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As the regenerative mechanisms of biological organisms, self-healing provides useful functions for soft electronics or associated systems. However, there have been few examples of soft electronics where all components have self-healing properties while also ensuring compatibility between components to achieve multifunctional and resilient bio-integrated electronics. Here, we introduce a stretchable, biodegradable, self-healing conductor constructed by combination of two layers: (i) synthetic self-healing elastomer and (ii) self-healing conductive composite with additives. Abundant dynamic disulfide and hydrogen bonds of the elastomer and conductive composite enable rapid and complete recovery of electrical conductivity (~1000 siemens per centimeter) and stretchability (~500%) in response to repetitive damages, and chemical interactions of interpenetrated polymer chains of these components facilitate robust adhesion strength, even under extreme mechanical stress. System-level demonstration of soft, self-healing electronics with diagnostic/therapeutic functions for the urinary bladder validates the possibility for versatile, practical uses in biomedical research areas.

Entamoeba histolytica is the parasite responsible for amoebiasis, which can result in amoebic colitis or amoebic liver abscess. Metronidazole has been the conventional treatment for intestinal amoebiasis, but concerns regarding resistance have emerged due to the identification of resistance pathways in E. histolytica. This study investigates a novel anti-amoebic approach targeting the CDP-choline pathway. Inhibition studies were conducted using potential choline kinase (CK) inhibitors to inhibit the EhCK enzyme, and RNA interference was employed to knock down the EhCK gene. K m and V max of purified EhCK and hCKa2 proteins were determined by pyruvate kinase-lactate dehydrogenase (PK-LDH) coupled assay. The IC 50 values for EhCK and hCKa2 were determined with several commercial CK inhibitors. Selected inhibitors were incubated with E. histolytica trophozoites for 48 hours to determine the EC50 for each inhibitor. Silencing of gene encoding EhCK was carried out using duplex siRNA and the gene expression level was measured by real-time qPCR. Based on the IC 50 values, three of the inhibitors, namely CK37, flavopiridol and H-89 were more potent against EhCK than hCKa2. Trophozoites growth inhibition showed that only HDTAB, H-89 and control drug metronidazole could penetrate and induce cell death after 48-hour incubation. siRNA concentration of 10 µg/mL was used for the transfection of positive control GAPDH, EhCK, and non-targeting GFP siRNAs. RNAi experiment concluded with positive control GAPDH downregulated by 99% while the level of EhCK mRNA was downregulated by 47%. In this study, potential inhibitors of EhCK and siRNA have been identified, paving the way for further refinement and testing to enhance their potency against EhCK while sparing hCK. The utilization of these specific inhibitors and siRNA targeting EhCK represents a novel approach to impede the growth of E. histolytica by disrupting its phospholipid synthesis pathway.

This manuscript presents the Microwave Temperature and Humidity Profiler (MTHP), a dual-band spectroradiometer designed for measuring multi-incidence angle temperature and humidity atmospheric profiles from an aircraft platform. The MTHP bands are at 60 GHz for measuring the oxygen complex lines, therefore at this band, MTHP has a hyperspectral radiometer able to provide 2048 channels over an 8 GHz bandwidth, and 183 GHz for measuring water vapor, which only uses four channels since this absorption band's spectral richness is simpler. The MTHP builds upon the Microwave Temperature Profiler (MTP) with the inclusion of the hyperspectral radiometer. The instrument's design, components, and calibration methods are discussed in detail, with a focus on the three-point calibration scheme involving internal calibration loads and static air temperature readings. Preliminary results from the Technological Innovation into Iodine and GV aircraft Environmental Research (TI3GER) campaign are presented, showcasing the instrument's performance during flights across diverse geographical regions. The manuscript presents successful antenna temperature measurements at 60 GHz and 183 GHz. The hyperspectral measurements are compared with a simulated antenna temperature using the Atmospheric Radiative Transfer Simulator (ARTS) showing an agreement better than R 2 > 0.88 for three of the flights analyzed. Additionally, the manuscript draws attention to potential Radio Frequency Interference (RFI) effects observed during a specific flight, underscoring the instrument's sensitivity to external interference. This is the first-ever airborne demonstration of a broadband and hyperspectral multi-incidence angle 60 GHz measurement. Future work on the MTHP could result in an improved spatial resolution of the atmospheric temperature vertical profile and, hence, help in estimating the Planetary Boundary Layer (PBL) with better accuracy. The MTHP and its hyperspectral multi-incidence angle at 60 GHz have the potential to be a valuable tool for investigating the PBL's role in atmospheric dynamics, offering insights into its impact on Earth's energy, water, and carbon cycles.

The enormous increase in heterogeneous wireless devices operating in real-time applications for Internet of Things (IoT) applications presents new challenges, including heterogeneity, reliability, and scalability. To address these issues effectively, a novel architecture has been introduced, combining Software-Defined Wireless Sensor Networks (SDWSN) with the IoT, known as the SDWSN-IoT. However, wireless IoT devices deployed in such systems face limitations in the energy supply, unpredicted network changes, and the quality of service requirements. Such challenges necessitate the careful design of the underlying routing protocol, as failure to address them often results in constantly disconnected networks with poor network performance. In this paper, we present an intelligent, energy-efficient multi-objective routing protocol based on the Reinforcement Learning (RL) algorithm with Dynamic Objective Selection (DOS-RL). The primary goal of applying the proposed DOS-RL routing scheme is to optimize energy consumption in IoT networks, a paramount concern given the limited energy reserves of wireless IoT devices and the adaptability to network changes to facilitate a seamless adaption to sudden network changes, mitigating disruptions and optimizing the overall network performance. The algorithm considers correlated objectives with informative-shaped rewards to accelerate the learning process. Through the diverse simulations, we demonstrated improved energy efficiency and fast adaptation to unexpected network changes by enhancing the packet delivery ratio and reducing data delivery latency when compared to traditional routing protocols such as the Open Shortest Path First (OSPF) and the multi-objective Q-routing for Software-Defined Networks (SDN-Q).

