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.

Experimental Study on Plasmodium knowlesi Normocyte Binding Protein Xa Region II (PkNBPXaII) for Erythrocyte Binding

@# Normocyte binding protein Xa (NBPXa) has been implied to play a significant role in parasite invasion of human erythrocytes. Previous phylogenetic studies have reported the existence of three types of NBPXa for Plasmodium knowlesi (PkNBPXa). PkNBPXa region II (PkNBPXaII) of type 1, type 2 and type 3 were expressed on mammalian cell surface and interacted with human and macaque (Macaca fascicularis) erythrocytes. The binding activities of PkNBPXaII towards human and macaque erythrocytes were evaluated using erythrocyte-binding assay (EBA). Three parameters were evaluated to achieve the optimal protein expression of PkNBPXaII and erythrocyte binding activity in EBA: types of mammalian cells, post transfection time and erythrocyte incubation time. COS-7, HEK-293, and CHO-K1 cells showed successful expression of PkNBPXaII, despite the protein expression is weak compared to the positive control. COS-7 was used in EBA. All three types of PkNBPXaII showed rosette formation with macaque erythrocytes but not with human erythrocytes. Future studies to enhance the PkNBPXaII expression on surface of mammalian cells is indeed needed in order to elucidate the specific role of PkNBPXaII in erythrocytes invasion.

Histopathology of Brugia pahangi and Plasmodium berghei ANKA co-infection in the Gerbil (Meriones unguiculatus)

@#Co-infection with multiple different parasites is a common phenomenon in both human and animals. Among parasites that frequently co-infect the same hosts, are the filarial worms and malaria parasites. Despite this, the mechanisms underlying the interactions between these parasites is still relatively unexplored with very few studies available on the resulting pathologies due to co-infection by filarial nematodes and malaria parasites. Hence, this study investigated the histopathological effect of Brugia pahangi and Plasmodium berghei ANKA (PbA) infections in gerbil host. Gerbils grouped into B. pahangi-infected, PbA-infected, B. pahangi and PbA-coinfected, and uninfected control, were necropsied at different time points of post PbA infections. Brugia pahangi infections in the gerbils were first initiated by subcutaneous inoculation of 50 infective larvae, while PbA infections were done by intraperitoneal injection of 106 parasitized red blood cells after 70 days patent period of B. pahangi. Organs such as the lungs, kidneys, spleen, heart and liver were harvested aseptically at the point of necropsy. There was significant hepatosplenomegaly observed in both PbA-infected only and coinfected gerbils. The spleen, liver and lungs were heavily pigmented. Both B. pahangi and PbA infections (mono and coinfections) resulted in pulmonary edema, while glomerulonephritis was associated with PbA infections. The presence of both parasites induced extramedullary hematopoiesis in the spleen and liver. These findings suggest that the pathologies associated with coinfected gerbils were synergistically induced by both B. pahangi and PbA infections.

Operative Treatment of Knee Cartilage Injuries: A Review of the Current Literature on Non-Cell-Based and Cell-Based Therapies.

