Placental involvement by non-Hodgkin lymphoma in a Crohn disease patient on long-term thiopurine therapy.

We report the first published case of aggressive diffuse large B-cell (non-Hodgkin) lymphoma in a 35-year-old pregnant woman who had Crohn disease and was taking long-term thiopurine therapy: the patient developed placental insufficiency, and there was intrauterine fetal death.

Indomethacin administered early in the postnatal period results in reduced glomerular number in the adult rat.

Indomethacin and ibuprofen are administered to close a patent ductus arteriosus (PDA) during active glomerulogenesis. Light and electron microscopic glomerular changes with no change in glomerular number were seen following indomethacin and ibuprofen treatment during glomerulogenesis at 14 days after birth in a neonatal rat model. This present study aimed to determine whether longstanding renal structural changes are present at 30 days and 6 mo (equivalent to human adulthood). Rat pups were administered indomethacin or ibuprofen antenatally on days 18-20 (0.5 mg·kg(-1)·dose(-1) indomethacin; 10 mg·kg(-1)·dose(-1) ibuprofen) or postnatally intraperitoneally from day 1 to 3 or day 1 to 5 (0.2 mg·kg(-1)·dose(-1) indomethacin; 10 mg·kg(-1)·dose(-1) ibuprofen). Control groups received no treatment or normal saline intraperitoneally. Pups were killed at 30 days of age and 6 mo of age. Tissue blocks from right kidneys were prepared for light and electron microscopic examination, while total glomerular number was determined in left kidneys using unbiased stereology. Eight pups were included in each group from 14 maternal rats. At 30 days and 6 mo, there were persistent electron microscopy abnormalities of the glomerular basement membrane in those receiving postnatal indomethacin and ibuprofen. There were no significant light microscopy findings at 30 days or 6 mo. At 6 mo, there were significantly fewer glomeruli in those receiving postnatal indomethacin but not ibuprofen (P = 0.003). In conclusion, indomethacin administered during glomerulogenesis appears to reduce the number of glomeruli in adulthood. Alternative options for closing a PDA should be considered including ibuprofen as well as emerging therapies such as paracetamol.

Effects of photobiomodulation and caffeine treatment on acute kidney injury in a hypoxic ischemic neonatal rat model.

Hypoxic ischemic encephalopathy (HIE) occurs in 2-5/1000 births, with acute kidney injury (AKI) occurring in 40%. AKI increases morbidity and mortality. Caffeine, an adenosine receptor antagonist, and photobiomodulation (PBM), working on cytochrome c oxidase, are potential treatments for AKI. To examine effects of caffeine and PBM on AKI in rats, Day 7 pups underwent a HIE intervention (Modified Rice-Vannucci model) replicating pathology observed in humans. Caffeine was administered for 3 days and/or PBM for 5 days following HIE. Weights and urine for biomarkers (NGAL, albumin, KIM-1, osteopontin) were collected prior to HIE, daily post intervention and at sacrifice. Both treatments reduced kidney injury seen on electron microscopy, but not when combined. HIE elevated urinary NGAL and albumin on Days 1-3 post-HIE, before returning to control levels. This elevation was significantly reduced by PBM or caffeine. KIM-1 was significantly elevated for 7 days post-HIE and was reduced by both treatments. Osteopontin was not altered by HIE or the treatments. Treatments, individually but not in combination, improved HIE-induced reductions in the enzymatic activity of mitochondrial complexes II-III. PBM and caffeine also improved weight gain. PBM and caffeine reduces AKI diagnosed by urinary biomarkers and confirmed by EM findings.

Efavirenz as a potential drug for the treatment of triple-negative breast cancers.

PURPOSE: In contrast to hormone receptor driven breast cancer, patients presenting with triple-negative breast cancer (TNBC) often have limited drug treatment options. Efavirenz, a non-nucleoside reverse transcriptase (RT) inhibitor targets abnormally overexpressed long interspersed nuclear element 1 (LINE-1) RT and has been shown to be a promising anticancer agent for treating prostate and pancreatic cancers. However, its effectiveness in treating patients with TNBC has not been comprehensively examined. METHODS: In this study, the effect of Efavirenz on several TNBC cell lines was investigated by examining several cellular characteristics including viability, cell division and death, changes in cell morphology as well as the expression of LINE-1. RESULTS: The results show that in a range of TNBC cell lines, Efavirenz causes cell death, retards cell proliferation and changes cell morphology to an epithelial-like phenotype. In addition, it is the first time that a whole-genome RNA sequence analysis has identified the fatty acid metabolism pathway as a key regulator in this Efavirenz-induced anticancer process. CONCLUSION: In summary, we propose Efavirenz is a potential anti-TNBC drug and that its mode of action can be linked to the fatty acid metabolism pathway.

