Altered microbial bile acid metabolism exacerbates T cell-driven inflammation during graft-versus-host disease.

Microbial transformation of bile acids affects intestinal immune homoeostasis but its impact on inflammatory pathologies remains largely unknown. Using a mouse model of graft-versus-host disease (GVHD), we found that T cell-driven inflammation decreased the abundance of microbiome-encoded bile salt hydrolase (BSH) genes and reduced the levels of unconjugated and microbe-derived bile acids. Several microbe-derived bile acids attenuated farnesoid X receptor (FXR) activation, suggesting that loss of these metabolites during inflammation may increase FXR activity and exacerbate the course of disease. Indeed, mortality increased with pharmacological activation of FXR and decreased with its genetic ablation in donor T cells during mouse GVHD. Furthermore, patients with GVHD after allogeneic hematopoietic cell transplantation showed similar loss of BSH and the associated reduction in unconjugated and microbe-derived bile acids. In addition, the FXR antagonist ursodeoxycholic acid reduced the proliferation of human T cells and was associated with a lower risk of GVHD-related mortality in patients. We propose that dysbiosis and loss of microbe-derived bile acids during inflammation may be an important mechanism to amplify T cell-mediated diseases.

Customization of a DADA2-based pipeline for fungal internal transcribed spacer 1 (ITS1) amplicon data sets.

Identification and analysis of fungal communities commonly rely on internal transcribed spacer-based (ITS-based) amplicon sequencing. There is no gold standard used to infer and classify fungal constituents since methodologies have been adapted from analyses of bacterial communities. To achieve high-resolution inference of fungal constituents, we customized a DADA2-based pipeline using a mix of 11 medically relevant fungi. While DADA2 allowed the discrimination of ITS1 sequences differing by single nucleotides, quality filtering, sequencing bias, and database selection were identified as key variables determining the accuracy of sample inference. Due to species-specific differences in sequencing quality, default filtering settings removed most reads that originated from Aspergillus species, Saccharomyces cerevisiae, and Candida glabrata. By fine-tuning the quality filtering process, we achieved an improved representation of the fungal communities. By adapting a wobble nucleotide in the ITS1 forward primer region, we further increased the yield of S. cerevisiae and C. glabrata sequences. Finally, we showed that a BLAST-based algorithm based on the UNITE+INSD or the NCBI NT database achieved a higher reliability in species-level taxonomic annotation compared with the naive Bayesian classifier implemented in DADA2. These steps optimized a robust fungal ITS1 sequencing pipeline that, in most instances, enabled species-level assignment of community members.

Rethinking the need for a platelet transfusion threshold of 50 × 109 /L for lumbar puncture in cancer patients.

BACKGROUND: Lumbar puncture (LP) is a frequently performed diagnostic and therapeutic procedure in oncology patients. Transfusing to a minimum preprocedural platelet threshold of 50 × 109 /L is widely upheld without good quality evidence. The objective was to compare the outcomes of LPs performed with platelets above and below this threshold. An increased risk of adverse events in patients with lower platelet counts was not expected. As a corollary, transfusion reaction rates incurred by transfusing to this recommended threshold are also reported. METHODS: A total of 2259 LPs performed on 1137 oncology patients (adult, n = 871, and pediatric, n = 266) were retrospectively analyzed between February 2011 and December 2017. The incidence of LP-related complications for groups above and below the minimum platelet threshold was compared. Traumatic tap was defined as 500 or more red blood cells per high-power field in the cerebral spinal fluid. Groups were compared using the 2-Proportion Z-test and Fisher exact test. RESULTS: At time of LP, the total number of events with platelets less than 50 × 109 /L and 50 × 109 /L or greater were 110 and 2149, respectively. There were no significant differences in LP-associated complications between patients with platelet counts above or below 50 × 109 /L (P = .29). Patients with a pre-LP platelet count of less than 50 × 109 /L had a higher proportion of traumatic taps (P < .001). Three patients developed transfusion-related adverse events. CONCLUSION: Patients with platelet counts less than 50 × 109 /L did not have a higher incidence of clinically significant post-lumbar puncture complications (P = .29).

IPEM code of practice for high-energy photon therapy dosimetry based on the NPL absorbed dose calibration service.

