Immobilised teicoplanin does not demonstrate antimicrobial activity against Staphylococcus aureus.

Antibacterial bone biomaterial coatings appeal to orthopaedics, dentistry and veterinary medicine. Achieving the successful, stable conjugation of suitable compounds to biomaterial surfaces is a major challenge. A pragmatic starting point is to make use of existing, approved antibiotics which are known to remain functional in a stationary, immobilised state. This includes the macrocyclic glycopeptide, teicoplanin, following the discovery, in the 1990's, that it could be used as a chiral selector in chromatographic enantiomeric separations. Importantly teicoplanin works at the level of the bacterial cell wall making it a potential candidate for biomaterial functionalisations. We initially sought to functionalise titanium (Ti) with polydopamine and use this platform to capture teicoplanin, however we were unable to avoid the natural affinity of the antibiotic to the oxide surface of the metal. Whilst the interaction between teicoplanin and Ti was robust, we found that phosphate resulted in antibiotic loss. Before contemplating the covalent attachment of teicoplanin to Ti we examined whether a commercial teicoplanin stationary phase could kill staphylococci. Whilst this commercially available material could bind N-Acetyl-L-Lys-D-Ala-D-Ala it was unable to kill bacteria. We therefore strongly discourage attempts at covalently immobilising teicoplanin and/or other glycopeptide antibiotics in the pursuit of novel antibacterial bone biomaterials.

Oncoplastic breast surgery: A guide to good practice.

Oncoplastic Breast Surgery has become standard of care in the management of Breast Cancer patients. These guidelines written by an Expert Advisory Group; convened by the Association of Breast Surgery (ABS) and the British Association of Plastic, Reconstructive and Aesthetic Surgeons (BAPRAS), are designed to provide all members of the breast cancer multidisciplinary team (MDT) with guidance on the best breast surgical oncoplastic and reconstructive practice at each stage of a patient's journey, based on current evidence. It is hoped they will also be of benefit to the wide range of professionals and service commissioners who are involved in this area of clinical practice.

Population-based study of the sensitivity of axillary ultrasound imaging in the preoperative staging of node-positive invasive lobular carcinoma of the breast.

BACKGROUND: Preoperative staging of the axilla is important to allow decisions regarding neoadjuvant treatment and the management of the axilla. Invasive lobular carcinoma metastases are difficult to detect because of the infiltrative pattern of the nodal spread. In this study the sensitivity of preoperative axillary staging between invasive lobular (ILC) and ductal (IDC) carcinoma was compared. METHODS: All women diagnosed with pure ILC or IDC in the West of Scotland in 2012-2014 were identified from a database maintained prospectively within the Managed Clinical Network. Pretreatment axillary ultrasound imaging (AUS), core biopsy and fine-needle aspiration cytology (FNAC) results were compared between ILC and IDC. RESULTS: Some 602 women with ILC and 4199 with IDC had undergone axillary surgery, of whom 209 and 1402 respectively had nodal metastases. Pretreatment AUS sensitivity was significantly lower in ILC than in IDC (32·1 versus 50·1 per cent respectively, P < 0·001; OR 0·47, 95 per cent c.i. 0·34 to 0·64). Core biopsy had equally high sensitivity of 86 per cent in both subtypes; however, FNAC was significantly less sensitive in both ILC (55 per cent; P = 0·003) and IDC (75·6 per cent; P = 0·006). Multivariable analysis revealed that cT3-4 status and symptomatic presentation were both significant in predicting nodal metastasis in patients with ILC and false-negative AUS findings (OR 3·77, 95 per cent c.i. 1·69 to 8·42, P = 0·001; and OR 1·92, 1·24 to 2·98, P = 0·003, respectively). CONCLUSION: AUS is inferior in detecting axillary node metastasis in ILC compared with IDC. Women with cT3-4 lobular carcinoma may benefit from ultrasound-guided axillary biopsy regardless of the ultrasonographic appearance of the nodes.

Alkaline phosphatase binds tenaciously to titanium; implications for biological surface evaluation following bone implant retrieval.