Author Correction: LSD1 activation promotes inducible EMT programs and modulates the tumour microenvironment in breast cancer.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Clinical role of flow cytometry in redefining bone marrow involvement in diffuse large B-cell lymphoma (DLBCL) - a new perspective.

AIMS: The clinical role of flow cytometry in staging bone marrow in diffuse large B-cell lymphoma (DLBCL), especially its impact on outcome, remains uncertain. The aim was to determine the contribution of flow cytometry to conventional staging, and to study the impact of this revised staging on survival. METHODS AND RESULTS: One hundred and thirteen cases of DLBCL diagnosed at The Canberra Hospital from 1996 to 2005 were identified. Blinded analysis of bone marrow (BM) morphology and flow cytometric data showed involvement on morphology (M) in 25 (22.1%) cases, on flow cytometry (F) in 21 (18.6%) cases and overall (M + F) in 32 cases (28.3%); discordance was noted in 16 cases (16.1%). Cases with and without marrow involvement on conventional staging alone (M) had no significant difference in survival (P = NS). However, when BM involvement was defined as positivity on morphology and/or flow cytometry (M + F), the median survival of patients with involvement was significantly worse than patients without involvement (P = 0.026). CONCLUSIONS: Flow cytometry-positive cases should be included with those positive on morphology in a summative model to define BM involvement in DLBCL, as it may have a potential impact on predicting outcome.

Histopathological examination of the placenta: key issues for pathologists and obstetricians.

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.

LSD1 activation promotes inducible EMT programs and modulates the tumour microenvironment in breast cancer.

Complex regulatory networks control epithelial-to-mesenchymal transition (EMT) but the underlying epigenetic control is poorly understood. Lysine-specific demethylase 1 (LSD1) is a key histone demethylase that alters the epigenetic landscape. Here we explored the role of LSD1 in global epigenetic regulation of EMT, cancer stem cells (CSCs), the tumour microenvironment, and therapeutic resistance in breast cancer. LSD1 induced pan-genomic gene expression in networks implicated in EMT and selectively elicits gene expression programs in CSCs whilst repressing non-CSC programs. LSD1 phosphorylation at serine-111 (LSD1-s111p) by chromatin anchored protein kinase C-theta (PKC-θ), is critical for its demethylase and EMT promoting activity and LSD1-s111p is enriched in chemoresistant cells in vivo. LSD1 couples to PKC-θ on the mesenchymal gene epigenetic template promotes LSD1-mediated gene induction. In vivo, chemotherapy reduced tumour volume, and when combined with an LSD1 inhibitor, abrogated the mesenchymal signature and promoted an innate, M1 macrophage-like tumouricidal immune response. Circulating tumour cells (CTCs) from metastatic breast cancer (MBC) patients were enriched with LSD1 and pharmacological blockade of LSD1 suppressed the mesenchymal and stem-like signature in these patient-derived CTCs. Overall, LSD1 inhibition may serve as a promising epigenetic adjuvant therapy to subvert its pleiotropic roles in breast cancer progression and treatment resistance.

Activation of LINE-1 Retrotransposon Increases the Risk of Epithelial-Mesenchymal Transition and Metastasis in Epithelial Cancer.

Epithelial cancers comprise 80-90% of human cancers. During the process of cancer progression, cells lose their epithelial characteristics and acquire stem-like mesenchymal features that are resistant to chemotherapy. This process, termed the epithelial-mesenchymal transition (EMT), plays a critical role in the development of metastases. Because of the unique migratory and invasive properties of cells undergoing the EMT, therapeutic control of the EMT offers great hope and new opportunities for treating cancer. In recent years, a plethora of genes and noncoding RNAs, including miRNAs, have been linked to the EMT and the acquisition of stem cell-like properties. Despite these advances, questions remain unanswered about the molecular processes underlying such a cellular transition. In this article, we discuss how expression of the normally repressed LINE-1 (or L1) retrotransposons activates the process of EMT and the development of metastases. L1 is rarely expressed in differentiated stem cells or adult somatic tissues. However, its expression is widespread in almost all epithelial cancers and in stem cells in their undifferentiated state, suggesting a link between L1 activity and the proliferative and metastatic behaviour of cancer cells. We present an overview of L1 activity in cancer cells including how genes involved in proliferation, invasive and metastasis are modulated by L1 expression. The role of L1 in the differential expression of the let-7 family of miRNAs (that regulate genes involved in the EMT and metastasis) is also discussed. We also summarize recent novel insights into the role of the L1-encoded reverse transcriptase enzyme in epithelial cell plasticity that suggest it might be a potential therapeutic target that could reverse the EMT and the metastasis-associated stem cell-like properties of cancer cells.

