Search

Background: Targeted protein degradation relies on inducing proximity between an E3 ubiquitin ligase and a target protein, and subsequent proteasomal degradation of the latter. Biophysical methods allow the measurement of the ternary complex formation by recombinant target and E3 ligase proteins in the presence of molecular glues and bifunctional degraders. The development of new chemotypes of degraders mediating ternary complex formation of unknown dimensions and geometries requires the use of different biophysical approaches., Methods: The TR-FRET and AlphaLISA platforms have been applied to study molecular glues and bifunctional degraders. The performance of the label-based proximity assays was compared with the BLI method, which is a label-free, sensor-based approach., Results: We present and compare two commonly used assays to monitor proximity induction, AlphaLISA and TR-FRET. The LinkScape system consisting of the CaptorBait peptide and the CaptorPrey protein is a novel method of protein labeling compatible with TR-FRET assay., Conclusions: The TR-FRET and AlphaLISA proximity assays enable detection of ternary complexes formed between an E3 Ligase, a target protein and a small molecule degrader. Experiments with different chemotypes of GSPT1 degraders showed that ALphaLISA was more susceptible to chemotype-dependent interference than TR-FRET assay., General Significance: The discovery and optimization of small-molecule inducers of ternary complexes is greatly accelerated by using biophysical assays. The LinkScape-based TR-FRET assay is an alternative to antibody-based proximity assays due to the CaptorPrey's subnanomolar affinity to the CaptorBait-tagged protein target, and the 10-fold lower molecular weight of the CaptorPrey protein compared to the antibody., Competing Interests: Declaration of Competing Interest K. Przytulski, P. Glaza, K. Brach, M. Sagan, G. Statkiewicz, J. Klajn, M. J. Walczak are employees and stockholders of Captor Therapeutics., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Purpose : The aim of this study was to evaluate the frequency of single nucleotide polymorphisms (SNP) of estrogen receptor genes ( ESR1 : rs12154178, rs1884054 and ESR2 : rs1268656, rs7159462) and to assess their possible influence on the clinical phenotype of primary open angle glaucoma (POAG). Methods : The study included 235 patients with POAG (143 patients with normal-tension glaucoma [NTG] and 92 patients with high-tension glaucoma [HTG]), and 165 healthy controls. DNA was isolated from peripheral blood, and SNP genotyping was performed using the Real-Time Polymerase Chain Reaction method to analyze the frequency of selected polymorphic variants of estrogen receptor genes. The clinical phenotype (best-corrected visual acuity, intraocular pressure [IOP], mean deviation [MD], cup to disc ratio, disc hemorrhages, notches, peripapillary atrophy, cold extremities) of participants were examined for association with the polymorphisms. Results : A similar frequency of the polymorphic variants of the studied genes was observed in patients with NTG, HTG and control group. Initial intraocular pressure was the lowest in NTG patients with GG variant of rs1268656 ( p = 0.044). The lowest maximal IOP in HTG patients was observed in CC variant of rs12154178 ( p = 0.039). Patients with HTG and CC variant of ESR1 polymorphism rs1884054 had the best visual acuity ( p = 0.009), similar tendency was also observed in the NTG group. This polymorphic variant of ESR1 gene in HTG was also related to earlier damage in visual field assessed according to MD values and higher percentage of notches. For rs12154178, homozygotic variant CC was related to earlier glaucoma damage according to MD in HTG patients ( p = 0.006). For polymorphism rs12154178, disc hemorrhages were found only for those with the AC variant. Cold extremities were most frequent in NTG patients with TT variant of rs1268656 comparing to other variants ( p = 0.021). Notches on optic disc were less frequent in patients with CC variant of rs12154178 of ERS-1 gene ( p = 0.022). Conclusions : The studied polymorphic variants of ESR1 and ESR2 genes may have an influence on the clinical phenotype of patients with POAG.

