ABSTRACT
BACKGROUND: Alternative forms of housing for persons with dementia have been developed in recent decades. These concepts offer small groups of residents familiar settings combined with efforts to provide normal daily life. The aim of this systematic review is to collate and analyze these more innovative forms of housing regarding residents' quality of life, behavioral aspects, as well as functional, cognitive and emotional aspects. METHODS: Searches were conducted in PubMed, EMBASE and PsycInfo in November 2020. Studies comparing traditional and more innovative living environments for persons with dementia were eligible. Concepts are described based on the results of additional searches. Risk of bias of included studies was assessed using checklists from the Joanna Briggs Institute. RESULTS: A total of 21 studies corresponding to 11 different concepts were included, namely Green Houses (USA), Group Living (Sweden), Cantou (France), Group Homes (Japan), Small-scale Group Living (Austria), Special Care Facilities (Canada), Shared-housing Arrangements (Germany), Residential Groups (Germany), Residential Care Centers / Woodside Places (USA/Canada), Small-scale Living (Netherlands/ Belgium), and Green Care Farms (Netherlands). The concepts are broadly similar in terms of care concepts, but partly differ in group sizes, staff qualifications and responsibilities. Several studies indicate that innovative forms of housing may encourage social behavior, preserve activity performance and/or positively influence emotional status compared to more traditional settings, while other studies fail to demonstrate these effects. Some studies also show increased behavioral and psychological symptoms of dementia (BPSD) in residents who live in more innovative housing concepts. The effect on cognition remains indistinct. DISCUSSION: The positive effects may be attributable to the inherent characteristics, including small group sizes, a stimulating design, and altered staff roles and responsibilities. Arguably, some of these characteristics might also be the reason for increased BPSD. Studies had variable methodological quality and results have to be considered with caution. Future research should examine these effects more closely and should investigate populations' preferences with regards to housing in the event of dementia.
Subject(s)
Dementia , Quality of Life , Humans , Quality of Life/psychology , Social Behavior , Housing , Cognition , Dementia/diagnosis , Dementia/epidemiology , Dementia/therapyABSTRACT
AIMS: Orthodontic care and its effectiveness have increasingly become the focus of political and public attention in the recent past. Therefore, this multicenter cohort study aimed to report about the effectiveness of orthodontic treatments in Germany and to identify potential influencing factors. METHODS: A total of 586 patients from seven German study centers were screened for this cohort study, of which 361 patients were recruited at the end of their orthodontic treatment. Of these, 26 patients had missing study models and/or missing treatment information. Thus, 335 participants were included. The severity of malocclusion was rated using the Peer Assessment Rating (PAR) Index at baseline (T0) retrospectively and-prospectively-after the retention period (T1). Practitioner-, treatment- and patient-related information were analyzed in order to detect potential predictive factors for treatment effectiveness. RESULTS: Study participants (202 female and 133 male) were on average 14.8 (standard deviation [SD]⯱ 6.1) years old at start of active treatment. Average PAR score at T0 was 25.96 (SD⯱ 10.75) and mean posttreatment PAR score was 3.67 (SD⯱ 2.98) at T1. An average decrease of total PAR score by 22.30 points (SD⯱ 10.73) or 83.54% (SD⯱ 14.58; pâ¯< 0.001) was detected. Furthermore, 164 treatments (49.1%) were categorized as 'greatly improved' but only 3 treatments (0.9%) as 'worse or no different'; 81.5% of all cases finished with a high-quality treatment outcome (≤5 PAR points at T1). Logistic regression analyses detected staff experience as a significant predictive factor for high-quality results (odds ratio 1.27, pâ¯= 0.001, 95% confidence interval 1.11-1.46). CONCLUSION: The improvement rate among this selected German cohort indicated an overall very good standard of orthodontic treatment. Staff experience proved to be a predictive factor for high-quality results.