A safety and feasibility study of the use of 670 nm red light in premature neonates.

OBJECTIVE: Retinopathy of prematurity (ROP) is a vasoproliferative disorder of the retina affecting extremely preterm or low birth weight infants The aim of this study was to assess the feasibility and safety of 670 nm red light use in a neonatal intensive care unit. STUDY DESIGN: Neonates <30 weeks gestation and <1150 g were enrolled within 48 h of birth. Data collected included cause of preterm delivery, Apgar scores and birthweight. 670 nm red light was administered for 15 min per day from a distance of 25 cm, delivering 9 J cm(-)(2), from the time of inclusion in the study until 34 weeks postmenstrual age. Infants were assessed daily for the presence of any skin burns or other adverse signs. RESULT: Twenty-eight neonates were enrolled, seven 24 to 26 weeks and twenty-one 27 to 29 weeks gestation. The most common cause for preterm delivery was preterm labor (14/28) with five of these having evidence of chorioamnionitis. There were no skin burns or other documented adverse events. Entry into the study was readily achieved and treatment was well accepted by parents and nursing staff. CONCLUSION: 670 nm red light appears to be a safe and feasible treatment for further research in respect to ROP.

Histologic-radiologic correlation of mammographically detected microcalcification in stereotactic core biopsies.

Core biopsy is an alternative technique to surgical excision for assessment of nonpalpable mammographically detected suspicious lesions. The pattern of radiologic calcification is often considered to have diagnostic importance. The aim of this study was to correlate radiologic and histologic features of calcification, with respect to appearance, distribution, and size, to determine the significance, if any, of different radiologic patterns of calcification. Core biopsy samples from 124 women who had 129 mammographically suspicious areas of calcification were examined. Core biopsy samples (five cores per procedure) were obtained stereotactically using a 14-gauge needle in an automated Biopty (Bard Australia, Chatswood, NSW, Australia) gun. In 30 lesions no histologic calcification was found. In the others, there was a poor correlation between radiology and histology with respect to the appearance and distribution of calcification. In a subgroup of 53 women, radiographs of biopsy cores were available to allow correlation with the size of histologic calcification. Calcification of <100 microm assessed histologically was not visible on core biopsy specimen radiographs and may not represent the calcification seen mammographically. Thus, radiography of core biopsy samples and histologic measurement of the size of calcification in core biopsy specimens is useful to reduce false-negative diagnoses in which a biopsy has been performed to evaluate mammographically suspicious calcifications.

Immunohistochemistry of omega class glutathione S-transferase in human tissues.

Omega class glutathione transferase (GSTO) has been recently described in a number of mammalian species. We used immunohistochemistry to determine the cellular and tissue distribution of GSTO1-1 in humans. Expression of GSTO1-1 was abundant in a wide range of normal tissues, particularly liver, macrophages, glial cells, and endocrine cells. We also found nuclear staining in several types of cells, including glial cells, myoepithelial cells of the breast, neuroendocrine cells of colon, fetal myocytes, hepatocytes, biliary epithelium, ductal epithelium of the pancreas, Hoffbauer cells of the placenta, and follicular and C-cells of the thyroid. These observations and the known activity of GSTO1-1 suggest biological functions that are not shared with other GSTs.

VACTERL with hydrocephalus: family with X-linked VACTERL-H.

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.

Electron microscopic assessment of smeared or imprinted specimens of solid tumours.