The membrane activity of some saponins, such as digitonin or alpha-hederin, is usually attributed to their interaction with membrane cholesterol (Chol). This contrasts with our recent publication showing that Chol, contrary to sphingomyelin (SM), can delay the cytotoxicity of the saponin ginsenoside Rh2, challenging the usual view that most saponins mediate their membrane effects through interaction with Chol. The aim of the present study was to elucidate the respective importance of Chol and SM as compared to phosphatidylcholine (PC) species in the membrane-related effects of Rh2. On simple lipid monolayers, Rh2 interacted more favorably with eggSM and DOPC than with Chol and eggPC. Using Large Unilamellar Vesicles (LUVs) of binary or ternary lipid compositions, we showed that Rh2 increased vesicle size, decreased membrane fluidity and induced membrane permeability with the following preference: eggSM:eggPC > eggSM:eggPC:Chol > eggPC:Chol. On Giant Unilamellar Vesicles (GUVs), we evidenced that Rh2 generated positive curvatures in eggSM-containing GUVs and small buds followed by intra-luminal vesicles in eggSM-free GUVs. Altogether, our data indicate that eggSM promotes and accelerates membrane-related effects induced by Rh2 whereas Chol slows down and depresses these effects. This study reconsiders the theory that Chol is the only responsible for the activity of saponins.

Under pathological conditions, cholesterol oxidation products (oxysterols) appear in enhanced concentration in blood and cerebrospinal fluid, which leads to cytotoxic effect, especially in central nervous system. However, the mode of action of oxysterols on the membrane level has not been fully resolved. In this paper we have investigated the interaction between 7α- hydroxycholesterol, 7α-OH (one of the most abundant oxysterol in human body) and two major membrane lipids: sphingomyelin, SM (basic component of lipid rafts and nerve membrane) and 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine, POPC (main phospholipid of mammalian cell membranes). 7α-OH/SM mixtures may mimic pathologically changed lipid raft (ordered phase, L O ) while the SM/POPC system can model its surrounding (liquid-disordered phase, L α ). For our study, the Langmuir monolayer technique (based on registration of the surface pressure/area, π/A isotherms), complemented with surface visualization technique (Brewster angle microscopy, BAM) and theoretical calculations, have been employed. The observed affinity of 7α-OH to SM, which appears to be stronger than in cholesterol/SM system, indicates that cholesterol might be partially replaced in lipid rafts by its oxidized derivative. Its incorporation significantly increases rigidity of the system in relation to normal (cholesterol-containing) raft, which can disturb its proper functioning. On the other hand, the poor effect of this oxysterol on the raft's environment was observed., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Secretion of renin juxtaglomerular cells into bloodstream initiates activation of an enzymatic-hormonal cascade known as the RAAS (renin - angiotensin - aldosterone system). As a result, blood pressure is increased by the means several interrelated mechanisms. Mechanism of Zjednoczoaction of this system has been known for decades, but a few previously unknown components were recently added, such as ACE-2 and Ang(1-7), and their role often seems to be opposite to that of the conventional components. Local tissue systems also have important biological functions. They operate largely independently of the systemic activity, and their activity is observed primarily in the kidney, heart, in blood vessels, adrenal gland and nervous system. Angiotensin-2 (Ang-2), the main RAAS effector, has a wide scope of action, and thus abnormalities in its functioning have many consequences. Excessive activation is accompanied by chronic inflammation, as Ang-2 stimulates inflammatory mediators. As a result, degenerative processes and atherosclerosis are initiated. RAAS imbalance is associated with the most common diseases of civilization, such as cardio-vascular diseases, diabetes, kidney diseases, preeclampsia, osteoporosis and even neurodegenerative diseases. Many of these pathological processes are attributed to the excessive activation of tissue RA system. Therapeutic strategies based on inhibition of the RAAS are commonly used mainly in the treatment of hypertension and other cardiovascular disorders. The benefits of this class of drugs is primarily a decrease in blood pressure, but also the suppression of inflammatory processes and other pathological phenomena resulting from excessive activation of the RAAS. For that reason, some consider to use RAAS inhibitors in other diseases, e.g. Parkinson's disease. Further studies give hope for the improvement of RAAS inhibitor therapy and the development of new therapeutic strategies.

