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Objectives: The objectives were to clarify if intraoral ultrasonography (USG) is: (1) more accurate than conventional periodontal examinations in detection of furcation involvement, and (2) comparable to conventional periodontal examinations in accurate horizontal classification of furcation involvement in comparison to cone beam computed tomography (CBCT). Methods: The buccal furcation in 61 lower first molars were evaluated with conventional periodontal examinations, intraoral USG and CBCT. The presence and classification of the horizontal depth of furcation involvement were defined clinically by assessment with a Nabers periodontal probe and a periapical radiograph with reference to the bone loss under the fornix. The horizontal depth of furcation involvement was measured in intraoral USG and CBCT images. Based on the measurements, presence diagnosis and horizontal classification were performed. Results from conventional periodontal examinationsand intraoral USG were compared with those from CBCT. Results: κ value (κ) for agreement of presence diagnosis of furcation involvement between intraoral USG and CBCT was 0.792, while agreement with conventional periodontal examinations was 0.225. Diagnostic accuracy of intraoral USG exhibited higher values (sensitivity: 98.3%, accuracy: 98.4 %) than conventional periodontal examinations (81.4% and 81.9 %). Weighted κ statistics showed substantial agreement in the classification between intraoral USG and CBCT (κ = 0.674). High agreement (ICC: 0.914) for the measurement of horizontal depth of furcation involvement was found between intraoral USG and CBCT. Conclusions: Intraoral USG may be a reliable diagnostic tool for assessment of furcation involvement of mandibular molars with a similar performance to CBCT, but without ionizing radiation.

Cycling consists of alternating periods of reduced and normal fetal heart variability, reflecting changes in fetal behavioral states. Occurrence of active and quiet sleep cycles is considered to be a hallmark of fetal autonomic nervous system integrity, demonstrating healthy interaction between the parasympathetic and sympathetic nervous systems. Cycling is an overlooked feature in most international cardiotocography (CTG) guidelines. The authors tested the hypothesis that fetuses showing no cycling in the intrapartum period have poorer outcomes. To investigate whether the absence of cycling at the commencement of intrapartum fetal monitoring is associated with poorer neonatal outcomes (umbilical arterial cord pH, Apgar scores and neonatal unit admission). Analysis of a database of sequentially acquired intrapartum CTG traces from a single center. Only cases of singleton pregnancies over 36 weeks gestation in cephalic presentation with recorded umbilical artery cord pH were considered. Neonatal outcomes were assessed based on umbilical cord artery pH, Apgar ≤7 at 5 min and unexpected admission to the neonatal unit. Intrapartum pyrexia, presence of meconium-stained amniotic fluid and mode of delivery were also recorded. A total of 684 cases were analyzed. Absence of cycling from the beginning of the intrapartum CTG recording was noted in 5% of cases. Cases with no cycling were more likely to have maternal pyrexia (≥37.8 °C) ( = .006) and Apgars ≤7 at 5 min ( = .04). There was an association between increasing baseline fetal heart rate and the proportion of cases with no cycling. There was no significant difference between the two groups with regard to the mode of delivery or umbilical cord arterial pH

Introduction Surgical site infections (SSIs) are a cause of considerable morbidity and mortality in healthcare. Increasingly, closed-incision negative pressure wound therapy (ciNPWT) is being studied as a potential method of reducing incidence of SSI with conflicting results in the literature. Few studies however have looked at its use in the field of gynecological oncology. Objectives We aimed to compare the incidence of SSI when using ciNPWT dressings versus conventional dressings in gynecological oncology patients undergoing midline laparotomies. Methods This was a pilot study involving 14 patients receiving the ciNPWT dressing and 26 control patients. All patients were followed up for a period of 30 days. We used the American College of Surgeons (ACS) risk calculator to estimate each patient’s risk of SSI in order to risk stratify the groups. Results The incidence of wound infection was 21% (3/14) in the ciNPWT group and 23% (6/26) in the control group (p=0.886). The ciNPWT group was found to be at significantly higher risk for SSI as calculated by the ACS tool (8.8% ciNPWT, 6% control, p=0.004). After stratifying for this difference in risk, still no significant difference in incidence of SSI was found between the two groups (27% (3/11) ciNPWT, 29% (2/7) control p=0.929). Conclusion The incidence of SSI does not appear to decrease by the prophylactic use of the closed-incision negative pressure wound dressing.

