Pulmonary Embolism Severity Index and troponin testing for the selection of low‐risk patients with acute symptomatic pulmonary embolism

Summary. Background: The combination of the Pulmonary Embolism Severity Index (PESI) and troponin testing could help physicians identify appropriate patients with acute pulmonary embolism (PE) for early hospital discharge. Methods: This prospective cohort study included a total of 567 patients from a single center registry with objectively confirmed acute symptomatic PE. On the basis of the PESI, each patient was classified into one of five classes (I–V). At the time of hospital admission, patients had troponin I (cTnI) levels measured. The endpoint of the study was all‐cause mortality within 30 days after diagnosis. We calculated the mortality rates in four patient groups: group 1, PESI class I–II plus cTnI < 0.1 ng mL^?1; group 2, PESI classes III–V plus cTnI < 0.1 ng mL^?1; group 3, PESI classes I–II plus cTnI ≥ 0.1 ng mL^?1; and group 4, PESI classes III–V plus cTnI ≥ 0.1 ng mL^?1. Results: The study cohort had a 30‐day mortality of 10% [95% confidence interval (CI), 7.6–12.5%]. Mortality rates in the four groups were 1.3%, 14.2%, 0% and 15.4%, respectively. Compared with non‐elevated cTnl, the low‐risk PESI had a higher negative predictive value (NPV) (98.9% vs. 90.8%) and negative likelihood ratio (NLR) (0.1 vs. 0.9) for predicting mortality. The addition of non‐elevated cTnI to low‐risk PESI did not improve the NPV or the NLR compared with either test alone. Conclusions: Compared with cTnl testing, PESI classification more accurately identified patients with PE who are at low risk of all‐cause death within 30 days of presentation.     

四环素类药物的流动注射分析研究

四环素类(主要包括四环素、金霉素、土霉素、强力霉素及甲烯土霉素等)是一类应用较广的抗生素药物。其含量测定,国内外药典均采用微生物法。该方法虽能反映其生物效价,但也存在一些问题,如分析周期较长,方法的准确度不高,且在测定条件下(pH6.6,     

Fragment E1 labeled with I-123 in the detection of venous thrombosis

Fragment E1, which has been shown to have specific binding affinity for thrombi in an animal model, was investigated in humans for its safety and ability to bind to venous thrombi. Human Fragment E1 was labeled with I-123 and administered intravenously to patients with proved or suspected deep vein thrombosis. The vascular distribution of radioactivity was documented by obtaining gamma camera images of the patients' legs for 30 minutes following administration of I-123-Fragment E1. All patients (n = 5) with documented venous thrombi had rapid localization of labeled Fragment E1 in the area of thrombus. Patients without evidence of thrombi (n = 5) showed no focal localization, although two of these patients showed diffuse uptake along the length of the veins, due to superficial phlebitis. Analysis of blood samples in four patients indicated that disappearance of Fragment E1 from the circulation was more rapid in individuals with thrombosis (t 1/2 = 20 min) than in individuals without thrombosis (t 1/2 = 90 min), and a radiolabeled species of high molecular weight was found in patients with thrombosis but was absent from patients without thrombosis. These early results suggest that radiolabeled Fragment E1 is a safe and potentially valuable agent for the rapid detection of venous thrombosis.     

The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

Ethnopharmacological relevance

Anti-inflammatory, anti-oxidant and effect of Zataria multiflora on Th₁/Th₂ balance were previously described. Different therapeutic effects of this plant have been described in Iranian traditional medicine. To evaluate the immune modulatory effects of Zataria multiflora on Th1/Th2 balance, which may be implicated in inflammatory disorders, in vitro and in vivo studies were carried out.

Materials and methods

The effects of three concentrations of the extract, dexamethasone, and saline on interleukin 4 (IL-4) and interferon γ (IFN-γ) gene expression were evaluated in phytohemagglutinin (PHA) stimulated and non-stimulated human peripheral blood mononuclear cells (hPBMCs). RNA was extracted from the hPBMCs to make cDNA for real time PCR relative quantification. Furthermore, the effect of the extract on serum level of IL-4 and IFN-γ was assessed in ovalbumin (OA) sensitized guinea pigs (n=6 for each group).

Results

Dexamethasone showed significant inhibitory effect on both IFN-γ and IL-4 gene expression and serum level of the cytokines and significantly enhanced IFN-γ/IL-4 ratio (p<0.05–p<0.001). The extract inhibited IL-4 and enhance IFN-γ gene expression and IFN-γ/IL-4 ratio too (p<0.05–p<0.001). In sensitized animals also serum level of IL-4 were significantly decreased after treatment with both dexamethasone and extract, but serum level of IFN-γ and IFN-γ/IL-4 ratio were significantly increased due to extract treatment (p<0.01 for medium and p<0.001 for high concentration).

