Search

Apocrine adenoma is a rare benign epithelial tumour of the breast that can radiologically present as a well-defined opacity. The clinical, radiological and pathological findings of apocrine adenoma of the breast in a 72-year-old woman are described. Histologically, the tumour is characterised by a circumscribed proliferation of metaplastic apocrine cells which may contain calcifications. Pathological assessment of the lesion is essential in making the diagnosis.

The aims of this study were to assess the value of specimen radiographs of stereotactic core biopsy, the usefulness of measuring size of calcifications on tissue sections, whether demonstration of calcifications in tissue sections alters the pathological diagnosis when specimen radiograph demonstrates calcifications, and to correlate these assessments with diagnostic outcome. A total of 301 core biopsies from 266 women with 274 mammographically suspicious areas of calcifications were examined. Core biopsies (five cores per procedure) were obtained stereotactically using a 14-gauge needle in an automated Biopty gun. Prior to processing of the tissue, 214 core biopsy specimens from 193 women with 197 lesions were radiographed. Of the 301 core biopsies, 56 (19%) were diagnosed as malignant, 15 (5%) were diagnosed as atypical ductal hyperplasia and 230 (76%) contained benign breast tissue. Of the core biopsies diagnosed as benign, 160 (70%) had specimen radiography prior to processing. Of these, 109 (69%) core biopsies showed calcifications on specimen radiographs. In 96 (88%) of these core biopsies, calcifications measuring100 microm were found on the initial tissue sections. In 11 (10%) further deeper sections were required to detect calcifications100 microm; however, this did not result in a change of the pathological diagnosis. Two of the 109 (1.8%) "benign" core biopsies, which contained tissue calcifications100 microm and at that time were considered representative of the mammographic lesion, have had a malignant outcome on clinical and mammographic follow-up ranging from 2.4 to 7.5 years. Of the 51 (31%) core biopsies where calcifications were not seen on specimen radiographs, histological calcifications were not found in 34 (67%) core biopsies, whereas in 17 (33%) core biopsies, calcifications measuring100 microm were found. All of these core biopsies were considered non-diagnostic and therefore not representative of the lesion targeted. Five (9.8%) of these cases had a malignant outcome with either immediate rebiopsy or excision. Accurate diagnosis of all mammographic lesions requires radiological-pathological correlation. This study shows that the presence of calcifications on the specimen radiograph and the demonstration of tissue calcifications100 microm are an essential and highly reliable part of core biopsy assessment for mammographically "suspicious" calcifications. Nevertheless, lesions with "highly suspicious" calcifications on mammography should be considered for excision even if the core biopsy diagnosis is benign and calcifications100 microm are present.

We describe a sporadic case of lethal prenatal onset infantile cortical hyperostosis (Caffey disease), which resulted in early postnatal death at 30 weeks gestation. The mother presented with antepartum haemorrhage and preterm labour. She was found to have polyhydramnios. The infant showed extensive symmetrical diaphyseal subperiosteal cortical thickening throughout the skeleton with short extremities. Hepatomegaly and lung hypoplasia were present. Currently, in the absence of a specific marker, diagnostic ultrasound offers the only prospect of prenatal diagnosis. This diagnosis should be considered in infants with short angulated long bones, where the diaphyses are irregular and echodense, and where there is no sign of fractures.

Ocular lymphoid tumours represent a spectrum of lymphoproliferative disease and can be subdivided into benign or reactive lymphoid hyperplasia, indeterminate or atypical lymphoid proliferations and malignant lymphoma. Treatment options include a wait and watch approach, systemic steroids, local radiotherapy or systemic chemotherapy. We describe a case of bilateral atypical lymphoid hyperplasia treated successfully with combination immunotherapy and radiotherapy. A 60-year-old lady presented with proptosis and left supra-orbital mass and was diagnosed to have bilateral atypical lymphoid hyperplasia. She had extensive extraocular facial infiltrates but no other sites of involvement on staging investigations. She was treated with eight doses of rituximab 375 mg/m² at weekly intervals with a good partial response, followed by consolidative radiotherapy. Rituximab may be an effective treatment adjunct/alternative for patients with atypical lymphoid hyperplasia of the orbit, particularly where widespread lesions preclude the use of initial radiotherapy.

Background Infiltrating syringomatous adenoma is a rare tumor of the breast that can radiologically mimic invasive duct carcinoma. Detailed fine needle aspiration cytology and needle core biopsy findings on this lesion have not been previously described. Case The clinical, radiologic and pathologic findings of an infiltrating syringomatous adenoma of the breast in a 71-year-old female who presented with a subareolar lump are described. The cytology of the tumor was characterized by a combination of a background of plump, fibroblastoid cells and cohesive sheets of bland epithelial cells. Histologically the tumor showed infiltrating, duct-like structures with squamous metaplasia and a desmoplastic stroma. Conclusion Fine needle aspiration cytology and needle core biopsy can distinguish infiltrating syringomatous adenoma from malignant disease of the breast.

OBJECTIVE: To compare the cytologic characteristics of gastric mucosal cell smears prepared by air drying and rehydration prior to alcohol fixing with cells wet fixed in alcohol. STUDY DESIGN: Gastric mucosal cells were obtained from 55 consecutive patients undergoing gastroscopy. Paired smears were made, one immediately fixed in 95% ethanol for 20 minutes (wet fixed [WF]) and the other air dried for at least 20 minutes prior to rehydration with normal saline for 30 seconds and fixation in 95% ethanol for 20 minutes (air dried/rehydrated/fixed [ARF]). Both slides were stained by the Papanicolaou method. Coded slides were examined blind and graded 1 (superior), 2 (satisfactory) or 3 (poor) with respect to staining of chromatin, nuclear membrane, nucleoli, cytoplasm/cell border and group morphology. Histology confirmed a benign disease process or normal mucosa. RESULTS: Comparing grade 1 versus grades 2 and 3, ARF slides were significantly better than WF slides for all cytologic features (P .05) (X 2 analysis). CONCLUSION: The cytologic features of ARF smears of gastric cells were equal or superior to those of WF smears. This method of preparing smears is simpler and avoids some of the problems of ethanol fixation of wet smears.

Antibiotic-associated haemorrhagic colitis is an uncommon cause of bloody diarrhoea in patients taking penicillin or penicillin-related antibiotics. Symptoms of abdominal pain and bloody diarrhoea occur within 1 week of antibiotic use and resolve without specific therapy within days of discontinuing the offending antibiotic. There is an apparent increased incidence of the disease in patients of Oriental ethnicity. The pathogenesis is unknown. We present two cases of haemorrhagic colitis in patients taking penicillin-related antibiotics who presented within 4 months of each other. One of the patients was being treated for Helicobacter pylori infection. The published literature is reviewed with particular emphasis on the histology and pathogenesis of the condition.