Eurycoma longifolia Jack (ELJ) is a herbal plant that has androgenic and antioxidant effects. We investigated the short-term effect of ELJ supplementation on muscle damage induced by eccentric exercise. Eighteen young (19-25 years), well-trained rugby 7s players were assigned to an ELJ or a placebo (PLA) group (n = 9/group). Each participant took four 100-mg capsules a day for seven days prior to performing a leg press eccentric exercise to failure in a double-blind fashion. Peak force, peak power and jump height in countermovement jump (CMJ), drop jump reactive strength index (RSI), muscle soreness assessed by a 100-mm visual analogue scale, plasma creatine kinase (CK) activity, and salivary hormones were measured at 24 h before and 0.5, 24, 48, 72, and 96 h after the exercise. Changes in the variables over time were compared between the groups by two-factor mixed-design ANOVA. The number of eccentric contractions performed was similar (P = 0.984) between the ELJ (21 ± 5) and PLA groups (21 ± 5). Salivary testosterone and cortisol concentrations did not change (P > 0.05) after the supplementation for both groups. CMJ peak power (-9.4 ± 5.6%) and height (-10.6 ± 4.9%), and RSI (-15.2 ± 16.2%) decreased at 24 h after exercise (P < 0.05), and muscle soreness (peak: 89 ±10 mm) and plasma CK activity (peak: 739 ± 420 IU/L) increased after exercise (P < 0.05) without significant differences between groups. These results showed that 7-day ELJ supplementation prior to the leg press eccentric exercise had no significant effects on hormones, performance and muscle damage markers for the athletes., Competing Interests: The authors declare there are no competing interests., (Copyright © Biology of Sport 2023.)

As unitised regenerative proton exchange membrane fuel cell (URPEMFC) is progressing in terms of its performance, more emphasis should be placed on the understanding of the interaction between multiphase reactants and products and its effect during the switching mode. A 3D transient computational fluid dynamics model was utilised in this study to simulate the supply of liquid water into the flow field when the system switched from fuel cell mode to electrolyser mode. Different water velocities were investigated to identify their effect on the transport behaviour under parallel, serpentine, and symmetry flow fields. From the simulation results, 0.5 m·s -1 water velocity was the best-performing parameter in achieving optimal distribution. Among different flow field configurations, the serpentine design achieved the best flow distribution due to its single-channel model. Modification and refinement in the form of flow field geometric structure can be performed to further improve the water transportation behaviour in URPEMFC.

Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The study aimed to determine how a teacher's competency at the university level related to professional commitment and job satisfaction. This systematic review intends to determine physical education teachers' competency and job satisfaction. One of the main objectives of physical education programs is to increase various activities of the student, especially physical activities and participation in it. Students who participate in physical activities feel a variety of emotions, and these may enhance their mental and physical well-being. The study concerns the teachers' satisfaction in effectively influencing and teaching the students. Job satisfaction refers to teachers' feelings and positive attitudes toward their work. The five components of job satisfaction are pay, promotion, supervision, co-workers, and work itself. Job satisfaction is broken down into these five components. Work as a teacher is one of the most important components of a teacher's or teacher's success at work and one of the key criteria for the school's achievement. Electronic libraries were explored, including Google Scholar, PubMed, Embase, Bing Academic, and Cochrane. Appropriate keywords were used for searching the literature. By applying certain inclusion and exclusion criteria, the final studies were selected for evaluation. The study's findings showed that physical education instructors had high levels of teaching competence and moderate levels of job satisfaction. The findings demonstrated that job satisfaction and competency rise when degree levels rise, and relatedness, skill, and autonomy self-motivation ensue. Students' motivation for physical education was greatly influenced by the teachers' support, motivation, and mastery climate. The self-determination theory positively enhances physical education from the teacher's perspective on PE, increasing instruction effectiveness., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Yan, Teo, Lim and Lin.)

Background: Mechanically assisted crevice corrosion (MACC) has been associated with the compromised durability and fixation of modular total hip implants, adverse reaction of local tissue, and other undesirable clinical outcomes in total hip arthroplasty (THA). MACC is primarily caused by the relative motion between the femoral head and stem. To minimize the relative motion the taper connection between the two components must be strong enough. The current study addressed the following questions: (1) Does increasing the mass of the femoral stem improve the taper connection strength intraoperatively? (2) Does increasing the mass of the femoral stem reduce the risk of periprosthetic tissue damage intraoperatively?, Hypothesis: Increasing the mass of the femoral stem improve the taper connection strength intraoperatively., Materials and Methods: During the experiment, femoral heads were impacted onto the stem tapers with and without an additional weight attached to the stem. The femoral heads were then pulled off to investigate the strength of the taper connection. The stem displacement and acceleration at impaction were also measured to evaluate the risk of periprosthetic tissue damage., Results: The results showed that the pull-off force was increased by 24% (p=0.011, n=6) when an additional weight was attached to the stem. The additional weight also reduced the maximum stem acceleration and maximum stem displacement by 37% (p<0.001, n=6) and 14% (p=0.094, n=6), respectively., Discussion: These findings suggest that the femoral head and stem taper connection strength can be significantly improved and the risk of periprosthetic tissue damage significantly reduced intraoperatively by attaching an additional weight to the stem to increase its mass., Level of Evidence: III, comparative in vitro mechanical investigation., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

To determine how brain oxygenation is stably maintained during advancing age, cerebral oxygenation and hemoglobin were measured real-time at 10 Hz using near-infrared spectroscopy (NIRS) at rest (30 seconds) and during a 10-repeated handgrip strength test (30 seconds) for 834 adults (M/F = 45/55%) aged 20-88 y. The amplitude of cerebral hemodynamic fluctuation was reflected by converting 300 values of % oxygen saturation and hemoglobin of each 30-second phase to standard deviation as indicatives of brain oxygenation variability (BOV) and brain hemodynamic variability (BHV) for each participant. Both BOV (+21-72%) and BHV (+94-158%) increased during the maximal voluntary muscle exertions for all age levels (α < 0.05), suggesting an increased vascular recruitment to maintain oxygen homeostasis in the brain. Intriguingly, BHV was >100 folds for both resting and challenged conditions (α < 0.001) in >80% of adults aged above 50 y despite similar BOV compared with young age counterparts, indicating a huge cost of amplifying hemodynamic oscillation to maintain a stable oxygenation in the aging brain. Since vascular endothelial cells are short-lived, our results implicate a hemodynamic compensation to emergence of daily deficits in replacing senescent endothelial cells after age 50 y.

The present work aims to study the influence of virtual reality (VR) technology on the teaching and curriculum of preschool physical education in colleges and universities and establish a virtual teaching model suitable for the college teaching system. The classroom teaching situation of using VR technology in physical training of preschool education major direction in colleges and universities is investigated using the questionnaire survey and teaching experiment. Firstly, the feasibility of applying VR technology to teaching is proved by analyzing the relevant education theories. Secondly, the experimental research method is designed to verify the application effect of VR technology in teaching behavior. Finally, the collected data is sorted out to judge the method's feasibility. The experimental results demonstrate that 88.0% of the respondents are curious about the application of VR, and 88.6% of the respondents can accept the application of VR in sports dance teaching. Besides, 89.1% claimed that VR technology could enhance students' understanding of knowledge, and 93.0% thought that VR applications would not interfere with teachers' explanations. In addition, 80.0% thought that virtual teaching could stimulate students' interest in learning, 75.0% said that VR application could attract students' attention, and 63.0% believed that VR application could improve learning efficiency. The preliminary investigation suggests that introducing VR technology can reduce the work intensity of teachers, and the students participating in the survey are optimistic about the application of VR technology in preschool physical education. The formal test based on a t -test indicates that the average score of the experimental group is 3.18 points higher than that of the control group, and there are significant differences in technology between the control group and the experimental group. VR technology can improve students' technical level, enhance self-confidence, and improve students' grades. To sum up, this study provides a reference for developing virtual teaching mode and applying VR technology to physical education., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Nina Wang et al.)

