Stability of enteroviruses on toys commonly found in kindergarten

@#Hand, foot, and mouth disease (HFMD) is a contagious childhood disease caused by enteroviruses including enterovirus A71 (EV-A71), coxsackievirus A6 (CV-A6) and CV-A16 transmitted via direct and indirect contact. Different types of toy surfaces can affect the stability of viruses. Understanding the stability of enteroviruses on toys provides insightful data for effective disinfection in kindergartens or homes. Porous (ethylene-vinyl acetate mat foam, paper, pinewood, polyester fabric, and squishy polyurethane foam) and non-porous (acrylonitrile butadiene styrene plastic and stainless-steel coin) surfaces were inoculated with EV-A71 at 4, 24, and 35°C, and coxsackieviruses at 24°C. Infectious enteroviruses were recovered and titred in median tissue culture infectious dose assay (TCID50). Atomic force microscopy (AFM) images were taken from surfaces to examine association of surface roughness with virus stability. Overall, infectious enteroviruses were persistent on all non-porous and porous surfaces. Virus persistence was longest at 4°C followed by 24°C and 35°C. EV-A71 half-lives ranged between 6.4-12.8 hours at 4°C, 2.4-6.7 hours at 24°C, and 0.13-2.7 hours at 35°C. At lower virus titres exposed to 24°C, half-lives of enteroviruses ranged from 0.1-1.4 hours. Surface roughness values from AFM suggested smooth surfaces of non-porous surfaces were associated with better virus stability. Temperature, enterovirus concentration, and type of surface affected persistence and stability of enteroviruses. Our findings suggest both porous and non-porous surfaces in kindergartens allow enterovirus persistence and should be frequently disinfected to curb HFMD outbreaks in kindergartens.

First molecular report of Babesia gibsoni infection in pet dogs in Hunan province, subtropical China

@#Canine babesiosis caused by Babesia spp. is a noteworthy tick-borne zoonotic disease of domestic dogs and wild canids. In present study, a total of 556 blood samples were randomly collected from pet dogs in eight cities of Hunan province, subtropical China. Genomic DNA was extracted and Babesia DNA was detected by amplification of partial 18S rRNA gene sequences. A total of 56 (10.1%) blood samples were tested positive for Babesia species. Sequence analysis showed that 29 dogs (5.2%) were positive for B. gibsoni, and other 27 dogs for B. vogeli (4.9%). The age and health status were considered as important risk factors for B. gibsoni and B. vogeli infections in pet dogs in this study (P<0.05). Phylogenetic analysis showed that the examined positive samples were highly clustered in the same branch with B. gibsoni and B. vogeli, respectively. This is the first molecular report of B. gibsoni infection in pet dogs in Hunan province, subtropical China. Our finding has provided a guide for the control of dog babesiosis in China and elsewhere.

In vitro antiviral activity of medicinal mushroom Ganoderma neo-japonicum Imazeki against enteroviruses that caused hand, foot and mouth disease

@# Hand, foot and mouth disease (HFMD) is a highly contagious viral disease that predominantly affects children younger than 5 years old. HFMD is primarily caused by enterovirus A71 (EVA71) and coxsackievirus A16 (CV-A16). However, coxsackievirus A10 (CV-A10) and coxsackievirus A6 (CV-A6) are being increasingly reported as the predominant causative of HFMD outbreaks worldwide since the past decade. To date, there are still no licensed multivalent vaccines or antiviral drugs targeting enteroviruses that cause HFMD, despite HFMD outbreaks are still being frequently reported, especially in Asia-Pacific countries. The high rate of transmission, morbidity and potential neurological complications of HFMD is indeed making the development of broad-spectrum antiviral drugs/agents against these enteroviruses a compelling need. In this study, we have investigated the in vitro antiviral effect of 4 Ganoderma neo-japonicum Imazeki (GNJI) crude extracts (S1-S4) against EV-A71, CV-A16, CV-A10 and CV-A6. GNJI is a medicinal mushroom that can be found growing saprophytically on decaying bamboo clumps in Malaysian forests. The antiviral effects of this medicinal mushroom were determined using cytopathic inhibition and virus titration assays. The S2 (1.25 mg/ml) hot aqueous extract demonstrated the highest broad-spectrum antiviral activity against all tested enteroviruses in human primary oral fibroblast cells. Replication of EV-A71, CV-A16 and CVA10 were effectively inhibited at 2 hours post-infection (hpi) to 72 hpi, except for CV-A6 which was only at 2 hpi. S2 also has virucidal activity against EV-A71. Polysaccharides isolated and purified from crude hot aqueous extract demonstrated similar antiviral activity as S2, suggesting that polysaccharides could be one of the active compounds responsible for the antiviral activity shown by S2. To our knowledge, this study demonstrates for the first time the ability of GNJI to inhibit enterovirus infection and replication. Thus, GNJI is potential to be further developed as an antiviral agent against enteroviruses that caused HFMD.

