Cells were then plated in a 96-well plate with 2 × 105 cells per well (106 cells/ml), allowed to incubate for 60–90 min at 37° (+5% CO2), and re-stimulated with soluble anti-CD3ε (2·5 μg/ml) antibody. Following the indicated stimulation time, culture medium was collected and spun down to remove any residual cells. The concentration of IL-4, IL-6, IL-10, IL-17A, IFN-γ and tumour necrosis factor-α (TNF-α) in the cell-free culture medium was analysed using

custom bead arrays from Millipore, and quantified on a Luminex 100 system (Austin, TX) with the Luminex XY plate handling platform. Assays were performed according to the manufacturer’s protocols. Duplicate wells were assayed for each sample, and data are representative of the average median

value for each sample. Analysis was performed PLX-4720 manufacturer using is 2.3 software (Luminex). A vehicle consisting of 90% emulsion solution (PBS + 0·9% Tween-20 + 0·9% BSA) and 10% ethanol was used. For delivery of compounds, E2 or G-1 was dissolved in ethanol Lumacaftor chemical structure and added at appropriate concentrations such that 100 μl per animal per injection was used. The compound was added to each injection as part of the 10% ethanol found in the vehicle, so it was diluted such that < 10 μl per animal per injection was required. Injections were administered in the afternoon, and to limit stress from the long series of injections inherent in this study, animals were sedated using isofluorane before injection. Compound was delivered Vitamin B12 subcutaneously on the dorsum adjacent to the hind limb, and the side of the injection was alternated every 2 days. To investigate the direct effects of G-1 on CD4+ T cells, we chose to use purified cultures of naive T cells activated by polyclonal stimulation with anti-CD3ε and anti-CD28 antibody. This eliminated secondary effects caused by the activity of G-1 on APCs within the culture. Furthermore, primary cells from male mice were used

throughout the study to avoid potential confounding effects of either; (i) varying estrogen levels in female mice, or (ii) the inflammatory effects of ovariectomy. We have also determined that CD4+ CD44loCD62Lhi naive T-cell and CD4+ Foxp3+ T-reg cell populations express the G-1 target GPER (R. L. Brunsing and E. R. Prossnitz, manuscript in preparation). Given that G-1 can protect mice from EAE38,39 and the importance of the the Th17 lineage to this model,3 we began by determining the effects of G-1 on naive T-cell differentiation under Th17-polarizing conditions (TGF-β/IL-6 ± IL-23). Hence, naive T cells from 7- to 11-week-old male C57BL/6 mice were collected by FACS and stimulated for 4 days ex vivo, supplemented with combinations of TGF-β, IL-6 and IL-23. Following 4 days of stimulation, cells were analysed for expression of IFN-γ, IL-17A and IL-10 by intracellular cytokine staining. Expression of IL-10 was present exclusively in cultures treated with IL-6 (Fig.

However, men with abnormal fasting plasma glucose (FPG) and waist circumference (WC) had larger prostates than normal groups. The logistic regression analysis showed that the FPG level and WC had a significantly positive correlation with prostate volume (odds ratios, FPG, 1.441 [95% CI: 1.303–1.643] and WC, 2.305 [95% CI: 1.470–3.614]). Unlike other studies they studied a younger age group (in their fourth to fifth decades) and concluded that obesity and DM could be more important factors than MS in prostate volume enlargement

in relatively young adults. Although there have been no regional or nationwide cohort studies observing the association of MS and LUTS in Korea, like previous studies conducted in other countries, several investigators have independently reported that MS patients had lower LUTS-related QoL, higher symptom scores or lower maximal selleck flow rate. Kim

et al.36 studied LUTS subfactors and flow rates in male patients with and without MS. Interestingly, MS patients had worse symptom scores, low QoL, lower maximal flow rate, higher residual urine volume, and larger prostate volume than non-MS patients. Like the BACH survey, which was a population-based cohort reporting the association of LUTS and MS, Kim et al. observed that MS patients also showed more significant symptom correlation in voiding symptom domains.36 Kim et al.37 also investigated those correlations in women. Patients with MS scored poorly for all the voiding factors of MS learn more patients compared to the control (non-MS) group. In both their male and female studies, they also extracted their Enzalutamide data according to presence of DM. Results showed that insulin resistance was the important factor for developing LUTS. IPSS, QoL score, and maximal flow rate in males; international prostate symptom score (IPSS), maximal flow rate, and postvoid residual urine volume were more correlated in female patients with DM for more than 5 years.36,37 Recently, Hong et al.38 evaluated 922 (538 men, 384 women) adults undergoing a health check. The overall prevalence of MS was 15.5%, 110 men (20.4%)

