Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique

Introduction: Liver perfusion has been the standard method to digest and isolate liver cells including liver sinusoidal endothelial cells (LSEC). Poor cannulating skills through portal vein results in a waste of animal resource. Familiarization of both liver perfusion technique and adhering strictly to aseptic technique during cell handling ensure high cell yield, minimum morphology disruption and cell contamination. We aimed to present a method of liver perfusion procedure followed by the isolation of LSEC. Materials and method: The study was conducted with the approval of IACUC committee. Seven Sprague Dawley rats underwent these procedures under anaesthesia. Liver perfusion was done as previously described. Briefly, LSEC were isolated by liberase enzyme perfusion of the liver, isopycnic sedimentation in a two- step Percoll gradient and selective adherence. The purification and cultivation of LSEC was evaluated by light and electron microscopy. Results: Purity and viability of LSEC after selective adherence was 80.5 ± 3.5% and ≥ 95 %, respectively. The average concentration of the cells ranged from 32 - 75 x 106 per 400 gm rat. After 8 hours of culture, LSEC monolayers were contaminated with less than 5% of other cells. Conclusion: This method is reliable and reproducible for the isolation of LSEC to enable the study of structure and function of these cells in vitro. However, improvement on the perfusion skills and isolation technique are vital to ensure better cell purity

Chronic organic arsenic induced liver ultra structural damage

Introduction: Inorganic arsenic is one of the environmental toxins that has been associated with increased risk of cardiovascular diseases and vascular contributions to liver diseases. It has generally been thought to be more toxic than organic arsenic. In human liver, inorganic arsenic promotes vascular remodelling, portal fibrosis and hypertension. The purpose of the current study was to determine whether chronic exposure to organic arsenic impair liver ultrastructure as in inorganic arsenic exposure. Materials and Methods: Twenty eight male SpragueDawley rats were divided into 2 groups with their own control group. They received oral intubation of monosodium- methylarsonate (MSMA) at 63.30 mg/kg body weight for 4 and 6 months duration respectively while the control groups received distilled water. The liver of euthanized rats were perfused- fixed with glutaraldehyde for transmission electron microscopy processing. Results: TEM revealed a marked reduction in the number of mitochondria in both treatment groups. Some typical features of apoptosis are present with pyknotic nuclei of hepatocytes and disintegrated hepatic cytoplasm in 4- month groups. The microvilli of hepatocyte are almost completely absent with the presence of many fibre bundles (collagen fibres) widening the space of Disse. In 6-month treatment group, disintegrated cytoplasms are more prominent with degrading mitochondria of varying stages. Liver sinusoidal endothelial cells (LSEC) of 6-month treatment group are noted to have chromatin condensation with few caveolae seen. Conclusion: Chronic exposure of MSMA leads to necrotic changes of hepatocytes and apoptotic changes in LSEC and collagenisation in the space of Disse

Topographical changes of rat's colonic mucosal after chronic arsenic exposure - A scanning electron microscopy study

Introduction: Human are exposed to arsenic threats in several ways. Our drinking water for instance, can be hazardous due to the contamination of arsenic-based pesticide and herbicide into our water supply. The most vulnerable part of our body due to ingestion of arsenic is our gastrointestinal system. Thus, the aim of this study was to determine the effects of chronic exposure to organic arsenic (Monosodium methylarsonate, MSMA) on the surface topography of rat's colonic mucosa by using scanning electron microscope (SEM). Materials and Methods: 30 Sprague Dawley rats were given daily oral gavage of MSMA 42.13 mg/kg, which is 1/30 LD50, and 30 Sprague Dawley rats acted as control. 10 exposed rats and 10 control rats were sacrificed at regular intervals (2 months, 4 months and 6 months) and their colon specimens were examined by SEM. Results: In the control group, the colonic mucosa appeared normal with uniform size individual glandular units and has a central crypt orifice. The goblet cells were located in between the absorptive cells to produce mucous. In the exposed group, the rat's colonic mucosa showed increasing features of surface alterations such as haphazard shape of glandular units, slit-like crypt opening and less goblet cells with reduced number of microvilli. Conclusion: There were topographical changes of colonic mucosa of rats exposed to chronic low dose of organic arsenic

Chronic low dose organic arsenic induced liver structural damage

Over the decades, organic arsenic has been thought to be less toxic than inorganic arsenic. Monosodium methylarsonate (MSMA) is a potent organoarsenical herbicide that is still being used in most Asian countries, even though in some countries the used has been restricted. Other organic studies reported the effects mainly on the gastrointestinal system. However, the evidence on the severity of it to the liver is still insufficient. The study objective was to investigate the effect of organic arsenic (MSMA) exposure on hepatocytes and liver sinusoidal endothelial cells (LSEC). Rats were exposed to MSMA at 63.20 mg/kg daily for 6 months duration through oral gavage daily. Control rats received distilled water ad libitum. At the end of the duration, they were sacrificed and underwent liver perfusion for tissue preservation. Liver tissue was prepared for light microscopy, scanning and transmission electron microscopy. Histopathological and ultrastructural comparison between control and treated rats were qualitatively described. Histopathological and ultrastructurally, MSMA has caused necrotic and apoptotic changes of the liver with a reduction of organelles in hepatocytes and capillarization of LSEC. Chronic low dose organic arsenic exposure showed evidence of toxicity to hepatocytes. Interestingly, LSEC demonstrated survival accommodation