Subject(s)
Malocclusion , Orthodontics, Corrective , Quality of Health Care , Cohort Studies , Female , Germany/epidemiology , Humans , Male , Malocclusion/diagnosis , Malocclusion/epidemiology , Malocclusion/therapy , Retrospective Studies , Treatment OutcomeABSTRACT
KEY POINTS: Diabetes is thought to induce neuropathic pain through activation of dorsal horn sensory neurons in the spinal cord. Here we explore the impact of hyperglycaemia on the blood supply supporting the spinal cord and chronic pain development. In streptozotocin-induced diabetic rats, neuropathic pain is accompanied by a decline in microvascular integrity in the dorsal horn. Hyperglycaemia-induced degeneration of the endothelium in the dorsal horn was associated with a loss in vascular endothelial growth factor (VEGF)-A165 b expression. VEGF-A165 b treatment prevented diabetic neuropathic pain and degeneration of the endothelium in the spinal cord. Using an endothelial-specific VEGFR2 knockout transgenic mouse model, the loss of endothelial VEGFR2 signalling led to a decline in vascular integrity in the dorsal horn and the development of hyperalgesia in VEGFR2 knockout mice. This highlights that vascular degeneration in the spinal cord could be a previously unidentified factor in the development of diabetic neuropathic pain. ABSTRACT: Abnormalities of neurovascular interactions within the CNS of diabetic patients is associated with the onset of many neurological disease states. However, to date, the link between the neurovascular network within the spinal cord and regulation of nociception has not been investigated despite neuropathic pain being common in diabetes. We hypothesised that hyperglycaemia-induced endothelial degeneration in the spinal cord, due to suppression of vascular endothelial growth factor (VEGF)-A/VEGFR2 signalling, induces diabetic neuropathic pain. Nociceptive pain behaviour was investigated in a chemically induced model of type 1 diabetes (streptozotocin induced, insulin supplemented; either vehicle or VEGF-A165 b treated) and an inducible endothelial knockdown of VEGFR2 (tamoxifen induced). Diabetic animals developed mechanical allodynia and heat hyperalgesia. This was associated with a reduction in the number of blood vessels and reduction in Evans blue extravasation in the lumbar spinal cord of diabetic animals versus age-matched controls. Endothelial markers occludin, CD31 and VE-cadherin were downregulated in the spinal cord of the diabetic group versus controls, and there was a concurrent reduction of VEGF-A165 b expression. In diabetic animals, VEGF-A165 b treatment (biweekly i.p., 20 ng g-1 ) restored normal Evans blue extravasation and prevented vascular degeneration, diabetes-induced central neuron activation and neuropathic pain. Inducible knockdown of VEGFR2 (tamoxifen treated Tie2CreERT2 -vegfr2flfl mice) led to a reduction in blood vessel network volume in the lumbar spinal cord and development of heat hyperalgesia. These findings indicate that hyperglycaemia leads to a reduction in the VEGF-A/VEGFR2 signalling cascade, resulting in endothelial dysfunction in the spinal cord, which could be an undiscovered contributing factor to diabetic neuropathic pain.
Subject(s)
Diabetes Complications/etiology , Diabetes Mellitus, Experimental/complications , Diabetic Angiopathies/etiology , Diabetic Neuropathies/etiology , Hyperalgesia/etiology , Neuralgia/etiology , Spinal Cord/pathology , Animals , Cells, Cultured , Diabetes Complications/metabolism , Diabetes Complications/pathology , Diabetes Complications/prevention & control , Diabetic Angiopathies/metabolism , Diabetic Angiopathies/pathology , Diabetic Angiopathies/prevention & control , Diabetic Neuropathies/metabolism , Diabetic Neuropathies/pathology , Diabetic Neuropathies/prevention & control , Endothelium, Vascular/metabolism , Endothelium, Vascular/pathology , Female , Humans , Hyperalgesia/metabolism , Hyperalgesia/pathology , Hyperalgesia/prevention & control , Male , Mice , Mice, Knockout , Mice, Transgenic , Microvessels/physiopathology , Neuralgia/metabolism , Neuralgia/pathology , Neuralgia/prevention & control , Rats , Rats, Sprague-Dawley , Spinal Cord/blood supply , Spinal Cord/metabolism , Vascular Endothelial Growth Factor A/administration & dosage , Vascular Endothelial Growth Factor Receptor-2/physiologyABSTRACT
Fetal calf serum (FCS) bears a potential risk for carrying diseases and eliciting immune reactions. Nevertheless, it still represents the gold standard as medium supplement in cell culture. In the present study, human platelet lysate (PL) was tested as an alternative to FCS for the expansion and subsequent chondrogenic differentiation of human adipose-derived stem cells (ASCs). ASCs were expanded with 10% FCS (group F) or 5% PL (group P). Subsequently, three-dimensional (3D) micromass pellets were created and cultured for 5 weeks in chondrogenic differentiation medium. Additionally, the de- and redifferentiation potential of human articular chondrocytes (HACs) was evaluated and compared to ASCs. Both HACs and ASCs cultured with PL showed strongly enhanced proliferation rates. Redifferentiation of HACs was possible for cells expanded up to 3.3 population doublings (PD). At this stage, PL-expanded HACs demonstrated better redifferentiation potential than FCS-expanded cells. ASCs could also be differentiated following extended passaging. Glycosaminoglycan (GAG) quantification and qRT-PCR of 10 cartilage related markers demonstrated a tendency for increased chondrogenic differentiation of PL-expanded ASCs compared to cells expanded with FCS. Histologically, collagen type II but also collagen type X was mainly present in group P. The present study demonstrates that PL strongly induces proliferation of ASCs, while the chondrogenic differentiation potential is retained. HACs also showed enhanced proliferation and even better redifferentiation when previously expanded with PL. This suggests that PL is superior to FCS as a supplement for the expansion of ASCs and HACs, particularly with regard to chondrogenic (re)differentiation.