Conclusions

These results indicated consistent in vitro and in vivo data for selective immune modulatory effect of the extract of Zataria multiflora which increased IFN-γ, decreased IL-4, and enhanced the ratio of IFN-γ to IL-4 (Th₁/Th₂ balance). Therefore, the extract of Zataria multiflora may have therapeutic value in inflammatory responses such as allergy, autoimmunity and infectious diseases associated with Th₁/Th₂ imbalance.     

Conserved TCP domain of Sas-4/CPAP is essential for pericentriolar material tethering during centrosome biogenesis

Pericentriolar material (PCM) recruitment to centrioles forms a key step in centrosome biogenesis. Deregulation of this process leads to centrosome aberrations causing disorders, one of which is autosomal recessive primary microcephaly (MCPH), a neurodevelopmental disorder where brain size is reduced. During PCM recruitment, the conserved centrosomal protein Sas-4/CPAP/MCPH6, known to play a role in centriole formation, acts as a scaffold for cytoplasmic PCM complexes to bind and then tethers them to centrioles to form functional centrosomes. To understand Sas-4’s tethering role, we determined the crystal structure of its T complex protein 10 (TCP) domain displaying a solvent-exposed single-layer of β-sheets fold. This unique feature of the TCP domain suggests that it could provide an “extended surface-like” platform to tether the Sas-4–PCM scaffold to a centriole. Functional studies in Drosophila, human cells, and human induced pluripotent stem cell-derived neural progenitor cells were used to test this hypothesis, where point mutations within the 9–10th β-strands (β9–10 mutants including a MCPH-associated mutation) perturbed PCM tethering while allowing Sas-4/CPAP to scaffold cytoplasmic PCM complexes. Specifically, the Sas-4 β9–10 mutants displayed perturbed interactions with Ana2, a centrosome duplication factor, and Bld-10, a centriole microtubule-binding protein, suggesting a role for the β9–10 surface in mediating protein–protein interactions for efficient Sas-4–PCM scaffold centriole tethering. Hence, we provide possible insights into how centrosomal protein defects result in human MCPH and how Sas-4 proteins act as a vehicle to tether PCM complexes to centrioles independent of its well-known role in centriole duplication.Centrosomes consist of a pair of centrioles surrounded by a protein network of pericentriolar material (PCM), the main sites for microtubule nucleation and anchoring and thus responsible for PCM’s role as the principle microtubule-organizing centers (MTOCs) of cells (1–4). When PCM is not recruited, centrioles are unstable, and thus no functional centrosomes are generated (5, 6). Although initial proteomic studies suggested PCM to be an amorphous cloud composed of more than a 100 different proteins (7), recent superresolution microscopy of fly and human centrosomes have indicated key centrosomal proteins essential for centrosome biogenesis to be organized into distinct spatial compartments before appearing as a PCM cloud surrounding the centriole (6, 8–11).Thus, there could be a protein providing an interface for mediating PCM tethering to a centriole, a suitable candidate of which is the conserved centrosomal protein Sas-4 (CPAP in human), forming a layer closely associated with the centriole wall and yet shown to interact with various PCM components (6, 12). Functional studies in various model organisms suggest that Sas-4 proteins are required for both centriole formation and PCM assembly (6, 12); in the absence of Sas-4, nascent centrioles form but fail to mature into centrosomes (6). Overexpression of Sas-4 in flies produces PCM-like structures (13), whereas reduced amounts of Sas-4 in worms result in centrosomes having proportionally less PCM (12). Thus, although it is clear that Sas-4 is essential for centrosome biogenesis, the mechanisms by which Sas-4 contributes to PCM assembly remains elusive.During the course of these studies, we and others have reported that Sas-4/CPAP, a protein essential for centriole formation was found to interact with several centrosomal and PCM proteins including Cnn, Asl, D-PLP, γ-TuRC, SIL, Cep135, Cep120, and tubulin dimers (5, 6, 14–16). In Drosophila, the N-terminal domain of Sas-4 provides a scaffolding site for cytoplasmic protein complexes (hereafter referred to as Sas-4–PCM scaffold) and tethers the components of Sas-4–PCM scaffold to a centrosome matrix via its C terminus (6).Interestingly, the C-terminal region of Sas-4 proteins contains a conserved TCP10c domain (Pfam: PF07202) (hereafter referred to as TCP for brevity) (Fig. 1A and SI Appendix, Fig. S1). An E1235V missense mutation within this domain in CPAP has been identified in patients with primary microcephaly (MCPH), resulting in a reduced interaction with STIL (Ana2 in Drosophila), a centriole duplication factor also implicated in MCPH (16–18). Accordingly, recent structural studies on CPAP-STIL complex revealed that CPAP-STIL interaction is required during centriole assembly (19, 20). The C-terminal domain of CPAP has also been shown to mediate an interaction with another MCPH protein Cep135 (Bld-10 in Drosophila) and that interaction is required for centriole assembly. Bld-10 is a core centriolar protein and is required to stabilize structural integrity of centrioles (21–23). Taken together, we therefore speculate that the TCP domain could mediate protein–protein interactions and might serve as a tethering site for Sas-4–PCM scaffold–centriole interactions.Open in a separate windowFig. 1.Crystal structure of Drosophila Sas-4–TCP domain. (A) Domain architecture of Drosophila Sas-4 and its human ortholog CPAP. The fragment used for crystallization is indicated by a black underline. (B) Cartoon view of the overall structure of Sas-4–TCP. The invisible part of β16–20 in the crystal structure is shown as dotted lines. (C) Side view of Sas-4–TCP along the longitudinal axis from the N to C termini. Twisting of the TCP β-strands is diagramed below. F_L, surface left to β1; F_R, surface right to β1. (D and E) Cross-strand ladder residues on F_L (D) and F_R (E) are shown in spheres and classified into different types by color (purple, positively charged residues; red, negatively charged residues; orange, polar residues; green, hydrophobic residues).Although it appears that Sas-4 plays pivotal roles in centriole formation, assembling protein complexes in the cytoplasm, and tethering them to a developing centrosome, the mechanisms by which Sas-4 accomplishes its tethering role have remained unclear. In this study, we therefore investigated the structural basis of Sas-4 and show that via its conserved C-terminal TCP domain, it could provide an “extended surface-like” platform by which Sas-4 could mediate the Sas-4–PCM scaffold–centriole interaction during centrosome biogenesis.     