and 33 women (8.6%), showing a significant gender difference. They evaluated LUTS by two symptom questionnaires: IPSS and Overactive Bladder Questionnaire Short Form (OABq-SF) in both men and women. Results showed some inconsistency between men and women. There were no significant differences in scores on the IPSS or OABq-SF with respect to the presence or absence of MS in men. However, in women, except for intermittency of IPSS (question 3) and severe urge incontinence (question 6) of OABq-SF, the remaining IPSS and OABq-SF items were significantly worse in the MS group. After regression analysis, in both genders, the IPSS total score was significantly correlated with age. Also, HDL cholesterol in males and triglyceride (TG) in women was significantly correlated with IPSS total score.38 Unfortunately we still do not have enough data about association of MS and LUTS in women.

Consistent with this, splenocytes from Camp−/− mice that had been administered with a T-cell-dependent antigen were also found to have increased IL-4 mRNA expression and increased numbers of CD4+IL-4+ T cells as compared with those from similarly treated WT mice. The connections between mCRAMP and IL-4 open up intriguing possibilities for the role of cathelicidins in adaptive

immunity. In the mice given TNP-OVA/alum and in the in vitro T cells, the responses indicate that mCRAMP suppresses both the development of a Th2 response and the Th2-mediated class switching to IgG1 through IL-4 17, 19. In contrast, selleck chemicals llc the results from isolated B cells stimulated with CD40L/IL-4 indicated that mCRAMP Small molecule library chemical structure promoted IgG1 production by increasing transcription 17. Kurosaka et al. 13 showed that mCRAMP administered as an adjuvant with OVA increases IL-4 and OVA-specific IgG1 in splenocytes, although the response

was not Th2-mediated. Similarly, An et al. 16 found that LL-37 acts as an effective adjuvant in a vaccine against tumor cells, while Davidson et al. 8 found a bias towards a Th1 response in human DCs. The conflicting reports may reflect methodological differences, such as using Camp−/− mice versus injecting cathelicidin into WT mice, or the timing and nature of other stimuli applied. Nonetheless, these studies indicate that mCRAMP likely mediates its effects on adaptive immunity through many other factors in addition to IL-4. The work by Kin et al. 17 shows that mCRAMP alters B- and T-cell responses, highlighting the novel role of mCRAMP in the T-cell-dependent activation of B cells, and thus providing evidence that mCRAMP and other cathelicidins have a greater role in the adaptive immune response than previously appreciated. However, many questions still remain, particularly whether

mCRAMP acts directly on components of the adaptive immune system or if intermediates are involved. It is also of interest to determine whether the changes seen by Kin et al. 17 in response to T-cell-dependent antigen are due to mCRAMP altering Isotretinoin both T and B cells or whether only one cell type is directly involved. The use of conditional knockouts or adoptive transfer to examine when Camp is absent from either T or B cells will help resolve these issues. Similar models could also be used to clarify the functions of APCs in shaping the Camp−/− effects on lymphocytes. Determining the specific cells and pathways altered by mCRAMP will provide further insight into the roles of cathelicidins in bridging innate and adaptive immunity. Funding from the Canadian Institutes for Health Research for the authors own peptide research is gratefully acknowledged. REWH holds a Canada Research Chair. Conflict of interest: The authors have declared no financial or commercial conflict of interest. See accompanying article: http://dx.doi.org/10.1002/eji.

