A cell-based high-throughput screen identifies drugs that cause bleeding disorders by off-targeting the vitamin K cycle.

Drug-induced bleeding disorders contribute to substantial morbidity and mortality. Antithrombotic agents that cause unintended bleeding of obvious cause are relatively easy to control. However, the mechanisms of most drug-induced bleeding disorders are poorly understood, which makes intervention more difficult. As most bleeding disorders are associated with the dysfunction of coagulation factors, we adapted our recently established cell-based assay to identify drugs that affect the biosynthesis of active vitamin K-dependent (VKD) coagulation factors with possible adverse off-target results. The National Institutes of Health (NIH) Clinical Collection (NCC) library containing 727 drugs was screened, and 9 drugs were identified, including the most commonly prescribed anticoagulant warfarin. Bleeding complications associated with most of these drugs have been clinically reported, but the pathogenic mechanisms remain unclear. Further characterization of the 9 top-hit drugs on the inhibition of VKD carboxylation suggests that warfarin, lansoprazole, and nitazoxanide mainly target vitamin K epoxide reductase (VKOR), whereas idebenone, clofazimine, and AM404 mainly target vitamin K reductase (VKR) in vitamin K redox cycling. The other 3 drugs mainly affect vitamin K availability within the cells. The molecular mechanisms underlying the inactivation of VKOR and VKR by these drugs are clarified. Results from both cell-based and animal model studies suggest that the anticoagulation effect of drugs that target VKOR, but not VKR, can be rescued by the administration of vitamin K. These findings provide insights into the prevention and management of drug-induced bleeding disorders. The established cell-based, high-throughput screening approach provides a powerful tool for identifying new vitamin K antagonists that function as anticoagulants.

Therapeutic exploitation of IPSE, a urogenital parasite-derived host modulatory protein, for chemotherapy-induced hemorrhagic cystitis.

Chemotherapy-induced hemorrhagic cystitis (CHC) can be difficult to manage. Prior work suggests that IL-4 alleviates ifosfamide-induced hemorrhagic cystitis (IHC), but systemically administered IL-4 causes significant side effects. We hypothesized that the Schistosoma hematobium homolog of IL-4-inducing principle from Schistosoma mansoni eggs (H-IPSE), would reduce IHC and associated bladder pathology. IPSE binds IgE on basophils and mast cells, triggering IL-4 secretion by these cells. IPSE is also an "infiltrin," translocating into the host nucleus to modulate gene transcription. Mice were administered IL-4, H-IPSE protein or its nuclear localization sequence (NLS) mutant, with or without neutralizing anti-IL-4 antibody, or 2-mercaptoethane sulfonate sodium (MESNA; a drug used to prevent IHC), followed by ifosfamide. Bladder tissue damage and hemoglobin content were measured. Spontaneous and evoked pain, urinary frequency, and bladdergene expression analysis were assessed. Pain behaviors were interpreted in a blinded fashion. One dose of H-IPSE was superior to MESNA and IL-4 in suppressing bladder hemorrhage in an IL-4-dependent fashion and comparable with MESNA in dampening ifosfamide-triggered pain behaviors in an NLS-dependent manner. H-IPSE also accelerated urothelial repair following IHC. Our work represents the first therapeutic exploitation of a uropathogen-derived host modulatory molecule in a clinically relevant bladder disease model and indicates that IPSE may be an alternative to MESNA for mitigating CHC.-Mbanefo, E. C., Le, L., Pennington, L. F., Odegaard, J. I., Jardetzky, T. S., Alouffi, A., Falcone, F. H., Hsieh, M. H. Therapeutic exploitation of IPSE, a urogenital parasite-derived host modulatory protein, for chemotherapy-induced hemorrhagic cystitis.

Efficacy of Cidofovir in Treatment of BK Virus-Induced Hemorrhagic Cystitis in Allogeneic Hematopoietic Cell Transplant Recipients.

