Immunochemical studies of Toxocara canis proteases.

Proteases of nematodes play a crucial role in larval molting and, in addition to their active role in egg hatching, proteases are also considered a crucial factor in tissue invasion and connective tissue remodeling. In Toxocara canis, proteases play important roles throughout the complex life cycle. They can degrade components of a model of extracellular matrix, basement membranes and different physiological substrates. In the present study, measurements of the proteolytic activity of the perivitelline fluid (PF) surrounding Toxocara canis embryos at different stages of development, the hatching fluid (HF) surrounding the infective larvae, as well as the excretory secretory (ES) products of the larvae in the culture media were performed. Measurements were made using casein as substrate following the Sigma non-specific protease activity assay. The results showed that enzyme activity increased as the embryo matured. The infective larvae were found to continuously produce proteases in the surrounding HF and ES products after in vitro cultivation indicating that Toxocara canis proteases might be important for the worm in the egg and the host. Optimal enzymatic activity was found at pH 8. Incubation of the antiserum from infected mice with the HF and ES products decreased their proteolytic activities, suggesting that there may be a link between the proteases present in these fluids and the immune response.

Immunochemical studies of Toxocara canis proteases

@#Proteases of nematodes play a crucial role in larval molting and, in addition to their active role in egg hatching, proteases are also considered a crucial factor in tissue invasion and connective tissue remodeling. In Toxocara canis, proteases play important roles throughout the complex life cycle. They can degrade components of a model of extracellular matrix, basement membranes and different physiological substrates. In the present study, measurements of the proteolytic activity of the perivitelline fluid (PF) surrounding Toxocara canis embryos at different stages of development, the hatching fluid (HF) surrounding the infective larvae, as well as the excretory secretory (ES) products of the larvae in the culture media were performed. Measurements were made using casein as substrate following the Sigma non-specific protease activity assay. The results showed that enzyme activity increased as the embryo matured. The infective larvae were found to continuously produce proteases in the surrounding HF and ES products after in vitro cultivation indicating that Toxocara canis proteases might be important for the worm in the egg and the host. Optimal enzymatic activity was found at pH 8. Incubation of the antiserum from infected mice with the HF and ES products decreased their proteolytic activities, suggesting that there may be a link between the proteases present in these fluids and the immune response.

The Impact of a Patient Education Package on Outcomes of Pain Management Following Orthopaedic Surgery in a Tertiary Hospital in Malaysia

Patient’s belief towards pain management may affect pain management outcomes and quality of life. The main aim of the present study was to determine the impact of a pre-operative pain education package towards pain belief among patients undergoing orthopaedic surgery in a tertiary hospital. A one-group pre-test post-test design study was conducted on orthopaedic surgery patients. Thirty respondents were recruited and pre-operative pain education was administered individually before surgery. Pre-operative and post-operative pain belief, management scores and side effects were measured using the Barrier Questionnaire (BQ-13). The results reported significant differences between pre-test scores (Mean = 41.87, Standard Deviation = 11.467) and post-test scores (Mean=34.80, Standard Deviation=13.026) of pain belief (t = 2.84, p = 0.004). There were also significant differences between pre-test scores (Mean = 37.10, Standard Deviation = 10.610) and post-test scores (Mean=30.80, Standard Deviation = 11.424) of pain management (t = 3.856, p = 0.0005). Respondent’s gender (t = -2.403, p = 0.023) and ethnicity (F = 5.038, p=0.014) reported significant differences with p value 0.05). There was positive impact of the pain education package towards pain belief and painmanagement among respondents who underwent orthopaedics surgery in a tertiary hospital. Reinforcement of pain educational program is pivotal in order to achieve optimal post-operative pain management.

Congenital insensitivity to pain with anhydrosis in a Malaysian family: a genetic analysis.

A Malaysian family with congenital insensitivity to pain with anhydrosis was diagnosed based on clinical symptoms of chronic ulcers, joint deformities, malunited fractures, anhydrosis, and learning disabilities. We detected a compound heterozygous mutation in exon 16: V709L from the mother and G718S from the father. Two novel mutations were identified: at amino acid 709, a change of G to C at nucleotide 2209 (approximately 2209G to C) causing a valine to leucine substitution (V709L), and at amino acid 718, a change of G to A at nucleotide 2236 (approximately 2236G to A) causing a glycine to serine substitution (G718S). Polymorphisms identified were at nucleotides approximately 2113G to C and approximately 2176T to C.