Subject(s)
Adipose Tissue/metabolism , Blood Platelets/chemistry , Cartilage, Articular/metabolism , Cell Differentiation , Chondrocytes/metabolism , Chondrogenesis , Stem Cells/metabolism , Adipose Tissue/cytology , Cartilage, Articular/cytology , Chondrocytes/cytology , Humans , Stem Cells/cytologyABSTRACT
Vascular endothelial growth factor-A (VEGF-A) is best known as a key regulator of the formation of new blood vessels. Neutralization of VEGF-A with anti-VEGF therapy e.g. bevacizumab, can be painful, and this is hypothesized to result from a loss of VEGF-A-mediated neuroprotection. The multiple vegf-a gene products consist of two alternatively spliced families, typified by VEGF-A165a and VEGF-A165b (both contain 165 amino acids), both of which are neuroprotective. Under pathological conditions, such as in inflammation and cancer, the pro-angiogenic VEGF-A165a is upregulated and predominates over the VEGF-A165b isoform. We show here that in rats and mice VEGF-A165a and VEGF-A165b have opposing effects on pain, and that blocking the proximal splicing event - leading to the preferential expression of VEGF-A165b over VEGF165a - prevents pain in vivo. VEGF-A165a sensitizes peripheral nociceptive neurons through actions on VEGFR2 and a TRPV1-dependent mechanism, thus enhancing nociceptive signaling. VEGF-A165b blocks the effect of VEGF-A165a. After nerve injury, the endogenous balance of VEGF-A isoforms switches to greater expression of VEGF-Axxxa compared to VEGF-Axxxb, through an SRPK1-dependent pre-mRNA splicing mechanism. Pharmacological inhibition of SRPK1 after traumatic nerve injury selectively reduced VEGF-Axxxa expression and reversed associated neuropathic pain. Exogenous VEGF-A165b also ameliorated neuropathic pain. We conclude that the relative levels of alternatively spliced VEGF-A isoforms are critical for pain modulation under both normal conditions and in sensory neuropathy. Altering VEGF-Axxxa/VEGF-Axxxb balance by targeting alternative RNA splicing may be a new analgesic strategy.
Subject(s)
Antibodies/therapeutic use , DNA, Recombinant/genetics , Neuralgia/metabolism , Neuralgia/therapy , RNA, Messenger/metabolism , Vascular Endothelial Growth Factor A , Animals , Antibodies/pharmacology , Benzofurans , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/therapeutic use , Ganglia, Spinal/cytology , Hyperalgesia/metabolism , Male , Mice , Mice, Transgenic , Neural Conduction/genetics , Pain Measurement , Pain Threshold/physiology , Quinolines , RNA, Messenger/genetics , Rats , Rats, Wistar , Sensory Receptor Cells/drug effects , Sensory Receptor Cells/metabolism , TRPV Cation Channels/deficiency , TRPV Cation Channels/genetics , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/immunology , Vascular Endothelial Growth Factor A/metabolismABSTRACT
Methylmethaqualone is a sedative designer drug created by adding a methyl group to the 3-phenyl ring of methaqualone, and is at present not subject to restrictive regulation in many countries. To our knowledge, no case of methylmethaqualone abuse has been published to date in the scientific literature, and the only sources of information are users' reports on Web discussion forums and data from preclinical animal studies. We report a case of oral methylmethaqualone abuse confirmed by liquid chromatography tandem mass spectrometry in a 24-year-old previously healthy Caucasian male. Observed symptoms and signs such as central nervous system depression alternating with excitation, psychomotor agitation, muscle hyperactivity, and tachycardia were compatible with methaqualone-induced adverse effects. Except for the mild tachycardia (115 beats/min), other vital signs were normal: blood pressure 134/89 mmHg, body temperature 36.2°C (97.16°F), and peripheral oxygen saturation 99% while breathing room air. The ECG showed no prolongation of the QT interval and the QRS duration was normal. Laboratory analysis revealed a slight increase in creatine kinase (368 U/L) and alanine aminotransferase (90 U/L) serum concentrations. Blood alcohol concentration was 0.32 g/L. Methylmethaqualone was identified in a serum sample collected on admission which was analyzed by a liquid chromatography tandem mass spectrometry toxicological screening method using turbulent flow online extraction. After a few days the patient ingested the same amount of substance with identical symptoms. Based on the chemical structure and animal data, and according to this case report and users' Web reports, methylmethaqualone appears to have a similar acute toxicity profile to methaqualone, with marked psychomotor stimulation. Symptoms of acute toxicity can be expected to resolve with supportive care.
