Altered Immunogenicity of Donor Lungs via Removal of Passenger Leukocytes Using Ex Vivo Lung Perfusion.

Passenger leukocyte transfer from the donor lung to the recipient is intrinsically involved in acute rejection. Direct presentation of alloantigen expressed on donor leukocytes is recognized by recipient T cells, promoting acute cellular rejection. We utilized ex vivo lung perfusion (EVLP) to study passenger leukocyte migration from donor lungs into the recipient and to evaluate the effects of donor leukocyte depletion prior to transplantation. For this purpose, female pigs received male left lungs either following 3 h of EVLP or retrieved using standard protocols. Recipients were monitored for 24 h and sequential samples were collected. EVLP-reduced donor leukocyte transfer into the recipient and migration to recipient lymph nodes was markedly reduced. Recipient T cell infiltration of the donor lung was significantly diminished via EVLP. Donor leukocyte removal during EVLP reduces direct allorecognition and T cell priming, diminishing recipient T cell infiltration, the hallmark of acute rejection.

A case series of paediatric hearing preservation cochlear implantation: a new treatment modality for children with drug-induced or congenital partial deafness.

OBJECTIVE: To investigate the use of hearing preservation cochlear implantation in children with partial deafness. PATIENTS AND METHODS: Five children with either drug-induced or congenital partial deafness were enrolled in a pilot study. The patients ranged in age from 13 months to 14 years. Implantation was performed using a hearing preservation technique. A Flex EAS electrode (MED-EL, Innsbruck, Austria) was used in all full insertions. RESULTS: Low frequency hearing was preserved in all patients with postoperative bone conduction within 10 dB of the preoperative hearing levels. These changes were preserved over the follow-up period of 12 months. There were significant improvements in speech perception. CONCLUSION: Hearing preservation cochlear implantation is a new effective modality in children with partial deafness.

Pulmonary Artery Catheter Seemingly Entrapped in the Skull.

A pulmonary artery catheter is an important tool for the monitoring of hemodynamics in patients. Unfortunately, misplacement of a catheter tip may occur in the vasculature local to the intended placement. Misplacement of the catheter can be further complicated by entrapment at the unintended destination. We present a case of a misplaced and entrapped pulmonary artery catheter in a patient with worsening pulmonary disease. After multiple unsuccessful attempts to float the catheter, it was partially retracted and found to be stuck. Imaging showed the tip terminating in the right internal jugular vein at the level of the jugular foramen. It was initially suspected that the catheter had become looped, knotted, or otherwise entangled within the vasculature of the skull and surgical removal would be necessary. Before surgical removal was performed, it was instead determined that the catheter had become kinked and entrapped at the end of the introducer sheath, and noninvasive removal was accomplished by first removing the introducer sheath.

Utilization of fresh human tympanic membranes for structural analysis and cytokeratin immunocytochemistry implementing resin techniques.

CONCLUSIONS: The results of this study have demonstrated for the first time that tympanic membrane (TM) structure is preserved following removal of fresh, normal tissue from patients undergoing surgery. Greater clarity has been demonstrated using resin sections than in previous studies on paraffin sections. Of particular note, cytokeratin (CK) immunocytochemistry was successfully performed on resin sections, which has not been previously reported. This may have potential applications for future work involving tissues that express CKs. OBJECTIVES: To analyse the structure of normal, fresh human TM specimens after surgical removal and to evaluate their CK immunocytochemistry using resin techniques, neither of which have been demonstrated previously. MATERIAL AND METHODS: Seven TM specimens were removed during surgery and then preserved in a modified Karnovsky's fixative. Semi-thin and thin sections were examined by means of light and electron microscopy, respectively. For comparison purposes, paraffin block-embedded specimens were also sectioned. CK immunocytochemistry was performed on semi-thin sections using standard immunoperoxidase techniques, with expression being demonstrated using light microscopy. RESULTS: The three-layer architecture of the TM was preserved. The morphology of the TM was vastly superior in the semi-thin resin sections than in the thicker paraffin sections. The outer, middle and inner layers were clearly demonstrated. The integrity of the outer epithelial layer was maintained, with an outer keratinizing stratum corneum and underlying stratum granulosum, stratum spinosum and stratum basale layers resting on the basal lamina. The thin inner mucosal layer was also viable, consisting of simple squamous or cuboidal cells. Preservation of the middle lamina propria was achieved, with demonstration of the outer radial and inner circular fibres. CK immunocytochemistry utilizing resin techniques provided excellent staining of CK 7 and 8 in the inner layer, with positive staining of CK 5 and 10 in the outer layer.

[Ignition and burning of biological tissue under simulated CO2-laser surgery conditions]. / Entflammungs- und Brandverhalten von biologischem Gewebe bei In-vitro-Bestrahlung mit dem CO2-Laser.

