Interleukin-11 Overexpression and M2 Macrophage Density are Associated with Angiogenic Activity in Proliferative Diabetic Retinopathy

ABSTRACT

Purpose

To investigate the expression of IL-11 and its receptor IL-11Rα and to quantify density of CD163⁺ M2 macrophages in proliferative diabetic retinopathy (PDR).     

Modulation of experimental autoimmunity: treatment of adjuvant arthritis by immunization with a recombinant vaccinia virus.

Live recombinant vaccinia viruses, expressing antigens from pathogenic microorganisms, are studied for their use as vaccines designed for the protection against infectious diseases. Infections with these vaccinia virus recombinants, expressing proteins or epitopes from viruses, parasites, or bacteria, have resulted in the development of specific neutralizing antibodies or cytotoxic T lymphocytes. Here, we describe the generation of a recombinant vaccinia virus expressing the mycobacterial 65-kDa heat shock protein (HSP65). A vaccinia recombinant virus was constructed by placing the gene for the Mycobacterium bovis BCG HSP65 under control of a vaccinia virus promoter and inserting this mycobacterial gene in the thymidine kinase locus of the vaccinia virus genome. Mycobacterial HSP65 is a critical antigen in the autoimmune model of adjuvant arthritis induced in Lewis rats by the immunization with Mycobacterium tuberculosis. We report the induction of immunity directed to this mycobacterial HSP65 by testing for the presence of specific antibodies and T-cell proliferation. Furthermore, induction of such immunity resulted in a reduction of arthritis severity when given to rats before or, even more interestingly, during development of arthritis. Disease reduction was not found after administration of HSP65 in the absence of vaccinia virus as a vector when given during arthritis development. Therefore, recombinant vaccinia virus may offer new prospectives for specific intervention in autoimmunity.     

Genotoxicity and physicochemical characteristics of traffic-related ambient particulate matter

Exposure to ambient particulate matter (PM) has been linked to several adverse health effects. Since vehicular traffic is a PM source of growing importance, we sampled total suspended particulate (TSP), PM(10), and PM(2.5) at six urban locations with pronounced differences in traffic intensity. The mutagenicity, DNA-adduct formation, and induction of oxidative DNA damage by the samples were studied as genotoxicological parameters, in relation to polycyclic aromatic hydrocarbon (PAH) levels, elemental composition, and radical-generating capacity (RGC) as chemical characteristics. We found pronounced differences in the genotoxicity and chemical characteristics of PM from the various locations, although we could not establish a correlation between traffic intensity and any of these characteristics for any of the PM size fractions. Therefore, the differences between locations may be due to local sources of PM, other than traffic. The concentration of total (carcinogenic) PAHs correlated positively with RGC, direct and S9-mediated mutagenicity, as well as the induction of DNA adducts and oxidative DNA damage. The interaction between total PAHs and transition metals correlated positively with DNA-adduct formation, particularly from the PM(2.5) fraction. RGC was not associated with one specific PM size fraction, but mutagenicity and DNA reactivity after metabolic activation were relatively high in PM(10) and PM(2.5), when compared with TSP. We conclude that the toxicological characteristics of urban PM samples show pronounced differences, even when PM concentrations at the sample sites are comparable. This implies that emission reduction strategies that take chemical and toxicological characteristics of PM into account may be useful for reducing the health risks associated with PM exposure.     

Cx43 hemichannel microdomain signaling at the intercalated disc enhances cardiac excitability

Cx43, a major cardiac connexin, forms precursor hemichannels that accrue at the intercalated disc to assemble as gap junctions. While gap junctions are crucial for electrical conduction in the heart, little is known about the potential roles of hemichannels. Recent evidence suggests that inhibiting Cx43 hemichannel opening with Gap19 has antiarrhythmic effects. Here, we used multiple electrophysiology, imaging, and super-resolution techniques to understand and define the conditions underlying Cx43 hemichannel activation in ventricular cardiomyocytes, their contribution to diastolic Ca²⁺ release from the sarcoplasmic reticulum, and their impact on electrical stability. We showed that Cx43 hemichannels were activated during diastolic Ca²⁺ release in single ventricular cardiomyocytes and cardiomyocyte cell pairs from mice and pigs. This activation involved Cx43 hemichannel Ca²⁺ entry and coupling to Ca²⁺ release microdomains at the intercalated disc, resulting in enhanced Ca²⁺ dynamics. Hemichannel opening furthermore contributed to delayed afterdepolarizations and triggered action potentials. In single cardiomyocytes, cardiomyocyte cell pairs, and arterially perfused tissue wedges from failing human hearts, increased hemichannel activity contributed to electrical instability compared with nonfailing rejected donor hearts. We conclude that microdomain coupling between Cx43 hemichannels and Ca²⁺ release is a potentially novel, targetable mechanism of cardiac arrhythmogenesis in heart failure.     

