. Here, we report a structural model of the pore-forming region of RyR1. Molecular dynamics simulations show high ion binding to putative pore residues D4899, E4900, D4938, and D4945, which are experimentally known to be critical for channel conductance and selectivity. We also observe preferential localization of Ca2+ over K+ in the selectivity filter of RyR1. Simulations of RyR1-D4899Q mutant show a loss of preference to Ca2+ in the selectivity filter as seen experimentally. Electrophysiological experiments on a central core disease mutant, RyR1-G4898R, show constitutively open channels that conduct K+ but not Ca2+. Our simulations with G4898R likewise show a decrease in the preference of Ca2+ over K+ in the selectivity filter. Together, the computational and experimental results shed light on ion conductance and selectivity of RyR1 at an atomistic level

An updated evaluation of the implementation of the sigmoid take-off landmark 1 year after the official introduction in the Netherlands

PURPOSE: The definition of rectal cancer based on the sigmoid take-off (STO) was incorporated into the Dutch guideline in 2019, and became mandatory in the national audit from December 2020. This study aimed to evaluate the use of the STO in clinical practice and the added value of online training, stratified for the period before (group A, historical cohort) and after (group B, current cohort) incorporation into the national audit.METHODS: Participants, including radiologists, surgeons, surgical and radiological residents, interns, PhD students, and physician assistants, were asked to complete an online training program, consisting of questionnaires, 20 MRI cases, and a training document. Outcomes were agreement with the expert reference, inter-rater variability, and accuracy before and after the training.RESULTS: Group A consisted of 86 participants and group B consisted of 114 participants. Familiarity with the STO was higher in group B (76% vs 88%, p = 0.027). Its use in multidisciplinary meetings was not significantly higher (50% vs 67%, p = 0.237). Agreement with the expert reference was similar for both groups before (79% vs 80%, p = 0.423) and after the training (87% vs 87%, p = 0.848). Training resulted in significant improvement for both groups in classifying tumors located around the STO (group A, 69-79%; group B, 67-79%, p &lt; 0.001).CONCLUSIONS: The results of this study show that after the inclusion of the STO in the mandatory Dutch national audit, the STO was consequently used in only 67% of the represented hospitals. Online training has the potential to improve implementation and unambiguous assessment.</p

Activation of BMP-Smad1/5/8 Signaling Promotes Survival of Retinal Ganglion Cells after Damage In Vivo

While the essential role of bone morphogenetic protein (BMP) signaling in nervous system development is well established, its function in the adult CNS is poorly understood. We investigated the role of BMP signaling in the adult mouse retina following damage in vivo. Intravitreal injection of N-Methyl-D-aspartic acid (NMDA) induced extensive retinal ganglion cell death by 2 days. During this period, BMP2, -4 and -7 were upregulated, leading to phosphorylation of the downstream effector, Smad1/5/8 in the inner retina, including in retinal ganglion cells. Expression of Inhibitor of differentiation 1 (Id1; a known BMP-Smad1/5/8 target) was also upregulated in the retina. This activation of BMP-Smad1/5/8 signaling was also observed following light damage, suggesting that it is a general response to retinal injuries. Co-injection of BMP inhibitors with NMDA effectively blocked the damage-induced BMP-Smad1/5/8 activation and led to further cell death of retinal ganglion cells, when compared with NMDA injection alone. Moreover, treatment of the retina with exogenous BMP4 along with NMDA damage led to a significant rescue of retinal ganglion cells. These data demonstrate that BMP-Smad1/5/8 signaling is neuroprotective for retinal ganglion cells after damage, and suggest that stimulation of this pathway can serve as a potential target for neuroprotective therapies in retinal ganglion cell diseases, such as glaucoma