Aims: Cartilage is frequently damaged through injury and disease but shows little or no capacity for repair. Injuries that extend to the subchondral level show some capacity for repair due to the release of bone marrow derived mesenchymal stem cells. Focal articular cartilage defects are challenging clinical problems that may progress to more generalised lesions. We reviewed the literature to analyse the results of available noncell- based and cell-based strategies for the repair of articular cartilage defects in the knee. Study design: Review Article Place and Duration of Study: University College London Institute of Orthopaedics and Musculoskeletal Sciences, Royal National Orthopaedic Hospital, Stanmore, Middlesex, HA7 4LP, United Kingdom Methodology: We reviewed the literature to identify studies on the use of non-cellbased and cell-based strategies for the repair of articular cartilage defects in the knee. Results: Repair techniques that do not utilise cell therapy include bone marrow stimulating techniques such as microfracture that is effective in small well-contained lesions and has the advantages of being performed arthroscopically as a single stage and cheaper costs compared to cell-based therapies. It also associated with no donor site morbidity unlike mosaicplasty, and perichondrial or periosteal grafting. The evidence suggests that none of the techniques described above consistently produce durable results. There are encouraging mid-term results with Autologous Matrix Induced Chondrogenesis (AMIC) procedures in small number of patients. Although microfracture is appropriate for smaller cartilage defects, Autologous Chondrocyte Implantation (ACI) and Matrix-carried Autologous Chondrocyte Implantation (MACI), as well as other cell carrier systems, are currently used to treat larger full thickness chondral defects in the knee. Although the results are fairly similar, MACI and procedures using other cell carrier systems are amenable to be performed arthroscopically or through a more limited approach. There are a small number of studies using mesenchymal stem cells with promising early results bur further in vitro and in vivo studies are needed before this treatment becomes more routinely available. Conclusion: Focal articular cartilage defects are challenging clinical problems that progress to more generalised lesions. Only cartilage injuries that extend to the subchondral level show some capacity for repair due to the release of bone marrow derived mesenchymal stem cells. Bone marrow stimulating techniques such as microfracture are effective in small well-contained lesions (<2cm2) and have the advantages of being performed arthroscopically as a single stage and cheaper costs compared to cell-based therapies. Mosaicplasty, and perichondrial or periosteal grafting are associated with donor site morbidity. Longer term studies on AMIC may help define the role for this procedure. The best form of non-cell-based treatment for focal articular cartilage defects in the long term is still unknown. ACI and MACI, as well as other cell carrier systems, are currently used in clinical practice to treat larger full thickness chondral defects in the knee. There are a small number of studies using mesenchymal stem cells and further in vitro and in vivo studies are needed before this treatment is optimised.

Scoring system for empyema thoracis and help in management.

OBJECTIVE: To evaluate the implications of a newly defined severity scoring of empyema in children for the prediction of surgical management and to compare the length of hospitalization as an outcome measure of patients treated using medical therapy, salvage video-assisted thoracoscopic surgery (VATS) vs early elective VATS. METHODS: A retrospective chart review of parapneumonic empyema of patients below 18 years of age admitted to a tertiary children's hospital in northern Taiwan from April 1993 to December 2002 was performed. Patients were categorized into a medical group who received antibiotic therapy, needle aspirations with/without tube thoracostomy; a salvage VATS group when the patients required surgery for the relief of persistent fever > 38 degrees C, chest pains or dyspneic respirations despite initial medical therapy; an early VATS group when the patients received elective surgery early after admission. The demographic data, clinical features, laboratory findings, and duration of hospitalization were compared using a severity score of empyema (SSE). RESULTS: Streptococcus pneumoniae was the most common infecting organism, followed by Staphylococcus aureus, Pseudomonas aeruginosa. No organisms were recovered in 39% of patients. A pleural pH < 7.1 increases the odds of requiring surgical intervention by 6 times among this cohort. Children who required decortication of empyema had a higher severity score (mean 4.8 vs 3.0, p < 0.005). The duration of hospitalization for patients having early VATS showed a shortening stay (mean 18 vs 28 days) as compared to salvage VATS. CONCLUSION: A pleural pH < 7.1 and a newly designed clinical severity score of empyema 4 are two predictors of surgical intervention for fibrinopurulent empyema in the present study. Early elective VATS may be adopted not later than 7 days after failure of appropriate antibiotic therapy and adequate drainage of empyema to decrease the length of stay and minimize morbidity.

