Kacip Fatimah also known as Labisia pumila (Myrsinaceae), is a traditional herbal medicine with a long history in the Malay community. It has been used by many generations of Malay women to induce and facilitate childbirth as well as a post-partum medicine. We tested the genotoxic potential of Kacip Fatimah in bone marrow cells obtained from Sprague-Dawley rats using micronuclei formation as the toxicological endpoints. Five groups of five male rats each were administered orally for two consecutive days with doses of 100, 700 and 2000 mg/kg body weight of Kacip Fatimah extract dissolved in distilled water. Micronucleus preparation was obtained from bone marrow cells of the animals following standard protocols. No statistically significant increase in micronucleated polychromatic erythrocytes (MNPCEs) was observed at any dose level and sacrifice/harvest time point (24, 48 and 72h). However, a significant decrease in polychromatic erythrocytes/normochromatic erythrocytes (PCE:NCE) ratio was observed from the highest dose level (2000 mg/kg of body weight) at 48h harvest time point. In this study, we investigated the effect of Kacip Fatimah on mammalian bone marrow cells using micronuclei formation to assess the genotoxicity of the herb.
Medicinal plants have many traditional claims including the treatment of ailments of infectious origin. In the evaluation of traditional claims, scientific research is important. The objective of the study was to determine the presence of antibacterial activity in the crude extracts of some of the commonly used medicinal plants in Malaysia, Andrographis paniculata, Vitex negundo, Morinda citrifolia, Piper sarmentosum, and Centella asiatica. In this preliminary investigation, the leaves were used and the crude extracts were subjected to screening against five strains of bacteria species, Methicillin Resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli, using standard protocol of Disc Diffusion Method (DDM). The antibacterial activities were assessed by the presence or absence of inhibition zones and MIC values. M. citrifolia, P. sarmentosum and C. asiatica methanol extract and A. paniculata (water extract) have potential antibacterial activities to both gram positive S. aureus and Methicillin Resistant S. aureus (MRSA). None of the five plant extracts tested showed antibacterial activities to gram negative E. coli and K. pneumoniae, except for A. paniculata and P. sarmentosum which showed activity towards P. aeruginosa. A. paniculata being the most potent at MIC of 2 g/disc. This finding forms a basis for further studies on screening of local medicinal plant extracts for antibacteria properties.
In the present study we examined the effect of E. longifolia methanol extract (TA164) on the GSH levels of P. falciparum infected erythrocytes and uninfected erythrocytes. Our study on parasite growth shows the IC50 and IC75 values of TA164 to be 0.17 g/ml and 6 g/ml respectively while for BSO was 25.5 g/ml and 46.5 g/ml respectively. About 95% to 100% growth inhibition of P. falciparum infected erythrocyte was observed when treated with TA164 and BSO at 16 g/ml and 64 g/ml respectively. The study on GSH contents indicated that non-infected erythrocytes treated with 6 g/ml (IC75 values) of TA164 at 24 hours incubation showed less GSH content as compared to non-treated erythrocytes. A similar observation was seen on treated trophozoite infected erythrocyte (10% parasitemia) when treated with 6 g/ml at 3 hours incubation. Analysis of the GSH contents of parasite compartments treated with TA164 at the same concentration (6 g/ml) for 3 hours incubation indicated a reduction of GSH contents. At the same concentration, TA164 did not affect the GSH contents of enriched trophozoite infected erythrocytes (60-70% parasitemia). TA164 did affect the GSH content of non-infected erythrocyte at 24 hours (accept IC50 value) as well as the parasite compartments (trophozoite infected erythrocyte and parasite itself) but fails to affect the GSH content of enriched trophozoite infected erythrocyte.