We describe in a five generation family four affected males with hydrocephalus (4 offspring/4 examined) due to aqueductal stenosis (3/3), symmetrical radial ray abnormalities (4/4), renal anomalies (2/3), anal atresia (3/4), hypoplastic penis/abnormal testes (2/3), and cardiac abnormalities (1/3). X-linked inheritance seems certain in this family. These abnormalities are characteristic of the rare X-linked VACTERL-H syndrome. In addition, one maternal female cousin had a severe tracheo-esophageal fistula. This may represent partial manifestation in a female carrier. Chromosomes were apparently normal (46XY) with no spontaneous or excess induced breakages in one of the affected offspring and his mother. In the absence of a genetic marker, diagnostic ultrasonography is the investigation of choice for early in utero detection of this syndrome. A confident ultrasonographic diagnosis was possible by 20 weeks in the 2 cases examined.

Prenatal and postnatal factors such as intrauterine growth restriction (IUGR) and high-fat (HF) diet contribute to type 2 diabetes. Our aim was to determine whether IUGR and HF diets interact in type 2 diabetes pathogenesis, with particular attention focused on pancreatic islet morphology including assessment for inflammation. A surgical model of IUGR (bilateral uterine artery ligation) in Sprague-Dawley rats with sham controls was used. Pups were fed either HF or chow diets after weaning. Serial measures of body weight and glucose tolerance were performed. At 25 weeks of age, rat pancreases were harvested for histologic assessment. The birth weight of IUGR pups was 13% lower than that of sham pups. HF diet caused excess weight gain, dyslipidemia, hyperinsulinemia, and mild glucose intolerance, however, this was not aggravated further by IUGR. Markedly abnormal islet morphology was evident in 0 of 6 sham-chow, 5 of 8 sham-HF, 4 of 8 IUGR-chow, and 8 of 9 IUGR-HF rats (chi-square, P = 0.007). Abnormal islets were characterized by larger size, irregular shape, inflammation with CD68-positive cells, marked fibrosis, and hemosiderosis. β-Cell mass was not altered by IUGR. In conclusion, HF and IUGR independently contribute to islet injury characterized by inflammation, hemosiderosis, and fibrosis. This suggests that both HF and IUGR can induce islet injury via converging pathways. The potential pathogenic or permissive role of iron in this process of islet inflammation warrants further investigation.

Summary The cardiac persistent sodium current (INaP) presents a novel target for cardiac ischaemic protection. Herein we investigated the effects of the INaP blocker riluzole in a pig model of regional myocardial ischaemia. Landrace or Large White pigs were subjected to 3 h ligation of the left anterior descending coronary artery (LAD). Pigs received either saline (500 mL/h, i.v.) throughout the experiment (control; n = 7) or riluzole (2 mg/kg in 2 mL propylene glycol in 100 mL saline, i.v.; RIL; n = 7) between 15 and 5 min prior to ligation. The arrhythmia score was calculated in 5 min epochs. Myocardial damage was assessed using epicardial image analysis and histological sectioning. In the control group, all seven pigs developed premature ventricular contractions (PVC), seven developed non-sustained arrhythmias and six of seven developed sustained arrhythmias. Of the sustained arrhythmias, 23 of 28 instances were initiated by R-on-T extrasytoles (extrasystoles within the vulnerable period that can trigger re-entrant arrhythmias). In the RIL group, all seven pigs developed PVC, six of seven developed non-sustained arrhythmias and only three developed sustained arrhythmias, of which two of five instances were R-on-T initiated. The riluzole-treated pigs exhibited less myocardial damage than pigs in the control group (65% smaller surface area (P = 0.008) on gross epicardial inspection, 51% less oedema (P = 0.01), 53% less fibre waviness (P = 0.029) assessed by haematoxylin and eosin staining and 79% fewer fragmented nuclei (P = 0.009) assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling). In conclusion, riluzole significantly reduced Phase 2 (the period associated with irreversible damage) ischaemic R-on-T triggered and non-R-on-T arrhythmias and myocardial damage occurring during the 3 h period of regional ischaemia.

Aim: There is debate as to what constitutes an adequate excision margin to reduce the risk of locoregional recurrence (LRR) after breast cancer surgery. We have investigated the relationship between surgical margin distance and LRR in women with invasive breast cancer (IBC). Methods: Tumour free margin distances were extracted from histopathology reports for women with IBC, treated by either breast conserving surgery or mastectomy, enrolled in the Breast Cancer Treatment Group Quality Assurance Project from July 1997 to June 2007. Cox proportional hazards regression analyses were conducted to compare the risk of LRR for involved margins compared with negative margins, measured in increments rounded to the nearest mm. Results: 88 of 2300 patients (3.8%) experienced an LRR after a mean follow-up of 7.9 years. An involved margin, or a margin of 1 mm was associated with an increased risk of LRR (HR 2.72, 95% CI 1.30–5.69), whilst margin distances of 2 mm or greater were not. Risk of LRR with margin distances

IntroductionTo investigate the validity of using 670nm red light as a preventative treatment for Retinopathy of Prematurity in two animal models of oxygen-induced retinopathy (OIR).Materials and methodsDuring and post exposure to hyperoxia, C57BL/6J mice or Sprague-Dawley rats were exposed to 670 nm light for 3 minutes a day (9J/cm²). Whole mounted retinas were investigated for evidence of vascular abnormalities, while sections of neural retina were used to quantify levels of cell death using the TUNEL technique. Organs were removed, weighed and independent histopathology examination performed.Results670 nm light reduced neovascularisation, vaso-obliteration and abnormal peripheral branching patterns of retinal vessels in OIR. The neural retina was also protected against OIR by 670 nm light exposure. OIR-exposed animals had severe lung pathology, including haemorrhage and oedema, that was significantly reduced in 670 nm+OIR light-exposed animals. There were no significance differences in the organ weights of animals in the 670 nm light-exposed animals, and no adverse effects of exposure to 670 nm light were detected.DiscussionLow levels of exposure to 670 nm light protects against OIR and lung damage associated with exposure to high levels of oxygen, and may prove to be a non-invasive and inexpensive preventative treatment for ROP and chronic lung disease associated with prematurity.

Preterm birth (birth prior to 37 completed weeks of gestation) may occur at a time when the infant kidney is very immature and nephrogenesis is often ongoing. In autopsied preterm human kidneys and in a baboon model of preterm birth it has been shown that nephrogenesis continues after preterm birth, with a significant increase in the number of glomerular generations and number of nephrons formed within the kidney after birth. Of concern, however, morphologically abnormal glomeruli (with a cystic Bowman's space) are often observed; the abnormal glomeruli are only located in the outer renal cortex, suggesting that it is the recently formed glomeruli (perhaps those formed in the extra-uterine environment) that are affected. The proportion of abnormal glomeruli within the renal cortex differs between infants with some kidneys appearing normal whereas others are severely affected. This suggests that it may be haemodynamic factors and/or factors in the neonatal care of the infant that lead to the glomerular abnormalities. Indeed, the haemodynamic transition at birth where there is a marked increase in systemic blood pressure and renal blood flow are likely to lead to injury of glomerular capillaries, although further studies are required to elucidate this. In order to optimize renal health at the beginning of life in the preterm infant, it is imperative in future studies to gain an understanding of the causes of the glomerular abnormalities in the preterm neonate.