Left ventricular mechanical dispersion (LVMD) is a novel speckle tracking parameter for prognostic assessment of arrhythmic risk prediction. There is growing evidence to support its use in a variety of cardiomyopathic processes. There is paucity of data addressing any presence of inter-vendor discrepancies for LVMD. The aim of this study was to assess inter-vendor variability of LVMD in vendor specific software (VSS) and vendor independent software (VIS) in subjects with preserved and reduced left ventricular function.Fifty-nine subjects (14 normal subjects and 45 subjects with cardiac disease) were recruited and 2D speckle tracking echocardiographic images were acquired on two different ultrasound machines (GE and Philips). LVMD was measured by two different VSS (EchoPac GE and QLAB Philips) and one VIS (TomTec Arena).There was significant bias and wide limits of agreement (LOA) in the overall cohort observed between two different VSS (17.6 ms; LOA: -29.6 to 64.8; r: .47). There was acceptable bias and narrower LOA with good agreement for LVMD between images obtained on different vendors when performed on VIS (-3.1 ms; LOA: -27.6 to 21.4; r: .75). QLAB LVMD was consistently higher than GE LVMD and TomTec LVMD in both preserved and reduced left ventricular function. LVMD measurements have high intra-vendor reproducibility with excellent inter and intra-observer agreement.There was acceptable bias and narrower LOA for LVMD assessment on a VIS. Inter-vendor variability exists for LVMD assessment between VSS. Serial measurements of LVMD should be performed using a single vendor for consistent and reliable results.

A simple method completely removes contaminating paraffin from samples for clinical Raman and enhanced immunohistological analysis., Incomplete removal of paraffin and organic contaminants from tissues processed for diagnostic histology has been a profound barrier to the introduction of Raman spectroscopic techniques into clinical practice. We report a route to rapid and complete paraffin removal from a range of formalin-fixed paraffin embedded tissues using super mirror stainless steel slides. The method is equally effective on a range of human and animal tissues, performs equally well with archived and new samples and is compatible with standard pathology lab procedures. We describe a general enhancement of the Raman scatter and enhanced staining with antibodies used in immunohistochemistry for clinical diagnosis. We conclude that these novel slide substrates have the power to improve diagnosis through anatomical pathology by facilitating the simultaneous combination of improved, more sensitive immunohistochemical staining and simplified, more reliable Raman spectroscopic imaging, analysis and signal processing.

Mitral regurgitation (MR) is a valvular heart disease associated with significant morbidity and mortality. Transcatheter mitral valve intervention (TMVI) repairs or replaces the mitral valve through small arterial and venous entry sites and so avoids risks associated with open heart surgery. Transcatheter devices targeting components of the mitral apparatus are being developed to repair or replace it. Numerous challenges remain including developing more adaptable devices and correction of multiple components of the mitral annulus to attain durable results. The mitral valve apparatus is a complex structure and understanding of the mechanisms of MR is essential in the development of TMVI. There will likely be a complementary role between surgery and TMVI in the near future.