Osteochondroma is a benign bone tumor composed of a bony protrusion with an overlying cartilage cap. Osteochondromas arise in the scapula in 3% to 5% of cases, making them the most common benign bone tumor in this location. Scapular osteochondromas cause various pathologies of the shoulder girdle, including snapping scapula syndrome, chest wall deformity, shoulder impingement, and bursa formation. Bronchogenic cyst is an exceedingly rare finding in the periscapular area. It is a congenital cystic mass lined by tracheobronchial structures and respiratory epithelium. To our knowledge, there are no reports of a contiguous osteochondroma and bronchogenic cyst. A 12-month-old boy presented with an incidentally noted mass on the spine of the scapula, which drained scant, clear fluid through an adjacent pinprick-sized hole. Imaging revealed an exostosis with an adjacent cystic mass. The mass and cyst were excised en bloc, and histopathologic examination confirmed the diagnosis of osteochondroma with contiguous bronchogenic cyst. In this case, we present the report of a novel dual lesion.

Objectives: First Nations communities in Manitoba were significantly affected by the pandemic H1N1 influenza virus (pH1N1) in 2009. Our objective was to conduct an epidemiologic investigation of a pH1N1 outbreak in one remote First Nations community (population 3,300) in northern Manitoba to inform a timely public health response and provide recommendations for preventing future outbreaks., Methods: Chart reviews were conducted at the nursing station for patients meeting the influenza-like illness (ILI) case definition during the study period (April 20 to June 11, 2009). Descriptive analyses examined age, gender, clinical presentation, management, outcomes and risk factors. Comparisons were made for hospitalized versus non-hospitalized cases and laboratory-confirmed versus possible cases using Pearson's chi-square test for gender and symptoms and using a t-test for age., Results: There were 180 ILI cases, including 23 laboratory-confirmed cases of pH1N1. Forty percent of children < 1 year old in the community and 9.4% of pregnant women presented to the nursing station with ILI. Most ILI cases were managed through the community nursing station, although 18.3% of cases (n = 33) were medically evacuated and 16.1% (n = 29) were hospitalized. There were no differences between hospitalized versus non-hospitalized or laboratory-confirmed versus possible cases. Risk factors identified in a subset of cases included exposure to an individual with ILI prior to illness onset, overcrowding and inadequate access to household water., Conclusions: Early arrival and rapid transmission of pH1N1 rendered usual non-pharmacological control measures largely ineffective. Recommendations for prevention of future outbreaks include an effective communications strategy and daily surveillance for disease detection and monitoring. Key determinants of health should be addressed in remote First Nations communities to prevent disease and protect the health of these populations.

The purpose of this study was to assess the needs of older case coordinated clients receiving community health services, by examining changes in cognitive status, physical and mental health status, social support, risk for institutionalization, and service use over a 6-month period from initial intake into home care. Significant predictors of professional and supportive home care, as well as emergency room use and hospital days, were also examined. Standardized interviews were conducted with 234 clients at the time of referral; follow-up interviews were conducted with 179 of these clients after 6 months of case coordination. Results reveal that physical and mental health improved, while cognitive status remained stable. Although social interaction and instrumental support decreased, subjective support remained stable. Risk of institutionalization decreased. Occupational therapy, nursing, and homemaking were the most frequently used services. The best predictor of professional and supportive home care was a risk of institutionalization score. The needs of this older adult population changed even within the relatively short span of 6 months. Frequent review of needs in some form may be warranted in order to maintain effective service plans.

This article describes the cost of developing and implementing an integrated care pathway (ICP) for Congestive Heart Failure. Costs were tracked prospectively and allocated to either development or implementation. The development took 1,980 hours of staff time at a salary cost of $67,800. The ICP implementation took 2,083 hours of personnel time at a salary cost of $66,900. Even though an available pathway was adapted for local use, a substantial amount of time and money was required. This article may be useful to others who are interested in monitoring costs of pathways.