Tendinopathy is a common, costly condition affecting both sporting and sedentary populations. Research into tendinopathy frequently involves the evaluation of tendinosis, a pathology characterized by a lack of inflammatory cells, collagen disruption, neovascularisation, altered cell numbers and morphology and increased glycosaminoglycans. Evaluation of these characteristics can be undertaken using the Bonar histopathology score, but the characteristics are heterogeneous throughout tendon specimens with no standardized method of determining the area to be evaluated. The objective of this study was to assess whether the Bonar score varies depending on the criteria used to define the area of evaluation.Case series.Two independent assessors, with a third to resolve disputes, evaluated 103 areas from 35 tendon specimens using the Bonar score. Specimens were scored once each in the area of worst collagen disruption, degree of vascularization, and cell morphological changes. The inter-tester reliability of the updated Bonar scale was good (r(2)=0.71) RESULTS: The Bonar score was highest in the areas of worst cell morphological (CM) changes, followed by collagen disruption (CD) and lowest for the area of most extensive vascular proliferation (VS) (regression: CD vs. CM, p=0.008, CM vs. VS, p0.001, CD vs. VS, p=0.013). Suggested modifications to the Bonar score include the addition of a cellularity domain, specific definitions of hypo- and hypercellularity, and changes to the vascularity score to include pathological avascularity.The updated Bonar score includes a standardized method of selecting the area of evaluation, which should provide increased reliability when assessing the extent of tendon degeneration.

Long interspersed nuclear element 1 (LINE-1) retrotransposons are mutagens that are capable of generating deleterious mutations by inserting themselves into genes and affecting gene function in the human genome. In normal cells, the activity of LINE-1 retrotransposon is mostly repressed, maintaining a stable genome structure. In contrast, cancer cells are characterized by aberrant expression of LINE-1 retrotransposons, which, in principle, have the potential to contribute to genomic instability. The mechanistic pathways that regulate LINE-1 expression remain unclear. Using deep-sequencing small RNA analysis, we identified a subset of differentially expressed endo-siRNAs that directly regulate LINE-1 expression. Detailed analyses suggest that these endo-siRNAs are significantly depleted in human breast cancer cells compared with normal breast cells. The overexpression of these endo-siRNAs in cancer cells markedly silences endogenous LINE-1 expression through increased DNA methylation of the LINE-1 5'-UTR promoter. The finding that endo-siRNAs can silence LINE-1 activity through DNA methylation suggests that a functional link exists between the expression of endo-siRNAs and LINE-1 retrotransposons in human cells.

Regulatory T (Treg) cell expansion is required for tolerance of the semi-allogeneic fetus in healthy pregnancy and impaired in preeclampsia in humans. However, the reasons remain unknown. Herein, we show that expansion of CD4 + Helios − Foxp3 + adaptive Treg (iTreg) cells, rather than CD4 + Helios + Foxp3 + natural Treg cells, accounts for this expansion in healthy pregnancy. This expansion is even more pronounced in the decidua, where there is an overrepresentation of iTreg cells. In preeclampsia, however, there is impaired systemic iTreg cell expansion, associated with a lack of iTreg cell overrepresentation in the decidua. Because decidual antigen-presenting cells (APCs) may be important for iTreg cell induction, we studied decidual CD14 + APCs using immunohistochemistry and flow cytometry. We show that decidual CD14 + DC-SIGN + APCs are closely associated with Foxp3 + Treg cells. Furthermore, CD14 + DC-SIGN + cells display a distinct phenotype compared with their CD14 + DC-SIGN − counterparts. In particular, they have increased expression of tolerogenic molecules, HLA-G, and immunoglobulin-like transcript 4. In vitro , CD14 + DC-SIGN + APCs from healthy pregnant women induced iTreg cells significantly more efficiently than CD14 + DC-SIGN − APCs. Conversely, in preeclampsia, both CD14 + DC-SIGN + and CD14 + DC-SIGN − APCs induced iTreg cells poorly. These results suggest that decidual CD14 + DC-SIGN + APCs may play important roles in iTreg cell induction, a process that is defective in preeclampsia and likely contributes to its pathogenesis.

Preterm infants are delivered while glomerulogenesis is ongoing and may be exposed to insults, including medications that may affect renal development. Podocytes detected in the urine are an indicator of glomerular injury. The aims of this study were to determine whether preterm and term infants excrete podocytes in their urine and whether exposure to gentamicin and indomethacin increase podocyte excretion in their urine. Preterm infants

Glutathione transferase kappa (GSTK1-1) is a highly conserved, mitochondrial enzyme potentially involved in redox reactions. GSTK1-1-deficient mice were generated to further study the enzyme's biological role. Reduced and total glutathione levels in liver and kidney were unchanged by GSTK1-1 deficiency and NADPH quinone oxidoreductase 1 expression was not elevated indicating that there is no general underlying oxidative stress in Gstk1(-/-) mice. Electron microscopy of liver and kidney showed no changes in mitochondrial morphology with GSTK1-1 deficiency. The death of a number of Gstk1(-/-) males with urinary tract problems prompted close examination of the kidneys. Electron microscopy revealed glomerular basement membrane changes at 3 months, accompanied by detectable microalbuminuria in male mice (albumin:creatinine ratio of 2.66±0.83 vs 1.13±0.20 mg/mmol for Gstk1(-/-) and wild-type (WT), respectively, P=0.001). This was followed by significant foot process effacement (40-55% vs 10% for Gstk1(-/-) and WT, respectively) at 6 months of age in all Gstk1(-/-) mice examined. Kidney tubules were ultrastructurally normal. Compared with human disease, the Gstk1(-/-) kidneys show changes seen in glomerulopathies causing nephrotic syndrome. Gstk1(-/-) mice may offer insights into the early development of glomerular nephropathies.

Liver biopsy with histological examination of liver tissue was for many years the cornerstone of the diagnosis of haemochromatosis, allowing assessment of the degree of iron overload and examination of liver histology for the acute and chronic effects of iron overload. In the past two decades the role of liver biopsy in haemochromatosis has changed dramatically. Liver biopsy is rarely requested for two main reasons: (1) genetic testing for human haemochromatosis (HFE) mutations has proved to be very reliable in the diagnosis of haemochromatosis in Caucasian populations, and (2) the majority of patients with haemochromatosis are now diagnosed at an early stage well before permanent tissue damage occurs, so the need to assess tissue and organ damage has diminished. Liver biopsy continues to have a very important role in a small number of haemochromatosis patients for whom it has both diagnostic and prognostic implications. Liver biopsy is essential for the accurate assessment of patients with non-HFE haemochromatosis and in patients who have dual pathology. It is also useful where there appears to be a discrepancy between HFE genotypes and iron studies, particularly in HFE heterozygotes. Finally, liver biopsy is currently the 'gold standard' for the diagnosis of fibrosis and cirrhosis, although this is changing as non-invasive methods for assessing fibrosis become more reliable and available. Therefore, it is important that pathologists maintain their knowledge and skills in the use of liver biopsy in haemochromatosis and other iron storage disorders.