BACKGROUND: Laser surgery in endoscopy has greatly enhanced the surgical armamentarium for treating various laryngeal and hypopharyngeal disorders, but harbours a risk of tissue ignition and inflammation of surgical and anaesthetic instrumentation. However, even if non-inflammable material is used, there is still a residual fire hazard from endogenous tissue that may develop an ignitable aerosol (so-called laser smog) as an effect of laser irradiation. The aim of this study was to investigate how tissue carbonisation and vaporisation contributes to the risk of airway fire. METHODS: For the simulation of gas accumulation in the hypopharynx and larynx following the European standard ISO-11990, a cylindrical steel chamber with an open and a closed end has been used to simulate the operative setting. Pork meat chunks with a tissue composition similar to the larynx and hypopharynx such as fat, muscle, cartilage and bone were introduced into this chamber. Ventilation was achieved through jet ventilation with disposable, non-inflammable laser jet catheters. The tissue was then repeatedly exposed to a laser beam in super-pulse mode (pulse rate 250 Hz) with various intensities and exposure lengths at an impact angle of 75 degrees. The laser intensity was varied from 2 to 15 W. The type, duration, intensity and incidence of tissue ignition were recorded and analysed. RESULTS: The degree of tissue ignition correlated with laser intensity. Low laser intensity caused spark formation whereas high intensity resulted in sustained tissue fire. The type of tissue had an impact on ignition intensity thereby showing lower ignition thresholds and higher ignition susceptibility in fat-containing tissue compared to muscle. The most important factor for occurrence of tissue ignition was the chamber oxygen concentration which displayed an inverse correlation with the time until tissue ignition. Oxygen concentrations of 35% led to tissue ignition in 42 s, 40% oxygen in 20 s. Oxygen concentrations higher than 60% resulted in immediate tissue ignition. CONCLUSIONS: Despite the use of non-inflammable materials in endoscopic laser surgery of the upper airway and hypopharynx, the risk of tissue ignition remains due to the inflammable laser smog which is easily ignited in an oxygen-rich environment. Hence to minimise this risk, we recommend using oxygen concentrations lower than 40%, low laser intensities (<6 W) and limiting continuous laser activation to periods shorter than 10 s.

Cochlear implantation in children with congenital unilateral deafness: Mid-term follow-up outcomes.

OBJECTIVES: Although cochlear implantation is widely used to treat unilateral deafness in adults, very little literature exists on its use and effects on the paediatric population. This report adds to the literature showing the mid-term follow-up outcomes achieved by these children. MATERIAL AND METHODS: Three children with congenital unilateral deafness were studied after implantation. Speech perception in noise, and sound localization ability were evaluated using age-appropriate materials. RESULTS: The preliminary data of our small group of 3 children with congenital unilateral profound hearing loss revealed that up to 3 years post-implantation, congenitally deaf children who received a cochlear implant after 4 years of age do not demonstrate binaural hearing benefits. CONCLUSION: Early intervention in the prelingual phase may be crucial for the development of binaural hearing.

Genome Wide Analysis of Chlamydia pneumoniae for Candidate Vaccine Development.

BACKGROUND: Chlamydia pneumoniae (C. pneumoniae) is a pathogen associated with respiratory tract infection of humans and its viable presence in atherosclerotic plaques is also assumed to play significant function in cardiac diseases. Unavailability of effective antimicrobial drugs has implicated the urgent need of some more disease associated vaccines that may provide relief efficiently. Thus, present study has been undertaken to analyse the whole proteome of C. pneumoniae in order to propose bacterial proteins as candidate vaccine for CAD by taking the aid of 'Reverse Vaccinology'. METHODS: Whole proteome of C. pneumoniae was downloaded and redundancy was removed by CD-HIT web server. Similarity search between proteins of C. pneumoniae and Homo sapiens was performed by BLASTP to avoid human similar proteins. Virulent proteins were selected by VirulentPred web server. Sub cellular localization of identified proteins was investigated by LocTree3 and pSORTb servers. Surface accessibility area (SAA) and antigenic epitopes prediction has been undertaken by prediction of protease specificity (POPS) and Ellipro, NetMHCpan 3.0 servers respectively. Functional significance of identified proteins was predicted through 3D model construction followed by the ligand binding site, active site and domain characterization. RESULTS: Three reference proteins RVOM1, RVOM2, RVEC1 were predicted with the crucial role in C. pneumoniae that may be capable of inducing B cell and T cell response. CONCLUSION: Detailed analysis of these proteins indicated that they may be utilized as the vaccine candidates during the chlamydial infection in near future.

Extra pulmonary tuberculosis presenting as fever with massive splenomegaly and pancytopenia.