From a pool of Medicago truncatula mutants--obtained by gamma-irradiation or ethyl methanesulfonate mutagenesis--impaired in symbiosis with the N-fixing bacterium Sinorhizobium meliloti, new mutants are described and genetically analysed, and for already reported mutants, complementary data are given on their phenotypic and genetic analysis. Phenotypic data relate to nodulation and mycorrhizal phenotypes. Among the five new mutants, three were classified as [Nod+ Fix- Myc+] and the mutations were ascribed to two loci, Mtsym20 (TRV43, TRV54) and Mtsym21 (TRV49). For the two other new mutants, one was classified as [Nod-/+ Myc+] with a mutation ascribed to gene Mtsym15 (TRV48), and the other as [Nod- Myc-/+] with a mutation ascribed to gene Mtsym16 (TRV58). Genetic analysis of three previously described mutants has shown that [Nod-/+ Myc+] TR74 mutant can be ascribed to gene Mtsym14, and that [Nod-/+ Myc-/+] TR89 and TRV9 mutants are ascribed to gene Mtsym2 (dmi2). Using a detailed analysis of mycorrhizal phenotype, we have observed a delayed typical arbuscular mycorrhizal formation on some mutants that present thick lens-shaped appressoria. This phenotype was called [Myc-/+] and mutants TR25, TR26, TR89, TRV9, P1 and Y6 were reclassified as [Myc-/+]. Mutant P1 was reclassified as [Nod-/+] because of a late nodulation observed on roots of this mutant.

The Regina Risk Indicator Tool (RRIT) is used to identify how at risk clients are for requiring admission to long-term care. This study examined the potential use of this tool by case managers of older community dwelling clients receiving home care. The RRIT exhibited moderate to good inter-rater reliability and good predictive validity as clients of varying degrees of risk differed in amount of case management and services used. Healthcare managers may wish to consider using the tool to support decision-making related to case management and home care service.

This study describes the development of a tool used to track the who, what, when, and how much of case coordination for elderly, community-based clients. The Case Coordination Activity Tracking Form measures phase of coordination (assessment, plan development, plan implementation, monitoring reassessient, and discharge), type of activity (in-person, telephone, documentation, research, travel, case conference), with whom contact took place (client, family, client/family together, supervisor/colleague, service provider, physician, program access committee), and special circumstances (e.g., infection control). Over a 6-month span, we used the tool to study case coordination of services for elderly, community-based clients. The average amount of coordination time was 5.15 hours (n = 234), with 71% of the clients requiring between 2 and 6hours of coordination, and 26% of the clients accounting for 49% of the total coordination time. Frequency and average amount of coordination declined greatly after the first month. Not all clients required coordination each month, and monitoring was the most common activity in later months. The tool was found to be reliable and easy to use, and of benefit to decision makers, managers, and case coordinators in measuring, analyzing, and describing case coordination. This tool could be readily used with other client groups.

In this study, we developed time-and-need-based community case management guidelines by tracking case management activity for 234 elderly clients over their first 6 months of care. Clients were interviewed at the time services began and again 6 months later to determine physical, emotional, and cognitive status. We found that the Regina Risk Indicator Tool (RRIT), a brief measure of risk for requiring institutional care, differentiated clients needing varying amounts of case management. Using these results, an expert panel devised guidelines for ranges of case management time for clients at different levels of need: low risk, some risk, and high risk. Quality measures, response times for initial contact and assessment, and service review parameters were also incorporated into the guidelines. Tools for workload measurement, quality review, client satisfaction, and service provider communication were developed. Focus group analysis of the guidelines with various stakeholders and a pilot implementation guided further revision. Future directions are outlined in the article, including the need to focus on outcomes of the guidelines and study of guideline implementation processes.