A 26-year-old Indian lady G2P0 presented at 33 weeks gestation complaining of itch. Liver function tests revealed an elevated alkaline phosphatase (ALKP) of 2983 U/L (normal range 20–110) which on fractionation was a placental or placental-like isoenzyme. All other investigations, including fetal ultrasound, were normal. Labour was induced at 37 weeks due to continued ALKP rise (peak level 4082 U/L). The baby and placental disc weighed 2740 g and 380 g, respectively. The ALKP level returned to normal over 8 weeks. Three years later the patient became pregnant again. The ALKP was monitored and rose to 1497 U/L. Fetal monitoring was increased. The baby was delivered at 38 weeks with evidence of IUGR (2400 g) with a placental disc weight of 356 g. Assessment of both placentas revealed moderate-massive chronic intervillositis associated with a mild chronic basal villitis and moderate chronic decidualitis. Placental tissue cultures grew no organisms. The features were those of chronic intervillositis of unknown etiology (CIUE). CIUE is a rare entity with an increased risk of poor pregnancy outcome including IUGR and stillbirth and a high recurrence rate. It is thought to be consequence of an abnormal maternal immune response to the fetal allograft. Serum ALKP can be a useful antenatal marker of CIUE to aid management.

A 33-year-old lady presented to her dentist with loose teeth and a 15×14 mm cavitating defect on the palatal mucosa adjacent to the teeth 22 and 23. Radiology showed a well demarcated radiolucent lesion with erosion of the roots of adjacent teeth. An initial biopsy showed features of odontogenic fibroma. Local excision confirmed a central odontogenic fibroma (COF). Light microscopy and immunohistochemistry of both specimens showed features of COF comprised of dense fibromyxoid stromal tissue with scattered bland epithelial nests which were immunoreactive for CK14, AE1/AE3, CK5/6, CK19 and 34βE12. S100 and CDla positive, CD68 negative Langerhans cells (LCs) were seen within epithelial islands. COF accounts for 0.1% of all odontogenic tumours. Researchers are uncertain whether it is a reactive process (hamartoma) or a neoplasm. It is diagnosed most frequently in patients between the second and fourth decades, and occurs with equal incidence in maxilla and mandible. A relationship between LCs and lower stage/prognosis has been published in a number of different tumours, raising the question of whether LCs have a role in the immunosurveillance against cancer. The pathological significance of LCs in COF is still unclear but perhaps they could have a role in local growth patterns.

Background There is a continuing need to improve the efficacy of anticancer agents and to reduce the toxicity associated with them. Recently, reversal of the glycolytic phenotype of cancer cells with novel agents such as DCA (dichloroacetate) has been recognised as an important new target for cancer therapy. DCA is currently being assessed as a novel chemotherapeutic in clinical trials. It is anticipated that DCA will be used in combination with other well-established chemotherapeutic drugs. It is therefore critically important to determine the effects of DCA on therapeutic efficacy, and off-target effects of existing frontline anti-cancer drugs. We have recently discovered that DCA induces glutamate cysteine ligase, the rate-limiting step in glutathione (GSH) synthesis, a major cellular antioxidant. We hypothesised that the ability of DCA to stimulate GSH-synthesising capacity could reduce cisplatin-induced nephrotoxicity, which is thought to be mediated in part by oxidative stress. Aims To determine whether DCA can attenuate cisplatin-induced nephrotoxicity, and to determine if DCA influences the anti-cancer properties of cisplatin. Methods 120 Balb/c mice were injected subcutaneously with the syngeneic 4T1 breast cancer cell line and then co-treated with DCA and cisplatin weekly for 1 month. Controls included treatment with cisplatin alone, DCA alone, or normal saline injections. Serum blood urea nitrogen (BUN) and creatinine were measured and kidney damage was assessed histologically. Tumour size was monitored throughout. Results and conclusions DCA prevented increases in serum creatinine, BUN, and renal proximal tubule apoptosis evident in cisplatin-only treated mice. The tumour size in mice in the co-treated group decreased at the same rate as the cisplatin-only treated mice. We have thus concluded that DCA largely prevents the development of cisplatin-induced nephrotoxicity, and does not attenuate its anti-cancer properties.

Purpose To examine mural cell differentiation and pericyte ensheathment during human choroidal vascular formation and into adulthood. Methods Triple- and double-labeled immunohistochemistry (alpha-smooth muscle actin [αSMA], desmin, NG2, calponin, caldesmon, CD44, CD34, and CD39) were applied to human fetal (8-32 weeks' gestation) and adult choroidal and retinal wholemounts and histologic cross-sections. Transmission electron microscopy (TEM) was also undertaken. Results Early in development CD44+ stem cells also stained with αSMA and CD39, suggesting a common precursor. At 12 weeks' gestation, αSMA+ mural precursor cells, confirmed by TEM, were found scattered and isolated over the primordial vascular tree. During development, αSMA+ cells formed a continuous sheath around large arterioles; in veins there were gaps in αSMA expression. The choriocapillaris had an extensive vascular bed but limited coverage by αSMA+ and NG2+ mural cells. Calponin was expressed only on large vessels, and no caldesmon was detected. Pericyte ensheathment of adult capillaries was 11% for choroid versus 94% for retina. Remarkably, choroidal pericytes had no visible intermediate filaments (IFs) on TEM, though IFs were present in retinal pericytes. Neither retinal nor choroidal pericytes stained with desmin. Conclusions CD44+ stem cells are involved in the formation of mural cells in the human choroidal vasculature. A marked reduction in pericyte ensheathment of human choroidal vessels suggests a permanently open "plasticity window" and a predisposition to vascular instability and poor autoregulatory ability.

Purpose: Uncertainty remains about the impact of bilateral breast cancer. Characteristics and outcomes of unilateral and bilateral breast cancer were compared within an Australian multi-institutional cohort.Methods: Demographic, tumour and treatment characteristics were compared among unilateral (n = 2336) and bilateral cases (52 synchronous, 35 metachronous) using descriptive analyses. Disease-specific outcomes were investigated using Cox regression modelling to adjust for prognostic and treatment factors.Results: Factors associated with increased risk of bilateral breast cancer included lobular histology (p = 0.046), family history (p = 0.025) and metropolitan residence (p = 0.006). Mastectomy was more common for bilateral cases (p = 0.001) while radiotherapy was less common (p = 0.015). Index metachronous cases were less likely to receive hormonal therapy (p = 0.001). Five-year survivals for metachronous, synchronous and unilateral cases were 79%, 88% and 94%, respectively. Poorer outcomes remained after adjusting for prognostic factors [HR = 2.26, 1.21–4.21].Conclusion: Our results confirm international findings indicating worse outcomes from bilateral compared with unilateral breast cancer. Refereed/Peer-reviewed

Background The International Prognostic Index (IPI) is used to determine prognosis in diffuse large B-cell lymphoma (DLBCL). One of the determinants of IPI is the stage of disease with bone marrow involvement being classified as stage IV. For the IPI, involvement on bone marrow is traditionally defined on the basis of histology with ancillary investigations used only in difficult cases to aid histological diagnosis. This study aimed to determine the effect of the routine use of flow cytometry, immunohistochemistry and molecular studies in bone marrow staging upon the IPI. Results Bone marrow trephines of 156 histologically proven DLBCL cases at initial diagnosis were assessed on routine histology, and immunohistochemistry using two T-cell markers (CD45RO and CD3), two B-cell markers (CD20 and CD79a) and kappa and lambda light chains. Raw flow cytometry data on all samples were reanalysed and reinterpreted blindly. DNA extracted from archived paraffin-embedded trephine biopsy samples was used for immunoglobulin heavy chain and light chain gene rearrangement analysis. Using immunophenotyping (flow cytometry and immunohistochemistry), 30 (19.2%) cases were upstaged to stage IV. A further 8 (5.1%) cases were upstaged using molecular studies. A change in IPI was noted in 18 cases (11.5%) on immunophenotyping alone, and 22 (14.1%) cases on immunophenotyping and molecular testing. Comparison of two revised IPI models, 1) using immunophenotyping alone, and 2) using immunophenotyping with molecular studies, was performed with baseline IPI using a Cox regression model. It showed that the revised IPI model using immunophenotyping provides the best differentiation between the IPI categories. Conclusion Improved bone marrow staging using flow cytometry and immunohistochemistry improves the predictive value of the IPI in patients with DLBCL and should be performed routinely in all cases.

