Tumor Cells and Tumor-Associated Macrophages: Secreted Proteins as Potential Targets for Therapy

Inflammatory pathways, meant to defend the organism against infection and injury, as a byproduct, can promote an environment which favors tumor growth and metastasis. Tumor-associated macrophages (TAMs), which constitute a significant part of the tumor-infiltrating immune cells, have been linked to the growth, angiogenesis, and metastasis of a variety of cancers, most likely through polarization of TAMs to the M2 (alternative) phenotype. The interaction between tumor cells and macrophages provides opportunities for therapy. This paper will discuss secreted proteins as targets for intervention

Topography-Mediated Myotube and Endothelial Alignment, Differentiation, and Extracellular Matrix Organization for Skeletal Muscle Engineering

Understanding the response of endothelial cells to aligned myotubes is important to create an appropriate environment for tissue-engineered vascularized skeletal muscle. Part of the native tissue environment is the extracellular matrix (ECM). The ECM is a supportive scaffold for cells and allows cellular processes such as proliferation, differentiation, and migration. Interstitial matrix and basal membrane both comprise proteinaceous and polysaccharide components for strength, architecture, and volume retention. Virtually all cells are anchored to their basal lamina. One of the physical factors that affects cell behavior is topography, which plays an important role on cell alignment. We tested the hypothesis that topography-driven aligned human myotubes promote and support vascular network formation as a prelude to in vitro engineered vascularized skeletal muscle. Therefore, we used a PDMS-based topography substrate to investigate the influence of pre-aligned myotubes on the network formation of microvascular endothelial cells. The aligned myotubes produced a network of collagen fibers and laminin. This network supported early stages of endothelial network formation.</p

Copy number variation in a hospital-based cohort of children with epilepsy

Objective: To evaluate the diagnostic yield of microarray analysis in a hospital-based cohort of children with seizures and to identify novel candidate genes and susceptibility loci for epilepsy. Methods: Of all children who presented with their first seizure in the University Medical Center Groningen (January 2000 through May 2013) (n = 1,368), we included 226 (17%) children who underwent microarray analysis before June 2014. All 226 children had a definite diagnosis of epilepsy. All their copy number variants (CNVs) on chromosomes 1-22 and X that contain protein-coding genes and have a prevalence of <1% in healthy controls were evaluated for their pathogenicity. Results: Children selected for microarray analysis more often had developmental problems (82% vs. 25%, p < 0.001), facial dysmorphisms (49% vs. 8%, p < 0.001), or behavioral problems (41% vs. 13%, p < 0.001) than children who were not selected. We found known clinically relevant CNVs for epilepsy in 24 of the 226 children (11%). Seventeen of these 24 children had been diagnosed with symptomatic focal epilepsy not otherwise specified (71%) and five with West syndrome (21%). Of these 24 children, many had developmental problems (100%), behavioral problems (54%) or facial dysmorphisms (46%). We further identified five novel CNVs comprising four potential candidate genes for epilepsy:MYT1L, UNC5D, SCN4B,andNRXN3. Significance: The 11% yield in our hospital-based cohort underscores the importance of microarray analysis in diagnostic evaluation of children with epilepsy

Changes in empowerment and anxiety of patients and parents during genetic counselling for epilepsy

Genetic testing and counselling are increasingly important in epilepsy care, aiming at finding a diagnosis, understanding aetiology and improving treatment and outcome. The psychological impact of genetic counselling from patients' or parents & rsquo; perspectives is, however, unknown. We studied the counseleereported outcome of genetic counselling before and after genetic testing for epilepsy by evaluating empowerment - a key outcome goal of counselling reflecting cognitive, decisional and behavioural control, emotional regulation and hope - and anxiety. We asked patients or their parents (for those (c) 2021 The Author(s). Published by Elsevier Ltd on behalf of European Paediatric Neurology Society. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

Effectiveness of percutaneous laser disc decompression versus conventional open discectomy in the treatment of lumbar disc herniation; design of a prospective randomized controlled trial

