Quantifying the ultrastructure changes of air-dried and irradiated human amniotic membrane using atomic force microscopy: a preliminary study.

Air-dried and sterilized amnion has been widely used as a dressing to treat burn and partial thickness wounds. Sterilisation at the standard dose of 25 kGy was reported to cause changes in the morphological structure as observed under the scanning electron microscope. This study aimed to quantify the changes in the ultrastructure of the air-dried amnion after gamma-irradiated at several doses by using atomic force microscope. Human placentae were retrieved from mothers who had undergone cesarean elective surgery. Amnion separated from chorion was processed and air-dried for 16 h. It was cut into 10 × 10 mm, individually packed and exposed to gamma irradiation at 5, 15, 25 and 35 kGy. Changes in the ultrastructural images of the amnion were quantified in term of diameter of the epithelial cells, size of the intercellular gap and membrane surface roughness. The longest diameter of the amnion cells reduced significantly after radiation (p < 0.01) however the effect was not dose dependent. No significant changes in the shortest diameter after radiation, except at 35 kGy which decreased significantly when compared to 5 kGy (p < 0.01). The size of the irradiated air-dried amnion cells reduced in the same direction without affecting the gross ultrastructure. At 15 kGy the intercellular gap decreased significantly (p < 0.01) with Ra and Rq, values reflecting surface roughness, were significantly the highest (p < 0.01). Changes in the ultrastructure quantified by using atomic force microscope could complement results from other microscopic techniques.

MULTILEVEL LINEAR REGRESSION ANALYSIS OF FACTORS INFLUENCING BODY MASS INDEX AMONG BANGLADESHI MARRIED NON-PREGNANT WOMEN.

The aim of this study was to investigate the socioeconomic and demographic factors influencing the body mass index (BMI) of non-pregnant married Bangladeshi women of reproductive age. Secondary (Hierarchy) data from the 2011 Bangladesh Demographic and Health Survey, collected using two-stage stratified cluster sampling, were used. Two-level linear regression analysis was performed to remove the cluster effect of the variables. The mean BMI of married non-pregnant Bangladeshi women was 21.60±3.86 kg/m2, and the prevalence of underweight, overweight and obesity was 22.8%, 14.9% and 3.2%, respectively. After removing the cluster effect, age and age at first marriage were found to be positively (p<0.01) related with BMI. Number of children was negatively related with women's BMI. Lower BMI was especially found among women from rural areas and poor families, with an uneducated husband, with no television at home and who were currently breast-feeding. Age, total children ever born, age at first marriage, type of residence, education level, level of husband's education, wealth index, having a television at home and practising breast-feeding were found to be important predictors for the BMI of married Bangladeshi non-pregnant women of reproductive age. This information could be used to identify sections of the Bangladeshi population that require special attention, and to develop more effective strategies to resolve the problem of malnutrition.

Comparison of in vitro developmental competence of cloned caprine embryos using donor karyoplasts from adult bone marrow mesenchymal stem cells vs ear fibroblast cells.

The aim of this study was to produce cloned caprine embryos using either caprine bone marrow-derived mesenchymal stem cells (MSCs) or ear fibroblast cells (EFCs) as donor karyoplasts. Caprine MSCs were isolated from male Boer goats of an average age of 1.5 years. To determine the pluripotency of MSCs, the cells were induced to differentiate into osteocytes, chondrocytes and adipocytes. Subsequently, MSCs were characterized through cell surface antigen profiles using specific markers, prior to their use as donor karyoplasts for nuclear transfer. No significant difference (p > 0.05) in fusion rates was observed between MSCs (87.7%) and EFCs (91.3%) used as donor karyoplasts. The cleavage rate of cloned embryos derived with MSCs (87.0%) was similar (p > 0.05) to those cloned using EFCs (84.4%). However, the in vitro development of MSCs-derived cloned embryos (25.3%) to the blastocyst stage was significantly higher (p < 0.05) than those derived with EFCs (20.6%). In conclusion, MSCs could be reprogrammed by caprine oocytes, and production of cloned caprine embryos with MSCs improved their in vitro developmental competence, but not in their fusion and cleavage rate as compared to cloning using somatic cells such as EFCs.

