Protein arginine methyltransferase 3 promotes glycolysis and hepatocellular carcinoma growth by enhancing arginine methylation of lactate dehydrogenase A.

BACKGROUND: Protein arginine methylation has emerged a pivotal role in cancer progression. However, the role of protein arginine methyltransferase 3 (PRMT3) in hepatocellular carcinoma (HCC) remains unknown. METHODS: The expression pattern of PRMT3 in HCC was analysed using quantitative real-time-polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry assays. Loss- and gain-of-function experiments were carried out to determine the oncogenic role of PRMT3 in HCC. Glucose consumption and lactate production assays, seahorse bioscience, mass spectrometry, co-immunoprecipitation, metabonomic analysis and site-specific mutation experiments were used to explore the underlying molecular mechanisms. Furthermore, a xenograft mouse model was established to investigate the effects of PRMT3 and its inhibitor, SGC707, treatment on tumour growth in vivo. RESULTS: The expression of PRMT3 was significantly upregulated in HCC, with high expression of which correlated with poor prognosis. PRMT3 knockdown led to the decrease in proliferation, glycolysis of HCC cells and tumour growth, whilst its overexpression showed opposite results. The catalytic activity of PRMT3 was important in mediating these biological processes. Mechanistically, our data showed that PRMT3 interacted with and mediated asymmetric dimethylarginine (ADMA) modification of lactate dehydrogenase A (LDHA) at arginine 112 (R112). Compared with LDHA-wild-type (LDHA-WT) cells, LDHA-R112K-mutant-expressing HCC cells exhibited a decrease in lactate dehydrogenase (LDH) activity, HCC cell glycolysis and proliferation. Furthermore, the administration of SGC707, a selective inhibitor of PRMT3, disrupted the PRMT3-mediated LDHA methylation and abolished PRMT3-induced HCC glycolysis and tumour growth. CONCLUSIONS: Our results suggested a novel oncogenic role of PRMT3 in HCC, and it could be a promising therapeutic target for HCC by linking post-translational modification and cancer metabolism.

Adverse Histological Features of Differentiated Thyroid Cancer Are Commonly Found in Autopsy Studies: Implications for Treatment Guidelines.

Background: While the popularity of lobectomy for differentiated thyroid cancer (DTC) has increased since the 2015 ATA (American Thyroid Association) guidelines, recent studies reported that adverse histological features (minimal extrathyroidal extension [mETE], multifocality, vascular invasion, and lymph node [LN] metastases) may be found in 30-60% of lobectomy specimens, questioning the validity of this approach. Aim: To assess the prevalence adverse histological features in occult DTC detected in autopsy studies. Methods: Meta-analysis of autopsy studies of the thyroid in subjects without known history of thyroid cancer. Results: Twenty-nine studies including 8750 subjects fulfilled the inclusion criteria, with incidentally discovered DTC in 740 autopsies (8.5%). Age was reported in 17 studies, with a median age of 61 years (range 41-68 years). Multifocality was reported in 27 studies with a calculated event rate of 28.2% ([CI 23.1-33.8], I2 = 46.3%), with bilateral involvement in 18% [CI 12.6-25.1]. mETE was reported in 5 studies, with an event rate of 24.5% ([CI 9.3-50.7], I2 = 88.5%), and the presence of LN metastases were reported in 13 studies with an event rate of 11% ([CI 6.1-19.1], I2 = 69.5%). Vascular invasion was reported in seven studies with an event rate of 16% ([CI 4-47], I2 = 86.8%). Of 25 studies with whole body autopsies (722 subjects), 3 cases of distant metastases were reported, of which 2 had fatal metastatic disease (where thyroid origin was not diagnosed before death), and 1 had occult disease. Conclusions: Adverse histological features including mETE, LN metastases, multifocality, and vascular invasion are common in occult DTC. When minimal in size, these adverse histological features do not seem to be markers of aggressive disease and may not be an indication for completion thyroidectomy or radioiodine therapy.

Reporting regression in primary cutaneous melanoma. Part 2: prognosis, evaluation and management.