Subject(s)
Designer Drugs/toxicity , Hypnotics and Sedatives/toxicity , Methaqualone/analogs & derivatives , Neurotoxicity Syndromes/blood , Adult , Chromatography, High Pressure Liquid , Designer Drugs/analysis , Designer Drugs/pharmacokinetics , Humans , Hypnotics and Sedatives/blood , Hypnotics and Sedatives/pharmacokinetics , Male , Methaqualone/blood , Methaqualone/pharmacokinetics , Methaqualone/toxicity , Methylation , Neurotoxicity Syndromes/physiopathology , Neurotoxicity Syndromes/therapy , Severity of Illness Index , Tandem Mass Spectrometry , Treatment Outcome , Young AdultABSTRACT
Vascular endothelial growth factors (VEGFs) regulate blood and lymph vessel formation through activation of the type V receptor tyrosine kinases VEGFR-1, -2 and -3. In addition, VEGFs interact with co-receptors such as neuropilins, integrins, semaphorins or heparansulfate glycosaminoglycans. Ligand binding dimerises the receptors and activates their intracellular tyrosine kinase domains, resulting in phosphorylation of tyrosine residues acting as docking sites for intracellular signalling molecules. Ligand-induced receptor is internalised and then transported through early, late, and recycling endosomes, and finally degraded by proteasomal or lysosomal pathways. Biological output by VEGF is mediated through distinct receptor/co-receptor complexes and generates signals in all cellular compartments triggering cellular responses such as cell migration, cell proliferation, vessel formation and maturation, as well as changes in vessel fenestration, constriction and permeability. Here we review recent experiments showing how VEGFR-2 is transported through intracellular vesicular compartments specified by Rab family GTPases, and discuss how different VEGF-A isoforms specify intracellular receptor trafficking. We also discuss how the biological consequences of aberrant receptor trafficking bear on the development of vascular disease.
Subject(s)
Cytoplasm/metabolism , Neuropilin-1/metabolism , Protein Transport/physiology , Vascular Endothelial Growth Factor Receptor-2/metabolism , rab GTP-Binding Proteins/metabolism , Animals , Humans , Signal TransductionABSTRACT
BACKGROUND AND OBJECTIVES: Molecular variations of the RHD gene may result in the reduced expression of the D antigen and altered Rh phenotypes. In many occasions, they cannot be typed reliably by standard serological methods. Sequence-based typing is the gold standard to determine rare and unknown RHD genotypes. For this pilot study, sequence-based typing by standard Sanger sequencing was compared to a newly established next-generation sequencing approach based on pyrosequencing. MATERIALS AND METHODS: Twenty-six DNA samples were selected after primary serological testing exhibiting a weak reaction in Rh phenotype. Parallel sequence analysis of the complete coding sequence including adjacent intronic sequences allowed a comparison of the methodical potency in mutation detection of Sanger with next-generation sequencing. RESULTS: Sanger sequencing revealed 39 RHD polymorphisms in 21 of 26 samples in the RHD coding region, while pyrosequencing detected all but two alterations resulting in a concordance rate of 94·9% and clearly revealed a heterozygous compound mutation in one sample with RHDψ and Weak D type 4 alleles. The resolution of cis/trans linkage of polymorphisms and exact characterization of a 37 bp duplication was achieved by next-generation sequencing. CONCLUSION: Our data suggest that next-generation sequencing offers a new development for high-throughput and clonal sequencing for molecular RHD genotyping. However, further attempts in the methodical set-up have to be undertaken prior to validation and introduction as a routine service.
