Assessment of Pathogenicity of Community-Acquired MRSA Isolates in Mice-Induced Peritonitis

@#Introduction: Methicillin-Resistance Staphylococcus aureus (MRSA) is known as a major nosocomial pathogen in healthcare. However, it has now spread in the community known as community-acquired MRSA (CA-MRSA). Thus, the survival and pathogenicity of CA-MRSA isolates were assessed using in vivo peritonitis model with comparison to ATCC-MRSA. Two CA-MRSA isolates; CA-MRSA1 and CA-MRSA2 that were isolated from healthy population, were studied and compared. Methods: Mice were assigned into 4 groups and injected intraperitoneally with ATCC-MRSA, CA-MRSA1 or CA-MRSA2, respectively. Sterile Dulbecco’s Phosphate-Buffered Saline (DPBS) represents negative control. Mice were observed twice daily, 0-72 hours of post-infection. Any signs of distress were recorded for severity score and survival analyses. Mice were euthanised at 72 hours post-inoculation or by referring to the Peritonitis Severity Scoring (PSS) system. Organs of interest, peritoneal lavage and abscess were processed for bacterial counts. Tissue samples were analysed for histopathological scores. Results: All mice inoculated with MRSA showed clear signs of illness with peritonitis symptoms of p<0.001 and comparable PSS scores were recorded in all infected mice groups. Intraperitoneal injection of lethal dose of MRSA resulted in significant death of ATCC-MRSA (p<0.05) and CA-MRSA-infected mice (p<0.01), compared to the un-infected. Bacterial burden was significantly high in all samples harvested from mice challenged with CA-MRSA2 compared to ATCC-MRSA except in abscess and lung. Significant liver necrosis and spleen inflammation were observed in CA-MRSA1, and lung inflammation in ATCC-MRSA-infected mice. Conclusion: Nasal carriage CA-MRSA isolates from a healthy population has the potential to cause peritonitis with comparable severity as ATCC-MRSA.

Cutaneous involvement of multiple myeloma

@#Cutaneous multiple myeloma (MM) is a rare disease. It can be primary or secondary in origin. The secondary type is further classified into specific and nonspecific types. The specific type is uncommon and is known as a secondary cutaneous plasmacytoma. We report a case of secondary cutaneous plasmacytoma in a 58-year-old man who had a history of plasma cell tumour of the lung and multiple myeloma. He achieved complete remission after the completion of chemotherapy and autologous stem cell transplant (ASCT). However, five months later, he developed multiple erythematous nodules on the whole body. Skin biopsy revealed diffuse neoplastic cells infiltrate in the reticular dermis with sparing of the upper papillary dermis and epidermis. The neoplastic cells were monotonous and homogenous with variable degrees of cytological atypia. Occasional cells showed distinctive plasma cell features. Plasma cell lineage was confirmed with CD138. The cells were immunoreactive to Kappa. Ki-67 was greater than 90%. They were non-immunoreactive to CD45, CD3, CD20, CD79 alpha and CK AE1/AE3. The findings were consistent with secondary cutaneous plasmacytoma. Our case illustrates that MM may present with nonspecific dermatological manifestations. As specific cutaneous involvement of MM is very uncommon; a high degree of clinical suspicion, detailed medical history and histopathological examination are required to arrive at an early diagnosis.

Correlation of E-Cadherin Expression with Clinicopathological Parameters and Its Outcomes among Prostate Cancer Patients in Hospital Kuala Lumpur

@#Introduction: Aberrant expression of E-cadherin has shown to have correlation with advanced disease of prostate cancer. In this study, we evaluated the potential of E-cadherin as a prostate cancer prognostic marker and determined its correlation with patient outcomes. Method: 46 prostate cancer specimens in the form of paraffin-embedded tissue blocks were retrieved from the Histopathology Unit, Department of Pathology, Hospital Kuala Lumpur. The expression patterns of E-cadherin were determined by immunohistochemistry staining. The E-cadherin expression was evaluated and scored as positive (3+) and negative or loss of expression (2+ and 1+). The correlations of E-cadherin expression with patient outcomes which included biochemical failure, disease-free, metastasis and local recurrence were determined. Correlations of E-cadherin expression with the currently used traditional clinicopathological parameters were also evaluated. Results: There were significant correlations between E-cadherin expression with biochemical failure (p=0.005) and local recurrence (p=0.003). However, there were no significant correlations between E-cadherin expression with disease-free (p=0.864) and tumour metastasis (p=0.430). Comparing the correlation of E-cadherin expression with the traditional clinicopathological parameters, there were significant correlations between E-cadherin expression with pathological staging (p=0.001), Gleason score (p=0.004) and perineural invasion (p=0.001). However, there was no significant correlation between E-cadherin expression with positive tumour margin (p=0.320). Conclusion: These results support the potential use of E-cadherin as a prognostic tool for prostate cancer as well as an additional marker along the currently available traditional clinicopathological parameters.