Eurycoma longifolia, locally known as 'Tongkat Ali' is a popular local medicinal plant that possess a lot of medicinal properties as claimed traditionally, especially in the treatment of malaria. The claims have been proven scientifically on isolated compounds from the plant. The present study is to investigate the anti malaria properties of Eurycoma longifolia standardized extract (root) (TA164) alone and in combination with artemisinin in vivo. Combination treatment of the standardized extract (TA164) with artemisinin suppressed P. yoelii infection in the experimental mice. The 4 day suppressive test showed that TA164 suppressed the parasitemia of P. yoelii-infected mice as dose dependent manner (10, 30 and 60 mg/kg BW) by oral and subcutaneous treatment. By oral administration, combination of TA164 at 10, 30 and 60 mg/kg BW each with artemisinin respectively showed a significant increase in the parasitemia suppression to 63, 67 and 80 percent as compared to artemisinin single treatment (31%). Using subcutaneous administration, at 10 mg/kg BW of TA164 in combination with 1.7 mg/kg BW of artemisinin gave a suppression of 80% of infection. This study showed that combination treatment of TA164 with artemisinin gives a promising potential anti malaria candidate using both oral and subcutaneous route, the later being the most potent.
Colorectal carcinoma ranks third among ten leading causes of cancer in Malaysia. The colorectal carcinoma tumourigenesis involves the inactivation of tumour suppressor genes, and activation of proto-oncogenes. The p53 is one of the tumour suppressor genes that is involved in the colorectal carcinogenesis. The p53 gene is located on human chromosome 17p13.1 and comprises of 11 exons. Deficiencies in the p53 gene can cause the cancerous cells to spread to distant organs such as liver, lungs, lymph nodes, spine and bone. The most common p53 abnormalities that can lead to the metastasis of colorectal tumours are mutation and deregulation of the gene. In this study, nine colorectal carcinoma samples were used to establish a simple and sensitive strategy in the study on in vivo p53 expression by using realtime LightCycler SYBR Green I technology.
A 14-months survey was carried out to identify the species composition of Anopheles mosquitoes from Kampung Bongor, Grik, Perak. Adding to that, a preliminary one month mosquito population screening was done at Kampung Tepin, Serian, Sarawak. Consequently, the insecticide susceptibility status of a pyrethroid was tested against two selected species of Anopheles collected from these two locations in Malaysia. A total of 4,497 Anopheles from 11 species were identified from collections in Kampung Bongor, whereas 2,654 An. letifer were collected from Kampung Tepin. The An. maculatus of Kampung Bongor and An. letifer of Kampung Tepin were then selected and tested using WHO standard diagnostic test kits and impregnated papers with 0.75% permethrin. The response values of KT50 and KT95 for An. maculatus were recorded at 28.09 minutes and 62.98 minutes respectively. Anopheles letifer recorded much slower response values of KT50 and KT95, which was at 35.09 minutes and 73.03 minutes respectively. Both An. maculatus and An. letifer showed 100% mortality after 24 hours holding period. The results indicate that both species were still susceptible to the tested pyrethroid. For effective vector control and resistance management, accurate and periodic insecticide resistance monitoring should be undertaken especially in rural areas with agricultural usage of insecticides.
Routine surveillance on resistant status of field mosquito populations is important to implement suitable strategies in order to prevent pest outbreaks. WHO test kit bioassay is the most frequent bioassay used to investigate the susceptibility status of field-collected mosquitoes, as it is relatively convenient to be carried out in the field. In contrast, the topical application of active ingredient is less popular in investigating the susceptibility status of mosquitoes. In this study, we accessed the susceptibility status of Aedes albopictus Skuse collected from two dengue hotspots on Penang Island: Sungai Dua and Persiaran Mayang Pasir. Two active ingredients: permethrin and deltamethrin, were used. WHO test kit bioassay showed that both wild strains collected were susceptible to the two active ingredients; while topical application assay showed that they were resistant. This indicated that WHO test kit bioassay less sensitive to low level of resistance compared to topical application assay. Hence, topical application is expected to be more indicative when used in a resistance surveillance programme.