Summary Animal models and archived human biobank tissues are useful resources for research in disease development, diagnostics and therapeutics. For the preservation of microscopic anatomical features and to facilitate long‐term storage, a majority of tissue samples are denatured by the chemical treatments required for fixation, paraffin embedding and subsequent deparaffinization. These aggressive chemical processes are thought to modify the biochemical composition of the sample and potentially compromise reliable spectroscopic examination useful for the diagnosis or biomarking. As a result, spectroscopy is often conducted on fresh/frozen samples. In this study, we provide an extensive characterization of the biochemical signals remaining in processed samples (formalin fixation and paraffin embedding, FFPE) and especially those originating from the anatomical layers of a healthy rat colon. The application of chemometric analytical methods (unsupervised and supervised) was shown to eliminate the need for tissue staining and easily revealed microscopic features consistent with goblet cells and the dense populations of cells within the mucosa, principally via strong nucleic acid signals. We were also able to identify the collagenous submucosa‐ and serosa‐ as well as the muscle‐associated signals from the muscular regions and blood vessels. Applying linear regression analysis to the data, we were able to corroborate this initial assignment of cell and tissue types by confirming the biological origin of each layer by reference to a subset of authentic biomolecular standards. Our results demonstrate the potential of using label‐free Raman microspectroscopy to obtain superior imaging contrast in FFPE sections when compared directly to conventional haematoxylin and eosin (H&E) staining.

Mitral regurgitation (MR) is a valvular heart disease associated with significant morbidity and mortality. Transcatheter mitral valve intervention (TMVI) repairs or replaces the mitral valve through small arterial and venous entry sites and so avoids risks associated with open heart surgery. Transcatheter devices targeting components of the mitral apparatus are being developed to repair or replace it. Numerous challenges remain including developing more adaptable devices and correction of multiple components of the mitral annulus to attain durable results. The mitral valve apparatus is a complex structure and understanding of the mechanisms of MR is essential in the development of TMVI. There will likely be a complementary role between surgery and TMVI in the near future.

The potential for Raman spectroscopy to provide early and improved diagnosis on a wide range of tissue and biopsy samples in situ is well documented. The standard histopathology diagnostic methods of reviewing H&E and/or immunohistochemical (IHC) stained tissue sections provides valuable clinical information, but requires both logistics (review, analysis and interpretation by an expert) and costly processing and reagents. Vibrational spectroscopy offers a complimentary diagnostic tool providing specific and multiplexed information relating to molecular structure and composition, but is not yet used to a significant extent in a clinical setting. One of the challenges for clinical implementation is that each Raman spectrometer system will have different characteristics and therefore spectra are not readily compatible between systems. This is essential for clinical implementation where classification models are used to compare measured biochemical or tissue spectra against a library training dataset. In this study, we demonstrate the development and validation of a classification model to discriminate between adenocarcinoma (AC) and non-cancerous intraepithelial metaplasia (IM) oesophageal tissue samples, measured on three different Raman instruments across three different locations. Spectra were corrected using system transfer spectral correction algorithms including wavenumber shift (offset) correction, instrument response correction and baseline removal. The results from this study indicate that the combined correction methods do minimize the instrument and sample quality variations within and between the instrument sites. However, more tissue samples of varying pathology states and greater tissue area coverage (per sample) are needed to properly assess the ability of Raman spectroscopy and system transferability algorithms over multiple instrument sites.

Despite the demonstrated potential as an accurate cancer diagnostic tool, Raman spectroscopy (RS) is yet to be adopted by the clinic for histopathology reviews. The Stratified Medicine through Advanced Raman Technologies (SMART) consortium has begun to address some of the hurdles in its adoption for cancer diagnosis. These hurdles include awareness and acceptance of the technology, practicality of integration into the histopathology workflow, data reproducibility and availability of transferrable models. We have formed a consortium, in joint efforts, to develop optimised protocols for tissue sample preparation, data collection and analysis. These protocols will be supported by provision of suitable hardware and software tools to allow statistically sound classification models to be built and transferred for use on different systems. In addition, we are building a validated gastrointestinal (GI) cancers model, which can be trialled as part of the histopathology workflow at hospitals, and a classification tool. At the end of the project, we aim to deliver a robust Raman based diagnostic platform to enable clinical researchers to stage cancer, define tumour margin, build cancer diagnostic models and discover novel disease bio markers.

