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Parameters for wound age estimation are essential in forensic pathology but difficult to assess. With this retrospective study, wound age parameters in canine skin wounds were assessed with archive material of known age. Haematoxylin and eosin (HE) staining as standard, Prussian blue, Van Gieson (VG) and multiple other special stains were used as well as various immunohistochemistry (IHC) tests. Collagen fibre formation examination included HE staining but also immunolabelling for elastin and collagen I. Collagen fibre formation was further assessed by polarization in HE and VG. HE staining, erythrophagocytosis and presence of haemoglobin decay products in the Prussian blue stained-slides proved to be useful tools in wound age estimation in the early phase of wound repair of less than 5 days. The earliest detection of newly formed collagen was possible in 5-day-old wounds with HE and VG staining. Collagen I reactivity by IHC was weak at this time point, moderate to strong in the lesions older than 10 days and up to 30 days and strong in the lesions older than 3 months. Very slight polarization of collagen fibres was observed at 6 days, becoming stronger in wound lesions of 10–14 days and up to 30 days and reaching the same intensity as normal tissue in a 3-month-old lesion. The morphology of newly formed collagen fibres, utilizing HE and VG staining and polarization as well as collagen I IHC, which proved to be the most important parameters, correlated with distinguishable time points in the available cases of known wound age. [ABSTRACT FROM AUTHOR]

Background: To use different cytological staining techniques to examine the cytomorphologic aspects of thyroid lesions. Materials and Methods: 100 individuals with adequate aspirates participated in an experimental study carried out at the cytopathology outpatient department. The assignment was finished when the Institutional Ethical Committee granted the necessary authorization. Results: Histopathological correlation was obtained for 25 of the 100 cases in this investigation. For seven cases of Hashimoto's thyroiditis, one case of lymphocytic thyroiditis, six cases of nodular colloid goitre, eight cases of nodular colloid goitre with cystic degeneration, seven cases of follicular neoplasm, and six cases of thyroid papillary carcinoma, histopathological linkage was available. Histopathological studies of those 25 instances showed a correlation with the cytological diagnosis. Three instances that were first diagnosed as follicular neoplasms based on cytology were later identified as follicular carcinomas. Conclusion: A combination of several stains and different fixation procedures will boost the diagnostic accuracy of thyroid lesions. Thus, in order to decrease false positive and false negative results in fine aspiration cytology of thyroid lesions, a range of cytological staining methods can be applied, given appropriate material availability. [ABSTRACT FROM AUTHOR]

Introduction: Prostate cancer (CaP) is a leading cancer diagnosis and cause of cancer-related deaths among Nigerian men, and the accuracy of the pathologic diagnosis is critical for optimal patient care. This study aims to define the relative proportional accuracy of the histological diagnosis of prostatic cancer with hematoxylin and eosin (H and E) slides at Federal Medical Centre (FMC) Owerri, Nigeria. It also seeks to determine Gleason's grading and grade group pattern of CaP in our hospital. Materials and Methods: This study is a 5-year retrospective study reviewing all the H and E slides of prostatic Tru-Cut biopsies that were already diagnosed as cancer at FMC Owerri. These diagnoses were read and confirmed using their morphological appearance on light microscopy. Those that were suspicious, especially those that were diagnosed with well-differentiated adenocarcinoma (Gleason 3 + 3) and poorly differentiated carcinoma (Gleason 5 + 5), were sent for immunohistochemical studies. The well-differentiated carcinomas were stained for basal cells using P63, whereas poorly differentiated carcinomas were stained with prostate-specific antigens to confirm tissue of origin. These are to differentiate them from benign mimickers and metastatic carcinomas, respectively. Results: The age range of the patients was between 50 and 99 years, with a mean age of 72.84 years and peak age in the 70–79 years of age group. Using immunohistochemistry on the specimens which fitted into the criteria for immunohistochemistry, this study showed the relative proportional accuracy of the histological diagnosis of prostatic cancer using H and E to be 95.55% and the positive predictive value of H and E to be 0.83. With respect to International Society of Urological Pathology (ISUP) group grading, high-grade cancers predominated, and the most predominant grade is 5. Conclusion: H and E stain has high diagnostic accuracy (95.55%), but a 4.45% diagnostic error using H and E is quite significant. Immunohistochemistry, therefore, should act as a most valuable adjunctive, which significantly increases the diagnostic accuracy of prostatic carcinoma diagnosis.