House flies were collected from April 2007-April 2008 from two poultry farms (Balik Pulau and Juru) in the state of Penang. The resistance level of the first generation offspring was evaluated against DDT, malathion, propoxur, and permethrin using the topical application method. The resistance ratio (RR) of the Balik Pulau strain house flies for propoxur, malathion and DDT ranged from 10.28 to 99.00, 7.83 to 47.01 and 6.05 to 31.10, respectively. Resistance to propoxur and malathion in house fly was attributed to cross resistance to organophosphate insecticides used in the farm. Increased metabolic detoxification might be the mechanism involved in DDT resistance due to excessive application of cypermethrin formulation. The RR of the Juru strain for propoxur, malathion and DDT was in a decreasing pattern throughout the study period, ranging from 5.58 to 83.38, 15.19 to 27.82, and 10.04 to 22.69, respectively. Permethrin appeared to be the most potent insecticide in controlling house fly in both the Balik Pulau (RR=0.50 to 1.96) and Juru poultry farms (RR=0.64 to 2.40). The fluctuations of insecticides resistance in house fly was also found to correlate with climatic factors due to its rapid breeding. Relative humidity exhibited positive correlation indices with the changes in the resistance level for DDT (r=0.481, p<0.05), malathion (r=0.698, p<0.01), and permethrin (r=0.580, p<0.05) in Balik Pulau. Similarly, relative humidity in Juru also showed strong correlation with the RR for DDT (r=0.900, p<0.01), malathion (r=0.762, p<0.05), permethrin (r=0.760, p<0.05), and propoxur (r=0.897, p<0.01).
Studies were carried out on the residual efficacy of Bacillus thuringiensis H-14 (water dispersible granule, VectoBac ABG 6511) as direct application in the control of Aedes larvae in the field. Field Aedes sp populations in the earthen and glass jars were predetermined before initiation of the trial. On confirmation of the presence of Aedes species in the designated area, Sungai Nibong Kecil, Penang Island, Malaysia, Bti was introduced in the 55L earthen and 3L glass jars). Two test designs were carried out. The first design had treated water replenished daily with 6L of seasoned water and the second design is without the replenishment of water but evaporated water was replenished. Bti was effective in the field for at least 35 days with more than 80% reduction in the Aedes larvae in the treated containers. For earthen jars with daily replenishment of water, 100% reduction was recorded for the first 3 days, while more than 80% reduction was recorded up to day 40. At day 60, Bti still provided an efficacy of 54.32 +/- 4.61 (%) of reduction. Whilst for earthen jars without daily replenishment of water, 100% reduction was recorded for the first 5 days, while more than 80% of reduction was recorded up to day 40. For the glass jars studied, similar efficacy was observed. In jars with daily replenishment of water a better larval control was observed. Percentage of reduction from day 50 to 60 for replenishment of water was between 50 to 70% compared to without replenishment of water with less than 40%.
Laboratory efficacy and residual activity of a water dispersible granule formulation of Bacillus thuringiensis israelensis (Bti) at the dosages of 3000, 6000 and 15000 ITU/L were conducted in this study. The study was conducted in two different size containers, earthen jar (45 L) and glass jar (3 L) with or without daily replenishment of 6 L and 0.3 L of water in the earthen and glass jars, respectively. Results indicate that for both earthen jar and glass jar evaluations, Bti at the tested dosages, performed effectively against Aedes aegypti, giving a minimum of 42 days effective killing activity. When the dosage was increased from 3000 ITU/L to 6000 ITU/L or 15000 ITU/L, the effective periods of the Bti increased by an additional one to three weeks. The Bti water dispersible granule provided better larvicidal activity with replenishment of water compared with non-replenishment of water especially for the higher dosage (15000 ITU/L).
There is a need for identification of new infection markers against common Leptospira isolates in Malaysia. To achieve this goal, seven-day-old cultures of Leptospira interrogans serogroup Icterohemorrhagiae (L44) and Leptospira interrogans serogroup Javanica (L55) were used for antigen preparation by sequential extraction method using 40 mM Tris, 8M Urea and 2M thiourea. Immunoblot analysis of the antigens were performed using serum samples from 46 local patients with confirmed acute leptospirosis, 28 patients with other infections and 14 healthy controls. The patients serum samples used in this study contained heterologous antibody against a number of different leptospira serovars. A strong IgM reactivity to a broad diffuse band of 10-15 kDa was observed. Combining results using L44 and L55 antigens showed sensitivity of 80.4% and specificity of 95.2% for detection of leptospirosis. Proteinase K and periodate treatment indicated that the band is likely to be lipopolysaccharide (LPS) in nature. This study showed that the 10-15 kDa antigen could potentially be useful for serodiagnosis of acute leptospirosis in Malaysia.