Geometric variable designs of cam/post mechanisms influence the kinematics of knee implants.

PURPOSE: Reproducing the femoral rollback through specially designed mechanism in knee implants is required to achieve full knee function in total knee arthroplasty. Most contemporary implants use cam/post mechanism to replace the function of Posterior Cruciate Ligament. This study was aimed to determine the most appropriate cam and post designs to produce normal femoral rollback of the knee. METHODS: Three different cams (triangle, ellipse, and circle) and three different posts (straight, convex, concave) geometries were considered in this study and were analysed using kinematic analyses. Femoral rollback did not occur until reaching 50° of knee flexion. Beyond this angle, two of the nine combinations demonstrate poor knee flexion and were eliminated from the study. RESULTS: The combination of circle cam with concave post, straight post and convex post showed 15.6, 15.9 and 16.1 mm posterior translation of the femur, respectively. The use of ellipse cam with convex post and straight post demonstrated a 15.3 and 14.9 mm femoral rollback, whilst the combination of triangle cam with convex post and straight post showed 16.1 and 15.8 mm femoral rollback, respectively. CONCLUSION: The present study demonstrates that the use of circle cam and convex post created the best femoral rollback effect which in turn produces the highest amount of knee flexion. The findings of the study suggest that if the design is applied for knee implants, superior knee flexion may be possible for future patients. LEVEL OF EVIDENCE: IV.

A comparative study on morphochemical properties and osteogenic cell differentiation within bone graft and coral graft culture systems.

The objective of this study was to compare the morphological and chemical composition of bone graft (BG) and coral graft (CG) as well as their osteogenic differentiation potential using rabbit mesenchymal stem cells (rMSCs) in vitro. SEM analysis of BG and CG revealed that the pores in these grafts were interconnected, and their micro-CT confirmed pore sizes in the range of 107-315 µm and 103-514 µm with a total porosity of 92% and 94%, respectively. EDS analysis indicated that the level of calcium in CG was relatively higher than that in BG. FTIR of BG and CG confirmed the presence of functional groups corresponding to carbonyl, aromatic, alkyl, and alkane groups. XRD results revealed that the phase content of the inorganic layer comprised highly crystalline form of calcium carbonate and carbon. Atomic force microscopy analysis showed CG had better surface roughness compared to BG. In addition, significantly higher levels of osteogenic differentiation markers, namely, alkaline phosphatase (ALP), Osteocalcin (OC) levels, and Osteonectin and Runx2, Integrin gene expression were detected in the CG cultures, when compared with those in the BG cultures. In conclusion, our results demonstrate that the osteogenic differentiation of rMSCs is relatively superior in coral graft than in bone graft culture system.

Adult anthropometric measures and socio-demographic factors influencing age at menarche of university students in malaysia.

Early onset of menarche has been shown to be associated with breast cancer and ischaemic heart disease. Studies on age at menarche of the Malaysian population are poorly documented. This study aimed to determine the influence of anthropometric and socio-demographic factors on the age at menarche of university students in Malaysia. Data were obtained in 2010-11 from 961 students between the ages of 18 and 25 years from the University of Malaya using stratified sampling, and multiple regression analysis was applied. Sixty-three per cent of students reached menarche at the age of 12 or 13 years, with the mean and median of age at menarche being 12.45 ± 1.17 and 12.01 years, respectively. Menarcheal age was positively associated with height (p<0.05) and negatively associated with BMI (p<0.001). Students from urban areas attained menarche earlier than those from rural areas (p<0.05). Students from small-sized families attained menarche earlier than those from larger families (p<0.05). First-born students experienced menarche earlier than those who were seventh-born or later. Obese and overweight students reached menarche earlier than students who were underweight or of normal weight (p<0.01). The variations in age at menarche among the Malaysian ethnic groups were statistically insignificant. The results suggest that heavier and first-born students from small families are more likely to attain menarche earlier than their counterparts.

Body mass index of married Bangladeshi women: trends and association with socio-demographic factors.