The effect of histological regression on patient prognosis for primary cutaneous melanoma is controversial. Some authors hypothesize that regression indicates a robust systemic immune response and may decrease risk of metastasis. Others argue that histological regression calls into question a T0 diagnosis because there may have been an invasive component of the melanoma that is no longer visible but is still active. The literature to date does not suggest that histological regression is associated with increased risk of positive sentinel lymph node status, metastasis or increased risk of mortality. Thus, the presence of histological regression should not change patient staging, evaluation or management. The criteria used for reporting regression have varied dramatically across studies, and standardized reporting is needed to foster evidence-based practices in the future.

Terminologia Histologica 10 years on: some disputable terms in need of discussion and recent developments.

At first sight, the issue of morphological terminology may seem to be a "closed and unchanging chapter", as many of the structures within the human body have been known for decades or even centuries. However, the exact opposite is true. The initial knowledge of the microscopic structure of the human body has been continuously broadening thanks to the development of new specialized staining techniques, discovery of the electron microscope, or later application of histochemical and immunohistochemical methods into routine tissue examination. Contrary to popular belief, histology has a status of constantly developing scientific discipline, with continuous influx of new knowledge, resulting in an unavoidable necessity to revise the histological nomenclature at regular intervals. The team of experts of the Federative International Programme on Anatomical Terminology, a working group of the International Federation of Associations of Anatomists, published in 2008 the First Edition of Terminologia Histologica. Terminologia Histologica (TH) is the best and most extensive of all the histological nomenclatures ever issued. However, here we suggest that several terms of important histological structures are still missing while several other terms are disputable. First, we present some clinically important terms of cells and tissue structures for inclusion in the next TH and, in a second part, we refer to some new terms in the current edition of the TH which are not yet mentioned in current histology textbooks (e.g., fusocellular connective tissue, bundle bone as the third type of bone tissue, spongy layer of vagina or arteria vaginata from the splenic white pulp). With this article we hope to start a wide scientific discussion which will lead to an inambiguous definition and demonstration of typical examples of all terms in the TH, with the result that the new edition of the Terminologia Histologica will become an internationally accepted communication tool for all practitioners and teachers of histology alike.

The changing face of adult posttransplant lymphoproliferative disorder: Changes in histology between 1999 and 2013.

Posttransplant lymphoproliferative disorder (PTLD) typically presents with either polymorphic or monomorphic histology. While both are the end result of immunosuppressive therapies, their origins are felt to be different with different prognoses and responsiveness to therapy, resulting in 2 different malignancies. We attempted to confirm reports suggesting that the relative frequency of these 2 histologies is shifting over time. We analyzed 3040 adult PTLD cases in the UNOS OPTN database from 1999 to 2013. Changes in PTLD cases over time were analyzed for histology, time from transplant to diagnosis, and patient EBV serostatus. We found that the relative proportion of polymorphic versus monomorphic histology has changed with an increase in the proportion of monomorphic cases with time (1999-2003, 54.9% vs. 45.1%; 2004-2008, 58.3% vs. 41.7%; 2009-2013, 69.7% vs. 30.3%; P = <.001). The change is driven by a gradual increase in the number of monomorphic PTLD with a steady number of polymorphic PTLD. The change is most strongly seen in transplant recipients who were EBV serostatus positive at the time of transplant. Potential causes are changes in immunosuppressive regimens with increased tacrolimus use (P = .009) and increased survival among transplant patients leading to later occurrence of PTLD (P = .001) that have occurred during the time frame analyzed. As organ transplantation has evolved over time, PTLD has coevolved. These changes in histology have important implications regarding the origin and clinical management of PTLD.

Tissue proteomics for the next decade? Towards a molecular dimension in histology.

The concept of tissues appeared more than 200 years ago, since textures and attendant differences were described within the whole organism components. Instrumental developments in optics and biochemistry subsequently paved the way to transition from classical to molecular histology in order to decipher the molecular contexts associated with physiological or pathological development or function of a tissue. In 1941, Coons and colleagues performed the first systematic integrated examination of classical histology and biochemistry when his team localized pneumonia antigens in infected tissue sections. Most recently, in the early 21(st) century, mass spectrometry (MS) has progressively become one of the most valuable tools to analyze biomolecular compounds. Currently, sampling methods, biochemical procedures, and MS instrumentations allow scientists to perform "in depth" analysis of the protein content of any type of tissue of interest. This article reviews the salient issues in proteomics analysis of tissues. We first outline technical and analytical considerations for sampling and biochemical processing of tissues and subsequently the instrumental possibilities for proteomics analysis such as shotgun proteomics in an anatomical context. Specific attention concerns formalin fixed and paraffin embedded (FFPE) tissues that are potential "gold mines" for histopathological investigations. In all, the matrix assisted laser desorption/ionization (MALDI) MS imaging, which allows for differential mapping of hundreds of compounds on a tissue section, is currently the most striking evidence of linkage and transition between "classical" and "molecular" histology. Tissue proteomics represents a veritable field of research and investment activity for modern biomarker discovery and development for the next decade.