The results suggest a complex mechanism of platelet aggregation ABT-888 chemical structure and P-selectin expression in sepsis, where generation of platelet-activating stimuli is required first, before platelet aggregation and adhesion in capillaries occur. The ability of ascorbate to reduce platelet aggregation and P-selectin expression could be an important mechanism by which ascorbate inhibits capillary plugging in sepsis. “
“Please

cite this paper as: Vachharajani, Wang, Mishra, El Gazzar, Yoza and McCall (2010). Curcumin Modulates Leukocyte and Platelet Adhesion in Murine Sepsis. Microcirculation17(6), 407–416. Objective:  Circulating cell–endothelial cell interaction in sepsis is a rate-determining factor in organ dysfunction, and interventions targeting this process have a potential therapeutic value. In this project, we examined whether curcumin, an active ingredient of turmeric and an anti-inflammatory agent, could disrupt interactions between circulating blood cells and endothelium and improve survival in a murine model of sepsis. Methods:  Mice were subjected to cecal ligation and puncture (CLP) to induce sepsis vs. sham surgery. We studied leukocyte and platelet adhesion in cerebral microcirculation using intravital fluorescent video microscopy technique,

blood–brain barrier (BBB) dysfunction using Evans Blue (EB) leakage method, P-selectin expression using dual radiolabeling technique, and survival in mice subjected Z-IETD-FMK supplier to Sham, CLP, and CLP with curcumin pre-treatment (CLP + curcumin). Results:  Curcumin significantly attenuated leukocyte and platelet adhesion in cerebral microcirculation, EB leakage in the brain tissue, and improved survival in mice with CLP. P-selectin expression in mice

with CLP + curcumin was significantly attenuated compared with CLP in various microcirculatory beds, including brain. Reduction in platelet adhesion was predominantly via modulation of endothelium by curcumin. Conclusion:  Curcumin pre-treatment modulates leukocyte and platelet adhesion and BBB dysfunction in mice with CLP via P-selectin expression and improves survival in mice with CLP. “
“We developed a model for direct assessment of BMC sequestration in the postischemic murine myocardium Tenoxicam after direct antegrade intracoronary injection. Modified syngeneic heterotopic heart transplantation was used as a basic model for global myocardial I/R injury in a total of n = 29 animals. IVM was employed to analyze the right ventricular subepicardial coronary microcirculation and for tracking fluorescently labeled BMCs. IVM allowed monitoring all segments of the coronary microcirculation including feeding arterioles, nutritive capillaries, and postcapillary venules. WI and generalized atherosclerosis induced profound reperfusion failure, particularly in nutritive myocardial capillaries.

RNA analysis indicated that mhuA and mhuB are each transcribed from individual Fur-regulated promoters. find more Moreover, RNA analysis of an mhuB deletion mutant and a promoter reporter assay coupled with β-galactosidase suggested that MhuB could function as an activator for mhuA transcription. Finally, the role of MhuA as the heme/hemoglobin receptor was confirmed by construction of an mhuA deletion mutant and its complemented strain followed by growth assay. Iron is an element integral to the growth of almost all bacteria.

However, the availability of iron for bacteria is limited because it is usually present as insoluble ferric hydroxide polymers in an aerobic environment or bound to iron-binding proteins such as transferrin and lactoferrin in mammalian hosts (1). Therefore, most bacteria have

evolved the ability to acquire iron under iron-restricted conditions. Numerous bacteria produce and secrete siderophores (low-molecular weight iron-binding chelators) which can remove ferric iron from iron-binding proteins. In Gram-negative bacteria, ferric ion complexed with siderophore (ferrisiderophore) is transported into cells via a TonB-dependent specific uptake system, consisting of an outer membrane receptor protein and an ABC transporter (2). In addition, certain bacteria acquire heme as a nutritional iron source by a TonB-dependent system, similar selleck inhibitor to those for ferrisiderophores, which includes the binding of heme or heme-containing proteins such as hemoglobin to the cell surface receptor, followed by transport of the intact heme moiety into the cell (3). Siderophores are unable to remove the iron from heme. Moreover, when intracellular iron concentrations are high, expression of those systems studied to oxyclozanide date is negatively regulated at the transcriptional level by a global iron-binding repressor protein called Fur (ferric uptake regulation) with ferrous ion as a corepressor, (4, 5). V. mimicus was first described as a group of biochemically atypical strains of V. cholerae

(6) but they share some pathogenic factors such as enterotoxins and hemolysins (7). V. mimicus, like other pathogenic Vibrio species, inhabits environmental water, including river, brackish, and sea water, and causes diarrhea through eating fish and shellfish contaminated with the bacterium (8). The present authors have previously reported that V. mimicus secretes the siderophore aerobactin in response to iron restriction (9), and that the iucABCD genes engage in aerobactin biosynthesis. They have also reported that the ferriaerobactin complex is incorporated into the cytosol via the 77-kDa IROMP, IutA, and the ABC transporter, MatCDB (10). V. mimicus also expresses 80-kDa IROMP under iron-restricted conditions (9). Hence, V. mimicus is expected to use at least one other iron source besides ferriaerobactin. Although many Vibrio species, including V.