Subject(s)
Blood Grouping and Crossmatching/methods , Polymorphism, Genetic , Rh-Hr Blood-Group System/genetics , Sequence Analysis, DNA/methods , Blood Grouping and Crossmatching/standards , Female , Humans , Male , Sequence Analysis, DNA/standardsABSTRACT
BACKGROUND: Amniotic membrane is a highly promising cell source for tissue engineering. Being part of the placenta, this tissue is abundantly available. It can be processed easily to yield large amounts of epithelial and mesenchymal cells that have shown broad differentiation potential. For tissue-engineering purposes, cells may be applied either directly after isolation from the tissue or after a period of in vitro expansion to obtain higher cell numbers. In order to investigate the advantages and drawbacks of these strategies we compared freshly isolated and cultivated human amniotic epithelial cells (hAEC) regarding their surface antigen (Ag) expression profile and osteogenic differentiation capacity. METHODS: Expression of surface Ag that are characteristic for mesenchymal stromal and embryonic stem cells was analyzed by flow cytometry. Different protocols for osteogenic and adipogenic differentiation were compared. RESULTS: We have demonstrated that expression of surface Ag changes dramatically during cultivation of hAEC. While not or only weakly expressed on primary isolates, the mesenchymal markers CD13, CD44, CD49e, CD54, CD90 and CD105 are strongly up-regulated during in vitro propagation. In contrast, expression of the embryonic markers TRA-1-60 and TRA-1-81, but not SSEA-4, rapidly decreases upon cultivation. This phenotypic shift is associated with a reduction in osteogenic differentiation. DISCUSSION: Our results suggest that phenotypic alterations of hAEC during in vitro cultivation might be responsible for a functional reduction of the differentiation potential, which has to be considered for the potential application of these cells in regenerative medicine.
Subject(s)
Amnion/cytology , Cell Differentiation , Epithelial Cells/cytology , Mesenchymal Stem Cells/cytology , Osteogenesis , Amnion/physiology , Cell Culture Techniques , Epithelial Cells/physiology , Female , Humans , Immunophenotyping , Mesenchymal Stem Cells/physiology , PregnancyABSTRACT
We report here the novel human leukocyte antigens (HLA)-Cw*0429 and HLA-DRB3*0223 alleles identified during routine cord blood characterisation by sequence-based typing.
Subject(s)
Exons/genetics , HLA-C Antigens/genetics , HLA-DR Antigens/genetics , Alleles , HLA-DRB3 Chains , HumansABSTRACT
BACKGROUND AND OBJECTIVE: Genotyping may be applied for rare blood group polymorphisms in a high-throughput mode as well for the molecular determination of blood groups due to unclear serological results. MATERIAL AND METHODS: We developed and validated a DNA typing method for the determination of KEL1/2, JK1/2, FY1/2, FY0, MNS1/2, MNS3/4, DO1/2, CO1/2 and LU1/2 alleles using a melting curve analysis downstream from a fully automated DNA extraction. All assays were validated in terms of specificity, sensitivity, assay variability and robustness. The usability was proven by a batch of 200 blood samples with partially known phenotype. RESULTS: Assays for all blood groups were within the range of specificity (100%), assay variability and robustness (coefficient of variance < 3%). Genotypes of 200 random platelet donors were fully consistent with existing phenotype data. The obtained genotype distribution is in complete concordance with existing data for the European population underlined by a complete absence of CO2 homozygous donors and the FY0 allele among the cohort. CONCLUSION: We introduce an approach for blood group genotyping of particular samples or gene loci in glass capillary format and for medium-throughput analysis in 96/384-well format. The advantages of this real-time polymerase chain reaction method are its automation potential, the flexibility regarding hardware and the rapid cycling time.
Subject(s)
Blood Group Antigens/genetics , Blood Grouping and Crossmatching/methods , Fluorescence Resonance Energy Transfer , Polymerase Chain Reaction/methods , Blood Donors , Computer Systems , DNA/blood , DNA/genetics , Genotype , Humans , Nucleic Acid Denaturation , Reproducibility of Results , Robotics , Sensitivity and Specificity , Serologic TestsABSTRACT
The new allele HLA-A*1129 showed one nucleotide difference form A*110101 at codon 172 (CTG>CAG).
