GP130 activation induces myeloma and collaborates with MYC

Multiple myeloma (MM) is a plasma cell neoplasm that results from clonal expansion of an Ig-secreting terminally differentiated B cell. Advanced MM is characterized by tissue damage that involves bone, kidney, and other organs and is typically associated with recurrent genetic abnormalities. IL-6 signaling via the IL-6 signal transducer GP130 has been implicated as an important driver of MM pathogenesis. Here, we demonstrated that ectopic expression of constitutively active GP130 (L-GP130) in a murine retroviral transduction-transplantation model induces rapid MM development of high penetrance. L-GP130–expressing mice recapitulated all of the characteristics of human disease, including monoclonal gammopathy, BM infiltration with lytic bone lesions, and protein deposition in the kidney. Moreover, the disease was easily transplantable and allowed different therapeutic options to be evaluated in vitro and in vivo. Using this model, we determined that GP130 signaling collaborated with MYC to induce MM and was responsible and sufficient for directing the plasma cell phenotype. Accordingly, we identified Myc aberrations in the L-GP130 MM model. Evaluation of human MM samples revealed recurrent activation of STAT3, a downstream target of GP130 signaling. Together, our results indicate that deregulated GP130 activity contributes to MM pathogenesis and that pathways downstream of GP130 activity have potential as therapeutic targets in MM.     

DSTP-27 Prevents Entry of Human Cytomegalovirus

Human cytomegalovirus (HCMV) can cause life-threatening diseases in neonates and immunocompromised patients. Due to multiple problems caused by the current available drugs, development of new antiviral compounds is urgently needed. In this study, we characterize the anti-HCMV spectrum and mechanism of action of the N-N′-(bis-5 nitropyrimidyl)dispirotripiperazine derivate 27 (DSTP-27). DSTP-27 exhibited strong antiviral activity against two laboratory HCMV strains with different cell tropism as well as ganciclovir (GCV)-sensitive and GCV-resistant clinical isolates in plaque reduction assays and viral growth kinetics experiments. Interestingly, neither infectious nor noninfectious viral particles were observed by electron microscopy. Pretreatment of cell-free virus with DSTP-27 prevented virus infection. The results from time of addition assays, in which DTSP-27 was added to cells (i) before infection, (ii) during virus adsorption, or (iii) after adsorption, demonstrated an inhibitory effect on early steps of the HCMV replication cycle. This observation was confirmed by immunofluorescence as well as Western blot analysis, whereby reduced levels of the immediate early protein IE1, the processivity factor pUL44, and the tegument protein pp28 were detected. Results from attachment and penetration analyses of prechilled human embryonic lung fibroblasts revealed that virus attachment is not blocked. In addition, DSTP-27 inactivated HCMV by stable binding. Taken together, these results demonstrate that DSTP-27 (i) blocks viral penetration by interacting with the host cell and (ii) inactivates HCMV by interacting with the virus.     

Management of Pneumothoraces Detected on Chest Computed Tomography: Can Anatomical Location Identify Patients Who Can Be Managed Expectantly?

Background

Pneumothorax (PTX) can be readily detected by computed tomography (CT) or ultrasound. However, management of PTX in hemodynamically stable patients remains controversial.

Study Objectives

We sought to investigate whether a distinct anatomical distribution of PTX along prespecified chest zones as detected by CT can be described in patients with or without subsequent chest tube thoracotomy (CTT), thus potentially allowing the extended focused assessment with sonography for trauma (EFAST) ultrasound examination to guide PTX management.

Methods

We performed a retrospective review of chest CT scans performed in the emergency department (ED) of a Level I trauma center. CT scans were analyzed for PTX distribution according to a chest zone model. Medical records of subjects with PTX were reviewed for subsequent CTT.

Results

Of 3636 chest CT scans performed, 183 PTX (156 patients) were detected without CTT at the time of CT scan (69% male, mean age 42 years). Of these, 66 subjects (40%) underwent CTT; 43 chest tubes (63%) were placed in the ED, 9 (13%) during hospitalization and 9 (13%) in the operating room. Median time to CTT was 140 min (interquartile range 52–199). Initial hemodynamic parameters, need for surgery, and need for mechanical ventilation were similar in both groups (p > 0.05 for all). Anatomical distribution and size of PTX were similar in the two groups.

Conclusion

Although the majority of patients with traumatic PTX could be managed conservatively, we did not identify a characteristic anatomical PTX pattern, which could identify subjects who may not require CTT.     

