Hubungan Positif antara Ulkus Kaki Diabetik dengan Persentase Sel Bermarkah Cd4+ Pembawa Malondialdehid

Tingginya angka kejadian ulkus kaki diabetik (UKD) dan luka di kaki yangsulit sembuh memberi petunjuk kemungkinan ada proses kematian sel imun yangsangat banyak dan belum jelas mekanismenya secara molekuler pada jaringan UKD.Telah diteliti hubungan antara derajat UKD dengan persentase sel bermarkahCD4+ pembawa malondialdehid (MDA). Penelitian ini adalah penelitianobservasional dengan rancangan cross sectional analytic study yang dilakukan diRumah Sakit pemerintah dan swasta di Denpasar, Badung, Tabanan, dan Gianyar.Parameter yang diukur dari bahan darah adalah kadar gula darah memakai metodeenzimatik (heksokinase), dan dari bahan jaringan kaki, dihitung sel bermarkah CD4+pembawa MDA memakai metode imunohistokimia (reagen dari Biodesign danAbcam ). Dari 80 sampel UKD didapatkan 49 (61,2%) penderita laki-laki dan 31(38.8%) penderita wanita, berdasarkan tingkat keparahan UKD, sampel dipilah lagimenjadi: 29 (31,9%) derajat 2; 20 (21,9%) derajat 3; 13 (14,3%) derjat 4; dan18(19,8%) derajat 5, rata-rata persentase sel bermarkah CD4+MDA adalah 75,0 ±20,5 %, Didapatkan korelasi positif kuat antara persentase sel bermarkah CD4+pembawa malondialdehid dengan derajat UKD (r = 0,71; p < 0,01). Pada penelitianini membuktikan ada mekanisme kematian sel imun dan sekaligus menjawabpermasalahan bahwa pada penderita UKD mudah terkena infeksi dan sulit untukdisembuhkan, dengan dibuktikan bahwa ada korelasi positif kuat antara derajat UKDdengan persentase pembentukan MDA dari sel bermarkah CD4+, ini menyatakanbahwa semakin berat derajat UKD semakin banyak mengalami kematian sel imun.

Mouse model of Schistosomiasis: infection with Schistosoma mansoni in CD-1 mice

Schistosomiasis is a parasitic disease that affects almost 240 million worldwide. CD1 mice were infected with cercariae of S. mansoni, after which infection developed for 8 weeks. Tissues were processed to immuno-histological techniques. It was performed H&E staining for overall analyses, Sirius Red for fibrosis and immunohistochemistry for inflammation biomarkers. The most infected organ was the liver, fibrosis decreased with egg development and Galectin-3 (Gal3) and Interleukin 6 (IL-6) were expressed inside granulomasThis work was also supported by FCT – Fundação para a Ciência e Tecnologia (REF UID/BIM/04293/2013) and by the project NORTE-01-0145-FEDER-000012 and by a scholarship to Carla Luís with the reference SAICT2016/FEDER/BIO4DIA/BTI under the supervision of Dr. Rúben Fernandes.N/

Virtual libraries of tissue and clinical samples: potential role of a 3-D microscope.

Our international innovative teaching group from different European Universities (De Montfort University, DMU, UK; and the Spanish University of Alcalá, University Miguel Hernández and University of San Pablo CEU), in conjunction with practicing biomedical scientists in the National Health Service (UK) and biomedical researchers, are developing two complete e-learning packages for teaching and learning medical parasitology, named DMU e-Parasitology (accessible at: http://parasitology.dmu.ac.uk), and biology and chemistry, named DMU e-Biology (accessible at: http://parasitology.dmu.ac.uk/ebiology/index.htm), respectively. Both packages will include a virtual microscope with a complete library of digitised tissue images, clinical slides and cell culture slides/mini-videos for enhancing the teaching and learning of a myriad of techniques applicable to health science undergraduate and postgraduate students. Thus, these packages include detecting human parasites, by becoming familiar with their infective structures and/or organs (e.g. eggs, cysts) and/or explore pathogenic tissues stained with traditional (e.g. haematoxylin & eosin) or more modern (e.g. immunohistochemistry) techniques. The Virtual Microscope (VM) module in the DMU e-Parasitology package is almost completed (accessible at: http://parasitology.dmu.ac.uk/learn/microscope.htm) and contains a section for the three major groups of human-pathogenic parasites (Peña-Fernández et al., 2018) [1]. Digitised slides are provided with the functionality of a microscope by using the gadget Zoomify®, and we consider that they can enhance learning, as previous studies reported in the literature have reported similar sensitivity and specificity rates for identification of parasites for both digitised and real slides. The DMU e-Biology’s VM, currently in development, will provide healthy and pathological tissue samples from a range of mammalian tissues and organs. This communication will provide a description of both virtual libraries and the process of developing them. In conjunction, we will use a three-dimensional (3D) super-resolution microscopy, 3D Cell Explorer (Nanolive, Lausanne, Switzerland), to incorporate potential 3D microscopic photographs/short videos of cells to provide students with information about the spatial arrangement and morphologies of cells that are essential for life