Human leukocyte antigen (HLA) class II association in chronic hepatitis B patients with hepatocellular carcinoma in a Malay population: A pilot study

@#Asian countries account for almost three quarter of hepatocellular carcinoma (HCC) reported globally and chronic hepatitis B infection is one of the main contributors. Clinical observations show that Malay patients with chronic hepatitis B and HCC tend to have a worse outcome, when compared to other two major races in Malaysia. The objectives of this study was to determine the frequency of human leukocyte antigen (HLA) class II alleles in chronic hepatitis B patients with HCC among Malays compared to the general population to identify potential associations of HLA alleles with this disease. HLA class II typing was performed in chronic hepatitis B patients with hepatocellular carcinoma (n=12) by -polymerase chain reaction, sequence specific primer (PCR-SSP) method. There were higher allelic frequencies of certain HLA-DQB1 and HLA-DRB1 alleles; HLA-DQB1*03 (07) (41.7%), and HLA-DRB1*12 (41.7% vs 28.6%) and compared to controls (41.7% vs 29.7%). However, there was no significant statistical correlation found when compared with the normal healthy general population. This study provides an insight into the HLA Class II association with chronic hepatitis B and hepatocellular carcinoma in Malays. However, findings from this study should be validated with a larger number of samples using a high resolution HLA typing.

First report of seroprevalence of Toxoplasma gondii in domestic geese in Hunan province, subtropical China

. Toxoplasma gondii is one of the most common protozoan parasites with widespread distribution globally. However, little information is available about the seroprevalence of T. gondii infection in geese (Anser domestica) in China. In the present study, the seroprevalence of T. gondii in geese were investigated in Hunan province, China. A total of 900 serum samples were collected from ten administrative regions in Hunan province, China, and assayed for T. gondii antibodies by Indirect Haemagglutination (IHA) test. Overall, 21.1% of the animals were positive for T. gondii antibodies. The results of the present survey indicated the presence of T. gondii infection in geese in Hunan province, China. Therefore, it is important to execute integrated control strategies and measures to prevent and control T. gondii infection in geese in this province. This is the first report seroprevalence of T. gondii in geese in Hunan province, China.

Effects of vitamin B fortified sucrose solution on the longevity and reproductive potentials of laboratory-bred Culex quinquefasciatus Say adult

Laboratory colonised mosquitoes are usually maintained on vitamin B complex fortified sucrose solution, however only few studies were conducted to evaluate the effects of such practice. This study aimed to determine the effects of different concentrations of sucrose solution fortified with and without 1% vitamin B complex on the longevity and reproductive potential of adult female of a local strain of Culex quinquefasciatus Say. Two arms of studies were carried out separately and each arm was triplicated. In one arm, concentrations of sucrose solution at 0%, 1%, 3%, 5% and 10% fortified with 1% Vitamin B complex were fed to the mosquitoes, while in the other arm, similar sucrose concentrations were used but without 1% vitamin B complex. Adult Cx. quinquefasciatus maintained on 5% sucrose solution fortified with 1% vitamin B complex exhibited significant extended vitality and longevity in stimulating ovarian development, compared with other vitamin fortified sucrose concentrations (p<0.05). The vitality and longevity of F0 and F1 males were 76.67±2.19 days and 57.67±8.19 days respectively. The F0 females survived the longest duration of 107.67±5.61 days and the F1 females survived 90.67±12.47 days with higher number of eggs laid, i.e. 1427.67±62.89 eggs at a higher hatchability rate of 57.05±8.39% or 814.49 eggs hatched. Thus, 5% sucrose solution fortified with 1% Vitamin B complex should be used to produce colonies of homogenous mosquitoes as this exerts positive biological effects on laboratory-bred Cx. quinquefasciatus.