Using SCID-Hu mouse models, Dick and colleagues showed that only 1/250 000 AML CD34+CD38– cells were capable of establishing leukaemic haematopoiesis in the recipient [21,22]. These cells could be targeted by alloreactive T cells recognizing minor antigens on the leukaemia stem Selleckchem Venetoclax cells [7,8]. These models should be interpreted with caution, as the

xenogeneic milieu of the recipient mouse underestimates the number of cells capable of self-renewal and do not provide clear evidence that long-lived AML progenitors are subject to the same degree of immune attack. Furthermore, they do not identify whether all subtypes of AML have comparable hierarchies of long-lived progenitors. Indeed, an alternative model of leukaemia cure is that a sustained T cell response to the progeny of the AML stem cell but not the small stem cell pool itself could contain the leukaemia at a minimal disease level, resulting in a functional cure [3]. Although the concept of immune surveillance is well accepted, evidence for IS specifically in AML is largely indirect, revealed through relationships between treatment outcome and PD-1/PD-L1 inhibitor drugs immune parameters and adaptive changes made by the leukaemia favouring immune evasion, unlike viral-induced malignancies. Perhaps the most compelling evidence for a significant role of immune control of AML comes from several observations indicating that

lymphocyte recovery following induction chemotherapy is strongly predictive for outcome. T cells are reduced after chemotherapy Unoprostone but have a rapid clonogenic potential which allows a swift T cell recovery [23]. Patients achieving the highest lymphocyte counts within 6 weeks of chemotherapy have the lowest relapse rates [24–26]. Long-term survival in AML is also favoured by normalized lymphocyte counts [27]. These data all suggest that an intact immune system can protect against relapse of disease, but do not define whether the effect is mediated through T cells or NK cells. There are diverse abnormalities in AML at presentation and relapse that suggest how the leukaemia may develop despite immunosurveillance and how an established leukaemia may acquire new characteristics to defeat immune control. Figure 1 depicts the interactions between AML cells and the immune environment. Genetic features are emerging that may favour the development of AML in the presence of an intact immune system. There is an increased frequency in AML of a particular genotype of the co-stimulatory molecule cytotoxic lymphocyte antigen -4 (CTLA-4) [28]. The inhibitory KIR molecule KIR 2DL2 is expressed more frequently in AML, again suggesting a predisposition for AML through some form of immune escape [29]. There is also strong evidence that an established AML can mutate to escape immune control.

However, activated neutrophils may also cause undesired tissue damage. Ample examples include small-vessel inflammatory diseases (vasculitis) that are associated with anti-neutrophil cytoplasmic autoantibodies (ANCA) residing in the patients’ plasma. In addition to being an important diagnostic tool, convincing evidence shows that ANCA are pathogenic. ANCA–neutrophil interactions induce important cellular responses that result in highly inflammatory necrotizing vascular damage. The find more interaction begins with ANCA binding to their target antigens on primed neutrophils, proceeds by recruiting transmembrane molecules to initiate intracellular signal transduction and culminates in activation of effector functions that ultimately

mediate the tissue damage. ANCA must recognize and bind their target antigens, proteinase 3 (PR3) or myeloperoxidase (MPO), in order to initiate signalling events and to subsequently activate the neutrophil. Thus, ANCA must either be internalized by the neutrophil or the antigens must be accessible on the cell surface,

or both may occur. Many studies exploring the membrane expression of ANCA antigens have been performed. MPO and the vast majority of PR3 antigens reside in azurophilic granules, which can be mobilized during activation in vitro and in vivo[1,2]. In contrast to MPO, PR3 is also stored in specific granules and in secretory vesicles that are mobilized more easily [3]. Moreover, significant PR3 amounts are already expressed on the surface of resting cells NVP-BEZ235 order with a strong increased expression after activation. Thus, there are major differences in PR3 and MPO membrane expression. Notably,