Body mass index (BMI) is a good indicator of nutritional status in a population. In underdeveloped countries like Bangladesh, this indicator provides a method that can assist intervention to help eradicate many preventable diseases. This study aimed to report on changes in the BMI of married Bangladeshi women who were born in the past three decades and its association with socio-demographic factors. Data for 10,115 married and currently non-pregnant Bangladeshi women were extracted from the 2007 Bangladesh Demographic and Health Survey (BDHS). The age range of the sample was 15-49 years. The mean BMI was 20.85 ± 3.66 kg/m(2), and a decreasing tendency in BMI was found among birth year cohorts from 1972 to 1992. It was found that the proportion of underweight females has been increasing in those born during the last 20 years of the study period (1972 to 1992). Body mass index increased with increasing age, education level of the woman and her husband, wealth index, age at first marriage and age at first delivery, and decreased with increasing number of ever-born children. Lower BMI was especially pronounced among women who were living in rural areas, non-Muslims, employed women, women not living with their husbands (separated) or those who had delivered at home or non-Caesarean delivery.

A preliminary study of the effects of glucosamine sulphate and chondroitin sulphate on surgically treated and untreated focal cartilage damage.

The effects of Glucosamine Sulphate (GS) and Chondroitin Sulphate (CS) on the healing of damaged and repaired articular cartilage were investigated. This study was conducted using 18 New Zealand white rabbits as experimental models. Focal cartilage defects, surgically created in the medial femoral condyle, were either treated by means of autologous chondrocyte implantation (ACI) or left untreated as controls. Rabbits were then divided into groups which received either GS+/-CS or no pharmacotherapy. Three rabbits from each group were sacrificed at 12 and 24 weeks post-surgery. Knees dissected from rabbits were then evaluated using gross quantification of repair tissue, glycosaminoglycan (GAG) assays, immunoassays and histological assessments. It was observed that, in contrast to untreated sites, surfaces of the ACI-repaired sites appeared smooth and continuous with the surrounding native cartilage. Histological examination demonstrated a typical hyaline cartilage structure; with proteoglycans, type II collagen and GAGs being highly expressed in repair areas. The improved regeneration of these repair sites was also noted to be significant over time (6 months vs. 3 months) and in GS and GS+CS groups compared to the untreated (without pharmacotherapy) group. Combination of ACI and pharmacotherapy (with glucosamine sulphate alone/ or with chondroitin sulphate) may prove beneficial for healing of damaged cartilage, particularly in relation to focal cartilage defects.

Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells.

This study investigates the feasibility of processed human amnion (HAM) as a substrate for chondrogenic differentiation of mesenchymal stem cells (MSCs). HAM preparations processed by air drying (AD) and freeze drying (FD) underwent histological examination and MSC seeding in chondrogenic medium for 15 days. Monolayer cultures were used as control for chondrogenic differentiation and HAMs without cell seeding were used as negative control. Qualitative observations were made using scanning electron microscopy analysis and quantitative analyses were based on the sulfated glycosaminoglycans (GAG) assays performed on day 1 and day 15. Histological examination of HAM substrates before seeding revealed a smooth surface in AD substrates, while the FD substrates exhibited a porous surface. Cell attachment to AD and FD substrates on day 15 was qualitatively comparable. GAG were significantly highly expressed in cells seeded on FD HAM substrates. This study indicates that processed HAM is a potentially valuable material as a cell-carrier for MSC differentiation.

COVID-19 in Singapore and Malaysia: Rising to the Challenges of Orthopaedic Practice in an Evolving Pandemic.

With the increasing number of COVID-19 cases and related deaths worldwide, we decided to share the development of this condition in Singapore and Malaysia. First few cases were diagnosed in the two countries at the end of January 2020, and the numbers have surged to thousands by end of March 2020. We will focus on strategies adopted by the government and also the Orthopaedic community of the two countries up till the beginning of April 2020. We hope that by sharing of relevant information and knowledge on how we are managing the COVID-19 condition, we can help other communities, and health care workers to more effectively overcome this pandemic.

Elevated plasma and synovial fluid interleukin-8 and interleukin-18 may be associated with the pathogenesis of knee osteoarthritis.