The parasitic protozoan fauna in sixty-six anurans comprising of Duttaphrynus melanostictus, Phrynoidis juxtaspera, Hylarana erythraea and Polypedates leucomystax collected from Zoo Negara Malaysia was investigated. The distribution and prevalence rate of parasitic species in the digestive tract and blood were examined. Seven species of intestinal protozoa (Opalina ranarum, Cepedea dimidiata, Nycthetorus cordiformis, Entamoeba ranarum, Iodamoeba butschlii, Endamoeba blattae, and Tritrichomonas sp.) and two species of blood protozoa (Lankesterella sp. and Trypanosoma sp.) were recorded. Opalina ranarum was the most common protozoan found in the rectum and intestine (prevalence rate: 34.8%) infecting all host species, with P. juxtaspera heavily infected with the parasite, whereas Tritrichomonas sp. was the least prevalent intestinal species infecting only D. melanostictus. Both Lankesterella sp. and Trypanosoma sp. were found in the blood of H. erythraea.
The use of Chrysomya megacephala larvae for detecting malathion for diagnosing the cause of death was investigated. This could prove useful when the visceral organs have become liquefied during decomposition and therefore cannot be sampled. A field experiment was conducted in which C. megacephala were allowed to colonise naturally the corpses of rabbits that had died of malathion poisoning. The concentration of malathion increased gradually during the larval stages of C. megacephala reaching the maximum concentration in the third instar larvae. The concentration of malathion declined during prepupal stage and reached its lowest level among tenerals. The average malathion concentrations in C. megacephala growing in poisoned rabbit corpses left in a sunlit habitat were significantly higher (p<0.05) than those growing on poisoned rabbits left in a shaded habitat. The concentrations of malathion in the different stages of development of C. megacephala were moderately correlated (r = 0.51-0.69) with the administered doses as well as with those estimated in visceral organs. Thus, it would not be reliable to suggest the formulation of mathematical algorithms for relating the concentration of malathion found in the different stages of development of C. megacephala with those found in the visceral organs. However, in the context of forensic investigation, the qualitative detection of malathion in C. megacephala may prove useful in diagnosing the cause of death, since malathion is a common cause of accidental and suicidal deaths.
Cryptosporidiosis is a particular concern in immunocompromised individuals where symptoms may be severe. The aim of this study was to examine the epidemiological and molecular characteristics of Cryptosporidium infections in HIV/AIDS patients in Malaysia in order to identify risk factors and facilitate control measures. A modified Ziehl-Neelsen acid fast staining method was used to test for the presence of Cryptosporidium oocysts in the stools of 346 HIV/AIDS patients in Malaysia. Standard coproscopical methods were used to identify infections with other protozoan or helminths parasites. To identify the species of Cryptosporidium, DNA was extracted and nested-PCR was used to amplify a portion of the SSU rRNA gene. A total of 43 (12.4%) HIV-infected patients were found to be infected with Cryptosporidium spp. Of the 43 Cryptosporidium-positive HIV patients, 10 (23.3%) also harboured other protozoa, and 15 (34.9%) had both protozoa and helminths. The highest rates of cryptosporidiosis were found in adult males of Malay background, intravenous drug users, and those with low CD4 T cell counts (i.e., < 200 cells/mm3). Most were asymptomatic and had concurrent opportunistic infections mainly with Mycobacterium tuberculosis. DNA sequence analysis of 32 Cryptosporidium isolates identified C. parvum (84.3%), C. hominis (6.3%), C. meleagridis (6.3%), and C. felis (3.1%). The results of the present study revealed a high prevalence of Cryptosporidium infection in hospitalized HIV/AIDS patients. The results also confirmed the potential significance of zoonotic transmission of C. parvum in HIV infected patients, as it was the predominant species found in this study. However, these patients were found to be susceptible to a wide range of Cryptosporidium species. Epidemiological and molecular characterization of Cryptosporidium isolates provides clinicians and researchers with further information regarding the origin of the infection, and may enhance treatment and control strategies.