Background: Placental vascular changes associated with maternal disease states may affect fetal vascular development. There is evidence suggesting that being born prematurely is associated with a higher blood pressure (BP) in later life. Aim: To determine whether maternal disease state affects BP in the early neonatal period. Methods: Cohort study of neonates admitted to neonatal intensive care unit with exposure to maternal hypertension and diabetes. Inclusion criteria were neonates greater than 27 weeks gestation not ventilated or requiring inotropes for more than 24 h, materna l hypertension (pregnancy induced or essential) or diabetes of any kind requiring treatment, and spontaneous delivery. Exclusion criteria included chromosomal or congenital anomaly and illicit maternal drug use. Oscillometric BP measurements taken until discharge on days 1, 2, 3, 4, 7, 14, 21 and 28. Placental histopathology was performed. Results: One hundred and ninety infants enrolled, 104 in the control and 86 in the study group. Sixty-five infants were born between 28–31 weeks and 125 infants between 32–41 weeks gestation. Those born between 28–31 weeks with a history of diabetes had a statistically higher systolic, mean and diastolic BP throughout the first 28 days of life (P = 0.001; P = 0.007; P = 0.02). Those born between 32–41 weeks gestation with placental pathology associated with altered uteroplacental perfusion had a higher systolic BP (P = 0.005). Conclusions: Maternal- or pregnancy-associated disease states appear to influence BP in the early neonatal period. Diabetes and altered placental perfusion were associated with higher BP readings. Clinical significance of these statistically elevated BPs in the early neonatal period is unknown.

The diabetic pregnancy is characterized by maternal hyperglycaemia and dyslipidaemia, such that placental trophoblast cells are exposed to both. The objective was to determine the effects of hyperglycaemia, elevated non-esterified fatty acids (NEFA) and their interactions on trophoblast cell metabolism and function. Trophoblasts were isolated from normal term human placentas and established in culture for 16 h prior to experiments. Glucose utilisation, fatty acid oxidation and fatty acid esterification were determined using radiolabelled metabolic tracer methodology at various glucose and NEFA concentrations. Trophoblast lipid droplet formation including adipophilin mRNA expression, viability, apoptosis, syncytialisation, secretion of hormones and pro-inflammatory cytokines were also assessed. Glucose utilisation via glycolysis was near maximal at the low physiological glucose concentration of 4mM; whereas NEFA esterification into triacylglycerol and diacylglycerol increased linearly with increasing NEFA concentrations without evidence of plateau. Culture of trophoblasts in 0.25 mM NEFA for 24h upregulated fatty acid esterification processes, inhibited fatty acid oxidation, inhibited glycerol release (a marker of lipolysis) and promoted adipophilin and lipid droplet formation, all consistent with upregulation of fatty acid storage and buffering capacity. NEFA also promoted trophoblast syncytialisation and TNFalpha, IL-1beta, IL-6 and IL-10 production without effects on cell viability, apoptosis or hormone secretion. Hyperglycaemia caused intracellular glycogen accumulation and reduced lipid droplet formation, but had no other effects on trophoblast metabolism or function. NEFA have effects on trophoblast metabolism and function, mostly independent of glucose, that may have protective as well as pathophysiological roles in pregnancies complicated by diabetes and/or obesity.

Interest in the presence and management of synchronous multiple ipsilateral breast cancer has been reported since the early 1920s. The demonstration of multiple foci of breast cancer has been reported in 9-75% of breast cancer related specimens. The large difference in reported incidence is multifactorial and related to the definitions applied, mode of detection and pathological assessment. However, randomised clinical trials comparing total mastectomy and segmental mastectomy with or without radiation over many years have shown no difference in distant disease-free survival or overall survival in patients with synchronous multiple ipsilateral breast cancer compared with unifocal breast cancer. This review examines the current definitions, incidence, pathological assessment, staging and surgical options of synchronous multiple ipsilateral breast cancer.

Examination of bone marrow (BM) trephines has been traditionally limited to histological analysis. With renewed interest in their use in molecular diagnostics, studies assessing the impact of duration of storage on DNA amplification from archived BM trephines are required.DNA extraction was performed on 169 formalin-fixed decalcified paraffin-embedded (FFDPE) trephine blocks with a mean duration of storage of 59.4 months (range 4-200 months). An amplification control master mix from the In Vivo Scribe Immunoglobulin Heavy Chain Clonality kit was used to determine DNA amplification at different amplicon sizes [96-600 base pairs (bp)].Amplification at 96 bp was noted in 145 of 169 specimens (85.8%). Progressive reduction in amplification was noted at higher base pairs with 99 of 169 (58.6%) amplifying at 200 bp, 56 cases (33.1%) at 300 bp, 35 (20.7%) at 400 bp and 19 (11.2%) at 600 bp. The impact of duration of storage on DNA amplification was not statistically significant.DNA was obtained from archival BM trephine specimens in approximately 86% of cases, indicating good potential for utilisation in polymerase chain reaction (PCR) techniques. The duration of storage of FFDPE tissue had no impact on the ability to amplify specimens.

Premature neonates are frequently administered indomethacin, ibuprofen and gentamicin during the period of active glomerulogenesis. These drugs are known to have nephrotoxic effects, but the morphological effect of these drugs is unknown. The purpose of this study was to determine whether administration of these drugs during the late stages of glomerulogenesis in the rat has an effect on glomerular endowment. Rat pups were given, intraperitoneally, indomethacin, ibuprofen or indomethacin and gentamicin for the first 5 days of their postnatal life. The pups were killed at 14 days of age at completion of glomerulogenesis. The total number of glomeruli in the left kidney was determined by the physical disector/fractionator stereological technique. There was no difference between treatment groups in total number of glomeruli per kidney (P = 0.45). There were significantly fewer glomeruli per gram of kidney in those rat pups that had received indomethacin or ibuprofen (P < 0.0001). The reduction in the number of glomeruli per gram of kidney may indicate augmented growth of nephron tubules and/or collecting ducts, and/or be a consequence of oedema secondary to drug exposure. Further study is required to determine whether reduced glomerular number is seen in older animals or following exposure to these drugs at different time-points in kidney development.