Background/aims: There have been few multicenter studies on colonic polyps conducted by primary medical institutions. This study examined the detection rate of colonic polyps in primary health care institutions and the related factors while following the guidelines., Methods: The medical records of 14,029 patients who underwent colonoscopy between January-June 2020 at 40 primary medical institutions in Korea were analyzed. High-risk adenoma was defined as advanced adenoma, carcinoma, or ≥3 adenomas., Results: Most patients (71.2%) aged ≥50 years underwent re-colonoscopy within 5 years (51.3%) for diagnostic purposes (61.3%) in Korean primary medical institutions. The detection rates of colon polyps, adenoma, advanced adenoma, high-risk adenoma, and carcinoma was 59.9%, 38.9%, 5.9%, 11.4%, and 0.3% in all subjects and 59.8%, 37.5%, 8.5%, 12.9%, and 0.3% in average-risk patients, respectively. The incidences of adenoma in average-risk patients increased significantly with age (30s/40s/50s: 20.1%/29.4%/43% for adenoma, 4.4%/6.7%/10.3% for advanced adenoma, and 5.6%/9.5%/14.6% for high-risk adenoma; p<0.05). Before 50 years of age, high-risk adenoma was detected in 9.1% of patients in the first-time screening group, and the significant risk factors were being male and ≥40 years of age. The detection rate of high-risk adenoma in the normal index colonoscopy group within 5 years was 9.0%. The significant risk factors included older age, male sex, positive fecal occult blood test, stool form changes, and nonspecific symptoms (gas and indigestion)., Conclusions: More colonic adenoma studies targeting real-world clinical practice will be needed to revise the Korean guidelines for colorectal cancer screening and surveillance.

Purpose: Many approaches have been described to accomplish tendon reattachment to the radial tuberosity in a distal biceps tendon rupture, with significant success, but each is associated with potential postoperative complications, including posterior interosseous nerve (PIN) injury. To date, there has been no consensus on the best approach to the repair. The purpose of this study was to evaluate the supination strength and the distance of drill exit points from the PIN in a power-optimizing distal biceps repair method and compare the findings with those of a traditional anterior approach endobutton repair method., Methods: Cadaveric arms were dissected to allow for distal biceps tendon excision from its anatomic footprint. Each arm was repaired twice, first with the power-optimizing repair using an anterior single-incision approach with an ulnar drilling angle and biceps tendon radial tuberosity wraparound anatomic footprint attachment, then with the traditional anterior endobutton repair. Following each repair, the arm was mounted on a custom-built testing apparatus, and the supination torque was measured from 3 orientations. The PIN was then located posteriorly, and its distance from each repair exit hole was measured., Results: Five cadaveric arms, each with both the repairs, were included in the study. On average, the power-optimizing repair generated an 82%, 22%, and 13% greater supination torque than the traditional anterior endobutton repair in 45° supination, neutral, and 45° pronation orientations, respectively. On average, the power-optimizing repair produced drill hole exit points farther from the PIN (23 mm) than the traditional anterior endobutton repair (14 mm)., Conclusions: The power-optimizing repair provides a significantly greater supination torque and produces a drill hole exit point significantly farther from the PIN than the traditional anterior endobutton approach., Type of Study/level of Evidence: Therapeutic III., (© 2021 The Authors.)

Purpose: Hardware prominence is a concern in the fixation of olecranon osteotomies. Staple fixation has provided low-profile secure fixation in other areas of orthopedics. Without insetting, staples still have subcutaneous prominence. This study examines whether nitinol staples, when inset into bone via cortical notching, in an olecranon osteotomy can provide fixation strength sufficient for daily activities., Methods: Olecranon osteotomies were created in 8 cadaver arms and fixed with 2 nitinol staples. For inset and juxtacortical (noninset) staples, a micrometer measured the displacement between preplaced proximal and distal wires for 3 increasing loads: 0 N, 15 N, and 150 N. This measurement reflected the loss of osteotomy compression. We placed each arm in a pneumatic machine that flexed the elbow from 0° to 90° for 500 cycles at each load. We performed a 2-tailed t test (α value 0.05, β value 0.2) to evaluate for differences in the loss of compression between inset and noninset nitinol staples., Results: We performed the displacement measurement procedure for both staple types at each of the 3 loads. At 0 N, the average displacement of inset was 0 mm and that of noninset was 0.02 mm. At 15 N, the average displacement of inset was 0.02 mm and that of noninset was 0.04 mm. At 150 N, the average displacement of inset was 0.05 mm and that of noninset was 0.09 mm. When comparing the displacement at the 3 force loads, there were no statistically significant differences between the staple types ( P = .323)., Conclusions: This study shows that inset staples do not considerably weaken osteotomy fixation with nitinol staples. Thus, nitinol staples may provide a low-profile, operatively-efficient fixation method compared with tension-band or screw-and-plate fixation methods for olecranon osteotomies. Future research can include comparing staples with plate constructs. Type of study/level of evidence : Therapeutic III., (© 2021 The Authors.)

The lemongrass plant, which is widely cultivated in Asia, Australia, and Africa, has been reported to have many significant health benefits such as antimicrobial, insecticide, anticancer, fight fever, and disinfection. Therefore, it is an added benefit to have lemongrass compounds in cooking oil. This study was aimed to compare the conventional (CSE), and ultrasound-assisted solvent extraction (UASE) for citral compounds from lemongrass ( Cymbopogon ) leaves and to optimize the best extraction method using the response surface methodology (RSM) and ANOVA. RSM design of experiments using three types of cooking oils; palm oil, sunflower oil, and corn oil. The effect of three independent variables, which are temperature (48.2-81.8°C), extraction time (4.8-55.2 min), and solvent to leaves ratio (5.3-18.7), was investigated. The characterization of lemongrass-infused cooking oil was evaluated by Fourier transform infrared spectroscopy (FT-IR), Gas Chromatography-Mass Spectrometry (GC-MS) and Scanning Electron Microscopy (SEM) analysis for confirmation of the citral compound extraction. This extraction process is optimized using Response Surface Methodology (RSM) for producing the lemongrass-infused cooking oil. After optimization, the UASE process gives 1.009 × 10 6 maximum citral area for palm oil and 1.767 × 10 6 maximum citral area for sunflower oil. CSE process only can give 2.025 × 10 5 and 2.179 × 10 5 citral area in the GC-MS spectrum for palm oil and sunflower oil respectively. For both the UASE and the CSE, the optimum operating conditions are 81.8°C of extraction temperature and 55.2 min of extraction time except for lemongrass-infused palm oil in the CSE process with 45 min extraction time. The optimum solvent to leaves ratio varies from 5.3:1 to 12.9:1. This study found that corn oil cannot be used as a solvent to extract lemongrass-infused cooking oil due to the insignificant changes and no citral peak. The lemongrass (Cymbopogon)-infused palm oil and sunflower oil extracted using the UASE have a higher maximum citral area than the CSE process., Competing Interests: The authors declare that they have no conflicts of interests., (© 2021 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