Here we present Fourier transform infrared (FTIR) and Raman images of Steinernema kraussei nematode worms and, in conjunction with chemometric analysis. We also distinguished the biochemical differences associated with different regions of these images. The nematodes are complex, multicellular organisms that have a simple, but defined body plan with distinct digestive, muscular and reproductive systems. As such, nematodes have often been used as model organisms for the study of biological processes in higher organisms. We show that FTIR spectra, collected in transmission mode, contain information from the entire thickness of the nematode, but still exhibit biochemical differences reflecting different tissues or anatomical structures, such as the digestive tract. Raman spectroscopy, in contrast, can be used to investigate the distribution of biological molecules on one plane within a constrained depth of the nematode, allowing imaging of fine details within the body of the worm, including the distribution of lipid, protein and collagen. Together these vibrational spectroscopic techniques provide complementary, non-invasive and label-free information on the spatial distribution of biomolecules within multicellular organisms and, therefore, have potential future applications in developmental studies of such model organisms.

Over the last decade Raman spectroscopy has become established as a physicochemical technique for the rapid identification of microbes. This powerful analytical method generates a spectroscopic fingerprint from the microbial sample, which provides quantitative and qualitative information that can be used to characterize, discriminate and identify microorganisms, in both bacteria slurry and at the single-cell level. Recent developments in Raman spectroscopy have dramatically increased in recent years due to the enhancement of the signal by techniques including tip-enhanced Raman spectroscopy and coherent anti-Stokes Raman spectroscopy and due to the availability of user-friendly instrumentation and software. The result of this has been reduced cost and rapid collection time, and it has allowed the nonspecialist access to this physical sciences approach for biological applications. In this article, we will briefly explain the technique of Raman spectroscopy and discuss enhancement techniques, including the recent application of tip-enhanced Raman spectroscopy to microbiology, as well as the move towards rapid microbial identification with Raman spectroscopy. Furthermore, recent studies have combined Raman spectroscopy with microfluidic devices, giving greater control of sample conditions, which will no doubt have an important impact in the future development of Raman spectroscopy for microbial identification.

The skin offers a perfect model system for studying the wound healing cascade, which involves a finely tuned interplay between several cell types, pathways and processes. The dysregulation of these factors may lead to wound healing disorders resulting in chronic wounds, as well as abnormal scars such as hypertrophic and keloid scars. As the contribution of stem cells towards tissue regeneration and wound healing is increasingly appreciated, a rising number of stem cell therapies for cutaneous wounds are currently under development, encouraged by emerging preliminary findings in both animal models and human studies. However, we still lack an in-depth understanding of the underlying mechanisms through which stem cells contribute to cutaneous wound healing. The aim of this review is, therefore, to present a critical synthesis of our current understanding of the role of stem cells in normal cutaneous wound healing. In addition to summarizing wound healing principles and related key molecular and cellular players, we discuss the potential participation of different cutaneous stem cell populations in wound healing, and list corresponding stem cells markers. In summary, this review delineates current strategies, future applications, and limitations of stem cell-based or stem cell-targeted therapy in the management of acute and chronic skin wounds.

The neuroanaesthesia management of patients contributing to successful surgical intervention is described. The authors focus on the principles of preoperative optimization, intraoperative anaesthetic management and postoperative intensive care. Subarachnoid haemorrhage and resection of arteriovenous malformations are discussed in detail. The management of ischaemic stroke is dealt with in brief.

TERS (tip-enhanced Raman scattering) provides exceptional spatial resolution without any need for labelling and has become a versatile tool for biochemical analysis. Two examples will be highlighted here. On the one hand, TERS measurements on a single mitochondrion are discussed, monitoring the oxidation state of the central iron ion of cytochrome c, leading towards a single protein characterization scheme in a natural environment. On the other hand, a novel approach of single molecule analysis is discussed, again based on TERS experiments on DNA and RNA, further highlighting the resolution capabilities of this method.