Advocacy and training on "Home care" for filarial lymphoedma (FLE) patients are provided through morbidity management and disability prevention (MMDP) clinic commonly known as filariasis clinic and clinical improvement is assessed by follow-up visits. While the physicians aim at reducing the recurrent ADL (coined as ADLA in 1997) episodes, the patients expect reduction in LE volume. The objective of the present study was to know whether the MMDP clinic serves the primary expectation of the FLE patients. LE patients who attended the clinic for at least four follow-up consultations and had LE volume measurements at three points of time during the one year period of observation were considered for analysis. Clinical assessment was done for LE grading and LE volume was measured by water displacement volumetry. Sixty-three patients who fulfilled the follow up criteria were included. It was observed that the median LE volume was 914ml (IQR 269 - 1935) at first visit of the observation period which reduced to 645ml (IQR 215- 1666) and 752ml (IQR 215 - 1720) at first and second follow-up visits respectively. Over all, in short span of one year, 21 of the 63 patients (33.3%) who visited MMDP clinic at least four times in a year were benefitted through the MMDP advocacy and the National filariasis control programme need to emphasise on the importance of follow up visits to FLE patients.
Dengue virus infects millions of people worldwide and there is no vaccine or anti-dengue therapeutic available. Screening large numbers of medicinal plants for anti-dengue activities is an alternative strategy in order to find the potent therapeutic compounds. Therefore, this study was designed to identify anti-dengue activities in nineteen medicinal plant extracts that are used in traditional medicine. Local medicinal plants Vernonia cinerea, Hemigraphis reptans, Hedyotis auricularia, Laurentia longiflora, Tridax procumbers and Senna angustifolia were used in this study. The highest inhibitory activates against dengue NS2B-NS3pro was observed in ethanolic extract of S. angustifolia leaves, methanolic extract of V. cinerea leaves and ethanol extract of T. procumbens stems. These findings were further verified by in vitro viral inhibition assay. Methanolic extract of V. cinerea leaves, ethanol extract of T. procumbens stems and at less extent ethanolic extract of S. angustifolia leaves were able to maintain the normal morphology of DENV2-infected Vero cells without causing much cytopathic effects (CPE). The percentage of viral inhibition of V. cinerea and T. procumbens extracts were significantly higher than S. angustifolia extract as measured by plaque formation assay and RT-qPCR. In conclusion, The outcome of this study showed that the methanolic extract of V. cinerea leaves and ethanol extract of T. procumbens stems possessed high inhibitory activates against dengue virus that worth more investigation.
Fructose-1,6-bisphosphate aldolase (FbA), a well characterized glycometabolism enzyme, has been found to participate in other important processes besides the classic catalysis. To understand the important functions of three fructose-1,6-bisphosphate aldolases from Clonorchis sinensis (CsFbAs, CsFbA-1/2/3) in host-parasite interplay, the open reading frames of CsFbAs were cloned into pET30a (+) vector and the resulting recombinant plasmids were transformed into Escherichia coli BL21 (DE3) for expression of the proteins. Purified recombinant CsFbAs proteins (rCsFbAs) were approximately 45.0 kDa on 12% SDS-PAGE and could be probed with each rat anti-rCsFbAs sera by western blotting analysis. ELISA and ligand blot overlay indicated that rCsFbAs of 45.0 kDa as well as native CsFbAs of 39.5 kDa from total worm extracts and excretory-secretory products of Clonorchis sinensis (CsESPs) could bind to human plasminogen, and the binding could be efficiently inhibited by lysine analog ε-aminocaproic acid. Our results suggested that as both the components of CsESPs and the plasminogen binding proteins, three CsFbAs might be involved in preventing the formation of the blood clot so that Clonorchis sinensis could acquire enough nutrients from host tissue for their successful survival and colonization in the host. Our work will provide us with new information about the biological function of three CsFbAs and their roles in hostparasite interplay.
