Identification Of Infectious Bronchitis Virus Isolates From Malaysia

Infectious bronchitis (IB) is a highly contagious respiratory, urogenital and reproduction disease of chickens and it is distributed worldwide. The disease is caused by infectious bronchitis virus (IBV). IBV is a member of the genus Coronavirus, family Coronaviridae and it has a single-stranded, positive-sense RNA genome of 27.6 kb. It possesses prominent surface spikes and has three major structural proteins; the spike (S) glycoprotein, the small integral membrane (M) glycoprotein and nucleocapsid (N) protein. In the commercial poultry industry, vaccination is used to control the disease. Despite vaccination program the disease continues to occur because IBV can exist in many serotypes. In many incidences, the existing vaccines are not able to provide full protection to the chickens against infectious bronchitis disease. The immune response stimulated to one serotype does not offer cross protection to another serotype. Moreover, the avian coronavirus capable of mutating and many IBV variants has been reported in many countries. Thus, it is crucial to develop a fast, sensitive and specific diagnostic technique to diagnose and identify the causative agent in order to control the disease spread. In recent years the reverse transcriptase-polymerase chain reaction (RT-PCR), reverse transcriptase-polymerase chain reaction-restriction fragment length polymorphism (RT-PCR-RFLP), cloning and genes sequencing had been used increasingly to detect and analyses IBV isolates. The objectives of these studies were to compare and optimize reverse transcriptase-polymerase chain reaction (RT-PCR) to diagnose IBV, to differentiate the Mass strain, and to characterize variant IBV isolated from this study. In this study, one-step and two-step RT-PCR techniques were used to amplify the conserved gene region of IBV by using universal and designed primers. This study was conducted on IBV isolates from year 1991 until 2003. In differentiation studies, isolates were group into serotype using reverse transcriptase-polymerase chain reaction (RT-PCR). Isolates recognized to be non Mass was further amplified their hypervariable region of S gene by using RT-PCR followed by RFLP technique to screen for nephropathogenic strain. Out of 31 IBV isolates, nine Mass strain were found. Following RT-PCR-RFLP, only one isolate showed different fragment pattern compared to nephropathogenic origin (MH5365/95). This particular isolates designated as V9/03 was neither Mass nor non-nephropathogenic serotype. The S1 region of V9/03 was further cloned, sequenced and its nucleotide and amino acid were compared to nephropathogenic MH5365/95 and Mass derivatives as well other references strains obtained from Gene bank. The V9/03 showed sequence homology to Taiwan (AY606321) and Korean (AY257060) strain with 82.5% and 81.6% identities respectively. The V9/03 has lower sequences homology (less than 80%) with nephropathogenic (MH5365/95) and Mass derivatives. The phylogenetic studies indicate that the strain V9/03 could be a local variant IBV which is different from local nephropathogenic MH5365/95 and vaccine strain. This study showed that variant IBV is circulating in the field as result of mutation of IBV due to the prolonged use of live virus vaccines and the immunological pressure of the virus to keep on survival in immune birds

The Descriptive Study of Knowledge and Awareness of Tuberculosis Among Students in Universiti Tun Hussein Onn Malaysia

Despite the disease management and controls introduced by Ministry of Health Malaysia, tuberculosis remains the major public threats among Malaysian. Enhancing the awareness among community is one of the strategy outlined by WHO in the Global Plan to Stop TB 2006-2015 program. Due to this, many programs have been introduced to increase awareness among the community. Universiti Tun Hussein Onn Malaysia is situated in the industrial radius area. The rate of migration is higher from neighboring countries such as Indonesia, Myammar, Bangladesh, Vietnam, Filipina and India which known to have high burden tuberculosis cases. Therefore, the anxious of the disease to spread is significant. This study employs a quantitative research approach by using semi structured questionnaires which have close ended questions. The questionnaire focused on tuberculosis knowledge on risk factors, clinical symptoms and treatments. About 270 questionnaires were randomly distributed amongst students at Universiti Tun Hussein Onn Malaysia and the results were analyzed using the Statistical Package of Social Sciences (SPSS) 20.0.The results demonstrated that, the majority of the respondents were aware of the infection, symptoms and curability of the disease. However, almost 90% of the respondents did not know the duration of treatments. The lack of awareness is associated with delays in seeking professional treatment. The study shows that awareness among students about TB is fairly good, but extensive survey is required especially on the risk factors