Subject(s)
HLA-A Antigens/genetics , Alleles , Austria , Blood Donors , Female , Fetal Blood/immunology , HLA-A Antigens/chemistry , Haplotypes , Humans , Infant, Newborn , MaleABSTRACT
This paper provides a brief review of recently published work on biophysical and biological aspects of Auger processes. Three specific questions have been considered. (1) Does charge neutralisation contribute to molecular damage such as DNA strand breaks? (2) How many DNA double strand breaks are produced by a single decay of DNA bound (125)I? (3) What is the correlation between number of gammaH2AX foci and number of double strand breaks (DSB)? The paper also gives preliminary reports on two new calculations: (a) calculation of the spectrum of Auger electrons released during decay of (124)I and (b) the use of Auger electrons in the decay of (125)I as a probing agent of novel DNA structures.
Subject(s)
DNA Damage , DNA/chemistry , DNA/radiation effects , Electrons , Iodine Radioisotopes/chemistry , Models, Chemical , Molecular Probe Techniques , Computer Simulation , Models, BiologicalABSTRACT
The development of functional blood and lymphatic vessels requires spatio-temporal coordination of the production and release of growth factors such as vascular endothelial growth factors (VEGFs). VEGF family proteins are produced in multiple isoforms with distinct biological properties and bind to three types of VEGF receptors. A VEGF-A splice variant, VEGF-A(165)b, has recently been isolated from kidney epithelial cells. This variant is identical to VEGF-A(165) except for the last six amino acids encoded by an alternative exon. VEGF-A(165)b and VEGF-A(165) bind VEGF receptors 1 and 2 with similar affinity. VEGF-A(165)b elicits drastically reduced activity in angiogenesis assays and even counteracts signaling by VEGF-A(165). VEGF-A(165)b weakly binds to heparan sulfate and does not interact with neuropilin-1, a coreceptor for VEGF receptor 2. To determine the molecular basis for altered signaling by VEGF-A(165)b we measured VEGF receptor 2 and ERK kinase activity in endothelial cells in culture. VEGF-A(165) induced strong and sustained activation of VEGF receptor 2 and ERK-1 and -2, while activation by VEGF-A(165)b was only weak and transient. Taken together these data show that VEGF-A(165)b has attenuated signaling potential through VEGF receptor 2 defining this new member of the VEGF family as a partial receptor agonist.
Subject(s)
Chorioallantoic Membrane/blood supply , Heparitin Sulfate/metabolism , Neuropilin-1/metabolism , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor Receptor-2/metabolism , Alternative Splicing , Animals , Cell Line , Chick Embryo , Chorioallantoic Membrane/metabolism , Humans , In Vitro Techniques , Mice , Neovascularization, Physiologic , Protein Binding , Protein Isoforms/pharmacology , Signal Transduction , Vascular Endothelial Growth Factor A/pharmacokinetics , Vascular Endothelial Growth Factor A/pharmacology , Vascular Endothelial Growth Factor Receptor-1/metabolismABSTRACT
Tumour-associated macrophages, TAMs, play a pivotal role in tumour growth and metastasis by promoting tumour angiogenesis. Treatment with clodronate encapsulated in liposomes (clodrolip) efficiently depleted these phagocytic cells in the murine F9 teratocarcinoma and human A673 rhabdomyosarcoma mouse tumour models resulting in significant inhibition of tumour growth ranging from 75 to >92%, depending on therapy and schedule. Tumour inhibition was accompanied by a drastic reduction in blood vessel density in the tumour tissue. Vascular endothelial growth factor (VEGF) is one of the major inducers of tumour angiogenesis and is also required for macrophage recruitment. The strongest effects were observed with the combination therapy of clodrolip and a VEGF-neutralising antibody, whereas free clodronate was not significantly active. Immunohistologic evaluation of the tumours showed significant depletion of F4/80+ and MOMA-1+ and a less pronounced depletion of CD11b+ TAMs. Blood vessel staining (CD31) and quantification of the vessels as well as TAMs and tumour-associated dendritic cells (TADCs) in the A673 model showed reduction rates of 85 to >94%, even 9 days after the end of therapy. In addition, CD11c+ TADCs, which have been shown to potentially differentiate into endothelial-like cells upon stimulation by tumour released growth and differentiation factors, were similarly reduced by clodrolip or antibody treatment. These results validate clodrolip therapy in combination with angiogenesis inhibitors as a promising novel strategy for an indirect cancer therapy aimed at the haematopoietic precursor cells that stimulate tumour growth and dissemination and as a tool to study the role of macrophages and dendritic cells in tumorigenesis.