Disseminated tuberculosis is an important differential diagnosis for fever of unknown origin (FUO) and it can present with hepatosplenomegaly and lymphadenopathy and may have meningitis and with hematological abnormalities including pancytopenia or a leukemoid reaction. We report the case of a 13-year old male who presented with fever, weight loss, pallor and massive splenomegaly with pancytopenia, in whom a bone marrow trephine biopsy showed caseating granulomata, who responded well to antituberculous treatment and has remained healthy on follow up after nine years.

Incidence and in-vivo relevance of anti-interferon antibodies during treatment of low-grade cutaneous T-cell lymphomas with interferon alpha-2a combined with acitretin or PUVA.

Interferon-alpha combined with retinoid or PUVA is used for the treatment of cutaneous T-cell lymphoma. Anti-IFN-alpha antibodies (IFN ab) occur regularly during IFN-alpha treatment. We investigated the incidence of neutralizing and binding IFN ab and analysed their relationship with clinical and immunological parameters. A group of 17 CTCL patients were treated with IFN alpha-2a three times weekly subcutaneously at a dose of 3 Mill. I.U. combined either with retinoid (acitretin, Neotigason; 0.5 mg/kg bodyweight) daily or with 5-methoxypsoralen (1.2 mg/kg bodyweight) plus UVA radiation three times weekly. Prior to and during treatment we monitored stage, skin involvement by a tumour burden index, serum levels of beta 2-microglobulin, neopterin, binding and neutralizing IFN ab, Interleukin-6 (IL-6), soluble IL-2 receptors (sIL-2r) and the CD4/CD8 ratio of peripheral blood mononuclear cells. We observed two complete, two partial and six minor responses, four patients with stable disease and three patients with progressive disease. Of the 17 patients, 7 developed binding IFN ab, but only 2 had neutralizing IFN ab which were associated with high titres of binding IFN ab. IFN ab formation was more frequent in patients with normal CD4/CD8 ratios and a high tumour burden index and showed a trend to be more frequent in PUVA-cotreated patients than in retinoid-cotreated patients. Responses were more frequently seen in IFN ab-negative patients. IFN ab developed in patients treated with PUVA or retinoid combined with IFN. Binding as well as neutralizing IFN ab may have an impact on the treatment success in CTCL patients.

Half pipe snowboarding: an (un)forgettable experience or an increasing risk for head injury?

The case is presented of a professional half pipe snowboarder with a large post-traumatic subdural haematoma, which allowed close to normal functioning and socialisation. It is an example of a potentially life threatening sports injury in an increasingly commercialised and popular winter sport. It highlights that a knowledge of injury patterns and a high level of suspicion should be maintained in the treatment of snowboarding injuries.

Waldenstrom's macroglobulinaemia.

Waldenstrom's macroglobulinaemia is a rare form of monoclonal plasma cell dyscrasia in which there is excess production of immunoglobulin M. Clinically the patient has features of hyperviscosity syndrome and the effects of bone marrow replacement by lympho-plasmacytoid cells. We report here two such cases seen within a period of two months.

Intravenous beta agonist in severe acute asthma.

STUDY OBJECTIVE: To determine whether salbutamol is more effective in treating severe asthma when given intravenously or by inhalation. DESIGN: Randomised trial of short term response to intravenous versus nebulised salbutamol in acute severe asthma. SETTING: District general hospital (secondary care centre). PARTICIPANTS: 76 patients aged 16-70 admitted to hospital with acute severe asthma (peak expiratory flow rate less than 50% of predicted) during study period. Five withdrawn because of adverse effects of treatment or non-response. Of remaining 71, 34 allocated to nebuliser group and 37 to intravenous treatment group. Patients with history of cardiovascular disease or recent corticosteroid or intravenous bronchodilator treatment excluded. Admission characteristics similar in the two groups. INTERVENTIONS: All patients given 5 mg nebulised salbutamol on admission before randomisation plus 200 mg hydrocortisone bolus intravenously and 35% inspired oxygen throughout. Nebuliser group received two more 5 mg doses of nebuliser salbutamol at 30 minutes and 2 hours; intravenous group received 4 hours' continuous salbutamol infusion (12 micrograms/min) starting at 30 minutes plus supplementary intravenous potassium chloride. No other bronchodilators used. ENDPOINT: Change in peak expiratory flow rate over 4 hours. MEASUREMENTS AND MAIN RESULTS: Peak expiratory flow rate improved more in intravenous group (25.2%) than in nebuliser group (14.3%) (p less than 0.01, 95% confidence interval 2.4 to 19.1%). Tachycardia caused two withdrawals from intravenous group; non-response caused three withdrawals from nebuliser group. CONCLUSIONS: Intravenous salbutamol is more effective than nebulised salbutamol in acute severe asthma but may have unacceptable cardiovascular effects.

A soccer player with a sore toe: recognizing subungual osteochondromas.