β-Cyclodextrin/lecithin: preparation and characterization

Lecithin is a fat which is well-known to be obtained from soybeans or egg yolks. It is also taken as a medicine and used as food additive that keep certain ingredients from separating out. Despite of its advantages, lecithin’s application in the industries is restricted due to its sensitivity to heat and light and can be easily oxidized. Hence, this study aims to investigate the effect of β-cyclodextrin (β-CD) in improving the properties of lecithin. The objectives of this study are to prepare and characterize β-CD/lecithin complex followed by study on the influence of β-CD on lecithin’s thermal stability and antioxidant property. Analytical methods employed are fourier transform infra-red (FTIR) spectroscopy for chemistry study, differential scanning calorimetry (DSC) for examination of its thermal stability, DPPH testing for its antioxidant property as well as scanning electron microscopy (SEM) for structural examination. Results from FTIR and SEM analyses confirmed the formation of β-CD/lecithin complex. Thermal stability and antioxidant property of the complexes on the other hand were found to be improved. Of the amount of β-CD studied, it was reported that the best combination of the complex was at β-CD/lecithin ratio of 2:1. The complex of the optimized ratio showed the greatest thermal stability as indicated by its highest melting point (177.8 ºC). The complex also exhibited the greatest antioxidant property as indicated by its highest percentage of DPPH scavenging activity that is 97.1%

Effect of cooking time on physical properties of almond milk-based lemak cili api gravy

One of the crucial elements in developing or reformulating product is to maintain the quality throughout its entire shelf life. This study aims to determine the effect of different cooking time on the almond milk-based of lemak cili api gravy. Various cooking times of 5, 10, 15, 20, 25 and 30 minutes were employed to the almond milk-based lemak cili api gravy followed by determination of their effects on physical properties such as total soluble solids content, pH and colour. pH was determined by using a pH meter. Refractometer was used to evaluate the total soluble solids content of almond milk-based lemak cili api gravy. The colours were determined by using spectrophotometer which expressed as L*, a* and b* values. Results showed that almond milk-based lemak cili api gravy has constant values of total soluble solids with pH range of 5 to 6, which can be classified as low acid food. Colour analysis showed that the lightness (L*) and yellowness (b*) are significantly increased while redness (a*) decreased. In conclusion, this study shows that physical properties of almond milk-based lemak cili api gravy changes by increasing the cooking time

Effect of Cooking Time on Physical Properties of Almond Milk-Based Lemak Cili Api Gravy

One of the crucial elements in developing or reformulating product is to maintain the quality throughout its entire shelf life. This study aims to determine the effect of different cooking time on the almond milk-based of lemak cili api gravy. Various cooking times of 5, 10, 15, 20, 25 and 30 minutes were employed to the almond milk-based lemak cili api gravy followed by determination of their effects on physical properties such as total soluble solids content, pH and colour. pH was determined by using a pH meter. Refractometer was used to evaluate the total soluble solids content of almond milk-based lemak cili api gravy. The colours were determined by using spectrophotometer which expressed as L*, a* and b* values. Results showed that almond milk-based lemak cili api gravy has constant values of total soluble solids with pH range of 5 to 6, which can be classified as low acid food. Colour analysis showed that the lightness (L*) and yellowness (b*) are significantly increased while redness (a*) decreased.  In conclusion, this study shows that physical properties of almond milk-based lemak cili api gravy changes by increasing the cooking time

Phytochemical analysis and in-vitro antiurolithiatic properties of selected Malaysian herbs