Enhancing the performance and longevity of titanium (Ti) implants continues to be a significant developmental theme in contemporary biomaterials design. Our specific focus pertains to the surface functionalisation of Ti using the bioactive lipid, lysophosphatidic acid (LPA) and certain phosphatase-resistant analogues of LPA. Coating survivorship to a plethora of testing regimens is required to align with due regulatory process before novel biomaterials can enter clinical trials. One of the key acceptance criteria is coating retention to the physical stresses experienced during implantation. In assessing coating stability to insertion into porcine bone we found that a subsequent in vitro assessment to confirm coating persistence was masked by abundant alkaline phosphatase (ALP) contamination adsorbed to the metal surface. Herein we report that ALP can bind to Ti in a matter of minutes by simply immersing Ti samples in aqueous solutions of the enzyme. We strongly discourage the in vitro monitoring of osteoblast and stromal cell ALP expression when assessing bioactive coating survivorship following Ti implant retrieval form native bone tissue.

Oncoplastic breast conservation surgery is oncologically safe when compared to wide local excision and mastectomy.

BACKGROUND: Support for the oncological safety of oncoplastic breast conservation surgery (OBCS) is mostly based on evidence comparing recurrence rates after OBCS to wide local excision (WLE). However, OBCS is often indicated for larger cancers and oncological results should also be compared to patients treated with mastectomy. In this study we compared recurrence and survival following OBCS, mastectomy and WLE. METHODS: Patients treated with OBCS between 2009 and 2012 were identified from a prospectively maintained database. For comparison, consecutive patients treated with WLE or mastectomy with or without immediate reconstruction (Ms ± IR) over the same time period were identified. Histological variables of patients were compared using Fisher Exact or Chi squared tests, and recurrence and survival were compared using Kaplan-Meier and Cox regression survival analysis. RESULTS: 980 patients' data were analysed (OBCS: n = 104; WLE: n = 558; Ms ± IR: n = 318). Tumour size, grade, nodal status, ER, and PR expression of patients treated with OBCS were all significantly more adverse compared with patients treated with WLE (p < 0.001). These histological variables were similar in patients treated with Ms ± IR and OBCS. 5-year local recurrence rates were similar in all three groups (WLE: 3.4 per cent, OBCS: 2 per cent, Ms ± IR: 2.6 per cent; log rank = 0.973), while distant recurrence rates were higher after Ms ± IR and OBCS (Ms ± IR:13.1 per cent, OBCS:7.5 per cent, WLE:3.3 per cent; log rank: p < 0.001). CONCLUSION: OBCS is oncologically safe in patients even when histological results are similar to patients treated with Ms ± IR.

High-Grade Myxoid Liposarcoma (Round Cell Variant) in a Dog.

A 10-year-old, neutered male, Basset hound had a 26 × 21 × 21 cm infiltrative mass on the left abdominal wall that did not extend into the peritoneal cavity based on radiographs and abdominal computed tomography. Cytological examination revealed moderate numbers of neoplastic round cells, which frequently contained numerous round, clear, cytoplasmic vacuoles. Histologically, the tumour was composed of two morphologically distinct cell populations forming a continuum of heterogeneously differentiated cells. The primary spindle-shaped population formed streams with abundant, lightly eosinophilic, alcian blue-positive, myxoid matrix. The second population was arranged in sheets and had a round cell appearance. Scattered within both populations were neoplastic cells containing variably sized, intracytoplasmic, osmium tetroxide-positive vacuoles (lipid). Multifocal large pools of mucin formed pseudocysts, and numerous small capillaries were present throughout the neoplasm. According to the current World Health Organization veterinary classification of liposarcomas, this neoplasm had morphological features of both the myxoid and pleomorphic variants of liposarcoma; however, it was analogous to the recently defined high-grade myxoid liposarcoma in man. Myxoid liposarcoma with round cells has not been described previously in dogs. This case highlights the need to potentially re-evaluate the current classification of liposarcomas in animals.

Chitinase 3-like 1 expression by human (MG63) osteoblasts in response to lysophosphatidic acid and 1,25-dihydroxyvitamin D3.