BK virus-associated hemorrhagic cystitis (BK-HC) is a common complication after allogeneic hematopoietic stem cell transplantation (allo-HCT), with incidences up to 70%. Cidofovir is an antiviral agent with growing evidence as a therapeutic intervention. To assess the safety profile and efficacy of intravenous and intravesical cidofovir in allo-HCT patients with BK-HC, a retrospective study was undertaken of the allo-HCT cohort who received cidofovir for symptomatic BK-HC (hematuria with BK viruria or viremia) from January 2010 until March 2017 in a single transplant center in Ontario, Canada. The primary outcome measure was a reduction in BK-HC severity (graded from 1 to 4); secondary outcomes included overall survival, BK virus titers, and the onset of acute kidney injury. Twelve allo-HCT patients received cidofovir for BK-HC, with pretreatment clinical severity of 3 (50%) or 4 (50%). Cidofovir was administered via intravenous (33%), intravesical (58%), or both modalities (8%). After a median cumulative dose of 10 mg/kg (range, 1 to 37), mean BK-HC grade decreased significantly by 1.8 (3.5 precidofovir, 1.7 postcidofovir, P < .01). Sixty-six percent of patients had at least partial response to cidofovir, with similar response rates between intravenous (66%) and intravesical (62%) administration. Sixty-seven percent of patients died, and 33% of patients experienced renal toxicity, including 2 patients receiving intravesical therapy. In this retrospective series, there was a significant reduction in BK-HC severity after cidofovir administration; most patients achieved at least partial response after cidofovir administration. Even with intravesical instillation, acute kidney injury remains a potential complication of cidofovir. Although cidofovir may be an efficacious therapy for BK-HC, albeit with potential demonstrated toxicities, further prospective trials are needed.

[Gastrointestinal bleeding]. / Hemorragia gastrointestinal.

In the Digestive Disease Week in 2015 there have been some new contributions in the field of gastrointestinal bleeding that deserve to be highlighted. Treatment of celecoxib with a proton pump inhibitor is safer than treatment with nonselective NSAID and a proton pump inhibitor in high risk gastrointestinal and cardiovascular patients who mostly also take acetylsalicylic acid. Several studies confirm the need to restart the antiplatelet or anticoagulant therapy at an early stage after a gastrointestinal hemorrhage. The need for urgent endoscopy before 6-12 h after the onset of upper gastrointestinal bleeding episode may be beneficial in patients with hemodynamic instability and high risk for comorbidity. It is confirmed that in Western but not in Japanese populations, gastrointestinal bleeding episodes admitted to hospital during weekend days are associated with a worse prognosis associated with delays in the clinical management of the events. The strategy of a restrictive policy on blood transfusions during an upper GI bleeding event has been challenged. Several studies have shown the benefit of identifying the bleeding vessel in non varicose underlying gastric lesions by Doppler ultrasound which allows direct endoscopic therapy in the patient with upper GI bleeding. Finally, it has been reported that lower gastrointestinal bleeding diverticula band ligation or hemoclipping are both safe and have the same long-term outcomes.

Relationship between hemorrhagic stroke hospitalization and exposure to fine particulate air pollution in Taipei, Taiwan.

This study was undertaken to determine whether there was a correlation between fine particle (PM2.5) levels and hospital admissions for hemorrhagic stroke (HS) in Taipei, Taiwan. Hospital admissions for HS and ambient air pollution data for Taipei were obtained for the period 2006-2010. The relative risk of hospital admissions was estimated using a case-crossover approach, controlling for weather variables, day of the week, seasonality, and long-term time trends. For the single-pollutant model (without adjustment for other pollutants), increased HS admissions were significantly associated with PM2.5 levels both on warm days (>23°C) and cool days (<23°C), with an interquartile range rise associated with a 12% (95% CI = 7-18%) and 4% (95% CI = 0-8%) elevation in admissions for HS, respectively. In the two-pollutant models, PM2.5 remained significantly high after inclusion of SO2 or O3 on both warm and cool days. This study provides evidence that higher levels of PM2.5 increase the risk of hospital admissions for HS.

Hemorrhagic diathesis in avian species following intramuscular administration of polysulfated glycosaminoglycan.