The large-conductance calcium-activated potassium channel (BK Ca channel) is expressed on various tissues and is involved in smooth muscle relaxation. The channel is highly expressed on urinary bladder smooth muscle cells and regulates the repolarization phase of the spontaneous action potentials that control muscle contraction. To discover novel chemical activators of the BK Ca channel, we screened a chemical library containing 8364 chemical compounds using a cell-based fluorescence assay. A chemical compound containing an isoxazolyl benzene skeleton (compound 1) was identified as a potent activator of the BK Ca channel and was structurally optimized through a structure-activity relationship study to obtain 4-(4-(4-chlorophenyl)-3-(trifluoromethyl)isoxazol-5-yl)benzene-1,3-diol (CTIBD). When CTIBD was applied to the treated extracellular side of the channel, the conductance-voltage relationship of the channel shifted toward a negative value, and the maximum conductance increased in a concentration-dependent manner. CTIBD altered the gating kinetics of the channel by dramatically slowing channel closing without effecting channel opening. The effects of CTIBD on bladder muscle relaxation and micturition function were tested in rat tissue and in vivo. CTIBD concentration-dependently reduced acetylcholine-induced contraction of urinary bladder smooth muscle strips. In an acetic acid-induced overactive bladder (OAB) model, intraperitoneal injection of 20 mg/kg CTIBD effectively restored frequent voiding contraction and lowered voiding volume without affecting other bladder function parameters. Thus, our results indicate that CTIBD and its derivatives are novel chemical activators of the bladder BK Ca channel and potential candidates for OAB therapeutics. SIGNIFICANCE STATEMENT: The novel BK Ca channel activator CTIBD was identified and characterized in this study. CTIBD directly activates the BK Ca channel and relaxes urinary bladder smooth muscle of rat, so CTIBD can be a potential candidate for overactive bladder therapeutics., (Copyright © 2021 by The American Society for Pharmacology and Experimental Therapeutics.)

Underactive bladder or detrusor underactivity (DUA), that is, not being able to micturate, has received less attention with little research and remains unknown or limited on pathological causes and treatments as opposed to overactive bladder, although the syndrome may pose a risk of urinary infections or life-threatening kidney damage. Here, we present an integrated expandable electronic and optoelectronic complex that behaves as a single body with the elastic, time-dynamic urinary bladder with substantial volume changes up to ~300%. The system configuration of the electronics validated by the theoretical model allows conformal, seamless integration onto the urinary bladder without a glue or suture, enabling precise monitoring with various electrical components for real-time status and efficient optogenetic manipulation for urination at the desired time. In vivo experiments using diabetic DUA models demonstrate the possibility for practical uses of high-fidelity electronics in clinical trials associated with the bladder and other elastic organs., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Background: This study reports the analytical sensitivity and specificity of a Loop-mediated isothermal amplification (LAMP) and compares its amplification performance with conventional PCR, nested PCR (nPCR) and real-time PCR (qPCR). All the assays demonstrated in this study were developed based on Serine-rich Entamoeba histolytica protein (SREHP) gene as study model., Results: A set of SREHP gene specific LAMP primers were designed for the specific detection of Entamoeba histolytica. This set of primers recorded 100% specificity when it was evaluated against 3 medically important Entamoeba species and 75 other pathogenic microorganisms. These primers were later modified for conventional PCR, nPCR and qPCR applications. Besides, 3 different post-LAMP analyses including agarose gel electrophoresis, nucleic acid lateral flow immunoassay and calcein-manganese dye techniques were used to compare their limit of detection (LoD). One E. histolytica trophozoite was recorded as the LoD for all the 3 post-LAMP analysis methods when tested with E. histolytica DNA extracted from spiked stool samples. In contrast, none of the PCR method outperformed LAMP as both qPCR and nPCR recorded LoD of 100 trophozoites while the LoD of conventional PCR was 1000 trophozoites., Conclusions: The analytical sensitivity comparison among the conventional PCR, nPCR, qPCR and LAMP reveals that the LAMP outperformed the others in terms of LoD and amplification time. Hence, LAMP is a relevant alternative DNA-based amplification platform for sensitive and specific detection of pathogens.

Purpose: In the developing world, bacillary dysentery is one of the most common communicable diarrheal infections. There are approximately 169 million cases of shigellosis reported worldwide. The disease is transmitted by a group of Gram-negative intracellular enterobacteria known as Shigella flexneri , S. sonnei , S. dysenteriae , and S. boydii . Conventional treatment regimens for Shigella have been less effective due to the development of resistant strains against antibiotics. Therefore, an effective vaccine for the long term control of Shigella transmission is urgently needed., Materials and Methods: In this study, a reverse vaccinology approach was employed to identify most conserved and immunogenic outer membrane proteins (OMPs) of S. flexneri 2a., Results: Five OMPs including fepA, ompC, nlpD&#95;1, tolC, and nlpD&#95;2 were identified as potential vaccine candidates. Protein-protein interactions analysis using STRING software (https://string-db.org/) revealed that five of these OMPs may potentially interact with other intracellular proteins which are involved in beta-lactam resistance pathway. B- and T-cell epitopes of the selected OMPs were predicted using BCPred as well as Propred I and Propred (http://crdd.osdd.net/raghava/propred/), respectively. Each of these OMPs contains regions which are capable to induce B- and T-cell immune responses., Conclusion: Analysis acquired from this study showed that five selected OMPs have great potential for vaccine development against S. flexneri infection. The predicted immunogenic epitopes can also be used for development of peptide vaccines or multi-epitope vaccines against human shigellosis. Reverse vaccinology is a promising strategy for the discovery of potential vaccine candidates which can be used for future vaccine development against global persistent infections., Competing Interests: No potential conflict of interest relevant to this article was reported., (© Korean Vaccine Society.)