Large Volume Osteosarcomas of the Femur Treated with Total Femoral Replacement

Patients with extensive malignancies involving the femur often require total femoral replacement when their limbs can be salvaged. Reported series are small and involve heterogeneity of tumours. We present nine patients with osteosarcomas of the femur treated at our institution between 2003 and 2010 with a mean follow-up of 27 (6 to 56) months. Their ages ranged from 9 to 17 (mean 14 years). They had large volume tumours (mean 911 cm3 ) and presented late with a mean of 5.5 months from the onset of symptoms to definitive treatment. All patients underwent resection and total femur replacement. Six patients have died and two are alive with good function at the time of this report. One was lost to follow-up. These patients require a high level of treatment care and have a guarded prognosis

Interleukin-6 promoter polymorphisms (-174 G/C) in Malaysian patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that involves the inflammation of various organs upon deposition of immune complexes and is characterized by uncontrolled B cell hyperactivity. Despite intensive research on the etiology of the disease, the exact cause of the onset of SLE is unknown. The pathogenesis of the disease has been proposed to be associated with the imbalance of T helper type 1 (Th1) and Th2 cytokine activities. Elevated serum levels of interleukin-6 (IL-6), a Th2 cytokine with various functions in the regulation of human biological systems, are observed in SLE patients. In the present study, 100 Malaysian SLE patients and 100 controls were evaluated in order to determine the association of polymorphisms existing in the promoter region of the IL-6 gene with the onset of SLE. The homozygous G genotype was found to be significant in SLE patients (χ² = 33.754; P = 0.00000000625), whereas the heterozygous G/C genotype was significant in the controls (χ²= 25.087; P = 0.000000548). We suggest that the C allele might have a masking effect on the G allele when both alleles are present in heterozygous individuals. However, we did not observe any significant association of the homozygous C allele with the onset of SLE or with protection from the disease (χ² = 1.684; P = 0.194366).

Changes in serum lipid profile and malondialdehyde following consumption of fresh or heated red palm oil

The effect of chronic consumption of fresh and heated red palm oil [RO] on lipid profile and lipid peroxidation was investigated. Thirty male rats were divided into 3 groups, each treated with the following prescribed food: [i] basal diet fortified with 15% weight/weight [w/w] fresh red palm oil [FRO], or [ii] heated once red palm oil [1H-RO] or [iii] heated 5 times red palm oil [5H-RO] for 20 weeks. There was a significant increase [p<0.05] in MDA concentration in all 3 groups compared to their respective baseline concentrations and a significant decrease [p<0.05] in total cholesterol [TC] concentration. However, the transient changes observed in serum triglyceride and HDL-cholesterol concentration did not attain significant values. The LDL-cholesterol concentration in 5H-RO group increased significantly [p<0.05] compared to pretreatment value, FRO and 1H-RO groups. Ratio of TC/HDL in 5H-RO group initially increased, but dropped to baseline level at the end of the study. There was no significant difference in the ratio of TC/HDL between the groups. In conclusion, both fresh and heated RO appeared comparable in their effect on serum cholesterol and lipid peroxidation. It appears that long term feeding with fresh and heated RO did not have an adverse effect on serum TG, HDL and TC/HDL ratio. However, it appears that prolonged heating increases LDL-cholesterol level. Further studies are required to ascertain whether the increase in LDLcholesterol and MDA with heated oil would render it more atherogenic

Isolation and polymerase chain reaction detection of Mycoplasma pneumoniae in Malaysian patients with respiratory tract infections.

Isolation and polymerase chain reaction (PCR) were performed for detection of Mycoplasma pneumoniae from respiratory tract specimens obtained from 200 adult and 200 pediatric patients. M. pneumoniae was isolated from bronchoalveolar lavage fluid of 1(0.5%) adult patient and 4(2.0%) tracheal aspirates of pediatric patients. PCR was positive for only one (0.5%) broncoalveolar lavage fluid of an adult patient and fifteen (7.5%) tracheal aspirates of pediatric patients. This study suggested that M. pneumoniae was more frequently detected in pediatric patients and PCR appears to have advantages over isolation, in terms of rapidity and sensitivity.