Superficial Angiomyxoma of the Eyelid. A Case Report and Review of the Literature

@#Superficial angiomyxoma (SA) is an angiomyxoid cutaneous tumour. It has predilection for the head and neck, torso, limbs and genital tract. Our case is a 27-year-old female, presented with painless right medial canthal mass for two years. It was associated with tearing when the lesion grew larger. We received a nodular brownish tissue measuring 25x20x15mm with homogenous brownish cut surface. Microscopically, the tumour was partially circumscribed, exhibiting bland stellate to spindle cells of moderate cellularity with pale eosinophilic cytoplasm, indistinct border, in a loose collagenous myxoid matrix with numerous blood vessels of different calibre. The lesional cells were present at the resected margin and were nonreactive towards CD34, SMA and S100. SA of the eyelid is sometimes mistaken clinically as dermoid cyst or lipoma. Reports have shown increased risks of local recurrence following incomplete excision. Close association with Carney’s complex is an important feature. Careful clinicopathologic correlation and proper investigations are needed for optimal patient care.

Fatal Non-traumatic Fat Embolism

@#Non-traumatic fat embolism (NTFE) is infrequently encountered in the clinical setting. The incidence of clinically detected fat embolism is less than 1%. It is diagnosed based on the evidence of fat emboli occluding the vessel lumen in a patient with no prior history of trauma. We report a case of NTFE in a lady who developed breathlessness and collapsed following home vaginal delivery. Post-mortem examination revealed extensive fat emboli in the pulmonary vessels. We elucidate the clinical challenges in diagnosing fat embolism (FE) in a patient without history of trauma.

Cardiac Related Mortality Trends in Forensic Department of Hospital Serdang, Selangor, Malaysia

@#Introduction: Cardiovascular disease (CVD) is the principal cause of admission and death in the Malaysian government hospitals. Method: The pattern of cardiac related mortality (CRM) cases in Hospital Serdang, Selangor was reviewed to determine the specific trends in ethnicity, age, gender and type of CRM. Data was drawn from the death registry records in Forensic Department of Hospital Serdang from January 2006 to December 2008. This preliminary project was carried out to assess the number of CRM cases and their characteristics to feed the decision to sensibly embark on another project related to atherosclerosis. Results: Out of 573 CRM cases, 84.5% were Malaysians out of which the majority were Malays (45.9%), followed by Chinese (30.2%), and Indians (22.1%). The majority were men (78.4%) with a mean age of death at 51.83 ± 14.10 (95% CI 50.52-53.14) years as compared to women (21.6%) with a mean age of death at 57.42 ± 16.92 (95% CI 54.41-60.43) years. Declining mortality trend was observed only in men. The 46-60 years old age group had the highest percentage of mortality and continually increase by year. Ischaemic heart disease (IHD) was the most common CRM type (69.1% in 2006, 66.2% in 2007, and 71.9% in 2008). Conclusion: We observed inconsistent trends of CRM in Hospital Serdang in terms of ethnicity, age, gender and type of CRM cases presented. Nevertheless, these findings do not necessarily represent the overall trends of CRM in Malaysia.

Identifying Common Mutations in Colorectal Cancer Using a 7-Gene Panel by Next Generation Sequencing

Abstract@#Introduction: Colorectal cancer (CRC) arises from the cumulative effects of genetic and epigenetic alterations. Current treatment of metastatic CRC relies on combination of chemotherapy and targeted therapies such as anti-EGFR therapies. The success of targeted therapies relies on the detection of actionable targets and predictive biomarkers of resistance. The study aims to determine mutations in common actionable targets and predictive biomarkers of resistance to anti-EGFR therapies in Malaysian CRC patients. Methods: Mutations in 10 CRC tissues were determined by next-generation sequencing with a panel of 7 cancer-related genes covering all exons in KRAS, BRAF, PIK3CA, PTEN, TP53, NRAS, and EGFR genes. Immunohistochemistry was used to determine mismatch repair (MMR) status. Results: Of the ten samples, 5 and 4 samples harboured two and one mutation, respectively and one had no mutation. All were missense mutations and were in five genes, namely, KRAS, PIK3CA, TP53, BRAF, and EGFR. They were, G12D, G12V, G12A, G13D, and V14I in KRAS, E545K, K733R, and D1056N in PIK3CA, G199V, D259Y, and R282W in TP53, V600E in BRAF and G696R in EGFR. Deficient mismatch repair (dMMR) was detected in three samples, of which two had KRAS mutation. Conclusion: Mutations in KRAS codon 12 and 13, BRAF and PIK3CA which predict resistance to anti-EGFR therapies and three TP53 mutations were found. This is the first report of EGFR mutation in Malaysian CRC patients. It is predicted to be a pathogenic variant. dMMR, one of the biomarkers for treatment with immune checkpoint inhibitor was also detected.