and in contrast to PR3, MPO is not detected on the plasma membrane of resting neutrophils. Furthermore, the membrane MPO that increases after cell activation is small compared to PR3. Neutrophils must be primed for subsequent ANCA-induced activation. Priming includes ANCA antigen translocation and can be achieved in vitro by various mediators, Ribose-5-phosphate isomerase including tumour necrosis factor (TNF)-α, interleukin (IL)-1, IL-6, IL-18, N-formyl-Met-Leu-Phe (fMLF) and complement 5a (C5a) [4–7]. In-vivo priming may occur during infections that frequently precede the clinical manifestation of ANCA vasculitis. Indeed, patients with active disease show increased neutrophil ANCA antigen membrane expression [5,8,9]. A synergistic effect for increased mPR3 expression by cytokines, adhesion and anti-PR3 antibodies was demonstrated that could become relevant when neutrophils leave the circulating blood [10]. Recently, α1-anti-trypsin polymers have been described to prime the neutrophil for ANCA activation, indicating that additional priming mechanisms exist [11]. An important observation established that PR3, but not MPO, has a bimodal membrane expression pattern. mPR3low- and mPR3high-expressing neutrophils can be distinguished with a percentage of mPR3high neutrophils ranging between 0 and 100% [12].

Conclusion: These results support a critical protective function for TIMP-1 expression on promoting survival and proliferation of liver cells and on regulating leukocyte recruitment and activation in liver PXD101 IRI. (HEPATOLOGY 2012;56:1074–1085) Hepatic ischemia/reperfusion injury (IRI) occurs during trauma, shock, orthotopic liver transplantation (OLT), and other surgical procedures where the blood supply to the liver is temporarily interrupted.1

Hepatic IR-related damage is the result of various factors that include leukocyte migration, release of cytokines, and free radicals.1, 2 Leukocytes migration across endothelial and extracellular matrix (ECM) barriers is dependent on cellular adhesion-release and focal matrix degradation mechanisms.3 Although adhesion molecules are important for the successful leukocyte transmigration by providing leukocyte attachment

to the endothelium, there is a growing body of evidence suggesting that matrix metalloproteinases (MMP) are critical for facilitating leukocyte movement across vascular barriers.3 In this regard, our previous studies showed an important role for leukocyte-expressed MMP-9, or gelatinase B, as a key mediator of leukocyte transmigration leading to liver injury.4 Tissue inhibitors of metalloproteinases (TIMPs) are a family G protein-coupled receptor kinase of naturally occurring inhibitors of MMPs. Alterations in the MMP-TIMP balance PD-0332991 supplier have been linked to pathological conditions that require disruption of the basement membrane, such as tumor invasion, angiogenesis, and wound healing.5 There are at least four identified members (TIMP 1-4)

in the TIMP family, varying in tissue-specific expression and in their ability to inhibit various MMPs.6 Among the different TIMPs, TIMP-1 is of particular interest; TIMP-1 is a 28.5-kDa soluble glycoprotein known to inhibit MMP-9 with high affinity, without interacting with MMP-2, or gelatinase A (the other member of the gelatinase family), as it lacks the required C-terminal MMP-2-interacting residues.7, 8 In addition to its ability to inhibit MMP activity, TIMP-1 possesses other biological activities, such as cell growth regulation, that are just beginning to be recognized and characterized.9 The specific effects of TIMPs likely depend on the cell context and on the pathological condition. Although TIMP-1 has been detected in the plasma of patients after liver transplantation,10 and in rat liver grafts after IRI,11 its role in liver IRI, or in OLT, remains to be established. Therefore, in the present study we used mice lacking TIMP-1 to examine the significance of TIMP-1 expression in hepatic IRI.

It appears that growth is inhibited with the concomitant degradation of the photosynthetic pigments and by a decrease in the photosynthetic capacity. Being an oxidative stress, we found an H2O2 burst within 15 min of menadione exposure, followed by an increase in antioxidant enzyme (superoxide dismutase [SOD], catalase [CAT], and ascorbate peroxidase [APX]) activities. In parallel, RT-PCR was performed for transcript Stem Cell Compound Library analyses of Mn-SOD, CAT, and APX. Our results clearly revealed that expression of these genes were up-regulated upon menadione exposure. Furthermore, classical hallmarks of PCD such as

alteration of mitochondrial membrane potential, significant increase in caspase-3-like DEVDase activity, PLX4032 in vivo cleavage of poly (ADP) ribose polymerase (PARP)-1-like enzyme, and DNA fragmentation as detected