Introduction: Prostate cancer (CaP) is a leading cancer diagnosis and cause of cancer-related deaths among Nigerian men, and the accuracy of the pathologic diagnosis is critical for optimal patient care. This study aims to define the relative proportional accuracy of the histological diagnosis of prostatic cancer with hematoxylin and eosin (H and E) slides at Federal Medical Centre (FMC) Owerri, Nigeria. It also seeks to determine Gleason's grading and grade group pattern of CaP in our hospital. Materials and Methods: This study is a 5-year retrospective study reviewing all the H and E slides of prostatic Tru-Cut biopsies that were already diagnosed as cancer at FMC Owerri. These diagnoses were read and confirmed using their morphological appearance on light microscopy. Those that were suspicious, especially those that were diagnosed with well-differentiated adenocarcinoma (Gleason 3 + 3) and poorly differentiated carcinoma (Gleason 5 + 5), were sent for immunohistochemical studies. The well-differentiated carcinomas were stained for basal cells using P63, whereas poorly differentiated carcinomas were stained with prostate-specific antigens to confirm tissue of origin. These are to differentiate them from benign mimickers and metastatic carcinomas, respectively. Results: The age range of the patients was between 50 and 99 years, with a mean age of 72.84 years and peak age in the 70–79 years of age group. Using immunohistochemistry on the specimens which fitted into the criteria for immunohistochemistry, this study showed the relative proportional accuracy of the histological diagnosis of prostatic cancer using H and E to be 95.55% and the positive predictive value of H and E to be 0.83. With respect to International Society of Urological Pathology (ISUP) group grading, high-grade cancers predominated, and the most predominant grade is 5. Conclusion: H and E stain has high diagnostic accuracy (95.55%), but a 4.45% diagnostic error using H and E is quite significant. Immunohistochemistry, therefore, should act as a most valuable adjunctive, which significantly increases the diagnostic accuracy of prostatic carcinoma diagnosis. [ABSTRACT FROM AUTHOR]

Background: Irisin, a polypeptide hormone that is released from skeletal muscle in response to exercise, has been found to improve endothelial functions, protect against endothelial injuries and change blood pressure, which also affected blood vessels. The aim of this study was to assess the histological changes of the rat thoracic aorta in response to irisin injection. Materials and methods: Twenty-four rats were used. They were divided into two groups: the control group without irisin injection and the irisin-injected group subdivided into three subgroups treated with different irisin concentrations (20, 40 and 160 nM, respectively) twice a week for 4 weeks. The control group and irisin-treated subgroups consisted of 6 rats each. After 4 weeks all rats were sacrificed, and the descending thoracic aorta was treated for histological evaluation. Sections were stained with haematoxylin and eosin and orcein stains. Morphometric measurement included: intima-media thickness, number of elastic lamellae and number of smooth muscle cells’ nuclei. Results: Histological study showed that intraperitoneal injection of different concentrations of irisin (20, 40 and 160 nM) in rats increased intima-media thickness, the number of smooth muscle cell’s nuclei and the number of elastic lamellae in media layer of the thoracic aorta in a dose-dependent manner. Conclusions: Irisin significantly affected the morphology of the wall of the rat thoracic aorta indicating a role of irisin in influencing the growth factors of the thoracic aorta walls and activating smooth muscle cells in the thoracic aorta layers. [ABSTRACT FROM AUTHOR]

Abstract In order to detect the distribution of Mycobacterium avium subsp avium (MAA) in naturally infected domestic Pekin ducks, immunohistochemistry (IHC) and Ziehl‐Neelsen (ZN) staining were used and compared. Six organs, the liver, spleen, lung, kidney, duodenum and pectoralis muscle, were collected from naturally infected Pekin ducks. Paraffin embedded tissues were examined, and the results were compared. Statistical analysis was performed using Chi‐Square test. The results showed that the detection rates by IHC were similar with ZN staining in liver, lung, spleen and pectoralis muscle, but the detection rates by IHC were much higher than ZN staining in kidney and duodenum (p = .013, p = .0044). The liver (87.5%) and lung (81.3%) had the highest detection rates. Acid‐fast bacilli (AFB) were primarily found intracellularly in six organs using ZN staining. Similarly, the MAA antigens in those selected organs were also detected in the cytoplasm with different cell types. Specifically, MAA antigen was distributed in epithelioid macrophages and necrotic centres within the liver, lung, spleen and kidney, while they were observed in macrophages of the lamina propria and duodenal glands and degenerative myocytes in the pectoralis muscle. This comparative study provides an important insight into the distribution of MAA in infected domestic ducks and indicates that the detection rate by IHC was higher than that of ZN staining.