Chitinase 3-like 1, otherwise known as YKL-40, is a secreted glycoprotein purported to have a role in extracellular matrix metabolism. The first mammalian cell type found to express YKL-40 was the human osteosarcoma-derived osteoblast, MG63. In that first study the active vitamin D3 metabolite, 1,25-dihydroxycholecalciferol (1,25D), stimulated YKL-40 expression, thereby indicating that a vital factor for skeletal health promoted YKL-40 synthesis by bone forming cells. However, when these MG63 cells were exposed to 1,25D they were also exposed to serum, a rich source of the pleiotropic lipid mediator, lysophosphatidic acid (LPA). Given that 1,25D is now known to co-operate with selected growth factors, including LPA, to influence human osteoblast differentiation we hypothesised that 1,25D and LPA may work together to stimulate osteoblast YKL-40 expression. Herein we report that 1,25D and LPA synergistically promote YKL-40 expression by MG63 cells. Inhibitors targeting AP1, MEK, Sp1 and STAT3 blunted the expression of both alkaline phosphatase and YKL-40 by MG63 cells in response to co-stimulation with 1,25D and LPA. Other ligands of the vitamin D receptor also co-operated with LPA in driving YKL-40 mobilisation. Collectively our findings highlight another important role of 1,25D and LPA in the regulation of human osteoblast function.

How to compare the oncological safety of oncoplastic breast conservation surgery - To wide local excision or mastectomy?

BACKGROUND: Comparative studies suggest that patients treated with oncoplastic breast conservation surgery (OBCS) have similar pathology to patients treated with wide local excision (WLE). However, patients treated with OBCS have never been compared to patients treated with mastectomy. The aim of this study was to identify which control group was comparable to patients undergoing OBCS. METHODS: Commonly reported histopathological variables of patients treated with OBCS, WLE or mastectomy ± immediate reconstruction (Ms ± IR) were compared using Fisher Exact or Chi squared tests. RESULTS: 1000 patients' data were analysed (OBCS: n = 119; WLE: n = 600; Ms ± IR: n = 281). Tumour size was significantly bigger after OBCS than WLE (p < 0.001), but similar to Ms ± IR (p = 0.138). Tumour grade was higher after OBCS than WLE (p < 0.001), but similar to Ms ± IR (p = 0.497). More axillary nodes were involved in patients with OBCS than WLE (p < 0.001), but comparable to Ms ± IR (p = 0.175). ER and PR expressions were lower after OBCS compared to WLE (p = 0.007, p = 0.009), but identical to Ms ± IR (p = 1, p = 0.904 respectively). Differences in application of systemic (neo)adjuvant therapy followed the above trend. CONCLUSION: Striking similarities found between OBCS and mastectomy patients' histopathological results are in sharp contrast with previously published data. This study suggests that oncological outcomes following OBCS should be compared to mastectomy besides WLE.

Immunohistochemical expression of ionized calcium binding adapter molecule 1 in cutaneous histiocytic proliferative, neoplastic and inflammatory disorders of dogs and cats.

Ionized calcium-binding adapter molecule 1 (Iba1) has been used widely as a marker for microglial cells and, recently, was also recognized as a 'pan-macrophage marker', as it is expressed by all subpopulations of cells of the monocyte/macrophage lineage. To determine the specificity of Iba1 as an immunohistochemical marker for canine and feline histiocytic proliferative, neoplastic and inflammatory disorders of the skin, we evaluated its expression in two types of histiocytic tumours, two non-neoplastic histiocytic proliferative conditions, one case of granulomatous dermatitis and four non-histiocytic tumours. Cells of the monocyte/macrophage lineage in all cases of canine cutaneous histiocytoma (9/9), reactive histiocytosis (9/9), histiocytic sarcomas (5/5), feline progressive dendritic cell histiocytosis (3/3) and macrophages in cutaneous mycobacteriosis (7/7) showed strong cytoplasmic expression of Iba1. Neoplastic cells of melanomas (10/10), lymphomas (7/7), mast cell tumours (7/7) and plasmacytomas (4/4) did not express Iba1. Iba1 is therefore a useful marker of cells of the monocyte/macrophage lineage in canine and feline inflammatory, proliferative and neoplastic conditions and can be used to identify these cells in formalin-fixed, paraffin wax-embedded tissues. Iba1 is not able to differentiate between macrophages and dendritic antigen presenting cells and expression does not allow classification of these histiocytic disorders.