Angiostrongylus cantonensis is the most frequent cause of eosinophilic meningitis in humans in Thailand and worldwide. Because of difficulty of recovering the Angiostrongylus larvae from infected patients, detection of parasite-specific antibodies is used to support clinical diagnosis. This study tested serum samples from eosinophilic meningitis patients and individuals at risk of infection with A. cantonensis to evaluate a recently developed simple and rapid dot-immunogold filtration assay (DIGFA) for detection of specific antibodies against A. cantonensis. Purified 31-kDa glycoprotein of A. cantonensis and protein A colloidal gold conjugate were employed to detect the 31-kDa anti-A. cantonensis antibody in patients sera from the parasite endemic areas of northeast Thailand. The results were compared with those obtained by dot-blot enzyme-linked immunosorbent assay (ELISA) with 31-kDa A. cantonensis antigen. The overall positivity rate of DIGFA and dot-blot ELISA for A. cantonensis infection in 98 clinically diagnosed cases from three highly endemic districts in Khon Kaen province were 39.79% and 37.75%, respectively. Among 86 sera of subjects at risk of infection with A. cantonensis, 24.41% were positive by DIGFA and 23.25% by dot-blot ELISA. There were good correlation between the visual grading of DIGFA and dot-blot ELISA in both groups of defined sera. DIGFA is as sensitive and specific as dot-blot ELISA for confirming eosinophilic meningitis due to A. cantonensis infection, with advantages of simplicity, rapidity and without the use of specific and expensive equipment, and can be used in field settings.
Understanding of antibody kinetics against SARS-CoV-2 and its vaccines is rapidly evolving. This study aims to (1) determine post-vaccination seroprevalence; (2) compare antibody levels between vaccine types and various clinical/demographic determinants; and (3) determine post-vaccination antibody concentrations against time. This is a retrospective cross-sectional study involving 148 healthcare employees all over Malaysia. IgG Spike (RBD), IgM Spike and IgG Nucleocapsid concentration medians were compared using Mann-Whitney U or Kruskal-Wallis tests. Chi Square and Spearman correlation coefficient tests were performed to identify variables associated with antibody titers. A scatter plot of IgG Spike (RBD) against time from last vaccine dose was also plotted. At 1-month post-vaccination, all employees successfully seroconverted regardless of vaccine type, health status and COVID- 19 history. Comirnaty, convalescent, female or Malay vaccinees had significantly higher IgG Spike (RBD) titers compared to their respective counterparts. No correlation was found between age and IgG Spike (RBD) levels. Concentration of all three antibodies waned with time post-vaccination, with IgM Spike and IgG Nucleocapsid waning faster than IgG Spike (RBD).
The hepatitis C virus (HCV) is a blood-borne pathogen that causes acute or chronic infection of the liver, sometimes leading to serious liver damage and fatality. The objective of this study was to evaluate HCV prevalence in patients attending the Regional Training and Research Hospital for Medical Examination and Surgery in Samsun Province of Turkey between 2014 and 2017. Blood specimens taken from 152 596 patients were screened for HCV infection by using the anti-HCV assay. Seropositive samples were subjected to polymerase chain reaction (PCR) testing in order to determine whether the HCV infection was active. Genotyping was then performed. Overall, HCV seropositivity and active HCV infection were 2.76% and 2.05%, respectively. Foreign nationals accounted for 5.61% of the seropositive samples and 1.37% of active HCV infective samples. We further report that 2017 was the year with the highest seroprevalence which was 3.64%. HCV genotype 1 was the most common genotype detected in residents of Samsun Province at 89.86%, followed by Genotype 3 at 4.54%. This study provides important information on the levels of HCV infection in the Samsun region of Turkey. The data indicate that there was a rising trend of HCV infection between 2014 and 2017.