Breast tumours are heterogeneous diseases that result from changes in the cells and connective tissues of the breast. Different histological and histochemical stains have been employed to aid in the detection of these changes in routine diagnosis. The aim of this study was to assess the usefulness of aqueous beetroot dye in the histochemical staining of breast tumours. Thirty breast tumour blocks from the Histopathology Laboratory of the University of Calabar Teaching Hospital, Calabar were retrieved, sectioned and stained with Cole's haematoxylin and eosin and aqueous beetroot. The histological staining and 3-D image characteristics were analysed. The histological types of breast tumours were 7(23.3%) benign, of which 5(16.7%) were grade II benign lesion, 1(3.3%) was fibrocystic disease and 1(3.3%) was fibroadenoma. Among the 23(76.7%) malignant tumours, 15(56.7%) were grade 3, 3(10%) were B4 carcinomas and 3(10%) were B5 types. The sections stained with the red beetroot dye gave distinct metachromatic staining of the connective tissues of the cell membranes of the epithelial cells and other connective tissue cells, cytoskeleton, cytoplasm, basement membrane, and collagen fibres of the stroma in shades of green, purple, and black. The histological staining and 3-D image characteristics of the beetroot-stained benign and malignant tumours were similar to the haematoxylin and eosin-stained counterparts in terms of nuclear, cytoplasmic and connective tissue staining, 3-D surface plot features, and histological details (X²=1.200; p=0.549). This finding has shown that betalain pigments in beetroot dye gave good metachromatic staining of epithelial and connective tissue changes in breast tumours. Beetroot dye and 3-D surface plots when combined can serve as a cost effective, health and environment-friendly alternative staining method to routine haematoxylin and eosin in diagnosis of breast tumours in low resource areas. [ABSTRACT FROM AUTHOR]

Introduction: Fine Needle Aspiration Cytology (FNAC) is an established diagnostic tool aiming at an early diagnosis of mass lesions which in turn aids in planning further management of the patients. The need for minimal turnaround time with good quality of cell morphology has encouraged the development of different staining techniques. Quality Index (QI) of cytochemical stains is evaluated with a scoring system based on morphological features. Aim: To find rapid staining methods which were economical without compromising on the quality of FNAC smears. QI of five stains namely Papanicolaou stain (Pap), Haematoxylin and Eosin (H&E), May Grunwald Giemsa (MGG), Modified Ultrafast Papanicolaou (MUFP) and Rapid Economic Acetic acid Pap stain (REAP) were studied and compared. Materials and Methods: This was a prospective study conducted in the Department of Pathology, Kempegowda Institute of Medical Sciences, Bengaluru, Karnataka, India, from August 2011 to July 2013. FNAC of 80 mass lesions including 44 of breast lesions and 36 lymph nodes were included in this study. A minimum of five smears were made in each case and stained with Pap, H&E, MGG, MUFP and REAP. The QI of stain was assessed by the scores given for background of the smear, overall staining, and preservation of cell morphology, nuclear characteristics, cytoplasmic details and air drying artifacts. Statistical analysis of the data was done using chi-square test and Statistical Package for the Social Sciences (SPSS) version 15.0. Results: Pap stain showed maximum QI score followed by H&E, REAP, MGG and MUFP in case of breast lesions. In cases of lymph node highest QI was again with Pap followed by H&E, REAP, MUFP and MGG. Good cytomorphology with lesser expenditure and turnaround time was obtained by REAP and MUFP stains. Conclusion: Pap stain is still the best for cell morphology whereas MUFP and REAP are good, economical and with lesser turnaround time and can be used in routine cytopathology.