The placenta is often not submitted for histopathological examination and obstetricians may be sceptical of the value of the examination. This article looks at the reasons for histopathological assessment of the placenta, examines what clinical information should be provided to pathologists and reviews what information can be gained from this 'diary of the pregnancy', especially for explaining adverse outcomes and potentially guiding the management of future pregnancies.

It is known that advanced stage disease in diffuse large B-cell lymphoma (DLBCL) confers a poor prognosis, and staging investigations are routinely performed at diagnosis, including a bone marrow (BM) biopsy. However, examination of the BM is usually limited to routine light microscopy, with the role of ancillary investigations remaining unestablished. The aim of this pilot study was to estimate the incidence of occult marrow involvement using flow cytometry, immunohistochemistry and immunoglobulin heavy chain (IgH) gene rearrangements, and to determine the impact on survival.Clinical and pathological data were obtained on 36 patients diagnosed with DLBCL. Immunohistochemistry using CD3, CD45RO, CD20 and CD79a, and polymerase chain reaction (PCR) to look for IgH gene rearrangements were performed on formalin fixed BM trephines.Nine patients had morphologically apparent BM involvement. Occult marrow involvement was found in seven of 36 (19.4%) patients using the additional diagnostic modalities. When these cases were included with morphologically apparent cases in a proposed new definition of marrow involvement, the median survival of patients with BM involvement was statistically worse (p=0.02) than those without involvement.The results indicate that use of additional tests on BM at diagnosis can upstage disease for a proportion of patients, which appears to correlate adversely with survival.

Given its prognostic value, there is renewed interest in molecular staging in non-Hodgkin's lymphoma (NHL) using immunoglobulin heavy and light chain (IgH, IgL) gene rearrangements.To compare the efficiency of DNA amplification from fresh frozen and formalin-fixed decalcified paraffin-embedded (FFDPE) bone marrow trephines for use in molecular staging using two methods.After manually extracting DNA from 13 FFDPE and 14 fresh frozen trephine biopsy specimens, two methods were used to test for amplifiability: use of the amplification control master mix supplied in the In Vivo Scribe immunoglobulin heavy chain (IgH) clonality kit, which creates 5 amplicons between 96-600 base pairs (bp); and real-time amplification of the beta-globin gene.Using the first method, the mean maximum length of amplicons generated from FFDPE trephines was statistically lower at 300 bp compared to fresh frozen samples, all of which generated amplicons up to 600 bp in size (p0.001). Real-time amplification of the beta-globin gene showed that the mean crossing threshold of fresh frozen samples was statistically lower than that of FFDPE samples (23.48 (95% CI 22.47 to 24.48) vs 33.64 (95% CI 32.15 to 35.12); p0.001).Although amplifiable DNA can be extracted from both fresh-frozen and FFDPE trephine samples for IgH/IgL analysis, freshly frozen specimens are superior as a source of template DNA, especially for higher base pair PCR products.

Objectives Pre-eclampsia is incompletely understood, however, maternal immune rejection of the fetus is a well-supported theory. In particular, maternal T cell studies have shown a reduction in regulatory T cells in pre-eclampsia, and a shift towards a pro-inflammatory state. Morphologic change of the fetal thymus, a primary center for T cell differentiation, has also been described, even preceding clinical disease. This might indicate a fetal role in disease pathogenesis, although histologic examination of T cell subsets in pre-eclamptic fetuses has not been performed. The aim of this study was to investigate thymic and splenic T cell subsets in pre-eclamptic fetuses. We hypothesized that pre-eclampsia would be linked to altered T cell populations, in both thymus and spleen. Methods This was a cohort study, with tissue collected from deceased fetuses at The Children’s Hospital at Westmead (Sydney Australia), and Canberra Hospital (Canberra, Australia). Six fetuses born to pre-eclampsia were included, with 7 fetuses in the control group. Immunohistochemistry techniques analysed FoxP3+, CD4+ and CD8+ T cell subsets, in fetal thymus and spleen. Each T cell subset was compared between groups using an exact interpretation of the Cochrane Armitage test for trend. Results FoxP3+ regulatory T cells were significantly reduced in the thymus (p = 0.02) and spleen (p = 0.02) of fetuses born to pre-eclampsia, versus controls. In both thymus and spleen, CD8+ T cells were also decreased in pre-eclamptic fetuses, versus controls (p = 0.04 and p = 0.004, respectively). Conversely, CD4+ T cells were comparable between groups in both thymus and spleen. Conclusions Pre-eclampsia is associated with an altered T cell profile in the fetal thymus and spleen. Altered fetal immune function might contribute to disease pathogenesis. Disclosures D.P. Eviston: None. M. Peek: None. P. Hsu: None. B. Santner-Nanan: None. M. Hu: None. J. Dahlstrom: None. S. Arbuckle: None. R. Nanan: None.

We report that natural killer receptors (NKR) for major histocompatibility complex (MHC) class I molecules (MHC-NKR), the inhibitory killer immunoglobulin-like receptors (KIR), and the CD94/NKG2A receptor are present on a small proportion of CD8 T cells in cord blood. On average, 1.67% of CD8 T cells in cord blood express KIR, and 0.74% expresses NKG2A, approximately fivefold less than in adult blood. CD8 T cells expressing MHC-NKR were present at similar levels in cord blood from preterm and term infants, and it is important that their presence was independent of placental pathology or infection. Cord blood CD8 T cells expressing MHC-NKR were relatively homogeneous and entirely CD27+, mostly CC chemokine receptor 7− and granzyme B−, with a majority being CD45RA+ and with no evidence for a skewed distribution of T cell receptor-Vβ when tested in KIR+ cells. This contrasted with adult blood, which was more heterogeneous, and where a majority of CD8 T cells expressing MHC-NKR was CD27− and granzyme B+. Functional studies revealed that cord blood KIR+ CD8 T cells were as capable as KIR− CD8 T cells in their ability to proliferate in response to CD3 ligation, yet it is interesting that they were more capable than KIR− CD8 T cells in their ability to secrete interferon-γ. These data suggest that cord blood CD8 T cells expressing MHC-NKR are a unique subset of cells, distinct from those in adult blood, and may represent a less-differentiated population.

Background: There is ongoing debate about the role of the posterior cruciate ligament (PCL) in total knee arthroplasty. Advocates of PCL retention cite better soft tissue balance and improved proprioception, whereas opponents report late flexion instability. The results of knee replacement are similar whether the PCL is retained or sacrificed. The aim of the present study was to examine the PCL for histological changes that would infer its competence and correlate these with changes easily observed by the operating surgeon. Methods: A prospective study of 50 osteoarthritic knees was performed. Results: Histology of the PCL showed changes secondary to degeneration and trauma. In most of the ligaments examined, arteriosclerosis and fibrosis were present. Half of the PCL examined showed perineural fibrosis, myxoid change and hyalinization. These changes, although very frequent, did not correlate well to the changes observed in either the anterior or PCL, or in the overall severity of osteoarthrosis. Conclusions: Posterior cruciate ligaments usually show degenerative and chronic traumatic change of varying degrees on histology. The changes are not predictable from inspection of the knee at surgery. The frequency of these changes suggests that many osteoarthritic PCL are of indifferent quality and the surgeon should consider this when choosing the style of knee replacement.

