ABSTRACT
Proteins provide essential functional regulation of many bioprocesses across all scales of life; however, new techniques to specifically modulate protein activity within living systems and in engineered biomaterials are needed to better interrogate fundamental cell signalling and guide advanced decisions of biological fate. Here we establish a generalizable strategy to rapidly and irreversibly activate protein function with full spatiotemporal control. Through the development of a genetically encoded and light-activated SpyLigation (LASL), bioactive proteins can be stably reassembled from non-functional split fragment pairs following brief exposure (typically minutes) to cytocompatible light. Employing readily accessible photolithographic processing techniques to specify when, where and how much photoligation occurs, we demonstrate precise protein activation of UnaG, NanoLuc and Cre recombinase using LASL in solution, biomaterials and living mammalian cells, as well as optical control over protein subcellular localization. Looking forward, we expect that these photoclick-based optogenetic approaches will find tremendous utility in probing and directing complex cellular fates in both time and three-dimensional space.
Subject(s)
Mammals , AnimalsABSTRACT
Given the critical role that proteins play in almost all biological processes, there is great interest in controlling their presentation within and release from biomaterials. Despite such outstanding enthusiasm, previously developed strategies in this regard result in ill-defined and heterogeneous populations with substantially decreased activity, precluding their successful application to fragile species including growth factors. Here, we introduce a modular and scalable method for creating monodisperse, genetically encoded chimeras that enable bioactive proteins to be immobilized within and subsequently photoreleased from polymeric hydrogels. Building upon recent developments in chemoenzymatic reactions, bioorthogonal chemistry, and optogenetics, we tether fluorescent proteins, model enzymes, and growth factors site-specifically to gel biomaterials through a photocleavable protein (PhoCl) that undergoes irreversible backbone photoscission upon exposure to cytocompatible visible light (λ ≈ 400 nm) in a dose-dependent manner. Mask-based and laser-scanning lithographic strategies using commonly available light sources are employed to spatiotemporally pattern protein release from hydrogels while retaining their full activity. The photopatterned epidermal growth factor presentation is exploited to promote anisotropic cellular proliferation in 3D. We expect these methods to be broadly useful for applications in diagnostics, drug delivery, and regenerative medicine.
Subject(s)
Biocompatible Materials , Immobilized Proteins/chemistry , Photochemical Processes , Proteins/chemistry , Drug Liberation , Models, Molecular , Protein ConformationABSTRACT
BACKGROUND: New and innovative concepts of care management have been developed to improve the health of older adults with dementia and depression. AIM: This article describes the American aging brain care (ABC) program and the possible transfer to the German healthcare system is discussed. MATERIAL AND METHODS: The ABC medical home model in Indianapolis incorporates a specialized geriatric healthcare center which is affiliated to the Eskenazi Hospital as well as a program involving home-based domestic visits by healthcare personnel to affected people. The diagnoses are made in the geriatric center where therapy and treatment are also planned. These stages are carried out in a multiprofessional team, which identifies the individual needs of the patients and relatives and discusses these in family conferences as well as in close consultation with the primary care center of the hospital. The care, diagnosis and therapy are coordinated using a self-developed software for the program and via predetermined pathways and procedural instructions on the approach in the healthcare center and in the domestic visit program. RESULTS AND CONCLUSION: From the perspective of the authors the core elements of the program include not only the use of a home-based care model but also the selection and training of a new type of front-line care provider. Models like the program presented here show great promise for meeting the demands of a rapidly expanding population of vulnerable older adults.
Subject(s)
Critical Pathways/organization & administration , Dementia/therapy , Depression/therapy , Health Services for the Aged/organization & administration , Home Care Services/organization & administration , National Health Programs/organization & administration , Aged , Aged, 80 and over , Dementia/diagnosis , Depression/diagnosis , Female , Humans , India , Interinstitutional Relations , International Cooperation , Male , Models, OrganizationalABSTRACT
The diagnostic value of radiological pelvimetry is subject to controversial interpretation. This examination is being conducted at the Department of Gynaecology in Aarau (Frauenklinik Aarau) mainly in pregnant women in pelvic presentation or during the puerperium after Caesarean section. Based on 164 radiological pelvimetric measurements in breech presentation and 112 X-ray films after Caesarean section an attempt was made to assess the prospective value of this examination. Abnormal shapes of the pelvis and narrow pelvic dimensions were found about equally often. Analysis of the indications of the section yielded both in breech presentation and in the entire group of the patients examined post partum an almost equally high percentage of about 20% of indications for a disproportionate head-pelvis relationship. Radiologically, considerably narrowed pelvic dimensions were seen in 10% of the examined women. The most frequent finding was a funnel-shaped pelvis, whereas an assimilation pelvis was never seen. Every pathologically narrowed funnel-shaped pelvis and generally narrow pelvis was detected also by digital palpation. All cases of conjugata vera masses below 10 cm concerned generally narrow cases of pelvic dimensions and were thus detectable by digital palpation. We recommended to abstain from routine performance of radiometric pelvimetry. Lateral pelvic X-ray can be an additional help in difficulties in pregnant women with regard to determining the mode of birth in breech presentation and after Caesarean section in case there is an urgent suspicion of malproportionate head-pelvis ratios.
Subject(s)
Pelvimetry/methods , Pregnancy Complications/diagnostic imaging , Cesarean Section , Embryonic and Fetal Development , Female , Humans , Obstetric Labor Complications/diagnostic imaging , Pregnancy , RadiographyABSTRACT
236 patients were reevaluated one year after vaginal or suprapubic continence surgery. Personal history, clinico-gynaecological status, morphology and urodynamics were considered. 70.6% respectively 82% of all patients considered themselves cured. These numbers correlate well with objective control parameters. The clinical examination shows that a suprapubic approach is better for an urethrocele whereas vaginal surgery is more successful for a cystocele or rectocele. The correction of an urethrocele has however a greater influence on the chance of cure. Obesity is a risk factor especially for vaginal surgery and in both groups oestrogen application improved the healing process. The morphological examination demonstrates the importance of urethro-vesical suspension giving better results after suprapubic than after vaginal operation. The urodynamic results depend on the choice of measurement parameters. Suprapubic surgery generally brings about a greater improvement in pressure conditions than vaginal surgery. The best parameter seems to be the Dep Q. Vaginal continence surgery needs a good indication with a best possible urethro-vesical suspension. This is not an operation for beginners.
Subject(s)
Urinary Incontinence/surgery , Abdomen/surgery , Female , Follow-Up Studies , Humans , Middle Aged , Postoperative Complications/etiology , Recurrence , Urodynamics , Vagina/surgerySubject(s)
Intrauterine Devices/adverse effects , Pelvic Inflammatory Disease/etiology , Female , Humans , RiskABSTRACT
The ion chelators picolinic acid, quinaldic acid, 1,10-phenanthroline, and 8-hydroxyquinoline, but not ethylenediaminetetraacetate, ethyleneglycol-bis-(beta-aminoethyl ether)-N,N-tetraacetate, or dipicolinic acid, rapidly but transiently arrest growth of Escherichia coli K-12. Cells adapt and become resistant to growth inhibition by these agents, a process which requires protein synthesis. Mn2+, at low concentrations, decreases the time required for resumption of growth. Proteins synthesized during the lag are quantitatively and qualitatively different from those synthesized during normal growth. Inhibition of growth can explained by an effect on RNA polymerase, a known metalloenzyme.