The Malaysian Orthopaedic Association humanitarian mission to Indonesia and Sri Lanka.

The tsunami which occurred off the west coast of North Sumatra on December 26, 2004 devastated the coastal areas of North Sumatra, South-West Thailand, South-East India and Sri Lanka killing more than a quarter of a million people. The destruction was enormous with many coastal villages destroyed. The other countries affected were Malaysia, Myanmar, Maldives, Bangladesh, Somalia, Kenya, Tanzania and the Seychelles. In January 2005, volunteers went in weekly rotation to Banda Aceh in collaboration with Global Peace Mission. These were Dr Hyzan Yusof, Dr Suryasmi Duski, Dr Sharaf Ibrahim, Dr Saw Aik, Dr Kamariah Nor and Dr Nor Azlin. In Banda Aceh, the surgical procedures that we could do were limited to external fixation of open fractures and debriding infected wounds at the Indonesian Red Crescent field hospital. In February, a team comprising Dato Dr K S Sivananthan, Dr T Kumar and Dr S Vasan spent a week in Sri Lanka. In Sri Lanka, Dato Sivananthan and his team were able to perform elective orthopaedic operations in Dr Poonambalam Memorial Hospital. We appealed for national and international aid and received support from local hospitals and the orthopaedic industry. International aid bound for Banda Aceh arrived in Kuala Lumpur from the Philippine Orthopaedic Association, the Chiba Children's Hospital in Japan and the Chinese Orthopaedic Association. The COA donated 1.5 tons of orthopaedic equipments. A special handing over ceremony from the COA to the Indonesian Orthopaedic Association was held in Putrajaya in March. Malaysia Airlines flew in the donated equipment to Kuala Lumpur while the onward flight to Aceh was provided by the Royal Malaysian Air Force. In April, Dr Saw Aik and Dr Yong Su Mei joined the Tsu-Chi International Medical Association for volunteer services on Batam Island, Indonesia. The MOA acknowledges the many individuals and organizations, both governmental and non-governmental, for their contributions in the humanitarian efforts.

Cardiopulmonary bypass in surgery for complex-combined vascular malformation of the lower limb: case report.

A 16-year-old boy was referred with features of Parkes Weber syndrome (PWS) involving the right lower limb. He had presented at birth with cutaneous vascular malformations (VM) in the right thigh and at the age of 7 years developed congestive cardiac failure, which was controlled with drugs. He received alpha interferon and steroids during this period without any benefit. He defaulted follow-up and at 12 years of age presented with further enlargement of the VM in the right thigh and leg with skin and soft tissue thickening. At this stage, embolization and subsequent excision of the VM were tried, but the surgery was abandoned because of massive hemorrhage. Over the next 4 years, the boy became totally bedridden because of massive increase in the size of the limb, repeated hemorrhages, and secondary infection of the VM. Right hip disarticulation was considered the best option to improve his quality of life. To prevent uncontrollable hemorrhage during surgery, the disarticulation was done under cardiopulmonary bypass with low circulatory flow. Postoperatively, the patient required intensive care nursing for a week. He is presently ambulatory with crutches. Cardiopulmonary bypass with low flow has been used for treating posttraumatic arteriovenous malformations. However, its use in surgery for PWS has not been reported earlier.

The effects of autologous human serum on the growth of tissue engineered human articular cartilage.

Culture media supplemented with animal serum e.g. fetal bovine serum; FBS is commonly used for human culture expansion. However, for clinical application, FBS is restricted as its carry a risk of viral or prion transmission. Engineering autologous cartilage with autologous human serum supplementation is seen as a better solution to reduce the risk of transmitting infectious diseases and immune rejection during cartilage transplantation. The purpose of this study is to establish and compare the effects of 10% autologous human serum (AHS) and 10% FBS on the growth of chondrocytes and the formation of tissue engineered human articular cartilage.

The significance of using pooled human serum in human articular cartilage tissue engineering.

Animal serum is commonly used in chondrocytes culture expansion to promote cell proliferation and shorten the time lag before new tissue reconstruction is possible. However, animal serum is not suitable for regeneration of clinical tissue because it has potential risk of viral and prion related disease transmission particularly mad cow disease and foreign protein contamination that can stimulate immune reaction leading to graft rejection. In this context, human serum as homologous supplement has a greater potential as growth promoting agents for human chondrocytes culture.