Eradication of chronic methicillin-resistant Staphylococcus aureus infection in cystic fibrosis patients. An observational prospective cohort study of 11 patients

BackgroundChronic airway infection with methicillin-resistant Staphylococcus aureus (MRSA) in patients with cystic fibrosis (CF) is an increasing clinical problem, and therapeutic options are limited. Because chronic infection with MRSA can be associated with accelerated decline in lung function, eradication of MRSA is attempted in most CF centres today. The aim of this observational prospective cohort study was to determine whether it is possible to eradicate MRSA from airways of CF patients using prolonged oral antibiotic combination therapy together with topical decolonization measures.ResultsEleven CF patients, (median age: 9 years (range 1–43); median FEV₁: 91%pred (95%CI 74%–100%pred)) who were chronically infected with MRSA, were treated daily for six months with rifampicin and fusidic acid orally. This study did not include a patient control group. Two patients had to switch to an alternative schedule, using rifampicin and clindamycin, due to the resistance pattern of MRSA. Topical decolonization measures were applied to all patients and included mupirocin-containing nasal ointment in both nostrils three times daily for five days and chlorhexidine hair and body wash once daily for five days. Microbiological eradication was achieved in all patients at the end of the six-month eradication protocol, even when significant time (range 18 months to 9 years) had elapsed since initial isolation. In only one patient MRSA reappeared in the six-month follow-up period after the initial study period. Side-effects, like nausea, vomiting and diarrhoea were seen in five out of eleven patients, but did not lead to therapy cessation.ConclusionChronic MRSA infection can be eradicated from respiratory tract samples using a six month dual antibiotic regimen and topical MRSA decolonization measures.     

Where is the neck?: Alpha angle measurement revisited

Background and purpose

The alpha angle is the most used measurement to classify concavity of the femoral head-neck junction. It is not only used for treatment decisions for hip impingement, but also in cohort studies relating hip morphology and osteoarthritis. Alpha angle measurement requires identification of the femoral neck axis, the definition of which may vary between studies. The original “3-point method” uses 1 single point to construct the femoral neck axis, whereas the “anatomic method” uses multiple points and attempts to define the true anatomic neck axis. Depending on the method used, the alpha angle may or may not account for other morphological characteristics such as head-neck offset.

Methods

We compared 2 methods of alpha angle measurement (termed “anatomic” and “3-point”) in 59 cadaver femora and 83 cross-table lateral radiographs of asymptomatic subjects. Results were compared using Bland-Altman plots.

Results

Discrepancies of up to 13 degrees were seen between the methods. The 3-point method had an “equalizing effect” by disregarding femoral head position relative to the neck: in femora with high alpha angle, it resulted in lower values than anatomic measurement, and vice versa in femora with low alpha angles. Using the anatomic method, we derived a reference interval for the alpha angle in normal hips in the general population of 30–66 degrees.

Interpretation

We recommend the anatomic method because it also reflects the position of the femoral head on the neck. Consensus and standardization of technique of alpha angle measurement is warranted, not only for planar measurements but also for CT or MRI-based measurements.Hip morphology variants may influence the development of osteoarthritis (OA) (Ganz et al. 2008). Femoral morphology variants may be best characterized by concavity, a compound measure determined by the sphericity and offset of the femoral head (determined from relative neck width and femoral head position on the neck). The most used concavity measure is the alpha angle, described initially by Nötzli et al. (2002) to diagnose cam deformity and increasingly used in cohort studies examining the risk of OA development (Johnston et al. 2008, Nicholls et al. 2011, Agricola et al. 2013). Nötzli et al. (2002) measured the alpha angle between 2 lines drawn between 3 points (“3-point method”). One line is called the femoral neck axis based on a single point at the center of the narrowest part of the femoral neck (Figure 1), but it is important to recognize that this line will only correspond to the anatomic femoral neck axis if the femoral head is positioned centrally on the neck.Open in a separate windowFigure 1.3-point and anatomic method compared in high alpha angle (A) and low alpha angle (B). 3-point method (A.1 and B.1) uses the midpoint of the femoral neck at its narrowest point. The anatomic method (B.2 and B.2) defines the femoral neck axis by connecting the centers of 3 circles projected over the neck contour. The axis is translated to the center of the femoral head if necessary, to measure the alpha angle. In this example, alpha angle A.1 = 64˚, A.2 = 73˚. Angle B.1 and B.2 are both 30˚, while the femoral head is positioned central on the femoral neck.However, in many human femora the position of the femoral head on the neck may not be central, but shifted or tilted posteriorly (Murray and Duncan 1971, Hogervorst et al. 2009). In such femora, use of a femoral neck axis line connecting the center of the femoral head and neck will decrease the alpha angle (Figure 1). Use of the anatomic center line (the “anatomic method”) rather than a single point for the femoral neck axis (the “3-point method”) probably more accurately represents femoral head-neck morphology, as it may also account for femoral head translation as measured by the anterior offset ratio (Eijer et al. 2001, Pollard et al. 2010). Furthermore, the increasing number of cohort studies using the alpha angle mandates consensus on measurement technique.Measurements in 155 cross-table radiographs