PURPOSE: Osteoarthritis (OA) of the knee is a multifactorial degenerative disease typically defined as the 'wear and tear' of articular joint cartilage. However, recent studies suggest that OA is a disease arising from chronic low-grade inflammation. We conducted a study to investigate the relationship between chronic inflammatory mediators present in both the systemic peripheral blood system and localised inflammation in synovial fluid (SF) of OA and non-OA knees; and subsequently made direct comparative analyses to understand the mechanisms that may underpin the processes involved in OA. METHODS: 20-Plex proteins were quantified using Human Magnetic Luminex® assay (R&D Systems, USA) from plasma and SF of OA (n = 14) and non-OA (n = 14) patients. Ingenuity Pathway Analysis (IPA) software was used to predict the relationship and possible interaction of molecules pertaining to OA. RESULTS: There were significant differences in plasma level for matrix metalloproteinase (MMP)-3, interleukin (IL)-27, IL-8, IL-4, tumour necrosis factor-alpha, MMP-1, IL-15, IL-21, IL-10, and IL-1 beta between the groups, as well as significant differences in SF level for IL-15, IL-8, vascular endothelial growth factor (VEGF), MMP-1, and IL-18. Our predictive OA model demonstrated that toll-like receptor (TLR) 2, macrophage migration inhibitory factor (MIF), TLR4 and IL-1 were the main regulators of IL-1B, IL-4, IL-8, IL-10, IL-15, IL-21, IL-27, MMP-1 and MMP-3 in the plasma system; whilst IL-1B, TLR4, IL-1, and basigin (BSG) were the regulators of IL-4, IL-8, IL-10, IL-15, IL-18, IL-21, IL-27, MMP-1, and MMP-3 in the SF system. CONCLUSION: The elevated plasma IL-8 and SF IL-18 may be associated with the pathogenesis of OA via the activation of MMP-3.

Autologous chondrocyte transplantation in the repair of full-thickness focal cartilage damage in rabbits.

PURPOSE: To compare the efficacy of autologous chondrocyte transplantation (ACT) versus non-operative measures for cartilage repair in rabbits. METHODS: Nine New Zealand white rabbits were used. Identical focal defects were created in the articular cartilage of both knees. One month later, the right knee was repaired via ACT, while the left knee was left untreated (control group). The quality of cartilage tissues in both knees was compared 3 months later, according to the quantitative analysis of glycosaminoglycan (GAG) in the cartilage and macroscopic examination of histology using the Brittberg/International Cartilage Research Society (ICRS) score. RESULTS: Microscopic examination showed enhanced regeneration following ACT repair. Quantification analysis revealed significantly higher cellular expression of GAG in the ACT-treated knees (1.12 vs 0.81 microgram GAGs/mg protein, p=0.008). The mean Brittberg/ICRS score was significantly higher in the treated knees (6.00 vs 1.89, p=0.007). CONCLUSION: ACT is superior to non-operative measures for repairing focal cartilage defects, as determined by favourable histological and immunohistological outcomes at the cellular level.

Tubercular synovitis mimicking rheumatoid nodules.

Tubercular tenosynovitis is an uncommon condition and usually affects the upper limb. We report a case of a patient with Systemic Lupus Erythematosus who presented with wrist swelling. The clinical findings were suggestive of rheumatoid nodules, but the radiographic finding of calcification associated with the nodules and marked erosive changes primarily of the radio-carpal joint with sparing of the metacarpal joints led the radiologist to believe that the nodules may not be rheumatoid nodules. The presence of solid and fluid nature of the nodule and hyperechoic small echogenic foci (matted rice bodies within thickened synovium) on ultrasound suggested the presence of chronic synovitis of tuberculous infection rather than rheumatoid nodule as in our case. We recommend the use of ultrasound to determine the nature of nodular swellings seen clinically in patients with arthropathy.

Fiber density of collagen grafts impacts rabbit urethral regeneration.