The flow distribution of a proton exchange membrane fuel cell within a manifold plays an important role on its performance. This study presents a numerical analysis of the flow distribution behavior within different manifold configurations. A two-dimensional model with 75 cells was employed to study the flow behavior. The variation in the stoichiometry and number of cells was also studied. Three different flow configurations were considered with different numbers of flow inlets and outlets. The flow characteristics, such as the pressure and velocity variations in the manifold and cells, were measured to determine the effects of the different flow configurations. The results indicated that the double inlet/outlet configuration had the best flow distribution when using 75 cells. Moreover, increasing the stoichiometry resulted in a better flow distribution to the cells in a stack.

The interleukin-21 (IL-21) protein was found to be expressed at an elevated level in clinical samples of colorectal cancer patients without or with a parasitic infection that were collected from Sudan in our previous study. The IL-21 gene in HT29 and HCT116 cells was then correlated to cell proliferation and cell migration, as well as the cellular mechanisms associated with gene expressions in our present study. Our results demonstrated that silencing the IL-21 gene in HCT116 cells increased the cytotoxic level and fibroblast growth factor-4 (FGF4) mRNA expression in the cancer cells. Moreover, specific gene silencing reduced the migration of cancer cells compared to non-silenced cancer cells. These events were not observed in IL-21-silenced HT29 cells. Neutralizing FGF4 in conditioned medium of IL-21-silenced HCT116 cells further increased the cytotoxic level and restored the migratory activity of HCT116 cells in the culture compared to silencing the IL-21 gene alone in the cancer cells. Our results indicate the importance of both silencing the IL-21 gene and co-expression of the FGF4 protein in HCT116 cells, which pave the way for the discovery of important factors to be used as biomarkers for the design of drugs or cost-effective supplements to effectively treat the patients having infectious disease and HCT116 cells of colorectal cancer simultaneously in the future.

Objectives: The Neural Tube Defects Research Group of University of Malaya was approached to analyze a tablet named TELSE, which may have resulted in a baby born with central nervous system malformation at the University of Malaya Medical Centre. In this animal experimental study, we investigated the content of TELSE and exposure of its contents that resulted in failure of primary neurulation., Results: Liquid Chromatography Tandem Mass spectrophotometry analysis of the TELSE tablet confirmed the presence of trimethoprim as the active compound. The TELSE tablet-treated females produced significant numbers of embryos with exencephaly (n = 8, 36.4%, *P < 0.0001), in all litters. The TELSE tablet-treated females subsequently given folic acid did not result in pregnancies despite there being evidence of possible resorption. Furthermore, after multiple rounds of mating which did not yield viable pregnancies, eventually, 2 embryos with exencephaly were harvested in a litter of 6 at 0.05% w/v pure trimethoprim once. The use of trimethoprim, a folic acid antagonist, peri-conceptionally increased the risk of exencephaly in the mouse.

Statins mediate vascular protection and reduce the prevalence of cardiovascular diseases. Recent work indicates that statins have anticonvulsive effects in the brain; however, little is known about the precise mechanism for its protective effect in kainic acid (KA)-induced seizures. Here, we investigated the protective effects of atorvastatin pretreatment on KA-induced neuroinflammation and hippocampal cell death. Mice were treated via intragastric administration of atorvastatin for 7 days, injected with KA, and then sacrificed after 24 h. We observed that atorvastatin pretreatment reduced KA-induced seizure activity, hippocampal cell death, and neuroinflammation. Atorvastatin pretreatment also inhibited KA-induced lipocalin-2 expression in the hippocampus and attenuated KA-induced hippocampal cyclooxygenase-2 expression and glial activation. Moreover, AKT phosphorylation in KA-treated hippocampus was inhibited by atorvastatin pretreatment. These findings suggest that atorvastatin pretreatment may protect hippocampal neurons during seizures by controlling lipocalin-2-associated neuroinflammation., Competing Interests: CONFLICTS OF INTEREST: The authors declare no conflicts of interest.

Immunoinformatics plays a pivotal role in vaccine design, immunodiagnostic development, and antibody production. In the past, antibody design and vaccine development depended exclusively on immunological experiments which are relatively expensive and time-consuming. However, recent advances in the field of immunological bioinformatics have provided feasible tools which can be used to lessen the time and cost required for vaccine and antibody development. This approach allows the selection of immunogenic regions from the pathogen genomes. The ideal regions could be developed as potential vaccine candidates to trigger protective immune responses in the hosts. At present, epitope-based vaccines are attractive concepts which have been successfully trailed to develop vaccines which target rapidly mutating pathogens. In this article, we provide an overview of the current progress of immunoinformatics and their applications in the vaccine design, immune system modeling and therapeutics.

Giardiasis is an intestinal infection caused by the protozoan parasite Giardia intestinalis, affecting hundreds of millions of people worldwide. This microaerophilic protozoan is capable of surviving in the host intestine in the presence of both oxygen and reactive oxygen species (ROS), despite the lack of conventional ROS-scavenging enzymes. The understanding of how G. intestinalis tolerates free radicals could help to identify essential biological processes that protect themselves against oxidative stress within the human gut. This review outlines the antioxidant mechanisms that are utilized by G. intestinalis, with an emphasis on the potential novel role of alpha giardins. The comprehensive understanding of the processes involved in oxidative stress management may provide new insights into improved treatments for giardiasis, and other medically important protozoan parasitic diseases.

Background: Scientific data on the performance of collegiate female tennis players during the menstrual phases are scarce., Trial Design: Double-blind, counter-balanced, crossover trials were conducted to examine whether tennis performance was affected during menstruation, with and without dehydroepiandrosterone sulfate (DHEA-S) supplementation., Methods: Ten Division 1 collegiate tennis players (aged 18-22 years) were evenly assigned into placebo-supplemented and DHEA-supplemented (25 mg/day) trials. Treatments were exchanged among the participants after a 28-day washout. Tennis serve performance was assessed on the first day of menstrual bleeding (day 0/28) and on days 7, 14 and 21., Results: Mood state was unaltered during the menstrual cycles in both trials. The lowest tennis serve performance score (speed times accuracy) occurred on day 14 (P=0.06 vs day 0; P=0.01 vs day 21) in both placebo and DHEA trials. Decreased performance on day 14 was explained by decreased accuracy (P=0.03 vs day 0/28; P=0.01 vs day 21), but not velocity itself. Isometric hip strength, but not quadriceps strength, was moderately lower on day 14 (P=0.08). Increasing plasma DHEA-S (by ~65%) during the DHEA-supplemented trial had no effects on mood state, sleep quality or tennis serve performance., Conclusion: We have shown that menses does not affect serve performance of collegiate tennis players. However, the observed decrement in the accuracy of serve speed near ovulation warrants further investigation., Competing Interests: Competing interests: The authors declare that they have no conflict of interest in any aspect.