Protein Modifications as Potential Biomarkers in Breast Cancer

A variety of post-translational protein modifications (PTMs) are known to be altered as a result of cancer development. Thus, these PTMs are potentially useful biomarkers for breast cancer. Mass spectrometry, antibody microarrays and immunohistochemistry techniques have shown promise for identifying changes in PTMs. In this review, we summarize the current literature on PTMs identified in the plasma and tumor tissue of breast-cancer patients or in breast cell lines. We also discuss some of the analytical techniques currently being used to evaluate PTMs

Targeting danger molecules in tendinopathy: the HMGB1/TLR4 axis

Objectives: To seek evidence of the danger molecule, high-mobility group protein B1 (HMGB1) expression in human tendinopathy and thereafter, to explore mechanisms where HMGB1 may regulate inflammatory mediators and matrix regulation in human tendinopathy. Methods: Torn supraspinatus tendon (established pathology) and matched intact subscapularis tendon (representing ‘early pathology’) biopsies were collected from patients undergoing arthroscopic shoulder surgery. Control samples of subscapularis tendon were collected from patients undergoing arthroscopic stabilisation surgery. Markers of inflammation and HMGB1 were quantified by reverse transcriptase PCR (RT-PCR) and immunohistochemistry. Human tendon-derived primary cells were derived from hamstring tendon tissue obtained during hamstring tendon anterior cruciate ligament reconstruction and used through passage 3. In vitro effects of recombinant HMGB1 on tenocyte matrix and inflammatory potential were measured using quantitative RT-PCR, ELISA and immunohistochemistry staining. Results: Tendinopathic tissues demonstrated significantly increased levels of the danger molecule HMGB1 compared with control tissues with early tendinopathy tissue showing the greatest expression. The addition of recombinant human HMGB1 to tenocytes led to significant increase in expression of a number of inflammatory mediators, including interleukin 1 beta (IL-1β), IL-6, IL-33, CCL2 and CXCL12, in vitro. Further analysis demonstrated rhHMGB1 treatment resulted in increased expression of genes involved in matrix remodelling. Significant increases were observed in Col3, Tenascin-C and Decorin. Moreover, blocking HMGB1 signalling via toll-like receptor 4 (TLR4) silencing reversed these key inflammatory and matrix changes. Conclusion: HMGB1 is present in human tendinopathy and can regulate inflammatory cytokines and matrix changes. We propose HMGB1 as a mediator driving the inflammatory/matrix crosstalk and manipulation of the HMGB1/TLR4 axis may offer novel therapeutic approaches targeting inflammatory mechanisms in the management of human tendon disorders

Occurrence and Regional Distribution of TRAIL and DR5 on Temporomandibular Joint Discs: Comparison of Disc Derangement with and without Reduction

Background Tumor necrosis factor (TNF)–related apoptosis-inducing ligand (TRAIL) is an apoptosis-inducing member of the TNF gene family which triggers apoptotic signals by interaction with its receptors. It has been suggested to be a major contributing factor to tissue degeneration. Objective The present study investigated, through immunohistochemistry, the regional expression of TRAIL and in temporomandibular joint (TMJ) disc of anterior disc displacement with reduction (ADDwR) and without reduction (ADDwoR) patients, to help determine the relationship between TMJ disc displacement and apoptosis. Study design We studied 18 TMJ diseased discs affected by disc displacement without or with reduction and 4 normal TMJ discs. Specimens were processed for immunohistochemistry to evaluate TRAIL and its receptor DR5 expression. Results Disc tissues from internal derangements (both ADDwR and ADDwoR) exhibited a much higher percentage of TRAIL- and DR5-positive cells as well as stain intensity compared with normal tissue though with regional variation according to the portion of the disc. There was a significantly higher percentage of stained cells in the posterior disc attachment compared with the anterior or intermediate bands of both ADDwR and ADDwoR discs for TRAIL and DR5. Conclusions TRAIL and DR5 are overexpressed in displaced human TMJ disc, especially in the posterior disc attachment. These results suggest a possible pivotal role of the TRAIL/DR5 system in TMJ disc degeneration