Synthetic organic food colouring agents and their degraded products: effects on human and rat cholinesterases.

Most synthetic coloured additives are carcinogenic; teratogenic and cause allergic reactions. In this study, the effects of synthetic azo dyes (sunset yellow FCF and carmoisine), as well as their degraded products (sulphanilic acid and naphthionic acid), on both true and pseudo-cholinesterases (ChEs) are studied. The results indicate that the synthetic azo dyes and their degraded products inhibit both human true and pseudo-ChE activities in vitro. The concentration of coloured additive that cause 50% inhibition (IC50) and enzyme inhibitor dissociation constant (Ki) show that sunset yellow FCF produces greater inhibition of both true and pseudo-ChEs than does carmoisine and sulphanilic acid, while naphthionic acid produces greater inhibition of pseudo-ChE only. Ki indicates that the affinity of sulphanilic acid for both true and pseudo-ChEs is higher than the other three inhibitors. Inhibition of both true and pseudo-ChEs by sunset yellow FCF is of mixed (competitive and non-competitive) type, but carmoisine and sulphanilic acid are non-competitive. Naphthionic acid produces a competitive inhibition kinetic with plasma ChE only. This inhibition is abolished by dialysis, indicating that their effects are reversible. The effects of sunset yellow FCF, carmoisine, sulphanilic acid and naphthionic acid on rat true and pseudo-ChEs are investigated. The data clearly show that there is a significant decrease in enzyme activity. Sulphanilic acid and sunset yellow FCF are the most potent in vivo inhibitors of true ChE and pseudo-ChE, respectively.

Ossifying fibromyxoid tumour in a child.

Ossifying fibromyxoid tumor (OFMT) is a rare benign tumor, most of which occurs in adults with localization in the subcutaneous tissue or muscle of the extremities. A five-year-old girl presented with a mass in her right upper thigh. Due to the large size of the mass (10 x 7 cm), our provisional diagnosis was a soft tissue sarcoma. A tru-cut biopsy showed that the lesion was benign. The mass was excised and has not recurred since. To the best of our knowledge, this patient is the youngest case of OFMT reported in the English literature.

Sleeve fracture of the patella in a child.

Sleeve fracture of the inferior pole of the patella is a rare and distinctive fracture in children with few published reports. These fractures are frequently misdiagnosed and neglected. We highlight a case of a neglected and misdiagnosed sleeve fracture of the patella in an eleven-year-old boy. This was initially diagnosed as an avulsion fracture of the tibial tubercle. A good outcome was achieved after open reduction and internal fixation.

The effects of autologous human serum on the growth of tissue engineered human articular cartilage

Culture media supplemented with animal serum e.g. fetal bovine serum; FBS is commonly used for human culture expansion. However, for clinical application, FBS is restricted as its carry a risk of viral or prion transmission. Engineering autologous cartilage with autologous human serum supplementation is seen as a better solution to reduce the risk of transmitting infectious diseases and immune rejection during cartilage transplantation. The purpose of this study is to establish and compare the effects of 10% autologous human serum (AHS) and 10% FBS on the growth of chondrocytes and the formation of tissue engineered human articular cartilage.

Influence of free radicals on cardiovascular risk due to occupational exposure to mercury.

This study was designed to investigate the relation between occupational exposure to mercury and cardiovascular risk, as well as free radicals. The study subjects included 30 male workers exposed to mercury from a fluorescent lamp plant in Alexandria after exclusion of 12 workers with conditions that may bias the results. A matched control group of 20 male subjects were selected from a food industrial plant. The participants were subjected to interview, clinical examination, electrocardiography and R-R deep breathing variability test as a measure of cardiac autonomic function. Spot urine and venous blood samples were collected to measure mercury levels. Free radicals related markers were measured in serum namely lipid peroxide, superoxide dismutase (SOD) activity, catalase activity and glutathione S-transferase. Results showed significant longer electrocardiographic Q-Tc interval, which is corrected to heart rate, in exposed workers than in controls (p=0.002). The R-R deep breathing variability indices were significantly increased among exposed workers in comparison to controls (maximal variation (MV) ratio, p=0.024 & MV rate, p=0.019). The mercury levels in both urine and blood of exposed workers were significantly higher than controls (p<0.0001), with mean levels exceeding the permissible limits. Free radicals related markers revealed significant higher level of lipid peroxide, as well as significant lower levels of SOD activity, catalase activity and glutathione S-transferase among exposed workers than among controls (p<0.0001 for all). Independent relations were observed between Q-Tc and lipid peroxide, and between MV rate and SOD and catalase activities among exposed workers after adjustment for the confounders by multivariate regression analyses. These findings may suspect a cardiac autonomic dysfunction probably related to free radicals in mercury exposed workers. Also, it may through light on some preventive implications.