Subject(s)
Angiogenesis Inhibitors/pharmacology , Antineoplastic Agents/pharmacology , Clodronic Acid/pharmacology , Macrophages/drug effects , Neoplasms/drug therapy , Angiogenesis Inhibitors/administration & dosage , Angiogenesis Inhibitors/therapeutic use , Animals , Antibodies, Monoclonal/pharmacology , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/therapeutic use , Cell Line, Tumor , Cell Proliferation/drug effects , Clodronic Acid/administration & dosage , Clodronic Acid/therapeutic use , Female , Humans , Immunohistochemistry , In Vitro Techniques , Liposomes , Macrophages/metabolism , Mice , Mice, Nude , Neoplasms/pathology , Tumor Cells, Cultured , Vascular Endothelial Growth Factor A/drug effects , Vascular Endothelial Growth Factor A/metabolism , Xenograft Model Antitumor AssaysABSTRACT
Vascular endothelial growth factors (VEGFs) regulate blood and lymphatic vessel development and homeostasis but also have profound effects on neural cells. VEGFs are predominantly produced by endothelial, hematopoietic and stromal cells in response to hypoxia and upon stimulation with growth factors such as transforming growth factors, interleukins or platelet-derived growth factor. VEGFs bind to three variants of type III receptor tyrosine kinases, VEGF receptor 1, 2 and 3. Each VEGF isoform binds to a particular subset of these receptors giving rise to the formation of receptor homo- and heterodimers that activate discrete signaling pathways. Signal specificity of VEGF receptors is further modulated upon recruitment of coreceptors, such as neuropilins, heparan sulfate, integrins or cadherins. Here we summarize the knowledge accumulated since the discovery of these proteins more than 20 years ago with the emphasis on the signaling pathways activated by VEGF receptors in endothelial cells during cell migration, growth and differentiation.
Subject(s)
Neovascularization, Physiologic/physiology , Receptor Protein-Tyrosine Kinases/metabolism , Receptors, Vascular Endothelial Growth Factor/physiology , Signal Transduction/physiology , Animals , Cadherins/metabolism , Humans , Vascular Endothelial Growth Factor A/metabolismABSTRACT
Protein transduction domains (PTDs) are used to enhance cellular uptake of drugs, proteins, polynucleotides or liposomes. In this study, functionalized Antennapedia (Antp, aa 43-58) and HIV Tat (aa 47-57) peptides were coupled to small unilamellar liposomes via thiol-maleimide linkage. Modified liposomes showed higher uptake into a panel of cell lines including tumor and dendritic cells than unmodified control liposomes. Liposome uptake was time and concentration dependent as analyzed by flow cytometry and live-cell microscopy. At least 100 PTD molecules per small unilamellar liposome (100 +/- 30 nm) were necessary for efficient translocation into cells. Cellular uptake of PTD-modified liposomes was 15- to 25-fold increased compared to unmodified liposomes and was inhibited by preincubation of liposomes with heparin. Glycosaminoglycan-deficient CHO cells showed dramatically reduced cell association of PTD-modified liposomes, confirming the important role of heparan sulfate proteoglycans in PTD-mediated uptake. Antp-liposomes used as carriers of the cytotoxic drug N4-octadecyl-1-beta-D-arabinofuranosylcytosine-(5'- 5')-3'-C-ethinylcytidine showed a reduction of the IC50 by 70% on B16F1 melanoma cells compared with unmodified liposomes. PTD-functionalized liposomes, particularly Antp-liposomes, represent an interesting novel carrier system for enhanced cell-specific delivery of a large variety of liposome-entrapped molecules.
Subject(s)
Heparan Sulfate Proteoglycans/metabolism , Liposomes/metabolism , Peptides/metabolism , Animals , Flow Cytometry , HeLa Cells , Humans , Liposomes/toxicity , Mice , Microscopy, FluorescenceABSTRACT
We prepared small unilamellar liposomes derivatised with single chain antibody fragments specific for the ED-B domain of B-fibronectin. This extracellular matrix associated protein is expressed around newly forming blood vessels in the vicinity of many types of tumours. The single chain antibody fragments were functionalised by introduction of C-terminal cysteines and linked to liposomes via maleimide groups located at the terminal ends of poly(ethylene glycol) modified phospholipids. The properties of these anti-ED-B single chain antibody fragments-liposomes were analysed in vitro on ED-B fibronectin expressing Caco-2 cells and in vivo by studying their biodistribution and their therapeutic potential in mice bearing subcutanous F9 teratocarcinoma tumours. Radioactively labelled ((114m)Indium) single chain antibody fragments-liposomes accumulated in the tumours at 2-3-fold higher concentrations during the first 2 h after i.v. injection compared to unmodified liposomes. After 6-24 h both liposome types were found in similar amounts (8-10% injected dose g(-1)) in the tumours. Animals treated i.v. with single chain antibody fragments-liposomes containing the new cytotoxic agent 2'-deoxy-5-fluorouridylyl-N(4)-octadecyl-1-beta-D-arabinofuranosylcytosine (30 mg kg(-1) per dose, five times every 24 h) showed a reduction of tumour growth by 62-90% determined on days 5 and 8, respectively, compared to animals receiving control liposomes. Histological analysis revealed a marked reduction of F9 tumour cells and excessive deposition of fibronectin in the extracellular matrix after treatment with single chain antibody fragments-2-dioxy-5-fluorouridylyl-N(4)-octadecyl-1-beta-D-arabinofuranosylcytosine-liposomes. Single chain antibody fragments-liposomes targeted to ED-B fibronectin positive tumours therefore represent a promising and versatile novel drug delivery system for the treatment of tumours.