Polysulfated glycosaminoglycans (PSGAGs) have been used for decades in a variety of species for the management of osteoarthritic pain. However, reports on the use of PSGAGs in avian species are scarce. In domestic cats and dogs, PSGAG injections have caused prolongation of clotting times but are considered to be an efficacious drug with a wide margin of safety. This publication documents four cases of fatal coagulopathies in different avian species (one coraciiforme, two raptors, and one psittacine) following the intramuscular administration of PSGAG. All affected birds received varying dosages and dosing intervals of PSGAG. Three of the four birds experienced fatal hemorrhage into the pectoral muscle, while the fourth bled continuously from the injection site. Only one bird had chronic, severe pre-existing disease; the remainder were being managed for osteoarthritis. This report highlights the importance of species-specific dosing of PSGAG and warrants further investigation into the etiopathogenesis of this process.

Diagnoses unveiled by early bronchoscopy in children with leukemia and pulmonary infiltrates.

Pulmonary complications in children with leukemia often display nonspecific clinical and radiologic manifestations that lead to a delay in diagnosis. The role of fiberoptic bronchoscopy (FOB) and the proper time for its performance are controversial. The aim of our study was to evaluate the frequency and nature of specific diagnoses revealed by FOB. Children with leukemia submitted to FOB because of suspicion of pulmonary involvement (mainly pneumonia) were retrospectively analyzed. A total of 33 FOB procedures performed in 31 patients (20 males) with an average age of 9.4 years (range, 3.5 to 15 y) were evaluated. Microorganisms isolated from 21 (63.6%) bronchoalveolar lavage samples were mainly fungi including Candida in 13 cases (39.4%) and Aspergillus in 3 cases (9.1%). Isolation rate in 10 procedures performed within the first 3 days was 90%. Tracheobronchitis was present in > 50% of patients, pulmonary hemorrhage was seen in 7 (21.0%) patients, and leukemic infiltration was demonstrated in 2 patients (6.1%), among other conditions visualized by FOB. Complications of FOB were minimal and transient. Our study suggests that FOB is a useful and safe procedure in patients with leukemia and pulmonary infiltrates. The earlier the FOB was performed, the higher the isolation rate of causative agents. In addition, this procedure allowed the identification of noninfectious airway comorbidities. Further studies in regard to this issue are warranted.

Hemorrhagic complications after systemic thrombolysis in acute stroke patients with abnormal baseline coagulation.

BACKGROUND AND PURPOSE: As patients with abnormal baseline coagulation were excluded from the large randomized trials, the safety of intravenous thrombolysis after ischaemic stroke in this patient population remains controversial. METHODS: We assessed the risk of symptomatic intracerebral hemorrhage (SICH) after systemic thrombolysis in patients with elevated baseline international normalized ratios (INRs) (≥1.3) or activated partial thromboplastin times (aPTT) (>37 s) using a prospectively recorded database from 2006 to 2010. An intracerebral hemorrhage leading to a deterioration of ≥4 points on the National Institutes of Health Stroke scale (NIHSS) was classified as symptomatic. RESULTS: Amongst 688 patients (mean age, 72 years; median NIHSS, 11, median onset-to-treatment time, 135 min), 36 patients (5%) had an abnormal baseline coagulation. Twenty-nine of these patients had taken oral anticoagulants leading to elevated baseline INRs (median INR: 1.5; IQR 1.4-1.9), whereas seven patients had elevated aPTTs because of heparin therapy (n = 2), a coagulation disorder (n = 2), or for unknown reasons (n = 3). The rate of SICH did not differ significantly between patients with abnormal and normal baseline coagulation (4.4% vs. 0%; P = 0.6). Moreover, the in-hospital mortality was not significantly different between both treatment groups (8.3% in patients with abnormal baseline coagulation vs. 8.7% in patients with normal baseline coagulation, P = 1.0). CONCLUSIONS: The risk of SICH following intravenous thrombolysis after ischaemic stroke does not appear to be increased in patients with abnormal baseline coagulation.

[Activated protein C, a protein at the crossroads between coagulation and inflammation]. / La protéine C activée: une protéine à l'interface de l'inflammation et de la coagulation.