Combating covid-19 : our management at the campus level

In Malaysia, Covid-19 has caused severe morbidity and mortality. Most people have been affected, including the university community. The spread of the virus has put the campus in peril. Covid-19 Kulliyyah (Faculty) Task Force was established to deal with covid-19 issues at our level. After the establishment of the task force, we faced difficulties in managing the students manually, especially in terms of contract tracing

Chronic exposure of low-dose organic arsenic induced oxidative stress injury on the rats kidney

Introduction: Monosodium methylarsonate (MSMA) is an organic arsenical herbicide widely used in agriculture. Humans are exposed to arsenic through contaminated drinking water and anthropogenic activities. Exposure to acute and high dose of inorganic arsenic induced oxidative stress injury in the tissue. However, studies focusing on effects of chronic low dose organic arsenic exposure on oxidative genes of kidneys are limited. Therefore, the objective of this study is to determine the effects of chronic low dose organic arsenic exposure on rats’ kidneys oxidative genes at 6-month duration. Materials and Methods: Twelve male Sprague-Dawley rats, aged 2-3 months old, with average weight of 200 to 250g divided into two groups (n=6); control and treatment respectively. The control group was given daily oral gavage of distilled water and the treatment group was given daily oral gavage of MSMA at 63mg/kg body weight for 6-months duration. Kidneys were harvested for histology and gene expression analysis. Results: Histologically, on H&E & PAS staining, the control group showed normal appearance of the glomerular and tubular structures. The treatment groups showed microscopic changes of the glomerular and tubular structures such as glomerular atrophy with widening of Bowman’s space, dilated tubular lumen, loss of brush border and presence of tubular cast. Gene’s analysis showed upregulation of Glutathione reductase (GSR) and downregulation of Catalase and nitrite oxide synthase 1 (NOS1). Conclusion: Chronic exposure to low dose organic arsenic induced kidney injury through oxidative stress mechanism

Ultrasonographic study on the arterial variations of the arm and forearm in Malaysian adults

Introduction: The information and incidence of arterial variations have been implicated in different clinical situations. In Malaysia no study on the variants has been done in vivo. Methodology: 400 subjects (staff and students of IIUM) were examined with Logiq P5 General Electric Ultrasound machine using 12L- RS linear probe. Data analysis was carried out with SPSS 19. Results: The variants were found in 11% of the subjects (6.1% of upper limbs). Out of 800 upper limbs, there were seven types of variants; (a) persistent median artery (PMA) (2.6%), (b) brachioradial artery (BRA) (1.7%), (c) superficial brachioulnar artery (SBUA) (1.2%), (d) double radial artery (DRA) (0.6%), (e) brachioulnar artery (BUA) (0.2%), (f) radial artery (RA) loop (0.1%), and (g) aberrant radial artery (0.1%). Variations found in the arm were the BRA, SBUA and BUA while in the forearm were PMA, DRA, RA loop, and aberrant RA. Among them four cases of PMA were associated with BRA, SBUA, BUA, and aberrant RA on the same side. One case of BRA was found together with DRA on the left side. Although the variations were more slightly common in females (6%) than males (5%), and on the right side (3.5%) than the left (2.6%), these differences were not statistically significant (p > 0.05). Conclusion: This study confirms that the arterial variations of the arm and forearm are not uncommon. The microsurgical techniques for reconstruction of upper extremity could be fulfilled by thorough knowledge of the structural variations between individuals in different population

Superficial Brachioulnar Artery (SBUA) & Persistent Median Artery (PMA) : a case report

Variations in the arterial pattern of the upper limbs are frequently observed. During ultrasonography with General Electric Ultrasound machine with 12L-RS linear probe on a 47-year old Malay man, superficial brachioulnar arteries (SBUA) were found bilaterally. While persistent median artery (PMA) was only found on the right forearm. Both SBUA originated 1.5 cm from the origin of the brachial arteries with diameter of 3 mm on the right and 2.8 mm on the left. On the left, it maintained its superficial position on the medial aspect of the arm and the forearm. On the right, initially it ran along the medial aspect of the arm and then coursed laterally to assume a midway position in the cubital fossa. At the elbow, it crossed anterior to the median nerve and coursed in a short distance deep to flexor carpi ulnaris muscle. Later it became superficial at the middle and distal forearm. At the wrist, both arteries ran just lateral to flexor carpi ulnaris and entered the hand with diameter of 1.6 mm. Both brachial arteries were present as their normal course and divided into radial and common interosseous arteries (CIA) approximately 2 cm distal to the elbow crease. The ulnar arteries were not found bilaterally. The right CIA gave off a PMA approximately 2.5 cm from its origin. This PMA descended medially towards the right median nerve and ran along the posterolateral aspect of the nerve. It terminated at the junction between proximal and middle thirds of forearm by fusing with the sheath of median nerve. Knowledge of these variations are clinically important in vascular surgeries and arterial flap plannings

Brachioradial Artery (BRA) & Superficial Brachioulnar Artery (SBUA): a case report

It is no longer uncommon to encounter arterial variants of upper extremities during routine dissection and clinical practice. It has been reported that arterial variations can occur up to 20%. Of these variants, radial and ulnar artery of high origin is quite commonly encountered. We reported a subject with high origin of radial (BRA) and ulnar artery (SBUA) on each side of the upper limb