There is a need for efficient and "off-the-shelf" grafts in urethral reconstructive surgery. Currently available surgical techniques require harvesting of grafts from autologous sites, with increased risk of surgical complications and added patient discomfort. Therefore, a cost-effective and cell-free graft with adequate regenerative potential has a great chance to be translated into clinical practice. Tubular cell-free collagen grafts were prepared by varying the collagen density and fiber distribution, thereby creating a polarized low fiber density collagen graft (LD-graft). A uniform, high fiber density collagen graft (HD-graft) was engineered as a control. These two grafts were implanted to bridge a 2 cm long iatrogenic urethral defect in a rabbit model. Histology revealed that rabbits implanted with the LD-graft had a better smooth muscle regeneration compared to the HD-graft. The overall functional outcome assessed by contrast voiding cystourethrography showed patency of the urethra in 90% for the LD-graft and in 66.6% for the HD-graft. Functional regeneration of the rabbit implanted with the LD-graft could further be demonstrated by successful mating, resulting in healthy offspring. In conclusion, cell-free low-density polarized collagen grafts show better urethral regeneration than high-density collagen grafts.

Hand grip strength in the adult Malaysian population.

PURPOSE: To measure the hand grip strength of Malaysians aged 18 to 65 years. METHODS: Between January and April 2003, 412 subjects (200 women and 212 men) were recruited from staff, students, and visitors of the University of Malaya Medical Centre. Socioeconomic, general health, and lifestyle data were collected from each subject using a standard questionnaire. Weight and height were measured prior to testing. Standardised positioning and instructions based on several hand grip protocols were used. Data were collected using the LIDO kinetic work set. RESULTS: 93% of the subjects were right-hand dominant and 7% were left-hand dominant. Hand grip strength was significantly correlated with hand dominance, gender, occupation, height, and weight, but not body mass index. No significant differences in grip strength were noted with regard to race or level of income. Men were stronger than women in all age-groups, with a ratio of 1.75:1. In both right- and left-hand dominant groups, the dominant hand was consistently stronger than the non-dominant side, with a ratio of 1.12:1 in the right-hand dominant group and 1.05:1 in the left-hand dominant group. The strongest hand grip strength in the right-hand dominant group occurred in the age-group of 25 to 34 years; in the left-hand dominant group it was in the age-group of 18 to 24 years. In western populations, the mean grip strength can be as much as 1.5 times greater than in the Malaysian population. CONCLUSION: Data derived from western populations cannot be applied to a comparable Malaysian population. Gender, hand dominance, age, occupation, weight, and height must be considered when establishing normal values for grip strength.

A novel and improved method of predicting hand grip strength in the adult Malaysian population.

Hand grip strength measurement is a recognized part of hand function assessment. The standard measurement using the Jamar dynamometer and comparing these results to the recommended normal values suggested by the manufacturers of the Jamar was questioned as these values were based on Western population. A study comparing a novel method of predicting grip strength using our software was conducted on 25 normal subjects using the LIDO kinetic workset (Group A and B ). These results were then compared against our predictive software (Group A) and the expected values supplied together with the Jamar Dynamometer (Group B). In another group, 22 normal subjects were tested using the Jamar (Group C and D) and then matched against the predicted values using their recommended chart (Group C). The last group (Group D) was tested using the Jamar but the values attained were compared to the results from our software. In group A, the predictability of our predictive method was 100% (both R & L) as compared to (R = 64%, L = 68%) in group B, (R = 27.3%, L = 59.1%) in group C and (R = 81.8%, L = 86.4%) in group D. The differences between the predictability of both methods were statistically significant. The data collected using both the Jamar and the LIDO kinetic workset correlated well to the data from our software but not to the values suggested by the manufacturers of Jamar. We conclude that our method of predicting hand grip values are superior to that suggested by the manufacturers of dynamometers. The standard reference for hand grip strength provided by the manufacturers is less accurate in predicting the grip strength of our local population.

A modified Blatt dorsal capsulodesis in the treatment of dynamic scapholunate instability.

We report a case of scapholunate dissociation which was initially missed and presented late. A modification of Blatt dorsal capsulodesis performed using dorsal intercarpal ligament (DICL) and extra tunnel appears not only to add to dorsal stability but also address the volar problem as well. This modification may be a better alternative to the current technique of using a single flap.