Cyclooxygenase-2 expression in clear cell renal cell carcinoma

@#Introduction: Cyclooxygenase-2 (COX-2) promotes carcinogenesis by inducing proliferation and angiogenesis while decreasing apoptosis and immunosuppressive activity. It is overexpressed in many malignancies including renal cell carcinoma (RCC). The aim of this study was to investigate COX-2 expression in clear cell RCC and its association with tumour grades and demographic parameters. Materials and Methods: Thirty-six clear cell RCC cases were selected. There were 21 (58.3%) men and 15 (41.7%) women with median age of 56.6 years (range: 16-74 years). Chinese constituted 16 (44.4%) of the cases; Malays 14 (38.9%) cases and Indian 6 (16.7%) cases. There were 6 (16.7%) grade 1, 20 (55.6%) grade 2, 10 (27.8%) grade 3 and none was grade 4. The paraffin embedded tissues were cut at 4 μm thick and stained with COX-2 monoclonal antibody. Results: Eighteen (50%) of the RCC cases were immunopositive, of which all showed strong positivity. The immunopositive cases showed cytoplasmic membrane positivity. Conclusion: There was no significant association between COX-2 expression with grade, age, sex and ethnicity (p=0.457, p=0.054, p=0.389 and p=0.568 respectively). Strong positivity of COX-2 suggest that COX-2 may play a role in cell proliferation and in carcinogenesis.

Vascular Endothelial Growth Factor Expression in Renal Cell Carcinoma

@#Introduction: Renal cell carcinoma (RCC) is the most common renal malignancies. In advanced stage, it is highly resistant to systemic therapies. RCC is a highly vascular tumour and angiogenesis pathway has been postulated in its carcinogenesis. Novel drug targeting Vascular Endothelial Growth Factor (VEGF) and advanced surgical interventions have shown to increase the overall patients’ survival. In this study, we evaluated the VEGF expression of RCC using immunohistochemistry technique and its potential correlation with the tumour grades. Methods: 40 RCC cases that underwent nephrectomy were selected. The archived samples of formalin fixed, paraffin-embedded (FFPE) were retrieved. The tumour tissue blocks were carefully chosen, sectioned and stained with VEGF using immunohistochemistry technique. The intensity of VEGF expression was scored as 0 (negative), 1+ (weak), 2+ (moderate) or 3+ (strong). Results: Majority of the RCC cases were male, with male to female ratio of 2.1:1. Mean patient age was 56.2 years (age ranged between 16 to 74 years). Most of the cases were Malays (42.5 %). VEGF was expressed in 36 (90%) of RCC cases. Among the 36 cases that were immunopositive, 8 (16.7%) were grade 1, 20 (55.6%) grade 2 and 8 (16.7%) grade 3. There was no significant association between VEGF expressions score and grades of RCC (p=0.39). Conclusion: VEGF was expressed in majority of RCC cases although there was no significant association with tumour grades.

Anti-ulcer activity of the aqueous extract of Melastoma malabathricum L. leaf in rats

Melastoma malabathricum L. Smith [Melastomaceae] has been used in the Malay traditional culture to treat ulcer-based ailments. The objective of the present study was to investigate the potential anti-ulcer effect of aqueous extract of M. malabathricum leaves [AEMM] using ethanol- and indomethacin-induced gastric ulcer models in rats. Rats were divided into ten groups [n=6] and received DMSO [10%; negative group], ranitidine [100mg/kg; positive group] or AEMM [50, 250 and 500mg/kg] orally for 7 days and on the 8[th] day subjected to the respective gastric ulcer models. The stomachs were collected and subjected to macroscopic and microscopic analysis. At all groups tested, the AEMM exerted significant [p<0.05] anti-ulcer effect only against the ethanol-induced gastric ulcer model. The percentage of anti-ulcer for the 50-500mg/kg AEMM ranging between 50-82%, respectively. The macroscopic observations were supported by histological findings. In conclusion, AEMM exhibits potential anti-ulcer activity attributed to its previously proven high flavonoids content and antioxidant activity

Comparison Of Invasion By Human Microvascular Endothelial Cell Lines In Response To Vascular Endothelial Growth Factor (Vegf) And Basic Fibroblast Growth Factor (Bfgf) In A Three-Dimensional (3d) Cell Culture System