[Impact of the pathology in modern medicine]. / Représentation en sciences du vivant (9) émergence d'une spécialité médicale nouvelle : la pathologie.

In this review, the authors emphasize the pivotal role of the pathology in the setting of a revolution which progressively transforms medical sciences into basic sciences. Several key aspects of this specialty will be discussed together with the main perspectives in the fields of oncologic disorders.

Real-time histology with the endocytoscope.

Endoscopic Imaging has progressed tremendously over the last few decades. Novel imaging technologies such as high-resolution and high-magnification white light endoscopy, narrow band imaging, optimal band imaging, autoflourescence imaging and optical coherence tomography not only aid the endoscopist in detecting malignant or pre-malignant lesions but also assist in predicting histology. Recently, the introduction of Endocytoscopy (EC) and Confocal Endomicroscopy has taken us into a new realm of diagnostic endoscopy. With the ability to magnify up to 1000 ×, cellular structures can be visualized in real-time. This advance in technology could potentially lead to a paradigm shift negating the need to obtain biopsies. EC is, however, still in the early stages of development and further research needs to be carried out before it can be accepted as standard practice. This review will focus on the diagnostic utility of the Endocytoscope.

Histology without xylene.

After the hazardous effects of xylene became indisputable in the 1970s, many potential substitutes became available, some with as many if not more hazards. This article discusses the inadequacy of 5 vegetable oils as substitutes, as well as the characteristics of 22 D-limonene-based substitutes, all less effective in their chemical role, some capable of inducing health problems, and costing more than twice as much as xylene. Some of the 35 alkane-based substitutes discussed are effective for tissue processing, less toxic, with a cost about the same as xylene, but are not very effective for dewaxing and other staining tasks. Isopropanol (2-propanol) alone or mixed with molten paraffin is a technically acceptable and cost-effective substitute for xylene for tissue processing, but in this study, we demonstrate that the best clearing agents from the sectioning quality and diagnostic value point of view, with automated or manual protocols, are mixtures of 5:1 and 2:1 isopropanol and mineral oil, followed by undiluted mineral oil, all at 50 degrees C, making them a safer and cheaper substitute than xylene. Using a 1.7% dishwasher soap aqueous solution at 90 degrees C to dewax before staining and oven drying the stained sections before coverslipping will eliminate xylene from the staining tasks. Tissue processors retorts and conduits can be dewaxed with a 2% solution of a strong glassware laboratory detergent. These 4 methodologies will make the histology laboratory xylene-free but, due to the natural resistance to change, many histotechs will be reluctant to adopt them if they think that their technical expertise could be jeopardized, and the only way these changes will succeed is if the pathologists, as stewards of the histology laboratory, commit to their implementation.

Histology aging workforce and what to do about it.

There are 28,400 histotechs now working in human surgical and forensic pathology, half of whom will reach retirement age within the next 4 years, and to compensate for those retiring and to increase their number to the 31,200 needed in 2015, a 3-pronged solution is required. Firstly, each individual medical laboratory has to implement a uniform salary scale for all its members with equivalent professional level that will probably delay the retirement plans of some histotechs and will allow the flow of personnel between its different areas. Secondly, a new certification of "qualification in histology" should be created so medical technologists can cover for retiring histotechs and also could find new working positions in laboratories that may be contemplating staff reductions because of more productive instruments and workflow schemes. Finally, given the historical examination passing rates, it is necessary to enroll at least 4,965 students in accredited histology programs (709 annually), and for this to happen, either the present number of 33 programs has to be increased by 17 (to a total of 50) with a similar capacity each to those now in operation or 2 to 3 distance learning programs with a capacity of 85 to 125 each have to be created after the example of some already existing. This solution will also require that local students soon to enter junior colleges are informed about these programs and work opportunities. To assure the success of any of the 3 approaches, it is imperative that the pathologists get involved, individually for the first one, and collectively for the other two; the pathologists should bring these solutions to their respective societies and college boards and meetings and sound the alarm on this problem that will adversely impact their work and their overall patient care. The pathologists have to be aware of this very grave situation-that the histotechs' retirement clocks are ticking and that the time to act is now!