Coadministration of alloxan and nicotinamide in rats produces biochemical changes in blood and pathological alterations comparable to the changes in type II diabetes mellitus.

BACKGROUND: In the present study, thirty six male Sprague Dawley rats were randomly divided into six groups and were injected with varying doses of alloxan (Ax) and nicotinamide (NA). The serum levels of glucose, insulin, and adiponectin were measured weekly up to 4 weeks. RESULTS: Elevated levels of glucose were observed in all groups on days 7, 14, 21, and 28, except in groups a and f (control). The serum insulin levels were significantly elevated in groups b and c on day 7, when compared with that in group f, whereas a decrease in the serum insulin levels was observed in groups d and e on days 21 and 28. The adiponectin levels showed inconsistencies on days 7 and 14. However, significant decrease in the adiponectin levels was observed on days 21 and 28. Histological section of the pancreas showed mild (group a), moderate (group b) to severe (groups c, d, and e) degenerative changes. Concomitant fatty changes in the liver and inflammatory infiltration of the kidney were markedly observed in all the treated groups, when compared to control. CONCLUSION: These results suggested that the use of selective combination of Ax120 + NA50 injection demonstrated type II diabetes mellitus in rats.

Potential of apoptotic pathway-targeted cancer therapeutic research: Where do we stand?

Underneath the intricacy of every cancer lies mysterious events that impel the tumour cell and its posterity into abnormal growth and tissue invasion. Oncogenic mutations disturb the regulatory circuits responsible for the governance of versatile cellular functions, permitting tumour cells to endure deregulated proliferation, resist to proapoptotic insults, invade and erode normal tissues and above all escape apoptosis. This disruption of apoptosis has been highly implicated in various malignancies and has been exploited as an anticancer strategy. Owing to the fact that apoptosis causes minimal inflammation and damage to the tissue, apoptotic cell death-based therapy has been the centre of attraction for the development of anticancer drugs. Increased understanding of the molecular pathways underlying apoptosis has enabled scientists to establish unique approaches targeting apoptosis pathways in cancer therapeutics. In this review, we reconnoitre the two major pathways (intrinsic and extrinsic) targeted cancer therapeutics, steering toward chief modulators of these pathways, such as B-cell lymphoma 2 protein family members (pro- and antiapoptotic), inhibitor of apoptosis proteins, and the foremost thespian of extrinsic pathway regulator, tumour necrosis factor-related apoptosis-inducing agent. Together, we also will have a look from clinical perspective to address the agents (drugs) and therapeutic strategies adopted to target these specific proteins/pathways that have entered clinical trials.

Effect of torsional loading on compressive fatigue behaviour of trabecular bone.

The present study reports the effects of combined torsional and compressive cyclic loading on trabecular bone in order to mimic true physiological conditions and thereby provides improved data that represents clinical and real life conditions. However, only compressive behaviour is evaluated in most previous studies of bone mechanics. From the monotonic evaluation, it is observed that lower stress is needed for the onset of microcrack in the sample under torsional loading, compared to the stress needed in compression. Trabecular bone samples were subjected to a combination of torsion and compression fatigue at different stress levels during which they were compared to compressive axial fatigue. The stress levels were determined by considering the monotonic strength at 25-50% for both compressive and shear stresses. Significant decrease in fatigue lifetime is observed in between samples of pure compression fatigue and those with superpositioned torsional loading (p<0.05). The reduction in fatigue lifetime became more evident at a high torsional stress level. In this case, the failure of the sample is said to be 'torsional dominant'. Fatigue behaviour of bovine trabecular bone begins with plastic deformation, followed by strain accumulation and modulus reduction. As the strain rate increases, more energy dissipates and the sample finally failed. Further, the analysis of fractograph revealed something on the trabeculae by bending in sample with superpositioned torsional loading. In conclusion, torsional loading decreases the quality of the trabecular properties in terms of stiffness, life and structural integrity. It is hoped that results from this study will improve the understanding of the behaviour of trabecular bone under combined fatigue and help to develop future assessments of trabecular failure.