Acute lymphangitic nocardiasis

The third case of lymphangitic nocardiasis caused by Nocardia brasiliensis to be recorded in Uruguay is presented. The clinical picture showed some of the features of sporotrichosis, but it was more acute, the nodules developing rapidly into abscesses. A review of eight similar cases, reported previously, reveals that N. brasiliensis was properly identified in five of them. All cases were localized on the upper limbs, and granules were not seen on direct examination. Mycetomas caused by N. brasiliensis have not been observed in Uruguay.     

Phase 1 human trial of autologous bone marrow-hematopoietic stem cell transplantation in patients with decompensated cirrhosis

AIM： TO evaluate safety and feasibility of autologous bone marrow-enriched CD34^＋ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.
METHODS： Four patients with decompensated cirrhosis were included. Approximately 200 mL of the bone marrow of the patients was aspirated, and CD34^＋ stem cells were selected. Between 3 to 10 million CD34^＋ cells were isolated. The cells were slowly infused through the hepatic artery of the patients.
RESULTS： Patient 1 showed marginal improvement in serum albumin and no significant changes in other test results. In patient 2 prothrombin time was decreased; however, her total bilirubin, serum creatinine, and Model of End-Stage Liver Disease （MELD） score worsened at the end of follow up. In patient 3 there was improvement in serum albumin, porthrombin time （PT）, and MELD score. Patient 4 developed radiocontrast nephropathy after the procedure, and progressed to type 1 hepatorenal syndrome and died of liver failure a few days later. Because of the major side effects seen in the last patient, the trial was prematurely stopped.
CONCLUSION： Infusion of CD34^＋ stem cells through the hepatic artery is not safe in decompensated cirrhosis. Radiocontrast nephropathy and hepatorenal syndrome could be major side effects. However, this study does not preclude infusion of CD34^＋ stem cells through other routes.     

Sorafenib targets dysregulated Rho kinase expression and portal hypertension in rats with secondary biliary cirrhosis

Background and purpose

Extrahepatic vasodilation and increased intrahepatic vascular resistance represent attractive targets for the medical treatment of portal hypertension in liver cirrhosis. In both dysfunctions, dysregulation of the contraction-mediating Rho kinase plays an important role as it contributes to altered vasoconstrictor responsiveness. However, the mechanisms of vascular Rho kinase dysregulation in cirrhosis are insufficiently understood. They possibly involve mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)-dependent mechanisms in extrahepatic vessels. As the multikinase inhibitor sorafenib inhibits ERK, we tested the effect of sorafenib on haemodynamics and dysregulated vascular Rho kinase in rats with secondary biliary cirrhosis.

Experimental approach

Secondary biliary cirrhosis was induced by bile duct ligation (BDL). Sorafenib was given orally for 1 week (60 mg·kg⁻¹·d⁻¹). Messenger RNA levels were determined by quantitative real time polymerase chain reaction, protein expressions and protein phosphorylation by Western blot analysis. Aortic contractility was studied by myographic measurements, and intrahepatic vasoregulation by using livers perfused in situ. In vivo, haemodynamic parameters were assessed invasively in combination with coloured microspheres.

Key results

In BDL rats, treatment with sorafenib decreased portal pressure, paralleled by decreases in hepatic Rho kinase expression and Rho kinase-mediated intrahepatic vascular resistance. In aortas from BDL rats, sorafenib caused up-regulation of Rho kinase and an improvement of aortic contractility. By contrast, mesenteric Rho kinase remained unaffected by sorafenib.

Conclusions and implications

Intrahepatic dysregulation of vascular Rho kinase expression is controlled by sorafenib-sensitive mechanisms in rats with secondary biliary cirrhosis. Thus, sorafenib reduced portal pressure without affecting systemic blood pressure.