Sepsis is defined as a systemic response to infection, characterized by an intense inflammatory response linked to coagulation activation and fibrinolysis inhibition, two processes which are intimately associated. In a field where mortality remains very high, administration of activated protein C, a physiological coagulation inhibitor with cytoprotective properties, has demonstrated its effectiveness and was able to reduce mortality. Protein C belongs to a system that involves plasma proteins and endothelial cell receptors. In addition to well documented effects on coagulation and fibrinolysis, activated protein C exhibits anti-inflammatory, anti-apoptotic but also anti-histone activities. Indeed, a recent study focusing on the cytoprotective effects of activated protein C showed that extracellular histones are released during severe sepsis and may participate in the pathophysiology of severe sepsis. These histones appear to be new targets of activated protein C.

Aplastic anemia as a cause of death in a patient with glioblastoma multiforme treated with temozolomide.

BACKGROUND: Standard treatment of glioblastoma multiforme consists of postoperative radiochemotherapy with temozolomide, followed by a 6-month chemotherapy. Serious hematologic complications are rarely reported. CASE REPORT AND RESULTS: The authors present the case of a 61-year-old female patient with glioblastoma multiforme treated with external-beam radiation therapy and concomitant temozolomide. After completion of treatment, the patient developed symptoms of serious aplastic anemia that eventually led to death due to prolonged neutro- and thrombocytopenia followed by infectious complications. CONCLUSION: Lethal complications following temozolomide are, per se, extremely rare, however, a total of four other cases of aplastic anemia have been reported in the literature so far.

Predictors of immediate bleeding during endoscopic submucosal dissection in gastric lesions.

BACKGROUND: Although there have been several reports regarding complications, especially immediate bleeding, of endoscopic mucosal resection for a gastric neoplasm, little is known about the predictors of complications of endoscopic submucosal dissection (ESD). Thus, this study was performed to evaluate the predictive factors for immediate bleeding during ESD procedures. METHODS: We analyzed 167 patients with 167 gastric lesions from June 2006 to June 2007. Patient-related variables (age, gender, history of aspirin or anti-platelet agents, triple therapy for H. pylori, and use of a proton pump inhibitor), endoscopic variables (lesion size, location, type, and mucosal ulceration), procedure-related variables (procedure time and volume of submucosal injection), and the pathology diagnosis were evaluated as potential risk factors. RESULTS: The mean age of the patients was 62 years. The mean size of the lesions was 15 mm. The overall en bloc resection rate was 98.2%. Immediate bleeding occurred in 20 out of 167 patients. Delayed bleeding was seen in only three patients within 24 h after the procedure. Older age and the location of the lesions (antrum) were associated with a lower frequency of bleeding (p = 0.006 and p = 0.007, respectively). On multivariate analysis, an older age (OR 0.931, 95% CI 0.88-0.98) and the location of the lesion (antrum; OR 0.254, 95% CI 0.09-0.69) were significant predictive factors for a successful ESD without bleeding. CONCLUSION: The results of this study demonstrated that age and lesion location were related to the ESD procedure outcome.

Coagulopathy induced by saw palmetto: a case report.

Saw palmetto is the most popular herbal supplement used to treat symptoms of benign prostatic hyperplasia (BPH). The safety and efficacy of saw palmetto has been established in the literature. While the majority of studies document the efficacy and safety of saw palmetto, some studies document the adverse side effects, including increased risk of bleeding. There are no reports in the literature about increased prothombin time (PT), partial thromboplastin time (PTT) or international normalized ratio (INR) while using saw palmetto. We present a case of hematuria and coagulopathy in a patient who was using saw palmetto.

"Falling down": a retroperitoneal catastropheC.

Hemoperitoneum due to ruptured retroperitoneal varices is an extremely rare condition and a poor prognostic sign with a catastrophic and life-threatening situation. Early recognition affords appropriate management and urgent surgical intervention in order to favor the survival rate. In this case report we accurately describe the complex clinical course of a 56-year old woman with retroperitoneal varices, who few months earlier had a chest trauma with multiple left lower rib fractures and 10 years earlier she underwent to ovarian hyperstimulation for an ovulation induction. She was taken to the emergency room for a fainting episode with signs of a clear hemodinamic shock without a present history of trauma. The intricacy of this case was mostly due to the choice of the correct management, where the damage control resuscitation turned out to have an important role.

