Treatment with PB125® Increases Femoral Long Bone Strength in 15-Month-Old Female Hartley Guinea Pigs.

Nuclear factor-erythroid 2-related factor-2 (Nrf2) is a transcription factor that serves as a master regulator of anti-inflammatory agents, phase I xenobiotic, and phase II antioxidant enzymes, all of which provide a cytoprotective role during disease progression. We hypothesized that oral administration of a purported phytochemical Nrf2-activator, PB125®, would increase long bone strength in aging Hartley guinea pigs, a model prone to musculoskeletal decline. Male (N = 56) and female (N = 56) guinea pigs were randomly assigned to receive daily oral treatment with either PB125® or vehicle control. Animals were treated for a consecutive 3-months (starting at 2-months of age) or 10-months (starting at 5-months of age) and sacrificed at 5-months or 15-months of age, respectively. Outcome measures included: (1) ANY-maze™ enclosure monitoring, (2) quantitative microcomputed tomography, and (3) biomechanical testing. Treatment with PB125® for 10 months resulted in increased long bone strength as determined by ultimate bending stress in female Hartley guinea pigs. In control groups, increasing age resulted in significant effects on geometric and structural properties of long bones, as well as a trending increase in ultimate bending stress. Furthermore, both age and sex had a significant effect on the geometric properties of both cortical and trabecular bone. Collectively, this work suggests that this nutraceutical may serve as a promising target and preventive measure in managing the decline in bone mass and quality documented in aging patients. Auxiliary to this main goal, this work also capitalized upon 5 and 15-month-old male and female animals in the control group to characterize age- and sex-specific differences on long bone geometric, structural, and material properties in this animal model.

Observing a dynamical skeleton of turbulence in Taylor-Couette flow experiments.

Recent work shows that recurrent solutions of the equations governing fluid flow play an important role in structuring the dynamics of turbulence. Here, an improved version of an earlier method (Krygier et al. 2021 J. Fluid. Mech. 923, A7 and Crowley et al. 2022 Proc. Natl Acad. Sci. USA 119, e2120665119) is used for detecting and analyzing intervals of time when turbulence 'shadows' (spatially and temporally mimics) recurrent solutions in both numerical simulations and laboratory experiments. We find that all the recurrent solutions shadowed in numerics are also shadowed in experiment, and the corresponding statistics of shadowing agree. Our results set the stage for experimentally grounded dynamical descriptions of turbulence in a variety of wall-bounded shear flows, enabling applications to forecasting and control. This article is part of the theme issue 'Taylor-Couette and related flows on the centennial of Taylor's seminal Philosophical Transactions paper (part 1)'.

Neurosurgical Study Design: Past and Future.

Clinical trials are performed to determine the safety, efficacy, or effectiveness of a medical or surgical intervention. A clinical trial is, by definition, prospective in nature with a uniform treatment of a defined patient cohort. The outcomes assessment should also be uniform. Often a control group is included. At present, the number of neurosurgical clinical trials is increasing, and the study designs have become more sophisticated. Historically, the standard of neurosurgical care has evolved from the findings from many case series and retrospective comparative studies. However, in the present report, we have focused exclusively on prospective clinical trials. An urgent need exists to understand how clinical trials have been performed in the past and how they can be improved to advance our neurosurgical practice. In the present review, we have discussed the barriers, successes, and failures regarding prospective clinical trials in neurosurgery with an outlook to the future.

Platelet SHARPIN regulates platelet adhesion and inflammatory responses through associations with αIIbß3 and LUBAC.