Hemiballismus is characterized by the abrupt onset of violent proximal flinging movements, affecting the limbs, neck and trunk on one side of the body. It is caused by the lesion in the region of the contralateral subthalamic nucleus of the Luys. Usually it is a self-limiting disease, lasting 6-8 weeks. A 49-year-old man has been admitted to the hospital after flinging movements of his right arm and the right side of the trunk occurred. A few days earlier he had undergone general anesthesia prior to a dental procedure. There was trouble in waking the patient afterwards. The movements lasted a few days. MRI of the brain revealed ischemic lesions areas in T2-weighted images localized in the region of globus pallidus bilaterally. EEG was abnormal, and showed slowed background activity with slow waves in left temporal lobe. He was treated with haloperidol, clonazepam and vasoactive medications. In spite of administered treatment, hemiballic movements reappeared occasionally. Due to increased frequency of the movements the patient was hospitalized again two years later. The second MRI revealed changes described earlier and a new ischaemic focus in left parietal lobe. Continuation of treatment with haloperidol was administered.

This study documents the development of a new tool to measure the quality of community case management for elderly clients. Information obtained from literature review, client focus groups, and pilot surveys with clients served as the basis for the development of a Case Management Quality Questionnaire (CMQQ). This measure, along with satisfaction, health status, and demographic questions, was administered to 174 home care clients and 78 family members of long term care residents for evaluation of case management services that clients had received in the last 2 to 6 months. Principal components analysis with oblique (Oblimin) rotation identified three subscales of the CMQQ: (1) accessibility, (2) efficiency, and (3) assessment/coordination skill. Study findings suggest that the CMQQ is reliable, valid, and of value in understanding satisfaction with case coordination. Based on client feedback and analysis of the results, some future modifications to the measure are worthy of study. Others interested in measuring and improving case management may similarly benefit from the use of the CMQQ.

Prevention of stroke and dementia seems to be a great and rising challenge nowadays. Increasing problem of cardiovascular diseases considers, apart from individuals' and state expenses, also important social effort. The aspect of prevention seems to be the most optimistic accent in that area. Based on the results of multiple randomized trials we can influence in both pharmacological and non-pharmacological way the major risk factors of stroke. Effective antihypertensive treatment, cholesterol lowering therapy, smoking cessation and antithrombotic treatment in subjects with atrial fibrillation result in noticeable reduction of stroke events in both primary and secondary prevention. The number of researches which aim the dementia prevention is still sparse, which makes the results of the recently presented PROGRESS trial even more interesting and valuable. It showed that it was possible to prevent dementia and cognitive impairment with therapy based on the long acting ACE inhibitor. An increasing number of data supporting the beneficial role of statins in the stroke and dementia prevention it is noteworthy. The multifactorial intervention both in prevention of stroke and dementia seems to be the most effective.

Several mutants defective in the nodulation process during rhizobial or endomycorrhizal endosymbiosis of pea have been identified previously. We have integrated the map positions of two such nodulation mutations, sym9 and sym10, into the molecular map of pea by applying molecular-marker techniques combined with bulked segregant analysis (BSA). Lines P2 and P54 were found to carry alleles of sym9, line P56 carried an allele of sym10. F2 populations were derived from crosses of P2, P54 and P56, to JI281 and JI15, two of the parental lines that have been used previously to generate a molecular map of pea. sym9 was located on linkage group IV by AFLP-BSA analysis and subsequently mapped by RFLP in both F2 populations, P2 x JI281 and P54 x JI281. RFLP-BSA analysis was applied to assign sym10 to linkage group I. The RFLP marker locus, chs2, co-segregates with sym10 in the F2 population of P56 x JI15.

In the present study the relationship between calcitonin and kappa-opioid system was analysed. The possibility that salmon calcitonin (S-CT) exerts analgesic activity in mice as well as the influence of salmon calcitonin on the analgesic effect of kappa-opioid agonist, U-50, 488H were examined. The writhing test in mice was used as analgesic test. S-CT given alone intraperitoneally at a high dose produced antinociceptive effect. S-CT given at a subanalgesic dose increased the antinociceptive effect of kappa-opioid agonist, U-50, 488H. The obtained results suggest that the increase in the effectiveness of kappa-opioid receptor agonist may be one of the mechanisms involved in the analgesia induced by salmon calcitonin.