Erythrocyte membrane fatty acid composition in cancer patients

Essential fatty acids (EFA) have an important role in complex metabolic reactions. The metabolism of essential polyunsaturated fatty acids (PUFA) appears to be one of the critical targets in the complex metabolic stages that lead to, or are associated with cancer. The goal of our research was to analyze the erythrocyte specific types of membrane fatty acid content, level and distribution in cancer patients as compared to non-cancer patients. Changes in fatty acid composition may affect different aspects of cell structure and function, including proliferation. Analyses of RBCs membrane fatty acids were performed for 255 patients with different types of cancer (breast, prostate, liver, pancreas, colon, and lung), 2,800 non-cancer patients and 34 healthy volunteers. Our research study demonstrated a lower level of stearic acid and an increased content of oleic acid in RBC of cancer patients in comparison with control and non-cancer patients. According to the results of this investigation, the ratio of Eicosa pentaenoic acid (EPA) and Decosa hexaenoic acid (DHA) to Alpha-linolenic acid (ALA) may be useful to estimate PUFA imbalances in cancer patients. EPA and DHA acid may be recommended as supplementation and in addition to current therapy during cancer treatment

Prediction of differential creatinine clearance in chronically obstructed kidneys by non-contrast helical computerized tomography

PURPOSE: We investigate the use of non-contrast helical computerized tomography (NCHCT) in the measurement of differential renal parenchymal volume as a surrogate for differential creatinine clearance (CrCl) for unilateral chronically obstructed kidney. MATERIALS AND METHODS: Patients with unilateral chronically obstructed kidneys with normal contralateral kidneys were enrolled. Ultrasonography (USG) of the kidneys was first done with the cortical thickness of the site with the most renal substance in the upper pole, mid-kidney, and lower pole of both kidneys were measured, and the mean cortical thickness of each kidney was calculated. NCHCT was subsequently performed for each patient. The CT images were individually reviewed with the area of renal parenchyma measured for each kidney. Then the volume of the slices was summated to give the renal parenchymal volume of both the obstructed and normal kidneys. Finally, a percutaneous nephrostomy (PCN) was inserted to the obstructed kidney, and CrCl of both the obstructed kidney (PCN urine) and the normal side (voided urine) were measured two 2 after the relief of obstruction. RESULTS: From March 1999 to February 2001, thirty patients were enrolled into the study. Ninety percent of them had ureteral calculi. The differential CrCl of the obstructed kidney ( percentCrCl) was defined as the percentage of CrCl of the obstructed kidney as of the total CrCl, measured 2 weeks after relief of obstruction. The differential renal parenchymal volume of the obstructed kidney ( percentCTvol) was the percentage of renal parenchymal volume as of the total parenchymal volume. The differential USG cortical thickness of the obstructed kidney ( percentUSGcort) was the percentage of mean cortical thickness as of the total mean cortical thickness. The Pearson's correlation coefficient (r) between percentCTvol and percentCrCl and that between percentUSGcort and percentCrCl were 0.756 and 0.543 respectively. The regression line was percentCrCl = (1.00) x percentCTvol - 14.27. The percentCTvol overestimated the differential creatinine clearance by about 14 percent, but the correlation is good. CONCLUSION: The differential renal parenchymal volume measured by NCHCT provided a reasonable prediction of differential creatinine clearance in chronically obstructed kidneys.

Paradoxical vocal cord adduction mimicking as acute asthma in a pediatric patient.

We report an adolescent girl with paradoxical vocal cord adduction who presented with acute onset of hyperventilation, wheezing and stridor that did not respond to bronchodilator and anti-inflammation therapy. The paradoxical vocal cord motion was confirmed by flexible fiberoptic bronchoscopic examination. We found the stridor was induced by hyperventilation, and was caused by paradoxical vocal cord movement. The abnormal cord motion may be psychogenic and could be misdiagnosed as asthma. It is important to investigate the underlying background and social history and to avoid unnecessary use of beta-agonists, steroids, and even endotracheal intubation or tracheostomy.

A profile of hypertension among rural elderly Malaysians.