Brain histology.

In this article we summarise nervous system histology in health and disease and acquaint the reader with developments in the staining techniques that are in current use, particularly immunostains. Although clinicians do not need to know the details of stain appearances, some familiarity with these aspects of neuropathology is invaluable in interpreting the reports they receive from the laboratory, as well as reminding them of the amazing beauty of the central nervous system's microscopic structure.

Histology safety: now and then.

Histology safety usually focuses on general laboratory issues, but this article concentrates on the hazards affecting the individual histotech and their evolution in the last half a century. Using the information from a survey especially designed for the occasion, the hazards were divided into 4 groups, and their prevalence was expressed as percentages for national and foreign laboratories. All the laboratories received a "safety index" (SI) with an average value of 0.77 +/- 0.11 for 63 national laboratories and 0.69 +/- 0.13 for 22 foreign laboratories, these 2 averages being statistically different (P < .02). The historical evolution of the SI required answering the same questionnaire retrospectively, and so it was done for 17 laboratories with an SI average of 0.27 +/- 0.12 for 1955/1989 and 0.77 +/- 0.13, almost 3 times larger for 1990/2007, with improvement of all safety issues. The technological, organizational, and regulatory advances before 1989 showed an unremarkable effect on the SI, and the only circumstance considered as the driving force behind the almost triple increment of the SI during 1990/2007 was the awareness that the AIDS epidemic instilled in the minds and consciences of the medical laboratory personnel in general. Even after almost tripling the average SI value in 2007, national histology laboratories obtained a grade average of "C+" only, leaving room for improvement.

Creating a histology-embryology free digital image database using high-end microscopy and computer techniques for on-line biomedical education.

The development of new technology and the possibility of fast information delivery by either Internet or Intranet connections are changing education. Microanatomy education depends basically on the correct interpretation of microscopy images by students. Modern microscopes coupled to computers enable the presentation of these images in a digital form by creating image databases. However, the access to this new technology is restricted entirely to those living in cities and towns with an Information Technology (IT) infrastructure. This study describes the creation of a free Internet histology database composed by high-quality images and also presents an inexpensive way to supply it to a greater number of students through Internet/Intranet connections. By using state-of-the-art scientific instruments, we developed a Web page (http://www2.uerj.br/~micron/atlas/atlasenglish/index.htm) that, in association with a multimedia microscopy laboratory, intends to help in the reduction of the IT educational gap between developed and underdeveloped regions.

Laboratory instruction in histology at the University at Buffalo: recent replacement of microscope exercises with computer applications.

Histology is a morphologic science in which the structure of the cells, tissues, and organs of the body are examined with a microscope. In the laboratory courses in histology at the School of Medicine of the University at Buffalo, histologic specimens had been used since the late 19th century to teach the principles of cell, tissue, and organ structure. Students also had to learn how to analyze or "read" slides with a microscope. Learning histology in this way, i.e., by direct examination of actual specimens, is time consuming and viewed by some as unnecessary. As a result of recent curricular reform at the School of Medicine that reduced contact time in histology, half of all laboratory exercises that would have been performed with a microscope were performed instead with interactive computer applications. By replacing some microscope exercises with more efficient computer applications, the histology course accommodated curricular change by both reducing contact time and continuing to offer valuable microscope laboratories for most of the organ systems of the body. To provide a basis for comparing traditional microscope exercises with computer-assisted instruction in histology, the nature of the laboratory experience between 1846 and 1998 is briefly reviewed. The instructional strategy behind the use of computers is presented, along with the nature of the computer applications and the means by which the computer applications were incorporated into the school's laboratory course in histology.

Will the "new biology" affect the future of histopathology?

The technologies used in histopathology are changing as a consequence of the current revolutionary progress in several areas of biology. It is likely that general cancer management will improve because of the impact of molecular techniques and immunohistochemistry on tumor diagnosis and classification and on the determination of prognosis and response to therapy. Moreover, as therapies are starting to be modelled after the distinctive molecular characteristics of a specific tumor, the availability of molecular tests to all patients will become a matter of great importance.