Introduction: Fine Needle Aspiration Cytology (FNAC) is an established diagnostic tool aiming at an early diagnosis of mass lesions which in turn aids in planning further management of the patients. The need for minimal turnaround time with good quality of cell morphology has encouraged the development of different staining techniques. Quality Index (QI) of cytochemical stains is evaluated with a scoring system based on morphological features. Aim: To find rapid staining methods which were economical without compromising on the quality of FNAC smears. QI of five stains namely Papanicolaou stain (Pap), Haematoxylin and Eosin (H&E), May Grunwald Giemsa (MGG), Modified Ultrafast Papanicolaou (MUFP) and Rapid Economic Acetic acid Pap stain (REAP) were studied and compared. Materials and Methods: This was a prospective study conducted in the Department of Pathology, Kempegowda Institute of Medical Sciences, Bengaluru, Karnataka, India, from August 2011 to July 2013. FNAC of 80 mass lesions including 44 of breast lesions and 36 lymph nodes were included in this study. A minimum of five smears were made in each case and stained with Pap, H&E, MGG, MUFP and REAP. The QI of stain was assessed by the scores given for background of the smear, overall staining, and preservation of cell morphology, nuclear characteristics, cytoplasmic details and air drying artifacts. Statistical analysis of the data was done using chi-square test and Statistical Package for the Social Sciences (SPSS) version 15.0. Results: Pap stain showed maximum QI score followed by H&E, REAP, MGG and MUFP in case of breast lesions. In cases of lymph node highest QI was again with Pap followed by H&E, REAP, MUFP and MGG. Good cytomorphology with lesser expenditure and turnaround time was obtained by REAP and MUFP stains. Conclusion: Pap stain is still the best for cell morphology whereas MUFP and REAP are good, economical and with lesser turnaround time and can be used in routine cytopathology. [ABSTRACT FROM AUTHOR]

In order to detect the distribution of Mycobacterium avium subsp avium (MAA) in naturally infected domestic Pekin ducks, immunohistochemistry (IHC) and Ziehl‐Neelsen (ZN) staining were used and compared. Six organs, the liver, spleen, lung, kidney, duodenum and pectoralis muscle, were collected from naturally infected Pekin ducks. Paraffin embedded tissues were examined, and the results were compared. Statistical analysis was performed using Chi‐Square test. The results showed that the detection rates by IHC were similar with ZN staining in liver, lung, spleen and pectoralis muscle, but the detection rates by IHC were much higher than ZN staining in kidney and duodenum (p =.013, p =.0044). The liver (87.5%) and lung (81.3%) had the highest detection rates. Acid‐fast bacilli (AFB) were primarily found intracellularly in six organs using ZN staining. Similarly, the MAA antigens in those selected organs were also detected in the cytoplasm with different cell types. Specifically, MAA antigen was distributed in epithelioid macrophages and necrotic centres within the liver, lung, spleen and kidney, while they were observed in macrophages of the lamina propria and duodenal glands and degenerative myocytes in the pectoralis muscle. This comparative study provides an important insight into the distribution of MAA in infected domestic ducks and indicates that the detection rate by IHC was higher than that of ZN staining. [ABSTRACT FROM AUTHOR]

Introduction: Quantification of Mitotic Figures (MFs) is a prognostic indicator in assessment of Oral Epithelial Dysplasia (OED), Oral Squamous Cell Carcinoma (OSCC) and is part of many histopathological grading systems. Evaluation of MFs is done as part of post treatment (chemoradiotherapy) to monitor any changes in the treated area. But identification and quantification of MFs in routine stains is a tedious process and subjective to errors. Aim: To stain, compare and analyse mitosis specific marker anti–phosphohistone H3 (anti-PHH3) with H&E stain and 1% crystal violet in OED and OSCC. Materials and Methods: Study sample included archival tissues embedded in paraffin blocks histopathologically diagnosed as OED (n=30) and OSCC (n=30). Three serial sections of each tissue specimen were stained separately with H&E stain, 1% crystal violet stain and anti-PHH3 (IHC stain). The stained sections were examined for identification and counting of MFs. The data were analysed using Chi-square test and Kruskal Wallis Test. Results: MFs were significantly increased in OSCC in comparison with OED. There was a significant increase in number of MFs in anti-PHH3 in comparison with crystal violet and H&E stained tissue sections. Conclusion: It was seen that anti-PHH3 is the most specific stain for identification of MFs amongst H&E, crystal violet and anti-PHH3.

Introduction : Oral cancers account for approximately 3% of all malignancies and is a significant worldwide health problem. Globally, oral cancers are one of the 10 most common cancers and accounts for almost 40% of all cancers in Indian subcontinent. A key factor in the lack of improvement in prognosis is the fact that a significant proportion of oral squamous cell carcinomas are not diagnosed and treated until they reach an advanced stage. Material and Method: The present study has been carried out on 50 patients suffering from oral cavity lesion. In all patients oral brush cytology using a toothbrush was done prior to a planned surgical biopsy. The study was done to assess the efficacy of oral brush cytology in detecting the premalignant and malignant lesions and its use as an early diagnostic aid. Result: 25 Cases were true positive on both histopathology and oral brush cytology. 20 Cases were true negative and 5 Cases were suspicious for malignancy on brush cytology, they were diagnosed as malignant on further histopathological examination. A sensitivity of 80%, specificity of 100%, negative predictive value was 83.3%, positive predictive value was 100% and accuracy was 90%. Conclusion: Oral brush cytology is an accurate diagnostic tool that plays a significant role in early cancer detection. It is also useful as a screening procedure for a high risk population or also for the clinical follow up. [ABSTRACT FROM AUTHOR]