Sonoclot analysis in elderly compared with younger patients undergoing coronary surgery.

BACKGROUND: The Sonoclot analyzer is a point-of-care method for assessment of the clotting mechanism in whole blood. The results are available within 20 min. The aim of the present study was to investigate whether repeated Sonoclot analyses could identify peri-operative differences in hemostatic function between elderly and younger patients undergoing coronary artery bypass grafting (CABG). In addition, we investigated whether Sonoclot analyses could identify disturbances in hemostatic function leading to post-operative bleeding. METHODS: Twenty-five elderly and 25 younger patients undergoing CABG were included. Blood samples for Sonoclot analyses were drawn pre-operatively, during surgery, and during the first 20 post-operative hours. The Sonoclot variables sonACT, clot rate, time-to-peak, amplitude of the peak, and R3 were analyzed, and the results were compared between the two groups. Post-operative blood loss volumes were recorded and correlated to the Sonoclot variables. The Sonoclot variables were also correlated to previously reported results on various hemostatic variables measured in the same patient population. RESULTS: There was a significant difference in sonACT between the two groups (P=0.018). There were no differences between the groups in any of the other Sonoclot variables. There were no significant correlations between any of the Sonoclot variables and post-operative bleeding, or between the Sonoclot variables and other hemostatic variables. CONCLUSIONS: The difference in sonACT between the two groups indicates a reduced hemostatic function in the elderly patients. However, repeated Sonoclot analyses were not able to identify more specific disturbances in hemostatic function, and did not predict increased post-operative bleeding.

Toxicity and gut associated lymphoid tissue translocation of polymyxin B orally administered by alginate/chitosan microparticles in rats.

Fluorescent calcium alginate/chitosan microparticles, prepared using a spray-drying technique followed by crosslinking reactions with calcium ions and chitosan, were assayed in-vivo for polymyxin B (PMB) oral toxicity, uptake by Peyer's patches and PMB oral absorption. A single PMB dose (300 mg kg(-1)), loaded in microparticles or dissolved in water, was administered to rats by oral gavage under fasted and fed conditions. By monitoring incidence of mortality, animal behaviour, clinical signs and abnormality in several organs, PMB in water solution was found lethal at a dose lower than the LD50 (790 mg kg(-1)) in the fasted state and toxic for the gastrointestinal tract in the fed state. However, no signs of acute toxicity at the level of the gastrointestinal tract were observed when animals were administered PMB loaded in microparticles under fasted and fed conditions. A lower PMB dose (125 mg kg(-1)), loaded in microparticles or dissolved in water, was given to rats in a fed state to determine PMB levels in Peyer's patches, urine and serum as well as to detect the loaded microparticles inside Peyer's patches for three days after dosing. Abnormalities were observed at gut level only when PMB was dosed in a water solution. Detectable antibiotic levels in Peyer's patches and urine as well as more constant PMB serum concentrations were provided by dosing PMB loaded in microparticles. Therefore, the use of alginate/chitosan microparticles to target the lymphatic system could improve safety when administering PMB orally.

S-allyl cysteine ameliorates cyclophosphamide-induced downregulation of urothelial uroplakin IIIa with a concomitant effect on expression and release of CCL11and TNF-α in mice.

BACKGROUND: The aim of this study was to evaluate the modulatory effect of S-allyl cysteine against cyclophosphamide-induced changes in uroplakin IIIa, CCL11 and TNF-α. METHODS: Mice were treated with cyclophosphamide (200mg/kg×7 d, ip). S-allyl cysteine (150mg/kg×7d, ip), and comparator compound mesna (40mg/kg×7d, ip) were administered 1h before and 4h after each cyclophosphamide dose. The urinary bladder was analysed for mRNA and protein changes in uroplakin IIIa (UPIIIa), CCL11 and TNF-α and histopathological findings. RESULTS: Cyclophosphamide caused hemorrhagic cystitis formation and downregulation of UPIIIa. These changes were accompanied by upregulation of CCL11 and TNF-α. S-allyl cysteine attenuated these changes including protection at histological level. Mesna which was used as a comparator drug also showed protection. However, relatively S-allyl cysteine showed a stronger protective effect than mesna. CONCLUSION: These findings highlight a correlation between downregulaion of UPIIIa and enhanced production of inflammatory biomarkers and protective effects of S-allyl cysteine which has been reported to be a potent uroprotective agent. The present study strengthens its role which could be clinically exploited in chemotherapy regimen.