Entamoeba species are commonly detected in stool samples of Orang Asli due to their substandard living conditions and poor hygiene. Among the Entamoeba spp., Entamoeba histolytica is the only known primary pathogenic species. This study determined the prevalence and distribution of anti-amoebic IgG antibody among Orang Asli in Peninsular Malaysia. The results would reflect the prevalence of amoebiasis in the population. This study analysed a total of 375 serum samples from archives of two Orang Asli projects conducted between 2011 and 2014. They were from six different states in Malaysia, namely Johor, Kedah, Kelantan, Pahang, Perak, and Selangor. Anti-amoebic IgG antibody was detected using an enzymelinked immunosorbent assay (ELISA) with crude soluble antigen produced from axenically grown E. histolytica trophozoites. From the analysis, the overall seropositivity was approximately 71% (266/375), while the seropositivity rates for each of the three Orang Asli tribes i.e. Senoi, Negrito and Proto-Malay, were 66% (137/208), 92% (103/112), and 43% (17/ 41) respectively. Orang Asli from Kedah [95% (52/55)] showed the highest seropositivity, followed by Kelantan [79% (54/68)], Perak [73% (78/107)], Pahang [60% (57/95)], Selangor [56% (14/25)], and Johor [48% (10/21)]. Orang Asli from rural [76% (192/254)] and peripheral urban [65% (69/106)] areas showed significantly higher seropositivity (p=0.002) than those from urban areas [36% (4/11)]. The high prevalences of anti-amoebic IgG antibody in these Orang Asli populations comprised both active and past infections. This study provides current insights of amoebiasis in selected Orang Asli settlements in Peninsular Malaysia. The high seropositivity of anti-amoebic IgG antibody suggests that the settlements are endemic for amoebiasis and there is a high risk of acquiring E. histolytica infection among the dwellers.

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a rate-limiting photosynthetic enzyme that catalyzes carbon fixation in the Calvin cycle. Much interest has been devoted to engineering this ubiquitous enzyme with the goal of increasing plant growth. However, experiments that have successfully produced improved Rubisco variants, via directed evolution in Escherichia coli, are limited to bacterial Rubisco because the eukaryotic holoenzyme cannot be produced in E. coli. The present study attempts to determine the specific differences between bacterial and eukaryotic Rubisco large subunit primary structure that are responsible for preventing heterologous eukaryotic holoenzyme formation in E. coli. A series of chimeric Synechococcus Rubiscos were created in which different sections of the large subunit were swapped with those of the homologous Chlamydomonas Rubisco. Chimeric holoenzymes that can form in vivo would indicate that differences within the swapped sections do not disrupt holoenzyme formation. Large subunit residues 1-97, 198-247 and 448-472 were successfully swapped without inhibiting holoenzyme formation. In all ten chimeras, protein expression was observed for the separate subunits at a detectable level. As a first approximation, the regions that can tolerate swapping may be targets for future engineering.

Background: Excretory-secretory (ES) proteins of E. histolytica are thought to play important roles in the host invasion, metabolism, and defence. Elucidation of the types and functions of E. histolytica ES proteins can further our understanding of the disease pathogenesis. Thus, the aim of this study is to use proteomics approach to better understand the complex ES proteins of the protozoa., Methods: E. histolytica ES proteins were prepared by culturing the trophozoites in protein-free medium. The ES proteins were identified using two mass spectrometry tools, namely, LC-ESI-MS/MS and LC-MALDI-TOF/TOF. The identified proteins were then classified according to their biological processes, molecular functions, and cellular components using the Panther classification system (PantherDB)., Results: A complementary list of 219 proteins was identified; this comprised 201 proteins detected by LC-ESI-MS/MS and 107 proteins by LC-MALDI-TOF/TOF. Of the 219 proteins, 89 were identified by both mass-spectrometry systems, while 112 and 18 proteins were detected exclusively by LC-ESI-MS/MS and LC-MALDI-TOF/TOF respectively. Biological protein functional analysis using PantherDB showed that 27% of the proteins were involved in metabolic processes. Using molecular functional and cellular component analyses, 35% of the proteins were found to be involved in catalytic activity, and 21% were associated with the cell parts., Conclusion: This study showed that complementary use of LC-ESI-MS/MS and LC-MALDI-TOF/TOF has improved the identification of ES proteins. The results have increased our understanding of the types of proteins excreted/secreted by the amoeba and provided further evidence of the involvement of ES proteins in intestinal colonisation and evasion of the host immune system, as well as in encystation and excystation of the parasite.

A 75-year-old woman with hypertension presented with acute suppurative cholangitis. Chest radiography revealed severe kyphosis. Abdominal computed tomography revealed a large stone impacted in the common bile duct (CBD). The patient underwent emergent endoscopic retrograde cholangiopancreatography, and cholangiography revealed a large stone (7×3 cm) in the CBD that could not be captured using a large basket. We could not use the percutaneous approach for stone fragmentation by using a cholangioscope because of severe degenerative kyphosis. Finally, we performed holmium laser lithotripsy under peroral cholangioscopy by using an ultraslim endoscope, and the large stone in the CBD was successfully fragmented and removed without complications.

Background. Orang Asli (aborigine) children are susceptible to soil-transmitted helminth (STH) infections due to their lifestyle and substandard sanitation system. Objectives. This study aimed to examine the helminthic and nutritional status of Orang Asli school children in Sekolah Kebangsaan Pos Legap, a remote primary school at Kuala Kangsar District in the state of Perak, Malaysia. In addition, the sensitivities of four STH stool examination techniques were also compared. Methods. Demography and anthropometry data were collected by one-to-one interview session. Collected stools were examined with four microscopy techniques, namely, direct wet mount, formalin ether concentration (FEC), Kato-Katz (KK), and Parasep™. Results. Anthropometry analysis showed that 78% (26/33) of children in SK Pos Legap were malnourished and 33% (11/33) of them were stunted. Stool examinations revealed almost all children (97%) were infected by either one of the three commonest STHs. FEC was the most sensitive method in detection of the three helminth species. Conclusion. This study revealed that STH infections and nutritional status still remain a health concern among the Orang Asli children. These communal problems could be effectively controlled by regular monitoring of STH infection loads, administration of effective antihelminthic drug regimen, and also implementation of effective school nutritional programs.