Platelets form hemostatic plugs to prevent blood loss, and they modulate immunity and inflammation in several ways. A key event during hemostasis is activation of integrin αIIbß3 through direct interactions of the ß3 cytoplasmic tail with talin and kindlin-3. Recently, we showed that human platelets express the adapter molecule Shank-associated RH domain interacting protein (SHARPIN), which can associate directly with the αIIb cytoplasmic tail and separately promote NF-κB pathway activation as a member of the Met-1 linear ubiquitination activation complex (LUBAC). Here we investigated the role of SHARPIN in platelets after crossing Sharpin flox/flox (fl/fl) mice with PF4-Cre or GPIbα-Cre mice to selectively delete SHARPIN in platelets. SHARPIN-null platelets adhered to immobilized fibrinogen through αIIbß3, and they spread more extensively than littermate control platelets in a manner dependent on feedback stimulation by platelet adenosine diphosphate (ADP) (P < .01). SHARPIN-null platelets showed increased colocalization of αIIbß3 with talin as assessed by super-resolution microscopy and increased binding of soluble fibrinogen in response to submaximal concentrations of ADP (P < .05). However, mice with SHARPIN-null platelets showed compromised thrombus growth on collagen and slightly prolonged tail bleeding times. Platelets lacking SHARPIN also showed reduced NF-κB activation and linear ubiquitination of protein substrates upon challenge with classic platelet agonists. Furthermore, the loss of platelet SHARPIN resulted in significant reduction in inflammation in murine models of colitis and peritonitis (P < .01). Thus, SHARPIN plays differential and context-dependent roles in platelets to regulate important inflammatory and integrin adhesive functions of these anucleate cells.

The pharmacist in same day emergency care: a service evaluation of pharmacy services on the ambulatory assessment unit at the John Radcliffe Hospital.

Same day emergency care (SDEC) is an increasingly important part of urgent care delivery in secondary care. This service evaluation examined the role of the pharmacy service on a busy SDEC unit over a 3 week period. A total of 634 patients were seen on the unit and 513 pharmacy interventions were made. Patients were taking a mean number of 6.7 medicines and the average age was 59.3. The most common medication type pharmacists intervened in were anticoagulants. To meet the demands of SDEC service, the pharmacy team is crucial for maintaining medication safety and ensuring patient flow through hospital pathways.

Optogenetics-based localization of talin to the plasma membrane promotes activation of ß3 integrins.

Interaction of talin with the cytoplasmic tails of integrin ß triggers integrin activation, leading to an increase of integrin affinity/avidity for extracellular ligands. In talin KO mice, loss of talin interaction with platelet integrin αIIbß3 causes a severe hemostatic defect, and loss of talin interaction with endothelial cell integrin αVß3 affects angiogenesis. In normal cells, talin is autoinhibited and localized in the cytoplasm. Here, we used an optogenetic platform to assess whether recruitment of full-length talin to the plasma membrane was sufficient to induce integrin activation. A dimerization module (Arabidopsis cryptochrome 2 fused to the N terminus of talin; N-terminal of cryptochrome-interacting basic helix-loop-helix domain ended with a CAAX box protein [C: cysteine; A: aliphatic amino acid; X: any C-terminal amino acid]) responsive to 450 nm (blue) light was inserted into Chinese hamster ovary cells and endothelial cells also expressing αIIbß3 or αVß3, respectively. Thus, exposure of the cells to blue light caused a rapid and reversible recruitment of Arabidopsis cryptochrome 2-talin to the N-terminal of cryptochrome-interacting basic helix-loop-helix domain ended with a CAAX box protein [C: cysteine; A: aliphatic amino acid; X: any C-terminal amino acid]-decorated plasma membrane. This resulted in ß3 integrin activation in both cell types, as well as increasing migration of the endothelial cells. However, membrane recruitment of talin was not sufficient for integrin activation, as membrane-associated Ras-related protein 1 (Rap1)-GTP was also required. Moreover, talin mutations that interfered with its direct binding to Rap1 abrogated ß3 integrin activation. Altogether, these results define a role for the plasma membrane recruitment of talin in ß3 integrin activation, and they suggest a nuanced sequence of events thereafter involving Rap1-GTP.

Left pulmonary arterial branch interruption with concurrent coarctation of the right pulmonary artery in a dog.