Modifications of low-molecular weight heparin use in a French university hospital after implementation of new guidelines.

BACKGROUND: Previous studies performed in 1999 and 2000 showed frequent misuse of low-molecular weight heparins (LMWHs) in France, leading to an increasing risk of bleeding. In 2002, the French Medicine Agency (Agence Française de Sécurité Sanitaire des Produits de Santé) released guidelines on the prescription and monitoring of LMWH. This study assesses LMWH use before and after the implementation of these guidelines. METHODS: We performed a 'pre and post' survey comparing data collected in 1999 (before guidelines) and in 2003 (1 year after guidelines) at a French university hospital. The same design and the same medical wards were used for both data-collection periods, and the data collected included patient characteristics, LWMH prescription information (daily dose, indication) and adverse drug reactions (ADRs). The main outcome was the frequency of prescription of LMWHs for curative treatment in patients with severe renal insufficiency, defined as having a creatinine clearance of

Propylthiouracil-related hemorrhagic diathesis: a case report and review of the literature.

We present an unusual case of a drug-related hemorrhagic diathesis. One month prior to admission, the patient was diagnosed at another medical center as having Graves' disease and propylthiouracil therapy (PTU) was initiated. Since clinical recovery was not achieved, the PTU was quickly increased to an unconventional daily dose of 1,000 mg. The patient was referred to our hospital because of spontaneous epistaxis, multiple ecchymoses and prolonged prothrombin time (PT) and activated partial thromboplastin time (aPTT), which developed soon after the increase in PTU dose. Drug-related hemorrhagic diathesis was considered, after other possible causes had been eliminated. To the best of our knowledge, this is the first reported case of spontaneous hemorrhage due to PTU use.

Bleeding Complications After Endoscopic Lung Volume Reduction Coil Treatment: A Retrospective Observational Study. / Complicaciones hemorrágicas tras el tratamiento endoscópico de reducción del volumen pulmonar con espiral: estudio observacional retrospectivo.

INTRODUCTION: Endoscopic lung volume reduction coil (LVRC) treatment is an option for selected patients with severe emphysema. This study was conducted to determine the incidence of bleeding complications after LVRC treatment, to identify risk factors and to discuss treatment options in case of hemoptysis which does not resolve spontaneously. METHODS: Retrospective observational study conducted in the Department of Respiratory Medicine at the University Medical Center Hamburg-Eppendorf in all subjects in whom LVRC treatment was performed between April 1, 2012 and September 30, 2015. RESULTS: During the study period, 101 LVRC procedures were performed in 62 subjects. Early post-procedural bleeding was encountered in 65.3% of cases. Hemoptysis was significantly more likely to occur in patients receiving acetylsalicylic acid (P=.005). Hemoptysis resolved spontaneously in 98.5% of cases. In the one case (1.5%) with persistent hemoptysis, bronchial artery embolization was successful in terminating bleeding. Hospital stay was significantly prolonged in subjects with hemoptysis (P=.01). No significant differences were found between subjects with or without hemoptysis in terms of chronic obstructive pulmonary disease exacerbations within four weeks after LVRC treatment (P=.18). Late bleeding complications were observed in 3 subjects (3.0%). In 2 of these cases, bronchial artery embolization was performed and bleeding was successfully terminated. CONCLUSIONS: Self-limiting low volume bleeding is a common finding in the first days after LVRC treatment. However, persistent bleeding may occur in the early post-procedural phase and late after LVRC treatment. In these cases, bronchial artery embolization was a feasible and successful approach to terminating bleeding.