Motivation has long been associated with sports engagement. However, to date no research has been performed to understand the domain of motivation among ten-pin bowlers. The purpose of this study was to investigate different types of motivation (i.e., intrinsic vs. extrinsic) based on self-determination theory from the perspective of gender and the bowler type (competitive vs. casual). A total of 240 bowlers (104 male, 136 female; 152 competitive, 88 casual) with a mean age of 16.61 ± 0.78 years were recruited in Kuala Lumpur. The Sport Motivation Scale, a 28-item self-report questionnaire measuring seven subscales (i.e., intrinsic motivation to know, intrinsic motivation to accomplish, intrinsic motivation to experience stimulation, extrinsic motivation to identify regulation, extrinsic motivation for introjection regulation, extrinsic motivation to external regulation, and amotivation) was administered. Results showed significant differences (t=10.43, df=239, p=0.01) between total scores of intrinsic and extrinsic motivation among ten-pin bowlers. There were significant gender differences with respect to intrinsic motivation to know, intrinsic motivation to accomplish, intrinsic motivation to experience stimulation, and extrinsic motivation to identify regulation. However, no significant bowler type differences were found for either the intrinsic (t=-1.15, df=238, p=0.25) or extrinsic (t=-0.51, df=238, p=0.61) motivation dimensions. In conclusion, our study demonstrated substantial intrinsic motivation for gender effects, but no bowler type effects among adolescent ten-pin bowlers.

Purpose: The purposes of the current study were to assess the early results of cementless hip arthroplasty (HA) for femoral neck fractures in elderly patients with severe osteoporosis and to compare the clinical outcomes between those who underwent total HA (THA) or bipolar hemiarthroplasty (BHA)., Materials and Methods: From April 2011 to May 2012, we performed 87 cementless HAs for displaced femoral neck fractures in elderly patients (≥65 years) with severe osteoporosis. Among them, we studied 70 hips that were able to be followed-up for >24 months. Of these, 34 underwent THA and 36 underwent BHA. Clinical results were evaluated using the Harris hip score (HHS), Koval classification, and radiographs., Results: Only one instance of femoral stem loosening was observed. Additionally, no dislocations were observed and no revision surgeries were required. The mean changes in the functional items of the HHS scores were 2.8 and 5.2 for those who underwent THA and BHA, respectively (P<0.05). According to the Koval classification used for the ambulatory status analysis, the mean perioperative change in the grade was 0.8 (0-4), with no significant differences noted between the THA and BHA groups., Conclusion: The early results of cementless HA for femur neck fractures in elderly patients with osteoporosis were satisfactory, and THA was found to have a functional advantage over BHA.

Background and Purpose: In 2008, the Ministry of Health and Welfare of South Korea initiated the Regional Comprehensive Stroke Center (CSC) program to decrease the incidence and mortality of stroke nationwide. We evaluated the performance of acute ischemic stroke management after the Regional CSC program was introduced., Methods: The Ministry of Health and Welfare established 9 Regional CSCs in different provinces from 2008 to 2010. All Regional CSCs have been able to execute the critical processes independently for stroke management since 2011. The Ministry of Health and Welfare was responsible for program development and financial support, the Regional CSC for program execution, and the Korea Centers for Disease Control and Prevention for auditing the execution. We analyzed prospectively collected data on the required indices from 2011 and repeated the analysis the following year for comparison., Results: After the Regional CSCs were established, the first brain image was taken within 1 hour from arrival at the emergency room for all patients with stroke; the length of hospital stay decreased from 14 to 12 days; for the rapid execution of thrombolysis, the first brain image was taken within 12 minutes; intravenous and intra-arterial thrombolysis were started within 40 and 110 minutes, respectively, after emergency room arrival; and the hospital stay of thrombolytic patients decreased from 19 to 15 days., Conclusions: The Regional CSC program has improved the performance of acute stroke management in South Korea and can be used as a model for rapidly improving stroke management., (© 2014 American Heart Association, Inc.)

Entamoeba histolytica causes amoebic diarrhoea, colitis and liver abscess (ALA). Diagnosis of ALA is difficult, as most patients do not have simultaneous intestinal amoebic infection. At Hospital Universiti Sains Malaysia (HUSM), diagnosis of ALA relies on a combination of clinical findings, ultrasound examination of the liver and serodiagnosis using a commercial kit. In this study, two in-house indirect ELISAs were developed and evaluated. One of the in-house assays utilises E. histolytica crude soluble antigen (CSA) to detect serum IgG specific to the parasite whereas the other uses E. histolytica ether extract antigen (EEA). Preparation of CSA requires a sonicator to lyse the amoeba whereas EEA was prepared by chemically solubilizing the trophozoites. Based on the cut-off value of mean optical density + 3SD, CSA-ELISA showed 100% (24/24) sensitivity and 93.33% (210/225) specificity; while EEA-ELISA showed 91.67% (22/24) sensitivity and 95.11% (214/225) specificity. In conclusion, both the in-house indirect ELISAs were found to be efficacious for diagnosis of ALA; and the EEA is easier to prepare than the commonly used CSA.

Objective: The ROBODOC system offers the theoretical advantage of providing better fit and mechanical stability of the stem in total hip arthroplasty. However, there has been no previous study on short metaphyseal-fitting stem implantation using the ROBODOC system. The aim of the present study was to compare the implant position and primary stability of short metaphyseal-fitting stems implanted by robotic milling and manual rasping in a human cadaveric femoral model., Methods: Eight matched pairs of human cadaveric femora were randomly assigned to a robotic milling group or manual rasping group. Operative time and intraoperative femoral fractures were monitored, and radiographic evaluation of stem alignment was performed by comparison of preoperative planning and postoperative CT data. Stability testing was performed on six matched pairs of femora, excluding two specimens in which intraoperative fractures occurred., Results: The robotic milling procedures took an average of 27 minutes longer than the manual rasping procedures (p < 0.001). The robotic milling group exhibited significantly better anteroposterior alignment and vertical seating, and also showed a significantly reduced variability in both alignment and vertical seating. No intraoperative femoral fracture was detected in the robotic milling group, whereas two femoral fractures and one femoral stem tip perforation were detected in the manual rasping group. Stability testing showed no significant difference in translational and rotational migrations between the two groups, although the robotic milling group showed a trend towards reduced variability of stability., Conclusions: Our cadaveric study suggests that the use of the ROBODOC system for short metaphyseal-fitting stem implantation may have advantages in improving implant fit and reducing the risk of intraoperative femoral fractures without compromising primary stability.

Entamoeba histolytica is the only Entamoeba species that causes amoebiasis in humans. Approximately 50 million people are infected, with 100, 000 deaths annually in endemic countries. Molecular diagnosis of Entamoeba histolytica is important to differentiate it from the morphologically identical Entamoeba dispar to avoid unnecessary medication. Conventional molecular diagnostic tests require trained personnel, cold-chain transportation and/or are storage-dependent, which make them user-unfriendly. The aim of this study was to develop a thermostabilized, one-step, nested, tetraplex PCR assay for the detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba species in cold-chain-free and ready-to-use form. The PCR test was designed based on the Entamoeba small subunit rRNA (SSU-rRNA) gene, which detects the presence of any Entamoeba species, and simultaneously can be used to differentiate Entamoeba histolytica from Entamoeba dispar. In addition, a pair of primers was designed to serve as an internal amplification control to help identify inhibitors in the samples. All PCR reagents together with the designed primers were thermostabilized by lyophilization and were stable at 24 °C for at least 6 months. The limit of detection of the tetraplex PCR was found to be 39 pg DNA or 1000 cells for Entamoeba histolytica and 78 pg DNA or 1000 cells for Entamoeba dispar, and the specificity was 100 %. In conclusion, this cold-chain-free, thermostabilized, one-step, nested, multiplex PCR assay was found to be efficacious in differentiating Entamoeba histolytica from other non-pathogenic Entamoeba species.

