Mechanistic Association of Hepatoblastoma with Cerebral Palsy: A Narrative Review.

Hepatoblastoma is a rare liver cancer that occurs most often in children who present with lower birth weight. Cerebral palsy (CP) is a neurodevelopmental disorder distinguished by irregularities in muscle tone, movement, and motor skills. CP is caused by damage to the developing brain and is often associated with secondary complications such as severe constipation. Clinicians must be aware of sudden worsening constipation occurring in CP children because it can also be a sign of hepatoblastoma. The aim of this review is to summarize the current understanding of the risks for hepatoblastoma development in children with CP. Cancer risks likely include dysfunction of the immune system surveillance in CP children. Elevated C-reactive protein and tumor necrosis factor-alpha levels may be higher in children with CP, which weakens their innate immune system. Metabolic disruption increases the risk of some cancers, and poor nutrition and reduced growth that occur in CP patients may have an impact on cancer development through a loss in immune function. Increased mobility and physical activity can increase the T-cell, natural killer cell, and neutrophil population. Children with CP tend to engage poorly in physical activity, and consequently, their immune system is affected. There are multiple factors associated with CP that increase the risk of childhood cancers such as hepatoblastoma.

Optimization of recycled rubber self-compacting concrete: Experimental findings and machine learning-based evaluation.

This research aims to assess the rheological and mechanical characteristics of Self-compacting concrete (SCC) incorporating waste tire rubber aggregates (WRTA) as an interim substitute for coarse aggregates. However, the standard experimental modeling approach has significant obstacles when it comes to overcoming the nonlinearity and environmental susceptibility of concrete parts. Therefore, linear regression (LR) and extreme gradient boosting (XGBoost) were used as two standard single machine learning (ML) models to predict the aforementioned rubberized SCC features. In this study, conventional coarse aggregates were supplanted with WRTA at 0%, 5%, 10%, and 20% to uncover the optimal proportion of coarse aggregates substituting rubber. To find the optimum amount of WRTA to use as a substitute, the study follows the impacts of rubber on the self-compacting rubberized concrete's (SCRC) rheological and mechanical characteristics. The consequences on fresh properties were investigated by the slump flow, J-ring, and V-funnel tests, while compressive and splitting tensile strengths tests were conducted to assess mechanical properties. Increasing WRTA test outputs indicated a deterioration in workability and hardened qualities. While a 10% swapping ratio is deemed feasible for producing SCRC, optimal results were achieved by reducing environmental impacts and efficiently managing a significant volume of rubber tire waste with a 5% substitution of rubber within the coarse aggregates. The research findings indicated a noticeable decrease in fresh properties as the WRTA content increased. Notably, after 28 days, a 10% WRTA substitution led to a 34% reduction in compressive strength and a 28% decrease in splitting tensile strength, satisfying ACI standards. Furthermore, XGBoost demonstrated superior predictive performance with the highest R2 values, outperforming the LR model and affirming its efficacy in delivering more accurate predictions.

Eco-friendly concrete incorporating palm oil fuel ash: Fresh and mechanical properties with machine learning prediction, and sustainability assessment.

Rising natural resource consumption leads to increased hazardous gas emissions, necessitating the concrete industry's focus on sustainable alternatives like palm oil fuel ash (POFA) to replace cement. Also, advanced machine learning (ML) techniques can uncover previously unreported insights about the effects of POFA that may be missing from the literature. Hence, this study investigates the influence of varying concentrations of POFA on fresh and mechanical characteristics with quantifying ML approaches and microstructural performance, as well as the environmental impact of structural concrete. For this, cement substitutions of 5 %, 15 %, 25 %, 35 %, and 45 % (by weight of cement) were utilized. POFA enhanced the overall concrete workability, with slump increments ranging from approximately 9 %-55 % and compacting factor increments of 4 %-12 %. Mechanical performance of POFA concrete improved up to 25 % replacement levels, with the highest enhancements observed in compressive (4.5 %), splitting tensile (36 %), and flexural (31 %) strength, for the mix containing 15 % POFA. The finer particle size of POFA improved microstructural performance by reducing porosity, aligning with the enhanced mechanical strength. The environmental impact of POFA was assessed by measuring eCO2 emissions, revealing a potential reduction of up to 44 %. Incorporating 5 %-15 % POFA yielded ideal mechanical performance results, significantly enhancing sustainability and cost-effectiveness. Regarding the ML approach, it can be observed that a low regression coefficient (R2) contrasts sharply with the higher R2 values for the random forest (RF) and the ensemble model, indicating satisfactory precision prediction with experimental results.

Short Term Side Effects Of Inactivated COVID-19 Vaccines Among Pakistani Population: A Survey-Based Cross-Sectional Study.