The 1990 code of practice (COP), produced by the IPSM (now the Institute of Physics and Engineering in Medicine, IPEM) and the UK National Physical Laboratory (NPL), gave instructions for determining absorbed dose to water for megavoltage photon (MV) radiotherapy beams (Lillicrap et al 1990). The simplicity and clarity of the 1990 COP led to widespread uptake and high levels of consistency in external dosimetry audits. An addendum was published in 2014 to include the non-conventional conditions in Tomotherapy units. However, the 1990 COP lacked detailed recommendations for calibration conditions, and the corresponding nomenclature, to account for modern treatment units with different reference fields, including small fields as described in IAEA TRS483 (International Atomic Energy Agency (IAEA) 2017, Vienna). This updated COP recommends the irradiation geometries, the choice of ionisation chambers, appropriate correction factors and the derivation of absorbed dose to water calibration coefficients, for carrying out reference dosimetry measurements on MV external beam radiotherapy machines. It also includes worked examples of application to different conditions. The strengths of the 1990 COP are retained: recommending the NPL2611 chamber type as secondary standard; the use of tissue phantom ratio (TPR) as the beam quality specifier; and NPL-provided direct calibration coefficients for the user's chamber in a range of beam qualities similar to those in clinical use. In addition, the formalism is now extended to units that cannot achieve the standard reference field size of 10 cm × 10 cm, and recommendations are given for measuring dose in non-reference conditions. This COP is designed around the service that NPL provides and thus it does not require the range of different options presented in TRS483, such as generic correction factors for beam quality. This approach results in a significantly simpler, more concise and easier to follow protocol.

Clinical Performance of (1,3) Beta-D Glucan for the Diagnosis of Pneumocystis Pneumonia (PCP) in Cancer Patients Tested With PCP Polymerase Chain Reaction.

BACKGROUND: Serum (1,3)-beta-D glucan (BDG) is increasingly used to guide the management of suspected Pneumocystis pneumonia (PCP). BDG lacks specificity for PCP, and its clinical performance in high-risk cancer patients has not been fully assessed. Polymerase chain reaction (PCR) for PCP detection is highly sensitive, but cannot differentiate between colonization and infection. We evaluated the diagnostic performance of serum BDG in conjunction with PCP PCR on respiratory samples in patients with cancer and unexplained lung infiltrates. METHODS: We performed a retrospective analysis of adult patients evaluated for PCP at our institution from 2012 to 2015, using serum BDG and PCP PCR. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the serum BDG at different thresholds were evaluated using PCP PCR alone or in conjunction with clinical presentation in PCP PCR-positive patients. RESULTS: With PCP PCR alone as the reference method, BDG (≥80 pg/mL) had a sensitivity of 69.8%, specificity of 81.2%, PPV of 34.6%, and NPV of 95.2% for PCP. At ≥200 pg/mL in patients with a positive PCR and a compatible PCP clinical syndrome, BDG had a sensitivity of 70%, specificity of 100%, PPV of 100%, and NPV of 52.0% for PCP. CONCLUSIONS: Patients negative by both BDG and PCR were unlikely to have PCP. In patients with a compatible clinical syndrome for PCP, higher BDG values (>200 pg/mL) were consistently associated with clinically-significant PCP infections among PCP PCR-positive oncology patients.

IPEM topical report 1: guidance on implementing flattening filter free (FFF) radiotherapy.

Flattening filter free (FFF) beams are now commonly available with new standard linear accelerators. These beams have recognised clinical advantages in certain circumstances, most notably the reduced beam-on times for high dose per fraction stereotactic treatments. Therefore FFF techniques are quickly being introduced into clinical use. The purpose of this report is to provide practical implementation advice and references for centres implementing FFF beams clinically. In particular UK-specific guidance is given for reference dosimetry and radiation protection.

Using evaluation to enhance educational support for dental teams in the UK.

Each United Kingdom regional postgraduate deanery offers courses and educational support for the NHS (National Health Service) primary care dental team, primarily through networks of tutors (dental educators). After reporting literature on effective continuing education and the role of evaluation, this article provides an analysis of five educational evaluations (2005-08), identifying key messages and distilling ways to enhance educational support for the UK dental team. These evaluations adopted case study design and principally employed interviews (n=51) and questionnaires (n=221). The studies' key messages and recommendations are as follows: clarify dental educator roles; provide strategic management and educational support for educators; ensure continuing education is matched to the learning needs of dental teams and specific groups; and use practice (office) visits to support practice (team) development plans. Evaluation enables initiatives to be evidence-informed, focus effort where most needed, argue for continued funding, and give voice to stakeholders. However, there is added value in a collective consideration of evaluation findings from related studies. Doing this has identified ways to enhance the strategic development of educational support for the UK dental team and is applicable in a wider context both nationally and internationally.