The aim of this study was to develop a reliable method for preparation of smeared or imprinted cytology specimens for electron microscopic examination. Ten different solid tumours were studied. In each case one air-dried (Diff-Quik stained) and one alcohol-fixed (Pap stained) smear was prepared for diagnostic purposes. Simultaneously, a third smear or imprint was prepared for electron microscopy on a coverslip that was coated with poly-L-lysine and attached to a glass slide using double-sided adhesive tape. The smear or imprint was primary fixed in 2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4, for 2 hours. The coverslip was removed from the slide, placed in a glass petri dish and processed for electron microscopy. The smear/imprint on the coverslip was embedded in resin on a silicon embedding mould and allowed to polymerise for 8-12 hours. The coverslip was removed using liquid nitrogen and the block sectioned using standard techniques as for a monolayer. The specimens collected for electron microscopy using this technique yielded sufficient material for assessment with excellent tissue preservation producing good ultrastructural detail. Focal mechanical damage was seen in some specimens but diagnostic areas were always found within the block. As the smear/imprint can be regarded as a monolayer, the processing time can be reduced compared with solid tissue specimens. This technique ensures that well-preserved tissue is available for electron microscopy even when the sample size is very limited.

Histological correlation of mammographically detected microcalcifications in stereotactic core biopsies.

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 measuring > 100 microm were found on the initial tissue sections. In 11 (10%) further deeper sections were required to detect calcifications > 100 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 calcifications > 100 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 measuring < 100 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 calcifications > 100 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 calcifications > 100 microm are present.

Joubert syndrome: an affected female with bilateral colobomata.

Joubert syndrome is an autosomal recessive disease characterised by hypoplasia or agenesis of the cerebellar vermis, a syndrome of episodic apnoea-hyperpnoea, rhythmic protrusion of the tongue, abnormal eye movements, hypotonia, ataxia, and psychomotor retardation. Extracerebral malformations include multicystic kidney disease, congenital hepatic fibrosis, sacral dermoid cyst and polydactyly. We report the clinical and pathological findings of a 15-year-old girl with Joubert syndrome diagnosed at autopsy. This patient had bilateral colobomata, which has not been previously described in females with Joubert syndrome.

Harlequin ichthyosis--a case report.

The Harlequin infant represents the most severe form of nonbullous ichthyosis. Although the clinical features of infants with Harlequin ichthyosis are generally similar, histological, ultrastructural, and biochemical analyses have not shown consistent findings. An unexpected case of Harlequin ichthyosis in a female infant born at 35 wks gestation is presented. Light microscopic and ultrastructural investigations of skin biopsies are detailed. The presence of extracellular lipid material in the stratum corneum has not been described in the previously reported cases of Harlequin ichthyosis.

Phyllodes tumor with lobular carcinoma in situ and liposarcomatous stroma.

We 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.

Apocrine adenoma of the breast: diagnosis on large core needle biopsy.

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.

Fine needle aspiration cytology of pulmonary lesions: a reliable diagnostic test.

The objective of this study was to determine the accuracy of image-guided fine needle aspiration cytology (FNAC) in the diagnosis of pulmonary lesions. A retrospective study was undertaken of 286 patients with 288 lesions, who underwent a total of 302 procedures. The FNAC diagnoses were reported as malignant, suspicious, atypical, benign or non-diagnostic. Subsequently the FNAC diagnoses were correlated with either the histological or clinical diagnoses. Of the 288 lesions, 64.6% were reported on FNAC as malignant, 2.1% suspicious, 2.4% atypical, 20.8% benign and 10.1% nondiagnostic. On review of the suspicious, atypical, selected benign cases and non-diagnostic FNAC by an independent pathologist there was agreement with the original FNAC diagnosis in all cases. All of 186 malignant FNAC diagnoses were confirmed malignant either clinically or on subsequent histology. Four of the six suspicious FNAC diagnoses had a malignant outcome, one patient had organising pneumonia on excision biopsy and one was lost to follow up. Six of the seven atypical FNAC diagnoses were confirmed on histology as malignant, while one lesion resolved spontaneously. Fifty-two of 60 benign FNAC diagnoses were confirmed benign either clinically or on histology. Seven of the lesions diagnosed as benign on FNAC were proven to be malignant. One patient with a benign FNAC diagnosis was lost to follow-up. Ten of the 29 non-diagnostic FNAC group were later shown on clinical or histological follow up to be malignant. This study shows that image guided FNAC for the diagnosis of malignant pulmonary lesions has a sensitivity of at least 92% and a specificity of at least 96%. It is a reliable diagnostic test although its accuracy is limited by technical difficulties in obtaining an adequate sample.