The aim of this study was to investigate the phytochemical content of selected Malaysian herbs and their potential antiurolithiatic effects using in-vitro method. The herbs involved are Ceiba pentandra, Cymbopogon citratus, Euphorbia hirta, Melastoma malabathricum and Ortosiphon stamineus. Aqueous extracts of each herbs were prepared through decoction while Standard drugs Cystone, was used as positive control in comparison. Qualitative analysis was carried out to detect phytochemical presence and nucleation assay to investigate their inhibition effects towards calcium oxalate crystallization urolithiasis in vitro. Based on results, the same trends were observed between phytochemical content and inhibition rate of calcium oxalate crystallization. O. stamineus extract (73.48%) which showed the highest inhibition rate hold the most phytochemical content. while the lowest inhibition rate was occupied by C. citratus extract (45.45%) with the least phytochemical content. The high amount of phytochemicals particularly saponin followed by steroid and terpenoid in O. stamineus extract might contributes to the high inhibition activities of calcium oxalate crystallization as compared to low amount of phytochemicals observed in C. citratus extract. It can be concluded that O. stamineus possesses highest inhibition percentage against calcium oxalate which could be attributed to its saponins, tannins, steroid and terpenoid content

Characterization of alginate–gelatin–cholesteryl ester liquid crystals bioinks for extrusion bioprinting of tissue engineering scaffolds

Tissue engineering (TE) is an innovative approach to tackling many diseases and body parts that need to be replaced by developing artificial tissues and organs. Bioinks play an important role in the success of various TE applications. A bioink refers to a combination of a living cell, biomaterials, and bioactive molecules deposited in a layer-by-layer form to fabricate tissue-like structures. The research on bioink attempts to offer a 3D complex architecture and control cellular behavior that improve cell physical properties and viability. This research proposed a new multi�material bioink based on alginate (A), gelatin (G), and cholesteryl ester liquid crystals (CELC) biomaterials, namely (AGLC) bioinks. The development of AGLC was initiated with the optimization of different concentrations of A and G gels to obtain a printable formulation of AG gels. Subsequently, the influences of different concentrations of CELC with AG gels were investigated by using a microextrusion-based 3D bioprinting system to obtain a printed structure with high shape fidelity and minimum width. The AGLC bioinks were formulated using AG gel with 10% weight/volume (w/v) of A and 50% w/v G (AG10:50) and 1%, 5%, 10%, 20%, and 40% of CELC, respectively. The AGLC bioinks yield a high printability and resolution blend. The printed filament has a minimum width of 1.3 mm at a 1 mL/min extrusion rate when the A equals 10% w/v, G equals 50% w/v, and CELC equals 40% v/v (AGLC40). Polymerization of the AGLC bioinks with calcium (Ca2+) ions shows well-defined and more stable structures in the post-printing process. The physicochemical and viability properties of the AGLC bioinks were examined by FTIR, DSC, contact angle, FESEM, MTT assay, and cell interaction evaluation methods. The FTIR spectra of the AGLC bioinks exhibit a combination of characteristics vibrations of AG10:50 and CELC. The DSC analysis indicates the high thermal stability of the bioinks. Wettability analysis shows a reduction in the water absorption ability of the AGLC bioinks. FESEM analysis indicates that the surface morphologies of the bioinks exhibit varying microstructures. In vitro cytotoxicity by MTT assay shows the ability of the bioinks to support the biological activity of HeLa cells. The AGLC bioinks show average cell viability of 82.36% compared to the control (90%). Furthermore, cultured cells on the surface of AGLC bioinks showed that bioinks provide favorable interfaces for cell attachment

Evaluation of Phyllanthus niruri L. from Malaysia for In-vitro Anti-Urolithiatic properties by different solvent extraction