Introduction: Quantification of Mitotic Figures (MFs) is a prognostic indicator in assessment of Oral Epithelial Dysplasia (OED), Oral Squamous Cell Carcinoma (OSCC) and is part of many histopathological grading systems. Evaluation of MFs is done as part of post treatment (chemoradiotherapy) to monitor any changes in the treated area. But identification and quantification of MFs in routine stains is a tedious process and subjective to errors. Aim: To stain, compare and analyse mitosis specific marker anti-phosphohistone H3 (anti-PHH3) with H&E stain and 1% crystal violet in OED and OSCC. Materials and Methods: Study sample included archival tissues embedded in paraffin blocks histopathologically diagnosed as OED (n=30) and OSCC (n=30). Three serial sections of each tissue specimen were stained separately with H&E stain, 1% crystal violet stain and anti-PHH3 (IHC stain). The stained sections were examined for identification and counting of MFs. The data were analysed using Chi-square test and Kruskal Wallis Test. Results: MFs were significantly increased in OSCC in comparison with OED. There was a significant increase in number of MFs in anti-PHH3 in comparison with crystal violet and H&E stained tissue sections. Conclusion: It was seen that anti-PHH3 is the most specific stain for identification of MFs amongst H&E, crystal violet and anti-PHH3. [ABSTRACT FROM AUTHOR]

In this study, high-fat diet (HFD)-induced obesity in mouse model was used to evaluate the dietary effect of saponins from stems and leaves of Panax ginseng (SLG), and to explore its mechanism of action in producing anti-obesity effects. The results indicate that SLG showed significant anti-obesity effects in diet-induced obese mice, represented by decreased serum levels of free fatty acids (FFA), total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL)-cholesterol, glucose, leptin and insulin, as well as a reduction in overall body and liver weight, epididymal adipose tissue weight, and food efficiency, and inhibition of abnormal increases in acyl carnitine levels normally caused by an HFD. Additionally, the down-regulated expression of PPARγ, FAS, CD36, FATP2 and up-regulated expression of CPT-1, UCP-2, PPARα, HSL, and ATGL in liver tissue was induced by SLG. In addition, the SLG groups showed decreased PPARγ, aP2 and leptin mRNA levels and increased expression of PPARα, PGC-1α, UCP-1 and UCP-3 genes in adipose tissues, compared with the HFD group. In short, SLG may play a key role in producing anti-obesity effects in mice fed an HFD, and its mechanism may be related to regulation of thermogenesis, lipogenesis and lipolysis. [ABSTRACT FROM AUTHOR]

Background: Tuberculous lymphadenitis is a common form of extrapulmonary tuberculosis in young adults and children which commonly involves superficial lymph nodes. The purpose of this study was to detect and confirm the presence of acid fast bacilli in lymph node biopsies using routine Ziehl Neelsen stain, Kinyoun stain and fluorescence staining and also to determine the best, rapid and more cost effective technique of staining for the detection of acid fast bacilli. Material & Methods: This descriptive cross-sectional study was conducted in University of Health Sciences, Lahore, Punjab, Pakistan, from November, 2013 to February 2014. Sample size was 97 patients with lymph node biopsies taken from through non probability, consecutive sampling technique. Demographic variables were; gender, age group. Research variables were; site of lymph node biopsy, type of granulomatous inflammation, positivity of AFB. The attributes of age group were; 1-15. 16-30, 31-45 and 46-75. The attributes of site of lymph node biopsy were; supra-clavicular, cervical, axillary, right mediastinal and submandibular. The attributes of type of granulomatous inflammation were; caseous and non-caseous and of positivity of AFB were; positive and negative. Data was collected in Gulab Devi Chest Hospital, Lahore. Lymph node biopsies from different sites. including supraclavicular, cervical, axillary, right mediastinal and Submandibular were included and stained with. Descriptive analysis was done. Results: Out of a total of 97 patients with granulomatous lymph node biopsies, 36 (37%) were males and 61 (63%) females. Caseous granulomas were 67(69%), while 30(31%) showed non-caseous granulomas. Lymph nodes were collected from different sites e.g. supraclavicular (20, 21%), cervical (69, 71%), axillary (4, 4%), right mediastinal (2, 2%) and Submandibular (2, 2%) lymph nodes. AFB were microscopically positive in 54 (56%) nodal tissue biopsy sections and forty three (44%) cases were negative. From a total of 54 positive cases routine ZN stain was positive in 45 (46%), Kinyoun stain was positive in 54 (56%) and fluorescent staining was positive in 54 (56%) cases. Conclusion: Caseous granulomas were detected in females especially. Kinyoun and Fluorescent staining using Auramine O and Rhodamine B are better staining techniques in the detection of AFB than Ziehl Neelsen Staining. [ABSTRACT FROM AUTHOR]