Background To visualise and characterise skin architecture, the tissue usually has to be destroyed and labelled. Objectives The use of Fourier transform infrared (FTIR) spectroscopy as a label-free, minimally sample destructive method to define hair follicular structure has been explored and demonstrated in this paper. Methods Human scalp skin cryosections were imaged using FTIR microscopy and the data was subsequently analysed with N-FINDR spectral unmixing algorithm. Results This resulted in an excellent distinction of known hair follicle tissue layers, which could be discerned based on their molecular structure. Conclusion The development of a minimally sample-destructive, label-free spectroscopy based technique that can differentiate layers of cells in the dermal papilla and connective tissue sheath in the mesenchyme of the hair follicle paves the way forward to identifying spectral markers important in wound healing and stem cell therapies.

CD8 antiviral activity is a subject of intense studies. Two very critical anti-viral arms of HIVspecific CD8+ T cells (cytolytic and non-cytolytic) have been shown to play a significant role in protection against HIV and appear to associate with asymptomatic survival and slower disease progression. The cytotoxic CD8+ T cells (CTL) recognize and eliminate HIV-infected cells displaying MHC class-I proteins. Along with recognition and elimination of HIV-infected cells, CD8+ T cells potentially influence various stages in HIV life cycle. For instance the chemokines produced by CD8+ T cells block the entry of the virus into T cells and macrophages. The activated CD8+ T cells also produce soluble factors that act at the transcriptional level. In contrast, the non-cytolytic CD8+T cells from HIV+ individuals suppress virus replication in CD4+ T cells in vitro, involving interplay of soluble factors such as chemokines and other antiviral factors such as unidentified soluble CD8 Antiviral Factor (CAF), urokinase-type plasminogen activator (uPA) and antiviral membrane-bound factor. In light of these factors, it is clear that the cell-mediated immune responses are critical in controlling HIV replication in vivo. Although the constituents of antiviral activity of CD8+ T cells remain largely elusive, the restoration of this activity during Highly Active Antiretroviral Therapy (HAART) and IL-2 treatment confers significant improvement in antiviral activity. The aim of this chapter is to provide a comprehensive and unbiased snapshot of antiviral potential of these cells and mechanisms involved in conferring this protective activity in HIV+ individuals and its overall relevance in HIV disease.

Institute of Photonic Technology, Albert-Einstein-Str. 9, Jena 07745, Germany. Institute for Physical Chemistry, F riedrich-Schiller-University, Lessing Str. 10, Jena 07743, Germany. Centre for Biospectroscopy School of Chemistry Monash University Wel lington Rd. Clayton, 3800 Victoria, Australia Department of Dermatology, University of Lubeck, Ratzeburger Allee 160, 23562 Lubeck, Germany School of Translational Medicine, University of Manchester, Stopford Building, Oxford Road, Manchester M13 9PL, UK

Recent studies have clearly demonstrated that Raman and surface enhanced Raman scattering (SERS) spectroscopies are information rich, non-destructive techniques for the monitoring of subtle intracellular changes. However, despite the demonstrated and sophisticated applications of these techniques in cell studies there still remains a lack of accompanying 3D images. Herein we demonstrate for the first time combined 3D Raman and SERS imaging for the simultaneous confirmation of the cellular inclusion and multiple component detection of SERS nanotags. We also report on the 3D elucidation of the cell nuclei by multivariate analysis methods. Imaging in 3D will be critical to understanding architectural changes between diseased and healthy cells, and tissues. It will also provide non-destructive definitive proof of cellular uptake whilst simultaneously confirming targeting of SERS nanotags to their intended destinations.

Invasion of melanoma cells from the primary tumor involves interaction with adjacent tissues and extracellular matrix. The extent of this interaction is not fully understood. In this study Raman spectroscopy was applied to cryo-sections of established 3D models of melanoma in human skin. Principal component analysis was used to investigate differences between the tumor and normal tissue and between the peri-tumor area and the normal skin. Two human melanoma cells lines A375SM and C8161 were investigated and compared in 3D melanoma models. Changes were found in protein conformations and tryptophan configurations across the entire melanoma samples, in tyrosine orientation and in more fluid lipid packing only in tumor dense areas, and in increased glycogen content in the peri-tumor areas of melanoma. Raman spectroscopy revealed changes around the perimeter of a melanoma tumor as well as detecting differences between the tumor and the normal tissue.