Background: Immortalized human endothelial cells are widely used as in vitro models for debilitating conditions such as cancer, cardiovascular and ocular diseases. Human microvascular endothelial cell (HMEC-1) is immortalized via stable transfection with a gene encoding SV40 large antigen whilst telomerase-immortalized human microvascular endothelial (TIME) cells is immortalized by engineering the human telomerase catalytic protein (hTERT) into primary microvascular endothelial cells. Here, we established a three-dimensional (3D) spheroid invasion assay with HMEC-1 and TIME and compared the difference in their ability to invade through the collagen matrix in response to exogenous growth factors, namely vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Methods: TIME and HMEC-1 spheroids were embedded in a collagen matrix. The spheroids were stimulated with exogenous growth factors, namely VEGF (50ng/mL) and bFGF (200ng/mL). Twelve points of invasion length from a spheroid was measured using image analysis software, Image J. Three independent experiments were conducted and data was analysis by GraphPad Instat software, version 3.05. Results: TIME spheroid invasion was 16.5 fold higher with exogenous VEGF (50ng/mL) and bFGF (200ng/mL) treatment as compared to those cultured in complete growth medium only. In contrast, no significant difference was observed between HMEC-1 spheroids stimulated with and without exogenous growth factors, VEGF and bFGF. Conclusions: This is the first report on the establishment of a 3D-spheroid invasion assay with TIME cells. The requirement of VEGF and bFGF for TIME spheroids invasion is a novel finding. In addition, this assay offers an advantage over HMEC-1 for testing novel angiogenic agents since it is not affected by endogenously secreted growth factors.

Effect of aqueous extract of Dicranopteris linearis leaves against paracetamol and carbon tetrachloride-induced liver toxicity in rats

The present study aimed to determine the hepatoprotective activity of Dicranopteris linearis L. [family Gleicheniaceae] leaf aqueous extract [DLAE] using two models of liver injury in rats. Rats were divided into ten groups [n=6] and received dH2O [negative control], 200 mg/kg silymarin [positive control] or DLAE [50, 250 and 500 mg/kg] orally once daily for 7 consecutive days and on the 8th day subjected to the hepatotoxic induction either using carbon tetrachloride [CCl[4]] or paracetamol [PCM]. The bloods and livers were collected and subjected to biochemical and microscopical analysis. From the data obtained, only the highest dose of DLAE significantly [p<0.05] reduced the ALP, ALT and AST levels in CCl[4]-and PCM-induced hepatotoxic rats while the other doses caused significant [p<0.05] reduction only in the levels of ALT and AST. The histological results obtained were in line with the biochemical analysis wherein reduction in the CCl[4]- and PCM-induced tissue formation of necrosis, steatosis and inflammation occurred in a dose-dependent manner. In conclusion, the DLAE possesses hepatoprotective activity, which could be attributed to its free radicals scavenging and antioxidant activities, and high flavonoids content. Thus, in-depth studies regarding the hepatoprotective activity of DLAE are warranted

Matrix metalloproteinase-9 and tissue inhibitors of metalloproteinases 1 and 2 as potential biomarkers for gestational hypertension

<p><b>INTRODUCTION</b>Gestational hypertension (GH) is a common disorder during pregnancy that can progress to preeclampsia and cause various subsequent fatal complications. A cluster of enzymes, called matrix metalloproteinases (MMPs), and its specific inhibitors, tissue inhibitors of metalloproteinases (TIMPs), have been reported to be involved in the pathophysiology of GH. The purpose of this study was to examine circulating levels of MMP-9, TIMP-1 and TIMP-2 in pregnant women who had GH and those who were normotensive.</p><p><b>METHODS</b>In a case-control study, the total levels of MMP-9, TIMP-1 and TIMP-2 in the sera of 108 pregnant patients were evaluated using enzyme-linked immunosorbent assays. 54 patients with GH (test group) and 64 normotensive pregnant women (control group) were included in the study.</p><p><b>RESULTS</b>While MMP-9 levels showed a high level of expression in the GH group (p = 0.085), TIMP-1 and TIMP-2 levels showed low levels of expression for the same. Weak positive correlations were found on correlation analysis between maternal age and TIMP-1 in the GH group (r = 0.278, p < 0.05), and between gestational age and TIMP-2 in the control group (r = 0.318, p < 0.05).</p><p><b>CONCLUSION</b>Our findings suggest that MMP-9 may be involved in the pathophysiology of GH. It may be of value to further evaluate MMP-9 as a potential biomarker for predicting preeclampsia in pregnant women.</p>