Background. The usual surgical treatment of refractory sciatica caused by lumbar disc herniation, is open discectomy. Minimally invasive procedures, including percutaneous therapies under local anesthesia, are increasingly gaining attention. One of these treatments is Percutaneous Laser Disc Decompression (PLDD). This treatment can be carried out in an outpatient setting and swift recovery and return to daily routine are suggested. Thus far, no randomized trial into cost-effectiveness of PLDD versus standard surgical procedure has been performed. We present the design of a randomized controlled trial, studying the cost-effectiveness of PLDD versus conventional open discectomy in patients with sciatica from lumbar disc herniation. Methods/design. The study is a randomized prospective multi-center trial, in which two treatment strategies are compared in a parallel group design. Patients (age 18-70 years) visiting the neurosurgery department of the participating hospitals, are considered for inclusion in the trial when sciatica due to lumbar disc herniation has lasted more than 8 weeks. Patients with disc herniation smaller than 1/3 of the spinal canal diameter, without concomitant lateral recess stenosis or sequestration, are eligible for participation, and are randomized into one of two treatment arms; either Percutaneous Laser Disc Decompression or conventional discectomy. The functional outcome of the patient, as assessed by the Roland Disability Questionnaire for Sciatica at 8 weeks and 1 year after treatment, is the primary outcome measure. The secondary outcome parameters are recovery as perceived by the patient, leg and back pain, incidence of re-intervention, complications, quality of life, medical consumption, absence of work and secondary costs. Discussion. Open discectomy is still considered to be the golden standard in the surgical treatment of lumbar disc herniation. Whether Percutaneous Laser Disc Decompression has at least as much efficacy as the standard surgical procedure, and is more cost-effective, will be determined by this trial. Trial registration. Current Controlled Trials ISRCTN25884790

Cervical high-intensity intramedullary lesions in achondroplasia:Aetiology, prevalence and clinical relevance

In achondroplastic patients with slight complaints of medullary compression the cervical spinal cord regularly exhibits an intramedullary (CHII) lesion just below the craniocervical junction with no signs of focal compression on the cord. Currently, the prevalence of the lesion in the general achondroplastic population is studied and its origin is explored. Eighteen achondroplastic volunteers with merely no clinical signs of medullary compression were subjected to dynamic magnetic resonance imaging (MRI). The presence of a CHII lesion and craniocervical medullary compression in flexed and retroflexed craniocervical positions was explored. Several morphological characteristics of the craniocervical junction, possibly related to compression on the cord, were assessed. A CHII lesion was observed in 39% of the subjects and in only one of these was compression at the craniocervical junction present. Consequently, no correlation between the CHII lesion and compression could be established. None of the morphological characteristics demonstrated a correlation with the CHII lesion, except thinning of the cord at the site of the CHII lesion. CHII lesions are a frequent finding in achondroplasia, and are generally unaccompanied by clinical symptoms or compression on the cord. Further research focusing on the origin of CHII lesions and their clinical implications is warranted. aEuro cent MRI now reveals exquisite detail of the cervical spinal cord. aEuro cent Cervical cord lesions are observed in one third of the achondroplastic population. aEuro cent These lesions yield high signal intensity on T2 weighted MRI. aEuro cent They are generally unaccompanied by clinical symptoms or cord compression. aEuro cent Their aetiology is unclear and seems to be unrelated to mechanical causes

Acidic drugs - intestinal solubility correlation fasted and fed in vitro vs ex vivo

A solid oral dosage form is the commonest method for self-administration, but solid drug must dissolve before absorption. Gastrointestinal solubility is a key parameter controlling oral absorption and fundamental to the Biopharmaceutics and Developability Classification Systems [1]. A reliable in vitro measurement of intestinal solubility is therefore critical to assess potential in vivo behaviour during development [2]. This group applied a five dimensional analysis (pH, bile salt, phospholipid, free fatty acid and cholesterol) to fasted and fed human intestinal fluid (HIF) composition to calculate nine simulated intestinal (SIF) fasted (FaSIM) and nine simulated fed (FeSIM) fluid recipes [3]. In each state the media recipes covered 95% of the HIF compositional variability. FaSIM and FeSIM could therefore be considered bioequivalent with the potential to measure HIF solubility in vitro covering 95% of the range. In this abstract we have compared FaSIM and FeSIM equilibrium solubility data for acidic drugs against published HIF values. MATERIALS AND METHODS Materials: Media components were from Merck Chemicals Ltd., UK, except soya bean phosphatidylcholine from Lipoid. Chloroform was from Rathburn Chemical Company, NaH2PO4⋅H2O was purchased from Fisher Scientific, acetonitrile (ACN) and methanol (MeOH) were HPLC quality from VWR. Water is ultrapure Milli-Q water. Furosemide, indomethacin, naproxen, and piroxicam were from Merck Chemicals Ltd., ibuprofen was obtained from BSAF and mefenamic acid from Sigma Aldrich. Method: Equilibrium Solubility Measurement The equilibrium solubility measurement method has been published for FeSIM [4], FaSIM [5] and DoE studies [6]. In brief, individual media recipes were prepared [4, 5], an excess of solid drug was weighed into a centrifuge tube, the media adjusted to final volume (4 mL) and pH adjusted using KOH or HCl. The tubes were shaken for 1 hour and the pH readjusted. Tubes were then shaken for 24 hours at 37 ºC and 240 rpm. The presence of solid drug was visually confirmed and 1 mL of each solution centrifuged (10,000 rpm for 15 minutes) and the supernatant was analysed by HPLC for drug content. Three replicate measurements of each media system were performed. HPLC Analysis HPLC analysis was performed using a Shimadzu High Performance Liquid Chromatography Prominence-I LC-2030C system [4, 5]. For each drug, calibration curves were constructed, and the line´s equation used to interpolate drug concentration. Data Analysis Data analysis and comparison was conducted using Graphpad Prism 9 for MacOSX. ESULTS Equilibrium Solubility Comparison: The FaSIM and FeSIM equilibrium solubility results for the six drugs and literature HIF values [7] are presented in Figures 1 and 2. For situations with three or more HIF values (3 fasted, 2 fed) a Mann-Whitney comparison with the nine simulated media was performed and did not detect a statistically significant difference (P < 0.05). The four additional available HIF data points (3 fasted, 1 fed) also lie within the FaSIM and FeSIM solubility range for the drugs concerned. For mefenamic acid no literature HIF values were located. The overall low number of HIF points available 23 (15 fasted, 8 fed) compared to 108 for the simulated media, highlights the difficulty of obtaining HIF and the consequent low measurement coverage available. Irrespective of this limitation all of the HIF values are statistically equivalent to or lie within the FaSIM or FeSIM solubility envelope. Equilibrium Solubility Correlation: The minimum and maximum FaSIM and FeSIM solubilities can be used to determine an upper and lower correlation boundary for both states (Figure 3A & B). Using these boundaries SIF and HIF solubility values for 10 additional acidic drugs [7] can be compared. For the 16 additional values only 1 is outside, indicating a 94% correlation between FaSIM, FeSIM and published SIF and HIF values. A close agreement with the original mathematical analysis of HIF composition. CONCLUSION Correlation is difficult due to restricted HIF data sets and also limited to the acidic drugs in this study. The available HIF data correlates to FaSIM and FeSIM when analysed as a direct comparison or using a correlation approach to cover additional drugs. This indicates that correlation is possible and further study is warranted, since a SIF = HIF correlation would allow QbD, modelling and prediction of multiple oral biopharmaceutical behaviours from simple in vitro measurements