This case report describes a rare disorder of a left pulmonary artery interruption with concurrent coarctation of the right pulmonary artery in a dog. A 5-year-old, male neutered, mixed-breed dog presented for evaluation of an asymptomatic heart murmur. Echocardiography and computed tomography revealed complete interruption of the proximal left pulmonary artery and coarctation of the right pulmonary artery. Collateral circulation to the left lung field was provided by a dilated bronchoesophageal artery with evidence of left pulmonary hypoplasia. Pulmonary artery interruption and coarctation is rarely reported in the veterinary literature.

Genetic Instruction of Megakaryocytes and Platelets Derived from Human Induced Pluripotent Stem Cells for Studies of Integrin Regulation.

Platelets are small, anucleate cells that play oversized roles in hemostasis, immunity, and inflammation. An important mediator of platelet function is integrin αIIbß3, which is required for fibrinogen-dependent platelet aggregation during hemostasis. This platelet response is dependent on conformational changes in the integrin induced by "inside-out" biochemical signals that are triggered by platelet agonists. In turn, fibrinogen binding to αIIbß3 initiates "outside-in" biochemical and mechanical signals that regulate the platelet cytoskeleton and help to promote full platelet aggregation and secretory responses. Without a nucleus, there is a limited range of experimental manipulations that are possible with human platelets to study the molecular basis of integrin signaling in these primary cells. Consequently, many studies of αIIbß3 function use genetic approaches that rely on heterologous expression systems or platelets from gene-targeted mice, sometimes with uncertain applicability to human platelets. This chapter will detail a method for genetic manipulation of megakaryocytes and platelets derived from human induced pluripotent stem cells for molecular studies of αIIbß3 signaling and for modeling of human platelet functions potentially relevant to hemostasis, immunity, and inflammation.

Underlying Immune Disorder May Predispose Some Transthyretin Amyloidosis Subjects to Inotersen-Mediated Thrombocytopenia.

Inotersen, a 2'-O-methoxyethyl (2'-MOE) phosphorothioate antisense oligonucleotide, reduced disease progression and improved quality of life in patients with hereditary transthyretin amyloidosis with polyneuropathy (hATTR-PN) in the NEURO-TTR and NEURO-TTR open-label extension (OLE) trials. However, 300 mg/week inotersen treatment was associated with platelet count reductions in several patients. Mean platelet counts in patients in the NEURO-TTR-inotersen group remained ≥140 × 109/L in 50% and ≥100 × 109/L in 80% of the subjects. However, grade 4 thrombocytopenia (<25 × 109/L) occurred in three subjects in NEURO-TTR trial, and one of these suffered a fatal intracranial hemorrhage. The two others were treated successfully with corticosteroids and discontinuation of inotersen. Investigations in a subset of subjects in NEURO-TTR (n = 17 placebo; n = 31 inotersen) and OLE (n = 33) trials ruled out direct myelotoxicity, consumptive coagulopathy, and heparin-induced thrombocytopenia. Antiplatelet immunoglobulin G (IgG) antibodies were detected at baseline in 5 of 31 (16%) inotersen-treated subjects in NEURO-TTR, 4 of whom eventually developed grade 1 or 2 thrombocytopenia while on the drug. In addition, 24 subjects in the same group developed treatment-emergent antiplatelet IgG antibodies, of which 2 developed grade 2, and 3 developed grade 4 thrombocytopenia. Antiplatelet IgG antibodies in two of the three grade 4 thrombocytopenia subjects targeted GPIIb/IIIa. Plasma cytokines previously implicated in immune dysregulation, such as interleukin (IL)-23 and a proliferation-inducing ligand (APRIL) were often above the normal range at baseline. Collectively, these findings suggest an underlying immunologic dysregulation predisposing some individuals to immune-mediated thrombocytopenia during inotersen treatment.

Treatment of horizontal silage bunker runoff using biochar amended vegetative filter strips.