SummaryWe describe a patient with a malignant phyllodes tumor with both lobular carcinoma in situ and liposarcomatous stromal differentiation. Although lobular carcinoma in situ and adipose differentiation have been reported as separate features rarely seen in the phyllodes tumor, we are unaware of any cases in which both of these features have been seen within the same tumor. The prognosis for this patient related not only to the malignant stromal component of the tumor but also to her increased risk of developing carcinoma in the remaining breast tissue.

Objective: To examine the effect of antenatal steroid exposure and in utero inflammation on the development of severe intraventricular haemorrhage, periventricular leukomalacia and long-term neurological outcome in infants less than 30 completed weeks gestation. Method: Infants less than 30 completed weeks gestation from January 1996 to July 2001 were identified from a prospectively managed database. Placental pathology was reviewed for the presence or absence of chorioamnionitis and funisitis. Infants were divided into three groups depending on the degree of exposure to fetal inflammation (no inflammation, chorioamnionitis only and chorioamnionitis and funisitis). Data relating to gestational age, birthweight, sex, antenatal steroid exposure, surfactant treatment, days of positive pressure ventilation and days of oxygen requirement were collected. Cerebral ultrasound studies were examined for evidence of intraventricular or intraparenchymal echodensity and periventricular leukomalacia. Long-term neurological outcome was assessed by neurological examination for cerebral palsy and by Griffiths Mental Developmental Assessment for general developmental quotient. Results: Two hundred and twenty infants were identified. The mean gestational age was 27.7 weeks and the mean birthweight 1092 g. Seventy-two per cent of mothers had received a complete course of antenatal steroids. The risk of Grade III intraventricular haemorrhage or intraparenchymal echodensity was associated with exposure to in utero inflammation if a complete course of antenatal steroids had not been received (P = 0.002). This association did not exist if a complete course of antenatal steroids was given (P = 0.62). Fourteen infants had cerebral palsy (7%). The presence of cerebral palsy was also associated with in utero inflammation in the absence of complete antenatal steroid cover (P = 0.03) and not in the presence of complete cover (P = 0.59). The mean general developmental quotient on Griffiths Mental Developmental Assessment at 12 months or 3 years was not affected by exposure to in utero inflammation regardless of antenatal steroid exposure. Conclusion: Risk of intraventricular haemorrhage or intraparenchymal echodensity and cerebral palsy was associated with in utero inflammation in the absence of a complete course of antenatal steroids. A complete course of antenatal steroids appeared to extinguish any association between in utero inflammation and adverse neurological outcome.

A generally healthy 66-year-old woman presented with a painless mass at the junction of the soft and hard palate, adjacent to the right maxillary third molar tooth (18) consistent with a draining abscess. Imaging showed a mass extending into the right maxillary antrum with a differential diagnosis of a salivary gland neoplasm or squamous cell carcinoma. There was no improvement with oral antibiotics. An incisional biopsy showed a dense infiltrate of predominantly plasma cells associated with fibrosis that showed no evidence of clonality. The mass was then excised with the tooth 18. Immunohistochemistry showed more than 70% of the plasma cells to be IgG4 positive. Her serum IgG4 level was elevated at 3.89 g/L (reference range 0.08–1.4 g/L). The patient has remained well with some asymptomatic abnormalities on her imaging studies but with no evidence of involvement of other organs. IgG4 related disease is an increasingly recognised entity characterised by a variety of clinical manifestations with the common features being an elevated serum IgG4 and infiltration of tissue by IgG4 positive plasma cells causing a tumour-like swelling which is accompanied by variable degrees of fibrosis. IgG4 related disease is of unknown aetiology and can involve many tissues. The majority of patients respond to treatment with glucocorticoids.

Background Retinopathy of prematurity is a vasoproliferative disorder that can cause blindness and adverse visual outcomes in extremely premature neonates. Red light at 670nm wavelength promotes cellular differentiation, proliferation and wound repair. Aims To determine whether 670nm light promotes normal retinal vessel development in a mouse model of Oxygen Induced Retinopathy of Prematurity (OIR) and whether it would affect organ development and growth. Methods Four groups of C57BL/6J mice were used: 1) Control; 2) OIR - 75% oxygen p7–12 days and normoxia p12–17 days; 3) OIR and 670nm light - 9 J/cm 2 daily from p7–17; 4) 670nm light - 9 J/cm 2 daily from p7–17. At p17 animals were sacrificed and retinal vasculature labelled with Lectin. Neovascularisation and vaso-obliteration were analysed using established protocols. Weight and length measurements were taken daily until the animals were sacrificed and all organs were harvested, weighed and examined macro- and microscopically. Results Neovascularisation was significantly reduced in the 670nm treated OIR group (P Conclusions Exposure to 670nm red light appears to promote normal retinal vessel development and may protect against ROP. 670nm treatment may also reduce oxygen induced lung injury.

Enterobius vermicularis (syn. oxyuriasis, pinworm, threadworm) live and reproduce in the terminal ileum, caecum, vermiform appendix, and proximal colon. The role of E. vermicularis in appendicitis has been disputed. The aims of this study were to determine the incidence of E. vermicularis in surgically removed appendices and to relate this to the age and sex of the patient, the time of the year, the presence of symptoms and the histological findings. The study included all appendices received in this laboratory during the 5 year period 1984 through 1988. They were fixed in 10% formalin, cut into blocks (one longitudinally from the apex, and at least 2 transversely) and stained with haematoxylin and eosin. There were 1867 appendices during this period of which 759 were acutely inflamed and 1108 were not inflamed (although 1U9 of these showed other pathological changes including fibrosis). E.vermicularis was identified in 63 appendices (3.4%). of these, 23 were acutely inflamed although E. vermicularis was not associated with mucosal invasion. Infestation was more frequent in female (4.6%) than in male patients (1.9%). The peak age was 12.8 years in females and 12.1 years in males, and the infestation was less common in summer. of 63 patients who had E. vermicularis, 98% presented with symptoms of acute or recurrent appendicitis, yet 40 had no histological evidence of appendicitis and only 4 had other possible explanations for abdominal pain. In an analysis of the subgroup of 147 patients who liad incidental appendectomy at the time of laparotomy for other reasons, only one had E.vermicularis. It is concluded that E.vermicular is occurred more frequently in uninflamed appendices. However, it may be a cause of symptoms resembling acute appendicitis although the mechanism for this does not involve mucosal invasion by the parasite.

We determined the presence or absence of and clinical significance of cytokeratin-positive cells in the lymph nodes of patients who had had mastectomies for ductal carcinoma-in-situ. Two pathologists independently assessed the axillary lymph nodes found. All patients had either a core or open biopsy performed before the time of mastectomy. The lymph nodes were assessed with hematoxylin and eosin stain and immunohistochemistry for cytokeratin marker AE1/AE3. The slides were assessed for the presence or absence of epithelial cells. As a control, axillary lymph nodes found in prophylactic mastectomies were assessed. None of these had had a previous biopsy performed. Lymph nodes from all patients demonstrated no obvious epithelial cells on hematoxylin and eosin stain. Peripheral sinuses of lymph nodes from six patients (23%) who had mastectomies for ductal carcinoma-in-situ contained a few cytokeratin-positive cells on immunohistochemistry. The lymph nodes of the control group demonstrated no cytokeratin-positive cells. The mean follow-up of patients was 5 years, and all patients were alive without recurrence at the time of the study. Epithelial cells may be present in the lymph nodes draining a site of recent breast biopsy in the absence of invasive carcinoma, indicating that these are an artifact of recent surgery and not of micrometastatic disease.