Neuronal antibodies in pediatric epilepsy:Clinical features and long-term outcomes of a historical cohort not treated with immunotherapy

OBJECTIVE: In autoimmune encephalitis the etiologic role of neuronal cell-surface antibodies is clear; patients diagnosed and treated early have better outcomes. Neuronal antibodies have also been described in patients with pediatric epilepsy without encephalitis. The aim was to assess whether antibody presence had any effect on long-term outcomes in these patients.METHODS: Patients (n = 178) were recruited between 1988 and 1992 as part of the prospective Dutch Study of Epilepsy in Childhood; none received immunotherapy. Healthy age-matched bone-marrow donors served as controls (n = 112). All sera were tested for serum N-methyl-d-aspartate receptor (NMDAR), alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor, leucine rich glioma inactivated 1, contactin associated protein like 2 (CASPR2), contactin-2, glutamic acid decarboxylase, and voltage gated potassium channel (VGKC)-complex antibodies by standard techniques. No cerebrospinal fluid (CSF) samples were available. Results were correlated with clinical data collected over 15 years.RESULTS: Seventeen patients (9.5%) were positive for VGKC complex (n = 3), NMDAR (n = 7), CASPR2 (n = 4), and contactin-2 (n = 3), compared to three (3/112; 2.6%) healthy controls (VGKC complex [n = 1], NMDAR [n = 2]; p = 0.03; Fisher's exact test). Titers were relatively low (≤1:100 for cell-surface antibodies), but 8 (47%) of the 17 positive samples bound to the surface of live hippocampal neurons consistent with a potential pathogenic antibody. Preexisting cognitive impairment was more frequent in antibody-positive patients (9/17 vs. 33/161; p = 0.01). Fourteen antibody-positive patients were treated with standard antiepileptic drugs (AEDs); three (17%) became intractable but this was not different from the 16 (10%) of 161 antibody-negative patients. In 96 patients with available follow-up samples at 6 and/or 12 months, 6 of 7 positive antibodies had disappeared and, conversely, antibodies had appeared for the first time in a further 7 patients.SIGNIFICANCE: Neuronal antibodies were found at low levels in 9.5% of patients with new-onset pediatric epilepsy but did not necessarily persist over time, and the development of antibodies de novo in later samples suggests they could be due to a secondary response to neuronal damage or inflammation. Moreover, as the response to standard AEDs and the long-term outcome did not differ from those of antibody-negative pediatric patients, these findings suggest that routine neuronal antibody testing is unlikely to be helpful in pediatric epilepsy. However, the higher incidence of preexisting cognitive problems in the antibody-positive group, the CASPR2 and contactin-2 antibodies in 7 of 17 patients, and the binding of 8 of 17 of serum samples to live hippocampal neurons suggest that neuronal antibodies, even if secondary, could contribute to the comorbidities of pediatric epilepsy.</p