Crossed-pin versus lateral-pin fixation in pediatric supracondylar fractures.

A retrospective study was done on 56 patients treated with percutaneous pinning of displaced supracondylar fractures of the humerus in the Paediatrics Institute of Hospital Kuala Lumpur between November 1999 and October 2000, to ascertain whether there is any significance clinically in the stability of a crossed pinning medial compared with lateral pinning method. There was equal number of patients in each group (28 patients). The radiographs were evaluated for change in Baumann's angle and Lateral Humero-capitellar angle from immediate post-op until the last follow-up. The changes in the angles did not reveal any statistically significant difference in the ability to maintain reduction of the fractures. There were 3 instances of iatrogenic ulnar nerve injury in the crossed pinning group; the lateral pinning group had 1 case each of anterior interosseous nerve and radial nerve injury post operatively. No vascular injury was noted. Two cases of superficial pin tract infection were present in each group. The lateral percutaneous pinning technique of displaced supracondylar fractures of the humerus therefore offers a viable alternative to the crossed pinning group as it offers the same stability without the incipient risk of iatrogenic ulnar nerve injury.

Screw osteosynthesis in the treatment of fracture lateral humeral condyle in children.

We reviewed the results of screw osteosynthesis for the treatment of fracture lateral condylar physis in twenty children whose average age was six years old (range, two to ten years) at the time of operation from January 1998 till December 2000. The average interval between the injury and the operation was three and half days (range, one day to two weeks). The average duration of follow up was one year (range, one year to two years). Osteosynthesis was revised in two patients due to anterior placement of screw and rotated distal fracture fragment. Osseous union was achieved in all twenty patients. The result was rated excellent in nineteen patients and good in one patient.

Acute haematogenous osteomyelitis: an unusual complication following a closed fracture of the femur in a child.

We report a rare case of an acute haematogenous osteomyelitis of the femur in a five-year-old boy following a closed fracture of the femur. Because of its rare occurrence, the diagnosis of osteomyelitis was missed initially. He presented with a groin abscess seven weeks after injury. He was treated with external fixation, repeated debridements and intravenous antibiotics. Culture grew Staphylococcus aureus. The latest follow-up one-year after the injury showed resolution of the infection and union of the fracture. The range of knee movement is limited from 0-90 degrees due to quadriceps contracture.

Compartment syndrome of the foot in a child.

We report a case of a 12-year-old boy with acute compartment syndrome of the foot following a road-traffic accident. Due to the rarity of the injury, there was a delay in diagnosing the injury. An emergency fasciotomy was performed 19 hours after the injury. The foot healed with a mild extension contracture of the second toe.

Ewing's sarcoma of the talus in a four-year-old child.

Ewing's sarcoma is a rare malignant tumor of the foot in children. We report a case of Ewing's sarcoma of the talus in a four-year-old Chinese girl to highlight the initial difficulty in diagnosis and the clinical course of the disease. She was initially diagnosed as osteomyelitis of the talus and died eight months after presentation with pleural and spinal metastases. To the best of our knowledge, Ewing's sarcoma of the talus in a young child has never been reported in Malaysia.

Metacyclogenesis of Leishmania major in an acidic medium.

An acidic medium was used to stimulate the production of a homogeneous stationary phase promastigotes, morphologically and functionally similar to the in vivo infective form. They possessed a short (< 8 Um) and narrow (< 1.5 Um.) cell body with a flagellum twice or more its length. They were PNA, highly infective to peritoneal macrophages in vitro and expressed high phosphatase activity. By elevating the incubating temperature, almost all of these promastigotes were transformed to amastigotes.