The aim of our study was to compare dilation forceps tracheostomy and sequential dilator tracheostomy in anaesthetized live adult sheep with respect to the characteristics of the stoma formed and the associated injury. We performed percutaneous tracheostomy on adult sheep randomly allocated to receive either dilation forceps or sequential dilators. Sheep were sacrificed immediately after insertion of the percutaneous tracheostomy and the tracheas dissected. Specimens were examined for site, shape and size of stoma, mucosal lacerations, and posterior wall trauma. Ten sheep had dilation forceps tracheostomy and ten had sequential dilator tracheostomy. All of the specimens were found to have cephalo-caudal mucosal tears, usually crossing tracheal rings. The dilation forceps technique was found to have a larger stoma (28.8 mm vs 24.0 mm, P=0.023). The incidence of posterior needle trauma and mucosal lacerations were common (35% and 50% respectively), but they were not statistically different between the two groups. The role of the mucosal tears in the development of tracheal stenosis is reviewed in the discussion.

Patients with longstanding extensive ulcerative colitis have an increased risk of developing colorectal cancer (CRC). There are significant differences in the early pathogenesis of colitis-associated tumors compared with common CRC, whereas the frequency, degree, and significance of microsatellite instability (MSI) as a marker of mismatch repair deficiency in colitis tumors remain unclear. Here we describe the application of the DSS model of chronic colitis to mice with a defect in the Msh2 mismatch repair gene to discern these early events. These mice do not develop CRC spontaneously without an external trigger. The aim of this study was to determine the effect of the Msh2 defect on the frequency and grade of colitis-associated colorectal dysplasia and adenocarcinoma in Msh2-/-, Msh2+/-, and wild-type (Msh2+/+) mice and on the MSI status of the tumors. We show that in mice with chronic colitis, 60% of the Msh2-/- and 29% of the wild-type mice developed high-grade dysplasia or adenocarcinoma, but heterozygosity for the Msh2 defect did not increase tumor susceptibility over wild-type genotype. The largest difference between genotypes was in the frequency of high-grade dysplasia, with 46.7, 8, and 12.5% in Msh2-/-, Msh2+/-, and Msh2+/+ mice, respectively. The Msh2-/- mice developed MSI-high tumors, whereas the majority of the Msh2+/- and wild-type tumors had no MSI. In the Msh2-/- mice, MSI appeared early in non-neoplastic colon tissue, presumably as a result of markedly increased epithelial cell proliferation associated with inflammation. These observations suggest that a homozygous mismatch repair defect predisposes to tumors triggered by chronic inflammation but is not the only factor involved because tumors also developed in the wild-type mice. This model of colitis offers opportunities to characterize the different molecular pathways of carcinogenesis operating in chronic colitis.

Normal pregnancy requires maternal immune tolerance towards the semi-allogeneic fetus. It is thought that preeclampsia might be due to breakdown of this tolerance. Aims and Methods To analyse the distribution and interaction between decidual dendritic cells (DCs) and T regulatory cells (Tregs) in healthy pregnancies and preeclampsia. The relevant demographic data were collected. Double staining immunohistochemistry [DC-SIGN (marker of immature DCs)/ Foxp3 (marker of Tregs) and CD83 (marker of mature DCs)/ Foxp3] was performed on paraffin blocks and the number of each cell type was assessed independently by two different observers. Results The preeclamptic cases had significantly raised blood pressure, proteinuria and low birth weight infants ( p p p = 0.013, respectively). There were higher numbers of DC-SIGN+, CD83+, Foxp3+ cells in preeclampsia ( p value = 0.0002, 0.004, 0.006, respectively). Of the Foxp3+ Tregs, 70–80% were attached to DC-SIGN+ DCs in normal pregnancy and preeclampsia. There was a significant positive correlation between the number of Foxp3+Tregs and DC-SIGN+ DCs in normal pregnancy (R 2 = 0.68, p Conclusions Foxp3+Tregs are closely associated with DC-SIGN+ DCs. The lack of correlation between Foxp3+Tregs and DC-SIGN+ DCs in preeclampsia implies that preeclampsia might be associated with ineffective tolerance induction at the decidual level.

Introduction Greater trochanteric pain syndrome (GTPS) is common and can cause significant pain and dysfunction. 3 While both trochanteric bursae and gluteal tendons have been implicated in the pathology, 2,6 the underlying pathology remains controversial. Studies reporting on pathology are limited by sample size or methods, including lack of comparison specimens. Thus the aim of this study was to clarify the role of tendon and bursa pathology in GTPS. Methods Tendon and bursa specimens were obtained from 64 participants: 35 with GTPS and 29 control participants. Specimens were evaluated by a minimum of two investigators via light microscopy for histopathological and morphological differences. Immunohistochemistry methods were used to evaluate macrophages (CD68), inflammatory cells (CD45) and substance P. Tendon was evaluated used the Bonar score3. Bursa specimens were evaluated via a semi-quantitative scoring system. Second harmonic generation microscopy, via multiphoton excitation fluorescence (MPEF) was used to illustrate morphological findings (Figure 1). Results Higher levels of pathology were found in specimens from participants with GTPS vs controls. The stroma of the bursa score, mean (SD): GTPS 4.18 (1.65) vs. control 2.53 (1.61), p = 0.051. Bonar (tendon) score, mean (SD): GTPS 12.65 (2.0) vs. control 10.43 (4.83), p = 0.04. There was a higher frequency of SubP in bursa (9/12 vs 6/16, p = 0.047), but not the tendon (8/12 vs. 8/15, p = 0.484) of specimens from participants with GTPS vs controls. There was no evidence of increased frequency of macrophages or other inflammatory cells in the bursa or the tendon specimens from either group. Intra adipose septa were identified and found to be present more frequently in the stroma of bursa specimens of participants with GTPS than controls (27/31 vs 15.27, Fisher exact: p = 0.008), (Figure 1). Discussion Tendon pathology present in both groups, with higher degree of pathology in GTPS. More extensive bursa pathology was seen in the GTPS group, along with a higher frequency of SubP in the bursae from the GTPS than the OA group. This may explain why bursectomy appears to be a successful treatment for some people with GTPS. 1 The IAS has previously reported in rats, 5 but not to our knowledge, in humans. This well organised structure may play a role in bursa stabilisation and shear mitigation. Pathologically it may reduce the ability of the bursa stroma to mitigate compression forces over the tendon. References Baker, et al . Arthroscopy. 2007;23:827–832 Connell, et al . Eur Radiol . 2003;13:1339–1347 Fearon, et al . J Arthroplasty . 2014;29:383–386 Fearon, et al . J Sci Med Sport. 2013;doi: 10.1016/j.jsams.2013.07.008 Shaw, et al . J Anat . 2007;211:436–443 Spear, et al . Surg Clin N Amer . 1952;32:1217–1224