P53 mutations in human adrenocortical neoplasms

The mechanisms of tumorigenesis of adrenocortical neoplasms have not been elucidated as yet. However, loss of heterozygosity at chromosomal locus 17p has been consistently observed in adrenocortical cancer. p53 is a recessive tumor suppressor gene located on chromosome 17p. Mutations in the p53 gene play an important role in the tumorigenesis of diverse types of human neoplasms including breast and colon cancers. More than 90% of all mutations discovered in such tumors have been detected in 4 hot spot areas that lie between exons 5 and 8. In contrast to wild-type p53, mutant p53 accumulates intracellularly and can be easily detected by immunohistochemistry. We therefore investigated the frequency of p53 mutations in human adrenocortical neoplasms using molecular biology and immunohistochemistry techniques. Five patients with adrenocortical adenomas (5 female; ages 39-72 yr), 11 patients with adrenocortical carcinomas (8 female, 3 male; ages 15- 50 yr), and two adrenocortical tumor cell lines were studied. After DNA extraction from frozen tumor tissue or paraffin-embedded material, exons 5 through 8 were amplified using the polymerase chain reaction and directly sequenced by the dideoxy termination method. Immunohistochemistry was performed on paraffin-embedded tumor specimens obtained during adrenalectomy using a monoclonal antibody reacting with both wild-type and mutant p53. Prevalence of mutations was adenomas, 0/5, carcinomas, 3/11, and adrenocortical cell lines, 2/2. Single point mutations were detected in 3 cases (exons 5, 6, and 7, respectively), and rearrangements of exon 7/8 and 8 were found in 2 cases. Immunohistochemistry detected strong nuclear and/or cytoplasmic p53 immunoreactivity in all adrenocortical carcinomas with point mutations of the p53 gene but not in adenomas and carcinomas with the wild-type sequence or with deletion/rearrangement of the p53 gene. We conclude that p53 plays a role in the tumorigenesis of adrenocortical carcinomas but is of less importance to benign adenomas

Effects of Methyl Cycle Substrate Availability on Epigenetic Stability of Human Embryonic Stem Cells

A link has been hypothesised to occur between suboptimal maternal nutrition and impaired foetal development leading to a predisposition to a range of adult pathologies. As a clear connection between dietary intake of methyl group donors and epigenetic defects has been demonstrated both *in vivo* and *in vitro*, this project had the purpose of generating a disruption into the methyl/folate cycle to investigate DNA methylation alterations during human preimplantation embryo development, using human embryonic stem cells (hESCs) as an in vitro model. In particular, HUES-7 stem cells were employed and cultured using either standard or methyl deficient media to test this hypothesis. After the treatments, that included an inhibitor of a key enzyme of the cycle, Differentially Methylated Regions (DMRs) of six imprinted genes were analysed and assessed for their methylation status at Cytosine-phospho-Guanosine (CpG) sites. As a consistent decrease of methylation was observed for the gene *H19* in treated cultures, its allelic expression was then investigated and an initial process of Loss Of Imprinting (LOI) was found. Additionally, global DNA MethylTransferase (DNMT) activity was examined and a statistically significant decrease in treated samples was detected. Finally, hESCs were differentiated into Embryoid Bodies(hEBs), which were compared and stained for pluripotency and germ-layer specific markers. Consistently different expression of *OCT-4* and *NANOG* was noticed for treated-culture derived hEBs

Differences in localization of P2X7 during epithelial wound healing in pre-type II diabetic models

Corneal injury, accompanied by improper wound repair, is the 4th highest cause of preventable blindness according to the World Health Organization. The cornea, which is the most densely innervated structure in the human body, serves to protect the delicate internal environment of the eye from damage. The integrity of this intricate nerve structure is critical in our ability to sense even the slightest insult to the corneal surface, with reduced sensitivity leading to increased susceptibility to trauma. In diabetes, decreased corneal sensitivity secondary to diabetic peripheral neuropathy can lead to increased corneal abrasion, ulceration, and even blindness. The P2X7 purinoreceptor is an ion channel that is expressed by the epithelium along with the intra-epithelial nerves and stromal nerves. The goal of our study was to use a type 2 diabetic mouse model (DIO) to characterize the changes in P2X7 localization and potentially correlate our results with changes in trafficking and sensory nerve loss. We hypothesized that the P2X7 receptor acts to sense changes at the leading edge and this fine tuned regulation is altered during the diabetic disease state. Further understanding of the corneal changes that occur in diabetes can help us better monitor progression of diabetic complications as well as develop new therapeutics for the treatment of diabetic corneal dysfunction

Imaging-guided chest biopsies: techniques and clinical results

Background This article aims to comprehensively describe indications, contraindications, technical aspects, diagnostic accuracy and complications of percutaneous lung biopsy. Methods Imaging-guided biopsy currently represents one of the predominant methods for obtaining tissue specimens in patients with lung nodules; in many cases treatment protocols are based on histological information; thus, biopsy is frequently performed, when technically feasible, or in case other techniques (such as bronchoscopy with lavage) are inconclusive. Results Although a coaxial system is suitable in any case, two categories of needles can be used: fine-needle aspiration biopsy (FNAB) and core-needle biopsy (CNB), with the latter demonstrated to have a slightly higher overall sensitivity, specificity and accuracy. Conclusion Percutaneous lung biopsy is a safe procedure even though a few complications are possible: pneumothorax, pulmonary haemorrhage and haemoptysis are common complications, while air embolism and seeding are rare, but potentially fatal complications