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

Objective: The aims of this study were: first, to evaluate the possible effects of a fructose rich diet (FRD)-induced metabolic syndrome (MS) on different aspects of long bone histomorphometry in young male rats; second, to investigate the effects of this diet on bone tissue regeneration; and third, to correlate these morphometric alterations with changes in the osteogenic/adipogenic potential and expression of specific transcription factors, of marrow stromal cells (MSC) isolated from rats with fructose-induced MS., Materials/methods: MS was induced in rats by treatment with a FRD for 28 days. Halfway through treatment, a parietal wound was made and bone healing was evaluated 14 days later. After treatments, histomorphometric analysis was performed in dissected femoral and parietal bones. MSC were isolated from the femora of control or fructose-treated rats and differentiated either to osteoblasts (evaluated by type 1 collagen, Alkaline phosphatase and extracellular nodule mineralization) or to adipocytes (evaluated by intracellular triglyceride accumulation). Expression of Runx2 and PPARγ was assessed by Western blot., Results: Fructose-induced MS induced deleterious effects on femoral metaphysis microarchitecture and impaired bone regeneration. Fructose treatment decreased the osteogenic potential of MSC and Runx2 expression. In addition, it increased the adipogenic commitment of MSC and PPARγ expression., Conclusions: Fructose-induced MS is associated with deleterious effects on bone microarchitecture and with a decrease in bone repair. These alterations could be due to a deviation in the adipogenic/osteogenic commitment of MSC, probably by modulation of the Runx2/PPARγ ratio., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background & Aims: Hirmi Valley liver disease was first reported in 2001 in Tigray, Ethiopia. 591 cases, including 228 deaths, were reported up to December 2009. The pyrrolizidine alkaloid acetyllycopsamine was detected in stored grain and residents reported adding the pesticide DDT (dichlorodiphenyldichloroethylene) directly to their food stores. We aimed to characterise the clinical features of the disease, and explore the role of these chemicals in its aetiology., Methods: 32 cases were examined and full clinical histories taken. Nine cases underwent liver biopsy in hospitals. Serum and urine samples were collected from cases and controls. Urine was analysed for acetyllycopsamine by UPLC-MS. Total DDT in serum was measured by ELISA. Hepatotoxicity of DDT and acetyllycopsamine alone or in combination was explored in C57BL/6J mice., Results: Clinical presentation included epigastric pain, abdominal swelling, bloody diarrhoea, hepatomegaly, splenomegaly, and ascites. Histology revealed acute injury characterised by centrilobular necrosis or chronic injury with bile ductular reaction, cytomegaly and fibrosis but no hepatic vein occlusion. Acetyllycopsamine was detected in urine samples taken in the affected area with significantly greater concentrations in 45 cases than in 43 controls (p=0.02). High levels of DDT (>125 ppb) were detected in 78% of serum samples. In mice, DDT (3 × 75 mg/kg) significantly increased the hepatotoxicity (plasma ALT, p=0.0065) of acetyllycopsamine (750 mg/kg), and in combination induced liver pathology similar to Hirmi Valley liver disease including centrilobular necrosis and cytomegaly., Conclusions: This novel form of disease appears to be caused by co-exposure to acetyllycopsamine and DDT., (Copyright © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Background & Aims: Carbamazepine (CBZ), a widely used anticonvulsant and mood stabilizer, activates multiple proliferative and pro-survival pathways. Here, we hypothesize that CBZ may promote hepatocellular proliferation and ameliorate liver regeneration., Methods: C57BL6/J mice were orally administered CBZ or vehicle and underwent a 70% partial hepatectomy (PHx), 85% PHx or treatment with carbon tetrachloride (CCl4). Liver regeneration was determined by liver to body weight ratio, hepatocyte proliferation markers, and activation of intracellular signalling pathways., Results: Two to 5days after the 70% PHx, the liver to body weight ratio was significantly higher in the CBZ-treated mice than in the vehicle-treated mice. CBZ treatment upregulated the number of proliferative hepatocytes following PHx or CCl4 treatment, as assessed by intrahepatic Ki-67 staining, BrdU uptake, and PCNA protein expression. PHx surgery induced the expression of several cyclins and activated Akt/mTOR signalling pathways, all of which were enhanced by CBZ treatment. The administration of the mTOR inhibitor temsirolimus abrogated the hepato-proliferative effect of CBZ. CBZ treatment significantly improved the survival rate of the mice that underwent lethal 85% massive hepatectomy., Conclusions: CBZ demonstrated