The pea mutant line P55 is defective in root nodule formation, and this phenotype is controlled by a single recessive gene. Complementation analysis revealed that the mutation in P55 is allelic to sym19, which has previously been mapped to linkage group I. Detailed mapping revealed that the sym19 and ENOD40 loci are separated by 2.7 cM. We identified four recombination events, demonstrating that the nodulation defect caused by mutation of the sym19 locus cannot be due to mutation of ENOD40. RT-PCR experiments showed that P55 expresses ENOD12A, but there was little or no increase in the level of its transcript in response to Nod factor or infection with Rhizobium. To investigate this expression pattern further, transgenic peas carrying a pENOD12A-GUS reporter construct were made. One transgenic line was crossed with line P55, to generate F2 progeny homozygous for sym19 and carrying pENOD12A-GUS. In both WT and sym19 mutant lines, ENOD12A-GUS expression was induced at sites of lateral root emergence in uninoculated plants. In Nod+ plants pENOD12A-GUS was induced in response to Rhizobium leguminosarumn bv. viciae, but no such induction was seen in the Nod- (sym19) mutants.

Within a set of 13 induced symbiotic pea (Pisum sativum L.) mutants, two distinct mutated genes sym25 and sym26, selected for ineffective nodulation have been counteracted in their effect by Rhizobium strains 254 and 94 respectively, thereby establishing an N 2 -fixing symbiosis. This suggests that mutagenesis can be a helpful tool in creating specificity systems such as those described in centres of origin and diversification of legume species.

In addition to central effects, which are the basis of their use in anesthesiology, alpha 2-adrenergic agonists have direct peripheral cardiovascular effects. Dexmedetomidine (DM) has been found to depress cardiac function in dogs, even after autonomic denervation. The present experiments evaluated the effects of DM on coronary flow, myocardial oxygen extraction, and cardiac function in intact, open chest dogs under enflurane anesthesia. Heart rate (HR), mean arterial pressure (MAP), left ventricular end-diastolic pressure (LVEDP), the first derivative of systolic left ventricular pressure (dP/dtmax), and flow in the left anterior descending coronary artery (CBF) were measured and continuously recorded. Cardiac output (CO), plasma catecholamines (CA), hemoglobin and oxygen saturation in arterial, mixed venous, and coronary sinus blood were measured at intervals. Cardiac index (CI), systemic vascular resistance index (SVRI), regional coronary vascular resistance (CVR), and oxygen concentration differences across the systemic [C(a-v)O2], and coronary [C(a-cs)O2] circulations were calculated. DM doses of 0.25, 0.5, 1.0, 2.0, and 4.0 micrograms/kg were given IV at 20-minute intervals. Measurements and samples were taken at peak drug effects and just prior to the next dose. The alpha 2-antagonist atipamezole, 0.5 mg/kg, was given after the last dose of DM. DM caused immediate dose-dependent increases in SVRI, CVR, LVEDP, C(a-v)O2, and C(a-cs)O2, and decreases in HR, and CI, with recovery between doses. DP/dtmax declined after the first two doses and stabilized thereafter, as plasma CA fell to minimal levels. Atipamezole completely reversed all changes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenomatoid hyperplasia of the mucous salivary glands is an uncommon benign tumor of the oral cavity. Two cases of palatal involvement are presented. Since clinical differentiation between benign and malignant growth of the palate cannot be established, microscopic examination is necessary. The palate is the most common location of this lesion, and as such it is useful for the periodontist to be aware of this tumor.

Cysts of the incisive papilla are uncommon variants of the nasopalatine duct cysts. They may be symptomatic or asymptomatic and usually are not seen radiographically. This case report presents a flat, darkly pigmented asymptomatic lesion which required differentiation from a possible malignant melanoma. Due to its location in the anterior palate, this lesion is of interest and clinical significance to the periodontist.