Lymphocytic ganglioneuritis secondary to intervertebral disc extrusion in a dog.

This study presents a case of lymphocytic ganglioneuritis in a dog secondary to intervertebral disc extrusion that mimicked a peripheral nerve sheath tumour on magnetic resonance imaging. A four-year-old spayed female dachshund with lumbar pain was imaged via magnetic resonance. A tubular, space-occupying, contrast-enhancing lesion was noted in the right intervertebral foramen at L6 to L7. This was presumed to represent focal enlargement of the right sixth lumbar spinal nerve. A right-sided haemilaminectomy was performed at L6 to L7 and material that grossly resembled extruded nucleus pulposus was removed. The right L6 dorsal root ganglion, dorsal nerve root and proximal spinal nerve were severely enlarged and a partial thickness biopsy was collected from the dorsal root ganglion. Results of histopathological examination of the submitted tissue samples were consistent with extruded disc material and lymphocytic ganglioneuritis. To the author's knowledge, this is the first published report of lymphocytic ganglioneuritis secondary to intervertebral disc disease in a dog.

Design of a simplified histopathologic model for gastrointestinal inflammation in dogs.

Significant interobserver variability in the diagnostic interpretation of endoscopic gastrointestinal (GI) specimens exists even with the use of World Small Animal Veterinary Association (WSAVA) standardization criteria. Chi-square analyses compared the extent of pathologists' agreement for microarchitectural features of inflammation in endoscopic specimens obtained from 253 animals of the original WSAVA study. Patterns of agreement between pathologists were classified as broad (3/4 pathologists agreed), dichotomous (2/4 pathologists agreed), or divergent (no agreement between pathologists). The simplified model for GI inflammation was based on those parameters for which the pathologists had either broad or minimally divergent opinions of histopathologic significance. In this model, the parameters chosen were as follows: gastric parameters (intraepithelial lymphocytes [IELs], lamina propria [LP] infiltrates, and mucosal fibrosis), duodenal parameters (villus atrophy, epithelial injury, IELs, crypt changes, and LP infiltrates), and colonic parameters (epithelial injury, crypt dilation, fibrosis, LP infiltrates, and goblet cell depletion). Preliminary data using this simplified model showed excellent correlation between pathologists in defining the presence and extent of GI inflammation in dogs.

Practice leadership and active support in residential services for people with intellectual disabilities: an exploratory study.

BACKGROUND: We hypothesised that a key factor determining the quality of active support was 'practice leadership' - provided by the first-line manager to focus staff attention and develop staff skills in providing direct support to enable people with intellectual disabilities to have a good quality of life. This exploratory study focused on what levels of practice leadership were found and its role in explaining variation in active support. METHOD: Relevant aspects of management, including practice leadership, were assessed by questionnaires administered to staff in residential settings alongside observational measures of active support and resident engagement in meaningful activity. Relationships between these variables were explored using regression and post hoc group comparisons. RESULTS: There was wide variation, with average levels of practice leadership being low, though improving over the period studied. Practice leadership had a significant impact on active support, but was fully mediated by the effect of quality of management. When the quality of management was higher better practice leadership did produce a significant difference in active support. However, higher quality of management on its own did not produce better active support. CONCLUSIONS: A number of limitations are acknowledged and further research is required. Practice leadership appears to be an important factor in enabling staff to provide active support but as part of generally good management. Given the rather low levels found, attention needs to be given to the training, career development and support of practice leaders and also to how to protect their time from their many other responsibilities.

Effect of endoscopic forceps on quality of duodenal mucosal biopsy in healthy dogs.