a novel hepato-proliferative effect through the activation of the mTOR signalling pathway in hepatectomised mice. CBZ has the potential to be a therapeutic option for facilitating efficient liver regeneration in patients subjected to liver surgery., (Copyright © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Background: Most cervical cancers are attributable to infection with one of fourteen types of human papillomavirus (HPV), but HPV types differ in oncogenic potential. Characterisation of cancers associated with specific HPV types is required to predict the likely impact of current prophylactic vaccines and the potential benefits of vaccine formulations including additional HPV types., Objective: The study aimed to correlate HPV type with histology and age at diagnosis, in Invasive Cervical Cancers (ICCs) from two of the devolved countries of the UK (Wales and Scotland)., Study Design: Centralised histopathology review and rigorously standardised HPV-DNA typing were applied to 592 ICC diagnosed 2001-2006. HPV status was analysed in relation to histology and age at diagnosis., Results: HPV infection was confirmed in 535/592 cases. Among the 497 tumours infected with single HPV types, the three most common types were HPV16 (62% 95%CI: 57.6-66.1), HPV18 (18.9% 95%CI: 15.7-22.6) and HPV45 (5.4% 95%CI: 3.7-7.8). HPV16 or 18 were present in 80.9% of HPV positive cases. Women with tumours associated with HPV types 16, 18 and 45 were on average 10.5 years younger at diagnosis than women with tumours associated with other HPV types., Conclusions: Prophylactic vaccines targeting HPV16 and 18 could potentially prevent up to 80.9% of ICC in the populations investigated. Cancers associated with HPV16, 18 and 45 were diagnosed at younger ages, supporting the hypothesis of faster progression than for tumours caused by other HPV types., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background & Aims: The use of livers from donors after cardiac arrest (DCD) is increasing in many countries to overcome organ shortage. Due to additional warm ischemia before preservation, those grafts are at higher risk of failure and bile duct injury. Several competing rescue strategies by machine perfusion techniques have been developed with, however, unclear effects on biliary injury. We analyze the impact of an end-ischemic Hypothermic Oxygenated PErfusion (HOPE) approach applied only through the portal vein for 1h before graft implantation., Methods: Rat livers were subjected to 30-min in situ warm ischemia, followed by subsequent 4-h cold storage, mimicking DCD-organ procurement and conventional organ transport. Livers in the HOPE group underwent also passive cold storage for 4h, but were subsequently machine perfused for 1h before implantation. Outcome was tested by liver transplantation (LT) at 12h after implantation (n=10 each group) and after 4 weeks (n=10 each group), focusing on early reperfusion injury, immune response, and later intrahepatic biliary injury., Results: All animals survived after LT. However, reperfusion injury was significantly decreased by HOPE treatment as tested by hepatocyte injury, Kupffer cell activation, and endothelial cell activation. Recipients receiving non-perfused DCD livers disclosed less body weight gain, increased bilirubin, and severe intrahepatic biliary fibrosis. In contrast, HOPE treated DCD livers were protected from biliary injury, as detected by cholestasis parameter and histology., Conclusions: We demonstrate in a DCD liver transplant model that end-ischemic hypothermic oxygenated perfusion is a powerful strategy for protection against biliary injury., (Copyright © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Islets form in the pancreas after the first endocrine cells have arisen as either single cells or small cell clusters in the epithelial cords. These cords constitute the developing pancreas in one of its earliest recognizable stages. Islet formation begins at the time the cords transform into a branching ductal system, continues while the ductal system expands, and finally stops before the exocrine tissue of ducts and acini reaches its final expansion. Thus, islets continuously arise from founder cells located in the branching and ramifying ducts. Islets arising from proximal duct cells locate between the exocrine lobules, develop strong autonomic and sensory innervations, and pass their blood to efferent veins (insulo-venous efferent system). Islets arising from cells of more distal ducts locate within the exocrine lobules, respond to nerve impulses ending at neighbouring blood vessels, and pass their blood to the surrounding acini (insulo-acinar portal system). Consequently, the section of the ductal system from which an islet arises determines to a large extent its future neighbouring tissue, architecture, properties, and functions. We note that islets interlobular in position are frequently found