The main objective of this study was to survey the prevalence of hypertension in Malaysians aged 55 years and above who responded to a public health screening campaign in Kuala Langat district, Selangor, Malaysia. The relationship between the detection, treatment and control in the study population was also examined. An epidemiological survey of the elderly was conducted in Kuala Langat, a rural district in Malaysia for a consecutive period of six weeks. The survey was conducted using a standardized questionnaire and interview by trained research assistants. The medical students and nurses were trained on the correct protocol of measuring blood pressure based on the recommendations of the British Hypertension Society. Newly detected persons with blood pressure readings 160/95 mmHg and above, were given a green card to return to the nearest health center for a further 2 visits to check his/her blood pressure. A total of 1,392 people with a mean age of 65 years and comprising of 53.1% males and 46.9% females responded to the health survey. Based on the history 335 (24.1%) of the respondents were known hypertensives and 82.1% of this hypertensive group were on regular treatment. However good control of hypertension was achieved only in 161 (48.1%) of them. Newly detected hypertension was found in 204 (14.6%) of the respondents. The overall prevalence of hypertension in our sample of respondents was 25.6%. There was no significant difference between the sexes and the races in terms of their detection, treatment and control. However there was a significant difference between the persons aged less than 65 years of age and those older than 65 years. There was a large proportion of previously known hypertensives among the survey population. Of those detected, not all were on regular medications and the majority on medications were not well controlled. Education of the public through intensive public health campaigns is vital to improve their knowledge on hypertension and the need for effective control. More community studies are needed to formulate better methods in the detection, treatment and control of hypertension.

Changing patterns of infectious keratitis: overview of clinical and histopathologic features of keratitis due to acanthamoeba or atypical mycobacteria, and of infectious crystalline keratopathy.

Acanthamoeba keratitis, infectious crystalline keratopathy and atypical mycobacterial keratitis have recently emerged as important types of infectious keratitis. These corneal infections have been associated with contact lens wear and with corneal surgical procedures such as radial keratotomy and penetrating keratoplasty, and the clinical setting of each of these infections is important in alerting the clinician to the possible diagnosis. There have been improvements in rapid diagnostic techniques for such infections in the last several years. Treatment has also improved, but remains a difficult problem, especially for Acanthamoeba. An overview of recent developments in the clinical and histopathologic methods for diagnosis and treatment options of these three corneal infections is provided.

Antinuclear antibody detection using streptavidin-biotin-peroxidase complex on HEp-2 cell substrate.

The streptavidin-biotin-peroxidase complex (SABC) technique was compared to conventional indirect immunofluorescence (IIF) for the detection of anti-nuclear antibody (ANA) on HEp-2 cell substrate. SABC showed higher specificity and predictive value and gave more reproducible titres and clearer staining patterns than IIF in sera from a series of rheumatic disease patients. Sera from 80 patients with various types of rheumatic diseases and 20 without rheumatic disease were further tested using the SABC method. All systemic lupus erythematosus (SLE) sera were positive. The overall sensitivity was 95%, specificity 90% and predictive value 97% for rheumatic disease. The rim pattern was associated with SLE and mixed connective tissue disease. The nucleolar/homogeneous pattern was associated with scleroderma and SLE in remission. ANA titre and staining pattern have limited value in the clinical assessment of rheumatic disease; however, ANA has very high sensitivity for SLE and remains an excellent screening test.

Danazol in treatment of lupus thrombocytopenia.

Seven patients with systemic lupus erythematosus (SLE), persistent thrombocytopenia (TP), in whom it was considered undesirable to institute an increase in steroid or immunosuppressive agents, were treated with danazol. Five patients completed the minimum period of 8 weeks. Two patients showed early response to danazol but were switched over to cyclophosphamide or azathioprine after 4 weeks because of systemic disease. Of the remaining five patients, four had complete responses. In one patient who failed treatment the TP was considered to be related to another drug (ranitidine). Other manifestations of SLE also improved with treatment. Side effects included amenorrhea in one patient, and hypoglycemia and hyponatremia in another. Infections were absent. Danazol can be a useful alternative treatment of lupus TP.

Lupus anticoagulant--a double misnomer.

The clinical features of thirteen Chinese patients with lupus anticoagulant were described. They were noted to conform to those reported among Caucasians and tend to suggest that the term 'lupus anticoagulant' is a double misnomer.