BACKGROUND: Limited data exist regarding influence of endoscopic forceps on duodenal mucosal biopsy quality and adequacy for histologic examination/assessment in dogs. HYPOTHESIS/OBJECTIVES: Hypothesizing that larger forceps would procure superior specimens, we evaluated effect of 6 disposable forceps on duodenal biopsy weight, depth, crush artifact, and adequacy for histologic examination/assessment. ANIMALS: Seventeen healthy adult dogs. METHODS: Prospective study. Two operators each obtained 4 duodenal specimens from each dog with each forceps. Lightest sample discarded. One pathologist evaluated blindly other 3 specimens. A total of 612 specimens evaluated. Results analyzed by one-way ANOVA of forceps effects with dog as blocking factor. Posthoc pairwise comparisons examined with Tukey's test when indicated. RESULTS: Biopsies performed with large capacity forceps heavier (10.56 ± 0.90 and 11.6 ± 0.62 mg (mean ± SD) versus 5.55 ± 0.53 to 8.61 ± 0.49; P < .0001) and adequacy for histologic examination/assessment superior to standard oval and 'pediatric' (scores 2.52 ± 0.41 and 2.58 ± 0.37 versus 2.08 ± 0.33 and 2.14 ± 0.29; P < .0001). No statistically significant difference in depth scores. Large capacity forceps with spike associated with less crush artifact than all smaller forceps (scores 1.19 ± 0.16 versus 1.38 ± 0.21 to 1.52 ± 0.21; P < .0001). In same size forceps, presence of spike had no effect on crush artifact and adequacy for histologic examination/assessment (P < .0001). CONCLUSIONS AND CLINICAL IMPORTANCE: Large capacity forceps are superior, providing higher quality and greater numbers of samples achieving adequacy for histologic examination/assessment. Choice of endoscopic biopsy forceps for duodenal samples influences sample volume and diagnostic utility.

Differential interactions of Streptococcus gordonii and Staphylococcus aureus with cultured osteoblasts.

The impedance of normal osteoblast function by microorganisms is at least in part responsible for the failure of dental or orthopedic implants. Staphylococcus aureus is a major pathogen of bone, and exhibits high levels of adhesion and invasion of osteoblasts. In this article we show that the commensal oral bacterium Streptococcus gordonii also adheres to and is internalized by osteoblasts. Entry of S. gordonii cells had typical features of phagocytosis, similar to S. aureus, with membrane protrusions characterizing initial uptake, and closure of the osteoblast membrane leading to engulfment. The sensitivities of S. gordonii internalization to inhibitors cytochalasin D, colchicine and monensin indicated uptake through endocytosis, with requirement for actin accumulation. Internalization levels of S. gordonii were enhanced by expression of S. aureus fibronectin-binding protein A (FnBPA) on the S. gordonii cell surface. Lysosomal-associated membrane protein-1 phagosomal membrane marker accumulated with intracellular S. aureus and S. gordonii FnBPA, indicating trafficking of bacteria into the late endosomal/lysosomal compartment. Streptococcus gordonii cells did not survive intracellularly for more than 12 h, unless expressing FnBPA, whereas S. aureus showed extended survival times (>48 h). Both S. aureus and S. gordonii DL-1 elicited a rapid interleukin-8 response by osteoblasts, whereas S. gordonii FnBPA was slower. Only S. aureus elicited an interleukin-6 response. Hence, S. gordonii invades osteoblasts by a mechanism similar to that exhibited by S. aureus, and elicits a proinflammatory response that may promote bone resorption.

Natural Heterobilharzia americana infection in horses in Texas.

The schistosome Heterobilharzia americana infects dogs, raccoons, and other mammals in the southeastern United States. Migration of eggs into the liver results in parasitic granulomas with varying degrees of fibrosis and inflammation. Recently, hepatic parasitic granulomas in horses were shown to be caused by H. americana infection. In the present study, samples of liver from 11 of 12 horses with hepatic granulomas identified at necropsy (n = 11) or surgical biopsy (n = 1) were used for DNA extraction, polymerase chain reaction amplification and sequencing using primers specific for a portion of the H. americana small subunit ribosomal RNA gene. A polymerase chain reaction amplicon of the correct size was produced from the extracted DNA in 8 of the 11 horses. Amplicons from 5 of the 8 positive horses were sequenced and had 100% identity with H. americana. In all but 2 of the 12 horses, Heterobilharzia was not responsible for the primary clinical disease, and the hepatic granulomas were considered an incidental finding.