in rodents (rats and mice), whereas intralobularly-located, peripheral duct islets prevail in humans and cattle. Also, we expound on bovine foetal Laguesse islets as a prominent foetal type of type 1 interlobular neuro-insular complexes, similar to neuro-insular associations frequently found in rodents. Finally, we consider the probable physiological and pathophysiological implications of the different islet positions within and between species., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Microparticles (MP) and fibres can be inhaled and cause inflammatory lung diseases. So far MP and fibres have not observed directly in the lung of living animals. A direct visualisation of particles and fibres would be important to study interactions with local and immigration host cells. In this methodical report latex beads were used as model particles for, e.g. nanoparticles, dusts, pollen or bacteria and were investigated using intravital fluorescence microscopy. Intravital fluorescence microscopy of the lung periphery is challenging because of the constant movement of the lung tissue and the heart. Chest window techniques have been described for investigation of lung vessels. For investigation of MP in larger areas of the lung surface this study presents an open chest-technique. Fluorescent MP were instilled into the trachea and could be observed in the alveoli of the right lung. Abundant numbers of MP were found within alveolar macrophages indicating that they are actively engulfed. Using the same setup also fluorescence labelled bacteria and its phagocytosis could be observed as shown in preliminary experiments. In conclusion, we present a method to analyse MP/fibres and its interaction with local and immigrating host cells in the living lung., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Background & Aims: Sinusoidal obstruction syndrome (SOS) following oxaliplatin based chemotherapy can have a significant impact on post-operative outcome following resection of colorectal liver metastases. To date no relevant experimental models of oxaliplatin induced SOS have been described. The aim of this project was to establish a rodent model which could be utilised to investigate mechanisms underlying SOS to aid the development of therapeutic strategies., Methods: C57Bl/6 mice, maintained on a purified diet, were treated with intra-peritoneal FOLFOX (n=10), or vehicle (n=10), weekly for five weeks and culled one week following final treatment. Sections of the liver and spleen were fixed in formalin and paraffin embedded for histological analysis. The role of oxidative stress on experimental-induced SOS was determined by dietary supplementation with butylated hydroxyanisole and N-acetylcysteine., Results: FOLFOX treatment was associated with the development of sinusoidal dilatation and hepatocyte atrophy on H&E stained sections of the liver in keeping with SOS. Immunohistochemistry for p21 demonstrated the presence of replicative senescence within the sinusoidal endothelium. FOLFOX induced endothelial damage leads to a pro-thrombotic state within the liver associated with upregulation of PAI-1 (p<0.001), vWF (p<0.01) and Factor X (p<0.001), which may contribute to the propagation of liver injury. Dietary supplementation with the antioxidant BHA prevented the development of significant SOS., Conclusions: We have developed the first reproducible model of chemotherapy induced SOS that reflects the pathogenesis of this disease in patients. It appears that the use of antioxidants alongside oxaliplatin based chemotherapy may be of value in preventing the development of SOS in patients with colorectal liver metastases., (Copyright © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Besides its classical role in bone and calcium homeostasis, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), the active form of vitamin D, has many non-classical effects; antiproliferative, anti-apoptotic and prodifferentiating effects of 1,25(OH)2D3 have been described in several tumour types in preclinical models. This review focuses on the insights gained in the elucidation of the role of 1,25(OH)2D3 in the normal thyroid and in the pathogenesis, progression and treatment of thyroid cancer, the most common endocrine malignancy. An increasing amount of observations points towards a role for impaired 1,25(OH)2D3-VDR signalling in the occurrence and progression of thyroid cancer, and a potential for structural analogues in the multimodal treatment of dedifferentiated iodine-resistant thyroid cancer. A role for vitamin D in thyroid-related autoimmunity is less convincing and needs further study. Altered 1,25(OH)2D3-VDR signalling does not influence normal thyroid development nor thyrocyte function, but does affect C-cell function, at least in rodents. If these findings also apply to humans deserves further study., (Copyright © 2013 Elsevier GmbH. All rights reserved.)