In contrast to usual osteochondromas, subungual osteochondromas may arise following trauma rather than from aberrations in bone development. The rarity of subungual osteochondromas frequently leads to misdiagnosis and undertreatment, especially as a sports injury. When patients have nail-bed bruising, x-rays can rule out or confirm bone involvement, as in our case of a 20-year-old soccer player who had increasing pain in his left great toe. Treatment consists of radical, anatomic resection of symptomatic osteochondromas with periosteal realignment to prevent recurrence. Functional outcome after resection is excellent.

Cancer stem-like cells from head and neck cancers are chemosensitized by the Wnt antagonist, sFRP4, by inducing apoptosis, decreasing stemness, drug resistance and epithelial to mesenchymal transition.

Cancer stem cells (CSCs) of head and neck squamous cell carcinoma (HNSCC) are defined by high self-renewal and drug refractory potential. Involvement of Wnt/ß-catenin signaling has been implicated in rapidly cycling cells such as CSCs, and inhibition of the Wnt/ß-catenin pathway is a novel approach to target CSCs from HNSCC. In this study, we found that an antagonist of FrzB/Wnt, the secreted frizzled-related protein 4 (sFRP4), inhibited the growth of CSCs from two HNSCC cell lines, Hep2 and KB. We enriched the CD44(+) CSC population, and grew them in spheroid cultures. sFRP4 decreased the proliferation and increased the sensitivity of spheroids to a commonly used drug in HNSCC, namely cisplatin. Self-renewal in sphere formation assays decreased upon sFRP4 treatment, and the effect was reverted by the addition of Wnt3a. sFRP4 treatment of spheroids also decreased ß-catenin, confirming its action through the Wnt/ß-catenin signaling pathway. Quantitative PCR demonstrated a clear decrease of the stemness markers CD44 and ALDH, and an increase in CD24 and drug-resistance markers ABCG2 and ABCC4. Furthermore, we found that after sFRP4 treatment, there was a reversal in the expression of epithelial to mesenchymal (EMT) markers with the restoration of the epithelial marker E-cadherin, and depletion of EMT-specific markers twist, snail and N-cadherin. This is the first report demonstrating that the naturally occurring Wnt inhibitor, sFRP4, can be a potential drug to destroy CSC-enriched spheroids from HNSCCs. The repression of EMT and the decrease in stemness profile further strengthen the use of sFRP4 as a potent therapeutic against CSCs.

MEK inhibition affects STAT3 signaling and invasion in human melanoma cell lines.

Elevated activity of the mitogen-activated protein kinase (MAPK) signaling cascade is found in the majority of human melanomas and is known to regulate proliferation, survival and invasion. Current targeted therapies focus on decreasing the activity of this pathway; however, we do not fully understand how these therapies impact tumor biology, especially given that melanoma is a heterogeneous disease. Using a three-dimensional (3D), collagen-embedded spheroid melanoma model, we observed that MEK and BRAF inhibitors can increase the invasive potential of ∼20% of human melanoma cell lines. The invasive cell lines displayed increased receptor tyrosine kinase (RTK) activity and activation of the Src/FAK/signal transducers and activators of transcription-3 (STAT3) signaling axis, also associated with increased cell-to-cell adhesion and cadherin engagement following MEK inhibition. Targeting various RTKs, Src, FAK and STAT3 with small molecule inhibitors in combination with a MEK inhibitor prevented the invasive phenotype, but only STAT3 inhibition caused cell death in the 3D context. We further show that STAT3 signaling is induced in BRAF-inhibitor-resistant cells. Our findings suggest that MEK and BRAF inhibitors can induce STAT3 signaling, causing potential adverse effects such as increased invasion. We also provide the rationale for the combined targeting of the MAPK pathway along with inhibitors of RTKs, SRC or STAT3 to counteract STAT3-mediated resistance phenotypes.

Towards a consensus on a hearing preservation classification system.

CONCLUSION: The comprehensive Hearing Preservation classification system presented in this paper is suitable for use for all cochlear implant users with measurable pre-operative residual hearing. If adopted as a universal reporting standard, as it was designed to be, it should prove highly beneficial by enabling future studies to quickly and easily compare the results of previous studies and meta-analyze their data. OBJECTIVES: To develop a comprehensive Hearing Preservation classification system suitable for use for all cochlear implant users with measurable pre-operative residual hearing. METHODS: The HEARRING group discussed and reviewed a number of different propositions of a HP classification systems and reviewed critical appraisals to develop a qualitative system in accordance with the prerequisites. RESULTS: The Hearing Preservation Classification System proposed herein fulfills the following necessary criteria: 1) classification is independent from users' initial hearing, 2) it is appropriate for all cochlear implant users with measurable pre-operative residual hearing, 3) it covers the whole range of pure tone average from 0 to 120 dB; 4) it is easy to use and easy to understand.