Microvascular blood flow and skin temperature changes in the fingers following a deep nspiratory gasp.

The aims of this study were to quantify the changes in finger pulp skin temperature, laser Doppler flow (LDF, microvascular flux) and photoplethysmogram (PPG, microvascular blood volume pulsatility), induced by a deep inspiration in healthy subjects, and to investigate the repeatability of these responses within a measurement session and between measurement sessions on separate days. A system comprising an electronic thermometer, a laser Doppler flowmeter and a PPG amplifier measured simultaneous vasoconstrictor responses to a deep inspiratory gasp from three adjacent fingers of one hand. Clearly defined responses were obtained in 15 of the 17 subjects studied. Skin temperature fell in all of these subjects after each gasp, with a median fall of 0.089 degrees C (P < 0.001). The median value of LDF flux reduction was 93% (P < 0.001) indicating a momentary almost complete shut-down of microvascular blood flow; and PPG also showed a large response relative to pulse amplitude of 2.6 (P < 0.001). The median times for waveforms to reach their minimum were 4.6 s (PPG), 6.3 s (LDF) and 29.1 s (skin pulp temperature), with median delays between minima of LDF and PPG of 1.6 s (P < 0.001) and skin temperature and PPG of 23.5 s (P < 0.001). The vascular responses of skin temperature, LDF and PPG to an inspiratory gasp were repeatable, with temperature change repeatable to within 10% of the median subject change.

Tissue changes adjacent to titanium plates in patients.

INTRODUCTION: Titanium miniplates are widely used for osteosynthesis in maxillofacial surgery. Titanium is considered to be well tolerated but the long-term effects of titanium retained within human tissues are unclear. AIMS: This study was designed to evaluate histomorphologically the soft tissues adjacent to titanium maxillofacial miniplates and screws in patients, and to determine the nature of pigmented, particulate debris found in the tissues. MATERIALS: Thirty-five soft tissue specimens were excised from the tissues adjacent to titanium miniplates which had been in situ for between 1 month and 13 years. METHODS: All of the soft tissue specimens were prepared for examination under the light microscope. Four specimens were examined under the scanning electron microscope and the transmission electron microscope. Energy dispersive X-ray analysis (EDX) was used to confirm the elemental composition of the particles under investigation. RESULTS: All of the soft tissues showed fibrosis. Pigmented debris was present in 70% of the specimens and titanium was identified by EDX analysis. The debris was predominantly extra-cellular and was not associated with any inflammatory response or giant cell reaction. Fibroblasts were the predominant cell with small aggregates of lymphocytes and scattered macrophages. CONCLUSION: Titanium is apparently well tolerated for up to 13 years.

Are Polish and Swedish dental graduates adequately prepared for dental practice in the UK? A discussion of the transferability of general dental practitioners in Europe.

In the context of European Union harmonization, this article compares the systems of training for general dental practice in three European countries, the UK, Sweden and Poland. A UK perspective is adopted and the question as to whether dentists who have qualified in Poland or Sweden are adequately prepared for practice in the UK is explored. The paper is a result of discussion between providers of dental training in the three study countries. Key similarities, strengths and weaknesses are identified and issues pertinent to the transferability of general dental practitioners within Europe are raised.

Surgical management of premalignant lesions of the oral cavity with the CO2 laser / Manejo cirúrgico de lesões prémalignas da cavidade oral com o laser de CO²

The management of patients with premalignant and malignant lesions of the oral cavity can present problems. The potentially invasive nature of premalignant lesions together with their large extent influences the treatment. The common modalities of treatment of these lesions are surgical excision, cryotherapy, electrosurgery and radiotherapy. Recently, Carbon dioxide laser surgery has become available. Less pain, litle bleeding, minimal post-operative edema, reduced risk of infection, and low recurrence rates were advantages observed following Carbon dioxide laser surgery in the mouth when compared to other modalities of treatment. Healing following Carbon dioxide laser surgery progressed well with litle post-operative scarring and re-epithelization was complete after 4-6 weeks. The newly formed epithelium appeared normal and was soft on palpation