by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and oligosomal DNA fragmentation were observed. Moreover, antibodies against a mammalian active caspase-3 shared epitopes with a caspase-3-like protein of ~17 kDa; its pattern of expression and activity correlated with the onset of cell death. To the best of our knowledge, this is the first report on menadione-induced PCD through a mitochondrian-caspase protease pathway in an algal species. “
“Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler is a cosmopolitan coccolithophore species that forms massive blooms in low phosphorus seawater, partly due to its ability to utilize organic phosphate via extracellular alkaline phosphatase (AP). A novel AP gene, ehap1, was identified from the strain CCMP374. In this study, we examined the expression of ehap1 in various E. huxleyi strains and Baf-A1 mouse its genetic diversity in those strains and field populations. Two EHAP1 proteins (EHAP1a, 75 kDa and EHAP1b, 110 kDa) with virtually identical

sequence were expressed under P limitation in all strains except one; a third protein (EHAP1c, 115 kDa) was expressed in a few strains. The correlation between AP activity and protein abundance suggests that EHAP1b is inactive and probably the precursor of EHAP1a. The transcript of ehap1 was induced by P depletion in all strains. The ehap1 gene sequence is highly conserved in these strains and field populations with <3% nucleic acid substitution. Most of the ehap1 sequences from one site in the English Channel and three sites in the Gulf of Alaska were essentially identical to one another. No EHAP1-like protein can be detected in other phytoplankton species tested via Western blot analysis. The rapid induction and high activity of EHAP1 in E. huxleyi suggest that it plays a significant role in P regeneration in the oligotrophic ocean where E. huxleyi is abundant. The EHAP1 antibody and gene-specific primers are well suited to study the dynamics of P limitation in field populations of E. huxleyi.

This includes vaccination against hepatitis B, avoidance of non-steroidal anti-inflammatory drugs, preservation of dental hygiene, correction of iron deficiency anemia and prenatal diagnosis when the mutation is known. (ii) Topical hemostatic measures. These can involve compression with gauze Buparlisib soaked with tranexamic acid, fibrin sealants containing tranexamic acid, acrylic splints for dental extraction, and packing for nose bleeds. (iii) Antifibrinolytic agents (epsilon aminocaproic acid or tranexamic acid). Such agents are useful for prevention of bleeding

following minor surgery, and can be employed as adjuncts of other treatment modalities such as recombinant factor VIIa (rFVIIa), 1-desamino-8D-arginine vasopressin (DDAVP), and platelet transfusion. Patients should be guided to start oral treatment with one of the antifibrinolytic agents whenever troublesome bleeding occurs and thereafter seek medical attention if necessary. Antifibrinolytic agents are essential for Quebec platelet syndrome. (iv) Management by DDAVP. This agent increases

the plasma concentrations of von Willebrand factor (VWF) and factor VIII. The mechanism of DDAVP action has been attributed to increased adhesiveness of platelets to the subendothelial matrix, and to augmented platelet aggregation at high shear rate resulting in shortening of bleeding time Saracatinib [40]. Several small series of DDAVP-treated patients with variable inherited platelet dysfunctions have been reported. Entities for which unequivocal evidence indicates that bleeding time shortens after DDAVP include delta-storage pool diseases, disorders of granule secretion, signal transduction disorders, thromboxane oxyclozanide receptor deficiency, May-Hegglin anomaly and patients with unexplained prolonged bleeding time. Equivocal evidence was provided for BSS, HPS and arachidonate metabolism

defects. Patients with GT do not respond to DDAVP [41]. Side effects of DDAVP administration include tachycardia, hypotension, facial flushing, headache, severe hyponatremia with seizures and arterial thrombosis. (v) rFVIIa. GT patients have been treated for bleeding episodes by rFVIIa with partial success [42,43]. The mechanism by which rFVIIa arrests bleeding is probably related to increased thrombin generation by a tissue factor-independent process, enhanced adhesion of platelets to extracellular matrix and restoration of platelet aggregation [44–46]. The use of rFVIIa in patients with inherited platelet dysfunction has not been examined by randomized controlled studies. Among 59 GT patients in an international survey, 75% of 108 spontaneous bleeding episodes and 94% of 34 surgical procedures were manageable by rFVIIa. However, two patients who received a high dose of rFVIIa developed pulmonary embolism and a ureteric clot, respectively [42]. (vi) Female hormones.