Comparative bone status assessment by dual energy X-ray absorptiometry, peripheral quantitative computed tomography and quantitative ultrasound in adolescents and young adults with cystic fibrosis

PurposeQuantitative ultrasound bone sonometry (QUS) might be a promising screening method for cystic fibrosis (CF)-related bone disease, given its absence of radiation exposure, portability of the equipment and low cost.The value of axial transmission forearm QUS in detecting osteopenia in CF was therefore studied.MethodsWe investigated the application of QUS in the evaluation of bone status in a group of 64 adolescents (> 12 years) and young adults (< 40 years) with CF in a comparison with a dual X-ray absorptiometry (DXA) of the whole body and peripheral quantitative computed tomography (pQCT) of the radius at 4% and 66% sites.ResultsMean (SD) Z-scores of speed of sound (SOS), whole body bone mineral content (BMC), radial trabecular bone mineral density (BMD), and radial cortical BMD were respectively − 0.31 (0.78), − 0.09 (1.28), 0.10 (1.16) and − 0.62 (2.88). The pQCT determined bone geometry values (cortical bone area and cortical thickness) were more depressed than the BMD data. QUS had a sensitivity and specificity of respectively 0% and 96% for diagnosing osteopenia (based on a whole body BMC Z-score < − 2).ConclusionsQUS cannot replace DXA, but can screen out patients with normal bone mass. Further and larger studies are needed to examine if QUS may reflect other aspects than bone mass, or if it is possible to improve its sensitivity by supplementing the SOS results with clinical risk factors.     

Guideline on the requirements of external quality assessment programs in molecular pathology

Molecular pathology is an integral part of daily diagnostic pathology and used for classification of tumors, for prediction of prognosis and response to therapy, and to support treatment decisions. For these reasons, analyses in molecular pathology must be highly reliable and hence external quality assessment (EQA) programs are called for. Several EQA programs exist to which laboratories can subscribe, but they vary in scope, number of subscribers, and execution. The guideline presented in this paper has been developed with the purpose to harmonize EQA in molecular pathology. It presents recommendations on how an EQA program should be organized, provides criteria for a reference laboratory, proposes requirements for EQA test samples, and defines the number of samples needed for an EQA program. Furthermore, a system for scoring of the results is proposed as well as measures to be taken for poorly performing laboratories. Proposals are made regarding the content requirements of an EQA report and how its results should be communicated. Finally, the need for an EQA database and a participant manual are elaborated. It is the intention of this guideline to improve EQA for molecular pathology in order to provide more reliable molecular analyses as well as optimal information regarding patient selection for treatment.     

The role of 3D structure and protein conformation on the innate and adaptive immune responses to silk-based biomaterials

We have investigated monocyte and T cell responsiveness to silk based biomaterials of different physico-chemical characteristics. Here we report that untransformed CD14+ human monocytes respond to overnight exposure to silk fibroin-based biomaterials in tridimensional form by IL-1β and IL-6, but not IL-10 gene expression and protein production. In contrast, fibroin based materials in bidimensional form are unable to stimulate monocyte responsiveness. The elicitation of these effects critically requires contact between biomaterials and responding cells, is not sustained and becomes undetectable in longer term cultures. We also observed that NF-κβ and p38 MAP kinase play key roles in monocyte activation by silk-based biomaterials. On the other hand, fibroin based materials, irrespective of their physico-chemical characteristics appeared to be unable to induce the activation of peripheral blood T cells from healthy donors, as evaluated by the expression of activation markers and IFN-γ gene.     

Tonic stimulation of the pharyngeal mucosa causes pain and a reversible increase of inflammatory mediators

Objective and design

To develop a model of the inflammatory component of non-infectious sore throat using tonic stimulation and quantification of inflammatory mediators in pharyngeal lavage fluid.

Material or subjects

Forty-five healthy volunteers.

Treatment

Cold dry air.

Method

Tonic stimulation of the pharynx was achieved using a constant stream of cold dry air to the back of the throat. Following optimization of stimulation conditions (phase 1), pharyngeal pain, irritation, and swallowing discomfort were assessed using visual analog scales, and the concentration of inflammatory markers were measured in pharyngeal lavage fluid (phase 2).

Results

Optimum conditions for tonic pharyngeal stimulation were cold dry air at 12 °C, relative humidity 20 %, at a flow rate of 12 L/min for 15 min. Analysis of pharyngeal lavage fluid collected 5 min after stimulation showed significant increases in prostaglandin E₂ (P = 0.018), thromboxane B₂ (P < 0.001), and substance P (P < 0.001), but no increase in peptidoleukotriene. When the stimulus was removed, the level of inflammatory markers in pharyngeal lavage fluid returned to baseline by 30 min post-stimulation. These objective measures mirrored subjective pain ratings.

Conclusions

Tonic stimulation of the pharyngeal mucosa with cold dry air causes pain and an increase of inflammatory mediators which are reversible.