The Phyllanthus niruri is traditionally used for curing of kidney disorders and urinary stones in Malaysia. Hence the current work was aimed to evaluate the effect of different solvents extract (n- hexane, ethyl acetate, methanol and water) of P. niruri for in vitro anti-urolithiatic properties in terms of inhibition activity on CaOx by using the rate of CaOx aggregation assay and dissolution of calcium oxalate (CaOx) crystal by using titrimetry method. Cystone was used as positive control. The effects of cystone on slope of nucleation and aggregation as well as growth of CaOx were evaluated spectrophotometrically. The highest yield percentage of P.niruri was occupied by methanol (5.74 %). The maximum inhibition against aggregation of CaOx crystals was also occupied by methanol (66.67 % ± 1.61) and was comprised with alkaloid, steroid, terpenoid and tannin. Dissolution effect on calcium oxalate crystals indicates that the aqueous extracts of P. niruri was found to be more effective in dissolution of CaOx with 63.33 % ± 1.44. P. niruri significantly (P < 0.05) inhibited the slope of nucleation and aggregation of CaOx crystallization, and reduced the crystal density. The results of the present study confirmed that P. niruri leaves can be used as remedial mediator for urolithiasis. However, further studies are required for isolation and identification of active constituents and their in-vivo confirmation

A rapid and sensitive system for recovery of nucleic acids from Mycobacteria sp. on archived glass slides

The field of diagnostics continues to advance rapidly with a variety of novel approaches, mainly dependent upon high technology platforms. Nonetheless much diagnosis, particularly in developing countries, still relies upon traditional methods such as microscopy. Biological material, particularly nucleic acids, on archived glass slides is a potential source of useful information both for diagnostic and epidemiological purposes. There are significant challenges faced when examining archived samples in order that an adequate amount of amplifiable DNA can be obtained. Herein, we describe a model system to detect low numbers of bacterial cells isolated from glass slides using (laser capture microscopy) LCM coupled with PCR amplification of a suitable target. Mycobacterium smegmatis was used as a model organism to provide a proof of principle for a method to recover bacteria from a stained sample on a glass slide using a laser capture system. Ziehl-Neelsen (ZN) stained cells were excised and catapulted into tubes. Recovered cells were subjected to DNA extraction and pre-amplified with multiple displacement amplification (MDA). This system allowed a minimum of 30 catapulted cells to be detected following a nested real-time PCR assay, using rpoB specific primers. The combination of MDA and nested real-time PCR resulted in a 30-fold increase in sensitivity for the detection of low numbers of cells isolated using LCM. This study highlights the potential of LCM coupled with MDA as a tool to improve the recovery of amplifiable nucleic acids from archived glass slides. The inclusion of the MDA step was essential to enable downstream amplification. This platform should be broadly applicable to a variety of diagnostic applications and we have used it as a proof of principle with a Mycobacterium sp. model system

β-Cyclodextrin/Lecithin: Preparation and Characterization

Lecithin is a fat which is well-known to be obtained from soybeans or egg yolks. It is also taken as a medicine and used as food additive that keep certain ingredients from separating out. Despite of its advantages, lecithin’s application in the industries is restricted due to its sensitivity to heat and light and can be easily oxidized. Hence, this study aims to investigate the effect of β-cyclodextrin (β-CD) in improving the properties of lecithin. The objectives of this study are to prepare and characterize β-CD/lecithin complex followed by study on the influence of β-CD on lecithin’s thermal stability and antioxidant property. Analytical methods employed are fourier transform infra-red (FTIR) spectroscopy for chemistry study, differential scanning calorimetry (DSC) for examination of its thermal stability, DPPH testing for its antioxidant property as well as scanning electron microscopy (SEM) for structural examination. Results from FTIR and SEM analyses confirmed the formation of β-CD/lecithin complex. Thermal stability and antioxidant property of the complexes on the other hand were found to be improved. Of the amount of β-CD studied, it was reported that the best combination of the complex was at β-CD/lecithin ratio of 2:1. The complex of the optimized ratio showed the greatest thermal stability as indicated by its highest melting point (177.8 ºC). The complex also exhibited the greatest antioxidant property as indicated by its highest percentage of DPPH scavenging activity that is 97.1%