Utility of endoscopic biopsies of the duodenum and ileum for diagnosis of inflammatory bowel disease and small cell lymphoma in cats.

BACKGROUND: Endoscopic duodenal biopsies are relatively convenient, minimally invasive tests for infiltrative intestinal disorders of cats. Ileal endoscopic biopsies might not be performed because of technical difficulty and effort required to prepare the colon. It is not known whether or not histopathology of feline duodenal and ileal biopsies for detection of inflammatory bowel disease (IBD) and small cell lymphoma (SC-LSA) provides comparable results. OBJECTIVES: To evaluate the agreement between endoscopic biopsies of duodenum and ileum in cats with IBD and SC-LSA. ANIMALS: Seventy client-owned cats with gastrointestinal disease and adequate duodenal and ileal tissue biopsies obtained endoscopically. METHODS: Retrospective study: Search of medical records of cats with enteropathy and endoscopy. Samples were blinded and re-evaluated by single pathologist (JM) for quality, number of biopsies, and diagnosis according to WSAVA standards. Agreement of IBD and SC-LSA diagnoses among biopsy sites assessed using Cohen's Kappa. RESULTS: Eighteen of 70 cats (26%) were diagnosed with SC-LSA in duodenum, ileum, or both. Of these 18 cats, 7 (39%) were diagnosed with only duodenal SC-LSA, 8 (44%) were diagnosed with only ileal SC-LSA, and 3 (17%) had SC-LSA in both duodenum and ileum. There was poor agreement on diagnosis between duodenal and ileal biopsies (kappa = 0.23). CONCLUSIONS AND CLINICAL IMPORTANCE: Although review by a single pathologist remains a limitation of this study, results suggest that there is a population of cats in which diagnosis of SC-LSA can be found only by evaluation of ileal biopsies. Clinicians should consider performing both upper and lower GI endoscopic biopsies in cats with infiltrative small bowel disease.

Lysophosphatidic acid and calcitriol co-operate to promote human osteoblastogenesis: requirement of albumin-bound LPA.

Lysophosphatidic acid (LPA), a pleiotropic signalling lipid is assuming growing significance in osteoblast biology. Although committed osteoblasts from several mammalian species are receptive to LPA far less is known about the potential for LPA to influence osteoblast formation from their mesenchymal progenitors. An essential factor for both bone development and post-natal bone growth and homeostasis is the active metabolite of vitamin D3, calcitriol (D3). Previously we reported how a combination of LPA and D3 synergistically co-operated to enhance the differentiation of immature human osteoblasts. Herein we provide evidence for the formation of human osteoblasts from multiple, primary human bone marrow derived stromal (stem) cells (hBMSCs). Importantly osteoblast development from hBMSCs only occurred when LPA was administered as a complex with albumin, its natural carrier. Collectively our findings support a co-operative role of LPA and D3 in osteoblastogenesis, findings which may aid the development of novel treatment strategies for bone repair.

Epithelioid haemangiosarcoma in the ocular tissue of horses.

Haemangiosarcomas (HSAs) are malignant tumours of endothelial cell origin. Epithelioid HSA is a variant of the histologically conventional HSA that has little or no morphological evidence of a vascular origin and has been reported rarely in domestic animals. The following report documents six cases of equine epithelioid HSA occurring in the ocular tissues of horses with a mean age of 19.8 years at the time of diagnosis. Microscopically, all of the lesions consisted of solid sheets or cords of epithelioid cells with rare narrow clefts or small spaces containing erythrocytes that were often the only feature indicating a vascular origin. On immunohistochemistry, the neoplastic cells expressed vimentin, CD31 and factor VIII-related antigen, but not cytokeratin, indicating an endothelial nature.