Horizontal silage bunkers produce leachate that contains contaminants that can be detrimental to the environment if released untreated. Vegetated filter strips are used to treat silage bunker runoff to prevent contamination of surface waters via infiltration, however increased infiltration poses risks to groundwater, particularly for nitrate (NO3-). Vegetated filter strip plots with a sandy loam soil, half of which are amended with biochar, were investigated to assess the treatment of silage bunker runoff over 20 application events. The subsurface effluent biological oxygen demand (BOD5), chemical oxygen demand (COD), and total phosphorus (TP) were reduced on average by 40%, 46%, and 75%, respectively, and there was no statistical difference between treatments. The total nitrogen (TN) was reduced by 49 and 64% for control and biochar plots, respectively, which was significantly different between treatments. Biochar significantly reduced nitrate nitrogen (NO3--N) leaching by 40% compared to the control, however, the NO3--N concentration in leachate was still high ranging from 0.19 to 191.04 mg NO3--N L-1 and 0.18-108.89 mg NO3--N L-1 for control and biochar plots, respectively. A mass balance suggests the primary mechanism for a decrease in TN and NO3--N leaching from biochar amended plots was greater retention of NO3--N and organic N (ORG-N) within the soil/biochar matrix. The development of oxygenated functional groups and/or formation of organomineral layer on the biochar surface likely enhanced N retention.

The effect of activated platelet-rich plasma (PRP) on tricalcium hydroxyapatite phosphate healing in experimental, partial defects of long bone shafts in animal models.

The purpose of the study was to assess the effect of autologous activated platelet-rich plasma on healing of a bone substitute - tricalcium phosphate hydroxyapatite in experimental long bone defects using an animal model. The experiment involved an animal model of femoral defect. 24 Termond white rabbits were used in the study. We evaluated the effect of autologous platelet-rich plasma on tricalcium phosphate using classical radiology, micro-CT studies, strength tests and histological evaluation. Radiological and histological assessment did not show a beneficial effect of PRP together with a bone substitute in comparison to filling the defects only with bone replacement material. The only benefit of adding platelet-rich plasma to a bone substitute was shown in microCT imaging. Autologous, activated platelet-rich plasma combined with hydroxyapatite tricalcium phosphate has a positive effect on the remodeling of the newly formed bone tissue, increasing its density.

Breast abscess in a competitive high school swimmer: a case of toxic shock syndrome.

Toxic shock syndrome (TSS) is a severe, acute, toxin-mediated disease process characterized by fever, diffuse erythroderma, hypotension, multisystem organ dysfunction and desquamation of skin. TSS represents the most severe form of disease caused by exotoxin-producing strains of Streptococcus pyogenes and Staphylococcus aureus. Menstrual and non-menstrual TSS become significant causes of morbidity and mortality. As a result of public awareness and various campaigns, the majority of TSS cases tend to be non-menstrual related. The clinical course is fulminant and can result in abrupt decompensation and death. Management within the emergency department (ED) includes removal of the potential foreign body, fluid resuscitation, appropriate antibiotics, potential vasopressor support and possible surgical intervention. We present the unique case of a 16-year-old female competitive swimmer who presented to the ED twice, demonstrating the fulminant course of TSS. She initially presented with non-specific symptoms with an unremarkable evaluation. She returned within hours of discharge with an abrupt onset of diffuse macular erythroderma, placed on norepinephrine and was diagnosed with TSS secondary to a breast abscess.

Nitrate sorption to biochar following chemical oxidation.