Detection of Mycoplasma pneumoniae from cerebrospinal fluid samples of pediatric patients.

Central nervous system manifestations are probably the most frequent extrapulmonary complications of infections due to Mycoplasma pneumoniae, occur mostly in children. In this study, we attempted to isolate M. pneumoniae and to detect the organism by polymerase chain reaction (PCR) from cerebrospinal fluid samples (CSF) of pediatric patients. Of the 244 CSF samples, no M. pneumoniae was isolated. Six (2.5%) of the CSF samples were positive by PCR amplification. More effort are necessary to isolate the organism from CSF samples in order to ascertain the role of M. pneumoniae in causing neurological complications.

Human leptospirosis: clinical and laboratory diagnosis

The findings in this study are of epidemiologic importance since the leptospires were isolated from human cases in 18 of 46 suspected cases (39%) which is a much better isolation rate than the previous local attemptsA clinical analysis of these patients was presented to enligthen the clinicians about this health problem and to help them confirm the diagnosis which is primarily the objective of this studyWith a wider realization of this health problem, we may in the future be able to institute adequate and necessary health sanitation and improvement to prevent and control such disease which should only be confined to animals. (Summary)

Disinfection: gaps between recommended and actual practice.

Basic practices on disinfection was surveyed in 6 hospitals using an observation and interview checklist. Two surveys were done, one pre-(first survey) and one post-intervention (second survey). The disinfection and sterilization policy of the Ministry of Health was not available in 66 (70.2%) and 12 (13%) of the units in the first and second survey respectively. In the second survey, staff in all the units washed disinfectant containers before refilling compared with 41.5% of the units in the first survey. Dilution of disinfectants not recommended was found to be used in the first survey. Storing cleaned and sterile items in disinfectants, using disinfectant as a substitute for sterilization of autoclavable items and not decontaminating spillages were some of the wrong practices observed. Considerable improvements were made in the second survey. Improper usage of disinfectants was also indicated by failure of the in-use test. Rate of failure of disinfectants in-use decreased from 11.6% in the first survey to 5.0% in the second survey. To ensure proper disinfection practices, a comprehensive training program on disinfection is required for nurses and attendants.

An evaluation of knowledge and awareness of disinfection and sterilization among health care workers.

Awareness of the disinfection and sterilization policy among hospital staff and their knowledge in basic principles and methods of disinfection and sterilization were studied before and after intervention using a self-administered questionnaire. Survey results showed that awareness (56.2%) before intervention was unsatisfactory. The nurses were more aware of the policy than other groups of medical personnel. Those unaware of the policy perform duties from memory or verbal instructions. A significant increase in awareness to 73.3% was observed after intervention (p < 0.05). Knowledge on methods of decontamination, disinfection and sterilization of equipment varies widely from 28.8% to 90.1%. 23.1% were unaware of the temperature used for sterilization while 72.4% did not know how containers of disinfectant should be refill. Only 14.7% knew the recommended method for washing containers. With education improvement was observed. The average knowledge improved from 44.4% to 57.3%. Our results indicated that continuous in-service education is needed to improve, supplement and update knowledge in this field after basic training. In addition orientation programs for new staff should also be aimed at creating awareness and providing information on guidelines and policies related to their duties.

Monitoring of diabetes in children.

There is increasing evidence to show that a high degree of metabolic control in diabetes mellitus delays and reduces the severity and incidence of the microvascular complications of diabetes. The goals of appropriate therapy for diabetes should hence include an all-out effort to achieve as near normal metabolism as possible but it should be individualized accordingly. In actual practice it is difficult to achieve "good" control especially in diabetic children without attendant hypoglycaemia and its sequelae. Diabetic control is difficult to define in clinical terms, and the maintenance of metabolic control in diabetes involves a complex interaction of pathological, physiological, psychological, familial, social and environmental variables. The aims in insulin-dependent diabetic children is to ensure optimal emotional and physical health of the child or adolescent. The various modalities available for monitoring or assessment of diabetic control shall be discussed.