BackgroundHepatitis C Virus (HCV), a single stranded RNA virus, affects millions of people worldwide and leads to chronic infection characterized by chronic inflammation in the liver and in peripheral immune cells. Chronic liver inflammation leads to progressive liver damage. MicroRNAs (miRNA) regulate inflammation (miR-155, -146a and -125b) as well as hepatocyte function (miR-122).MethodsHere we hypothesized that microRNAs are dysregulated in chronic HCV infection. We examined miRNAs in the circulation and in peripheral monocytes of patients with chronic HCV infection to evaluate if specific miRNA expression correlated with HCV infection.ResultsWe found that monocytes from chronic HCV infected treatment-naïve (cHCV) but not treatment responder patients showed increased expression of miR-155, a positive regulator of TNFα, and had increased TNFα production compared to monocytes of normal controls. After LPS stimulation, miR-155 levels were higher in monocytes from cHCV patients compared to controls. MiR-125b, which has negative regulatory effects on inflammation, was decreased in cHCV monocytes compared to controls. Stimulation of normal monocytes with TLR4 and TLR8 ligands or HCV core, NS3 and NS5 recombinant proteins induced a robust increase in both miR-155 expression and TNFα production identifying potential mechanisms for in vivo induction of miR-155. Furthermore, we found increased serum miR-155 levels in HCV patients compared to controls. Serum miR-125b and miR-146a levels were also increased in HCV patients. Serum levels of miR-122 were elevated in cHCV patients and correlated with increased ALT and AST levels and serum miR-155 levels.ConclusionIn conclusion, our novel data demonstrate that miR-155, a positive regulator of inflammation, is upregulated both in monocytes and in the serum of patients with chronic HCV infection. Our study suggests that HCV core, NS3, and NS5 proteins or TLR4 and TLR8 ligands can mediate increased miR-155 and TNFα production in chronic HCV infection. The positive correlation between serum miR-155 and miR-122 increase in cHCV may be an indicator of inflammation-induced hepatocyte damage.

There has been a worldwide increase in the incidence of asthma, and the disease has greatly impacted the public health care system. Chlamydia pneumoniae has been reported as a possible contributing factor in asthma. The organism has been detected by polymerase chain reaction (PCR) in bronchial tissue, but there has been no direct evidence of viability. To determine the frequency of viable Chlamydia in children, blood and bronchoalveolar lavage were collected from 70 pediatric patients undergoing flexible fiberoptic bronchoscopy. Forty-two of these patients had asthma, whereas the remaining patients had various respiratory disorders. Fifty-four percent (38) of the bronchoalveolar lavage samples were PCR-positive for Chlamydia, and 31% (22) of the PCR-positive samples were positive when cultured on macrophages. Twenty-eight samples (40%) and 14 samples (20%) of the PCR- and culture-positive samples, respectively, were from patients with asthma. Culture of the blood samples revealed that 24 (34.3%) of 70 were positive for Chlamydia compared with 8 (11%) of 70 matched nonrespiratory control subjects (p < 0.01); 17 (24%) of the positive blood cultures from the respiratory group were from patients with asthma. Elevation of total IgE was strongly associated with lavage culture positivity for Chlamydia. We therefore conclude that viable Chlamydia pneumoniae organisms are frequently present in the lung lavage fluid from this cohort of predominantly asthmatic pediatric patients.

An emerging body of evidence suggests that half of asthma in both children and adults is associated with chronic lung infection. The aim of the present study was to determine the frequency of viable Chlamydia pneumoniae (Cp) and C. trachomatis (Ct) in the respiratory tracts of paediatric patients with chronic respiratory diseases.Bronchoalveolar lavage fluid (BALF) samples obtained from 182 children undergoing bronchoscopy for clinical reasons were assayed using PCR analysis, in vitro tissue culture and immunofluorescence staining for the presence of Cp and Ct.Chlamydia-specific DNA was detected by PCR in 124 (68%) out of 182 patients; 79 were positive for Cp, 77 positive for Ct and 32 for both organisms; 75 patients had cultivable Chlamydia. Ct DNA prevalence decreased, whereas Cp positivity generally increased with age. A total of 59 out of 128 asthma patients and 16 out of 54 nonasthmatics were Chlamydia culture positive. When the patients were divided into inflammatory versus noninflammatory airway disease, there were 69 (46%) out of 150 and six (18%) out of 32 BALF samples with cultivable Chlamydia, respectively.Viable Chlamydia pneumoniae and Chlamydia trachomatis occur frequently in children with chronic respiratory diseases and may be more prevalent in asthma patients. To the current authors' knowledge, this is the first report of viable Chlamydia trachomatis in the lungs of children.

Navigation for surgery is an important tool that can provide surgeons with enhanced information when operating on fixed anatomic targets [1]. The use of augmented reality for improved surgical precision [2] has recently been applied to pelvic surgery [3], and the use of these approaches could help improve resection quality as digital information from various source data can aid surgeons by providing real-time, usable and relevant data [4][5][6].Stereotactic navigation for surgery utilizes computed tomography (CT) or magnetic resonance imaging (MRI) images that are rendered in a multiplanar format such that the data points can be interpreted by the navigation software [6]. Using infrared trackers and a ceiling mounted infrared camera, 3D data from body imaging can be used in real time to perform 'digital surgery' [4].In this video, the technique of real-time navigation is applied to the laparoscopic excision of a pelvic neoplasm. The video emphasizes how the operating surgeon is able to interpret and utilize stereotactic navigation during surgery to localize and then excise a pelvic neoplasm in the deep perirectal space.In this clinical example, a 79-year-old female presented with chronic pelvic pain. Workup included physical examination-including anoscopy, proctoscopy and colonoscopy which were all unremarkable. However, a pelvic MRI demonstrated a complex, mixed cystic and solid neoplasm in the left perirectal space measuring 2 9 2.3 9 2.9 cm in dimension with an ill-defined solid, internal component measuring 1.1 cm. Malignancy could not be excluded by radiographic criterion. Surgical intervention using a minimally invasive, laparoscopic technique was utilized to perform a complete excision of the neoplasm, which was removed intact. The navigation accuracy (mean deviation) measured ±1.8 mm. Operative time was 124 min which included 38 min of navigation setup time. The setup time included time for intra-operative CT scan with placement of skin fixed fiducials, navigation software and instrument calibration.The patient was discharged on the first postoperative day. There was no perioperative morbidity. At 10-week follow-up, the patient reported complete resolution of her pelvic pain. Finally, pathology revealed a 50 % solid and 50 % cystic neoplasm measuring 3.0 9 2.0 9 1.5 cm and was consistent with a serous cyst adenofibroma of the ovary with an atypical placement beneath the peritoneal reflection.The video forum provides a description of the operation as seen from the surgeon's point of view to help viewers understand how augmented reality can aid in the localization of fixed anatomic targets with precision.