Mast cells in laser and surgical wounds / Mastócitos em feridas cirúrgicas e à laser

Precooling of tissues was investigated as a possible means of reducing thermal damage during CO² laser surgery of the oral mucosa. The changes in mast cells in scalpel, and in non-cooled and precooled (tissue temperature lowered to approximately l0ºC) CO² laser wounds were studied. Standard wounds five mm in length were created with the CO² laser or scalpel on the dorsum of the tongues of 32 Sprague-Dawley rats under general anesthesia with fentanyl/fluanisone and midazolam. Animals were killed with excess anesthetic immediately or six hours after surgery, their tongues were removed, trimmed, fixed in neutral formalin and processed to paraffin wax. Acid (pH 1.4) toluidine blue stained sections were used to count normal and degranulated mast cells in five fields (0.1mm²) located at defined positions immediately adjacent to the wound site. At both 0 and 6 hours normal mast cell numbers were significantly different between treament groups (P<0.045; ANOVA) with mean numbers highest in scalpel wounds and lowest in uncooled laser wounds. Similary, at 0 time, there were significant differences in degranulated mast cells between treatment groups (P=0.004; ANOVA) but highest numbers were detected in uncooled laser wounds and lowest in scalpel wounds. There were no significant differences in degranulated mast cell counts six hours although there was a similar distribution in numbers between groups. Total numbers of mast cells (normal + degranulated) did not differ between treatment groups. These results demonstrated that i) laser wounds are associated with greater levels of mast cell degranulation than scalpel wounds and ii) precooling of tissues prior to laser treatment decreases the level of mast cell degranulation. It is concluded that tissue damage in CO² laser surgery may be reduced by precooling of tissue

An audit of CO2 laser surgery in the mouth / Avaliação do laser CO2 cirurgico de boca

The use of the carbon dioxide laser to perform surgical procedures in the oral cavity has been described the last few years. This study involved the clinical audit of 83 patients who had undergone laser surgery in the oral cavity during 1985 to 1990 in Birmingham, England. The diagnosis of the 83 lesions treated were benign, premalignant or malignant. The most common sites of the lesions were the floor of the mouth/ventral surface of the tongue and other parts of the tongue, followed by upper alveolus and/or palate, angle of the mouth and/or cheeks, lower alveolus, and lips. The resuts showed that the use of carbon dioxide laser in treating oral soft-tissue pathology presented advantages over conventional techniques. Although trismus, numbness and early signs of infection were observed after surgery, local discomfort and pain were the most common complaints after laser surgery. The carbon dioxide laser does not offer any enhanced cure-rate for oral pathology, but rather it is a precise means of removing soft tissue lesions in selected patients with little upset afterwards

Laser em Odontologia / Lasers in Dentistry

Os autores apresentam um trabalho de atualização sobre o uso clínico do "laser" na Odontologia, com documentação fotográfica e especificações por especialidade

O uso do laser CO2 no tratamento de lesöes de tecidos moles da cavidade oral / The use of the CO2 laser in the treatment of injuries of soft tissue oral cavity

O uso do lase CO2 em cirurgia buco-maxilo-facial tem sido largamente descrito nos últimos cinco anos. Neste estudo, 70 pacientes com diversos tipos de patologias da cavidade oral tratados com o laser CO2 no período de 1985 a 1990, em Birminghan (Inglaterra), tiveram seus prontuários revistos. Os resultados deste estudo mostram que o uso do laser CO2 no tratamento de patologias da cavidade oral apresenta algumas vantagens sobre as técnicas convencionais. A efetiva reduçäo das complicaçöes pré e pós-operatórias resultaram em um menor período de internaçäo e reduçäo de custos. Desconforto local e dor moderada ou severa foram as queixas mais freqüentes após as cirurgias. Embora um caso de trismo, um caso de aprestesia transitória do lábio inferior e dois casos de infecçäo terem sido observados, parece-nos improvável que estas complicaçöes tenham sido um resultado direto da irradiaçäo laser. Esses resultados indicam que o uso do laser CO2 na cavidade oral é muito promissor, porém estudo adicionais säo ainda necessários, principalmente no tocante a respostas teciduais e procedimentos cirúrgicos envolvendo tecido ósseo