Biochar amendments can reduce nitrate (NO3) leaching in agricultural soil. It has been hypothesized that functional groups on the biochar surface from oxidation can increase NO3 sorption. This study evaluates the effect of chemical oxidation of biochar on NO3 sorption characteristics. Eight biochars, made from wood and corn cobs, underwent sodium hypochlorite (NaClO) and hydrogen peroxide (H2O2) oxidation and then assessed for NO3 sorption capacity using batch isotherm methods. The unoxidized and oxidized biochar produced at low temperatures (400 °C) had no significant NO3 sorption. Oxidized biochars produced at higher temperatures (600 °C and 700 °C) had calculated maximum NO3 sorption capacities (Smax) ranging from 0.50 to 3.97 mg NO3-N g-1. Biochar oxidations with 50 mmol NaClO g-1 (N50) in combination with an acid wash (AW) had the largest estimated sorption capacities of 3.68, 3.97, and 1.46 mg NO3-N g-1 for CTN50,AW, BW3N50,AW, and CC3N50,AW, respectively. Sorption capacity of wood-based biochars was higher than corn cob biochars due to increased oxidation as measured by total acid group content (TAGC). Wood biochar Smax values were correlated with ΔTAGC (R2 = 0.86), with a slope of 1.2 µmol NO3-N µmol TAGC-1 suggesting that cationic bridging of NO3 to oxidized sites is the primary mechanism for NO3 sorption.

Autologous activated platelet-rich plasma (PRP) in bone tissue healing - does it work? Assessment of PRP effect on bone defect healing in animal models.

INTRODUCTION: Platelet-rich plasma (PRP) preparations can be used in bone tissue healing but there are numerous doubts among clinical orthopedists about effectiveness of this method. MATERIALS AND METHODS: The studies were carried out in 12 rabbits of white termond breed. In operating room we operationally generated cylindrical, unicortical defects of the diameter of 4 mm in the middle of the shafts of both femurs. The defects in the left bones were left without filling and served as controls, and 0.7 ml of the ready-to-use PRP was administered to the defects in the right bones (experimental group). We evaluated the usefulness of the diagnostic methods applied: biomechanical tests, micro-CT tests, densitometry, typical radiology, macroscopic measurements, histopathological examinations. RESULTS: The macroscopic measurements showed a statistically significant increase in the dimension in the area of the right defect filled with PRP in relation to the control group. In experimented group, the assessment of the X-ray images showed the formation of a callus cuff around the defects. Densitometric examinations showed no statistically significant differences between defects in the experimental and control group. The analysis of the micro-CT examina- tions showed an increase in the total volume of the tissue examined (Vb) and the low density tissue fraction (Vb2) in the experimental group. The biomechanical examinations revealed signi- ficant decrease in the maximum breaking force (F max) necessary to break the bone in the experi- mental group in relation to the control group. CONCLUSIONS: Platelet-rich plasma (PRP) stimulates bone formation in the area of bone defects and may accelerate bone regeneration.

uPAR isoform 2 forms a dimer and induces severe kidney disease in mice.

Soluble urokinase plasminogen activator receptor (suPAR) is an immune-derived circulating signaling molecule that has been implicated in chronic kidney disease, such as focal segmental glomerulosclerosis (FSGS). Typically, native uPAR (isoform 1) translates to a 3-domain protein capable of binding and activating integrins, yet the function of additional isoforms generated by alternative splicing is unknown. Here, we characterized mouse uPAR isoform 2 (msuPAR2), encoding domain I and nearly one-half of domain II, as a dimer in solution, as revealed by 3D electron microscopy structural analysis. In vivo, msuPAR2 transgenic mice exhibited signs of severe renal disease characteristic of FSGS with proteinuria, loss of kidney function, and glomerulosclerosis. Sequencing of the glomerular RNAs from msuPAR2-Tg mice revealed a differentially expressed gene signature that includes upregulation of the suPAR receptor Itgb3, encoding ß3 integrin. Crossing msuPAR2-transgenic mice with 3 different integrin ß3 deficiency models rescued msuPAR2-mediated kidney function. Further analyses indicated a central role for ß3 integrin and c-Src in msuPAR2 signaling and in human FSGS kidney biopsies. Administration of Src inhibitors reduced proteinuria in msuPAR2-transgenic mice. In conclusion, msuPAR2 may play an important role in certain forms of scarring kidney disease.