Background and Purpose: We conducted this study to define the relationship between the hyperventilation-induced EEG changes (HV-EEG changes) and PCO2., Methods: In consecutive EEG recordings of 190 patients, we gathered data on PCO2 during the hyperventilation procedure. The data included baseline PCO2 (B-PCO2), PCO2 after 5 min of hyperventilation (5 min-PCO2), the mean value of the PCO2 (M-PCO2), and the difference between B-PCO2 and 5 min-PCO2 (ΔPCO2). We divided the enrolled patients into two groups by hyperventilation response (response group and no-response group), presence of epilepsy (epileptic group and non-epileptic group) and age (child-adolescent group and adult group) repeatedly. We compared the four variables between the two groups in each pair., Results: ΔPCO2 was 14.2±5.0 mmHg (mean±SD) in response group (n=48) and 12.4±5.0 in no-response group (n=142; p=0.033) for all the patients. For adult patients only, 5 min-PCO2 was 24.3±3.4 in response group (n=30) and 26.2±4.6 in no-response group (n=115; p=0.048), and ΔPCO2 was 15.8±4.0 and 12.9±5.0, respectively (p=0.004). In non-epileptic patients, 5min-PCO2 was 23.4±2.2 in response group (n=7) and 26.3±3.8 in no-response group (n=44; p=0.026), and ΔPCO2 was 15.9±4.3 and 12.7±3.9, respectively (p=0.053)., Conclusions: In adults and non-epileptic patients, ΔPCO2 and 5 min-PCO2 may be crucial to the induction of EEG changes by hyperventilation. PCO2 could be a crucial factor for provoking HV-EEG changes in a limited group of patients.

Serodiagnosis of amoebiasis remains the preferred method for diagnosis of amoebic liver abscess (ALA). However, the commercially available kits are problematic in areas of endemicity due to the persistently high background antibody titers. Human serum samples (n = 38) from patients with ALA who live in areas of endemicity were collected from Hospital Universiti Sains Malaysia during the period of 2008 to 2010. Western blots using excretory-secretory antigen (ESA) collected from axenically grown Entamoeba histolytica were probed with the above serum samples. Seven antigenic proteins of ESA with various reactivities were identified, i.e., 152 kDa, 131 kDa, 123 kDa, 110 kDa, 100 kDa, 82 kDa, and 76 kDa. However, only the 152-kDa and 110-kDa proteins showed sensitivities above 80% in the Western blot analysis. All the antigenic proteins showed undetectable cross-reactivity when probed with healthy human serum samples (n = 30) and serum samples from other infections (n = 33). From the matrix-assisted laser desorption ionization-two-stage time of flight (MALDI-TOF/TOF) analysis, the proteins were identified as heavy subunits of E. histolytica lectin and E. histolytica pyruvate phosphate dikinase, respectively. Use of the E. histolytica lectin for diagnosis of ALA has been well reported by researchers and is being used in commercialized kits. However, this is the first report on the potential use of pyruvate phosphate dikinase for diagnosis of ALA; thus, this molecule merits further evaluation on its diagnostic value using a larger panel of serum samples.

The Influenza A H1N1 pandemic (A H1N1) occurred between June 2009 and August 2010. Although the pandemic is now over, the virus has emerged as the predominant strain in the current seasonal influenza phase in the northern hemisphere. The A H1N1 influenza is a novel strain of the influenza A virus and is widely known as swine flu. The virus contains a mixture of genetic material from human, pig and bird flu virus. It is a new variety of flu which people have not had much immunity to. Much has been learnt from the Pandemic of 2009/2010 but the messages about vaccination and treatment seem to be taken slowly by the clinical profession. Most people affected by the virus, including pregnant women, suffer a mild viral illness, and make a full recovery. The median duration of illness is around seven days. This influenza typically affects the younger age group i.e. from the ages of 5-65 years. Current experience shows that the age group experiencing increased morbidity and mortality rates are in those under 65 years of age. Pregnant women, because of their altered immunity and physiological adaptations, are at higher risk of developing pulmonary complications, especially in the second and third trimesters. In the United Kingdom, twelve maternal deaths were reported to be associated with the H1N1 virus during the pandemic and clear avoidable factors were identified (Modder, Review of Maternal Deaths in the UK related to A H1N1 2009 influenza (CMACE). www.cmace.org.uk, 2010). The pregnancy outcomes were also poor for women who were affected by the virus with a fivefold increase in the perinatal mortality rate and threefold increase in the preterm delivery rate (Yates et al. Health Technol Assess 14(34):109-182, 2010). There continues to be a low uptake of the flu vaccine and commencement of antiviral treatment for pregnant women.

Vibrio cholerae has caused severe outbreaks of cholera worldwide with thousands of recorded deaths annually. Molecular diagnosis for cholera has become increasingly important for rapid detection of cholera as the conventional methods are time-consuming and labour intensive. However, traditional PCR tests still require cold-chain transportation and storage as well as trained personnel to perform, which makes them user-unfriendly. The aim of this study was to develop a thermostabilized triplex PCR test for cholera which is in a ready-to-use form and requires no cold chain. The PCR test specifically detects both toxigenic and non-toxigenic strains of V. cholerae based on the cholera toxin A (ctxA) and outer-membrane lipoprotein (lolB) genes. The thermostabilized triplex PCR also incorporates an internal amplification control that helps to check for PCR inhibitors in samples. PCR reagents and the specific primers were lyophilized into a pellet form in the presence of trehalose, which acts as an enzyme stabilizer. The triplex PCR was validated with 174 bacteria-spiked stool specimens and was found to be 100 % sensitive and specific. The stability of the thermostabilized PCR was evaluated using the Q10 method and it was found to be stable for approximately 7 months at 24 °C. The limit of detection of the thermostabilized triplex PCR assay was 2×10(4) c.f.u. at the bacterial cell level and 100 pg DNA at the genomic DNA level, comparable to conventional PCR methods. In conclusion, a rapid thermostabilized triplex PCR assay was developed for detecting toxigenic and non-toxigenic V. cholerae which requires minimal pipetting steps and is cold chain-free.