Subject(s)
Antibodies, Neoplasm/immunology , Antineoplastic Agents/pharmacology , Cytarabine/analogs & derivatives , Cytarabine/pharmacology , Fluorodeoxyuridylate/analogs & derivatives , Fluorodeoxyuridylate/pharmacology , Immunoglobulin Fragments/immunology , Teratocarcinoma/immunology , Testicular Neoplasms/immunology , Animals , Female , Fibronectins , Immunoglobulin Fragments/pharmacology , Injections, Intravenous , Liposomes , Male , Mice , Mice, Nude , Teratocarcinoma/drug therapy , Teratocarcinoma/pathology , Testicular Neoplasms/drug therapy , Testicular Neoplasms/pathology , Tissue Distribution , Tumor Cells, CulturedABSTRACT
Using venous occlusion plethysmography, Sunahara et al. reported that Coriolis-induced nausea was accompanied by an increase in forearm blood flow, suggesting a decrease in sympathetic activity to this vascular bed. No significant blood pressure and heart rate changes were observed. Vasodilation of the limbs theoretically impairs orthostatic tolerance, particularly if blood flow is shown to increase simultaneously in the lower limbs. This study examined the latter possibility. Seventeen subjects were exposed to the Coriolis cross-coupling effects induced by 20 RPM yaw rotation, and a simultaneous 45 degrees pitch forward head movement in the sagittal plane every 12 s. Forearm and calf skin blood flow were monitored in real-time using laser Doppler flowmetry (PeriFlux 4001). Our results indicated a significant (p < 0.001) simultaneous forearm and calf skin blood flow increase as a result of Coriolis cross-coupling across all 15 susceptible subjects. No significant changes in blood pressure and heart rate were observed. Coriolis-induced cardiovascular changes may confound previous reports on reduced G tolerance using ground-based centrifuges that invariably evoke cross-coupling effects.
Subject(s)
Coriolis Force , Forearm/physiopathology , Leg/physiopathology , Motion Sickness/physiopathology , Regional Blood Flow/physiology , Skin Physiological Phenomena , Adult , Blood Pressure/physiology , Female , Heart Rate/physiology , Humans , Hypotension, Orthostatic/physiopathology , Laser-Doppler Flowmetry , Male , Middle Aged , Motion Sickness/psychology , Respiratory Physiological Phenomena , Vasodilation/physiologyABSTRACT
Chinese hamster ovary (CHO) K1 and radiosensitive CHO irs-20 cells were synchronized in S phase and labeled for 10 min with 5-[(125)I]-iodo-2'-deoxyuridine ((125)IdU). The cells were washed, incubated in fresh medium for 1 h for incorporation of the intracellular radionucleotides into DNA, and then frozen (-80 degrees C) for accumulation of (125)I decays. At intervals after freezing, when the cells had accumulated the desired number of decays, aliquots of the frozen cells were thawed and plated to determine survival. The survival curves for K1 and irs-20 cells were similar from 100% to 30% survival. At higher (125)I doses (more decays/cell), the survival of K1 cells continued to decline exponentially, but the survival of X-ray-sensitive irs-20 cells remained at approximately 30% even after the cells had accumulated 1265 decays/cell. The results contradict the notion that increased DNA damage inevitably causes increased cell death. To account for these findings, we propose a model that postulates the existence of a second radiation target. According to this model, radiation damage to DNA may be necessary to induce cell death, but DNA damage alone is not sufficient to kill cells. We infer from the survival response of irs-20 cells that damage to a second (non-DNA) structure is involved in cell death, and that this structure directly affects the repair of DNA and cell survival.