Objectives: To assess the short-term adverse effects of two inactivated coronavirus disease-2019 vaccines, and the demographic factors associated with such events. METHODS: The cross-sectional study was conducted in Karachi from August to October 2021 after approval from the ethics review board of Dow University of Health Sciences, Karachi, and comprised adults of either gender who had received at least one dose of either Sinopharm or CoronaVac vaccine. Data was collected using online and printed survey forms. The questionnaire investigated the symptoms experienced by the participants after the administration of the vaccine dose. Data was analysed using SPSS 22. RESULTS: Of the 1000 survey forms filled, 896 were analysed; 505(56.4%) women and 391(43.6%) men were included in the study. Most of the participants were aged 18-30 years 644(71.9%). Overall, 581(64.8%) subjects had received Sinopharm vaccine, and 315(35.2%) received CoronaVac. The incidence of side effects after the first and second dose respectively was 63.3% (368 out of 581) and 55.2% (239 out of 433) for Sinopharm and 65.4% (206 out of 315) and 61.4% (89 out of 145) for CoronaVac. The factors associated with a higher risk of side effects were female gender and young age (p<0.05). CONCLUSIONS: Most of the reported symptoms were minor in nature, like pain at the injection site, and women and those of young age reported such symptoms more than men and the elderly.

Clinical evaluation of cemented and adhesively resin-bonded monolithic and partially layered zirconia and lithium disilicate crowns.

Purpose: Zirconia and lithium disilicate crowns are very popular dental restorations. The cement type and layering technique used can profoundly affect the clinical performance of these crowns.This retrospective study is designed to investigate the success rate of cemented and adhesively bonded monolithic and partially layered zirconia and lithium disilicate crowns placed in faculty practice settings. Materials and Methods: Patients who had received zirconia or lithium disilicate crowns at the faculty practice were invited for clinical examination. The examiner used the modified United States Public Health Service (MUSPHS) evaluation criteria to evaluate the crowns. The crowns were either glass ionomer cemented zirconia (GIC-Zr), resin-bonded zirconia (Adh-Zr), or resin-bonded lithium disilicate (Adh-LD). The crowns were also divided into monolithic and layered groups. Inferential analysis was used to examine the differences through bivariate analysis using t-testing and one-way ANOVA. Results: Thirty-five patients, with a combined total of 218 single crowns, agreed to participate in the study. No statistically significant differences in the quality outcome variables considered were found between the groups, except for marginal adaptation, where Adh-Zr achieved significantly higher scores compared to GIC-Zr and Adh-LD. Layered zirconia and lithium disilicate crowns have significantly higher quality outcomes in terms of anatomic form, marginal adaptation, and color match compared to monolithic zirconia and lithium disilicate crowns. Conclusions: Conventionally cemented zirconia and adhesively bonded zirconia and lithium disilicate crowns are reliable treatment options with high short-term success rates. Clinical studies with longer follow-up times are needed to investigate their long-term success rates.

Integration of Rice Husk Ash as Supplementary Cementitious Material in the Production of Sustainable High-Strength Concrete.

The incorporation of waste materials generated in many industries has been actively advocated for in the construction industry, since they have the capacity to lessen the pollution on dumpsites, mitigate environmental resource consumption, and establish a sustainable environment. This research has been conducted to determine the influence of different rice husk ash (RHA) concentrations on the fresh and mechanical properties of high-strength concrete. RHA was employed to partially replace the cement at 5%, 10%, 15%, and 20% by weight. Fresh properties, such as slump, compacting factor, density, and surface absorption, were determined. In contrast, its mechanical properties, such as compressive strength, splitting tensile strength and flexural strength, were assessed after 7, 28, and 60 days. In addition, the microstructural evaluation, initial surface absorption test, = environmental impact, and cost-benefit analysis were evaluated. The results show that the incorporation of RHA reduces the workability of fresh mixes, while enhancing their compressive, splitting, and flexural strength up to 7.16%, 7.03%, and 3.82%, respectively. Moreover, incorporating 10% of RHA provides the highest compressive strength, splitting tensile, and flexural strength, with an improved initial surface absorption and microstructural evaluation and greater eco-strength efficiencies. Finally, a relatively lower CO2-eq (equivalent to kg CO2) per MPa for RHA concrete indicates the significant positive impact due to the reduced Global Warming Potential (GWP). Thus, the current findings demonstrated that RHA can be used in the concrete industry as a possible revenue source for developing sustainable concretes with high performance.

Challenging ecogeographical rules: Phenotypic variation in the Mountain Treeshrew (Tupaia montana) along tropical elevational gradients.

Bergmann's and Allen's rules were defined to describe macroecological patterns across latitudinal gradients. Bergmann observed a positive association between body size and latitude for endothermic species while Allen described shorter appendages as latitude increases. Almost two centuries later, there is still ongoing discussion about these patterns. Temperature, the common variable in these two rules, varies predictably across both latitude and elevation. Although these rules have been assessed extensively in mammals across latitude, particularly in regions with strong seasonality, studies on tropical montane mammals are scarce. We here test for these patterns and assess the variation of several other locomotory, diet-associated, body condition, and thermoregulatory traits across elevation in the Mountain Treeshrew (Tupaia montana) on tropical mountains in Borneo. Based on morphological measurements from both the field and scientific collections, we found a complex pattern: Bergmann's rule was not supported in our tropical mountain system, since skull length, body size, and weight decreased from the lowest elevations (<1000 m) to middle elevations (2000-2500 m), and then increased from middle elevations to highest elevations. Allen's rule was supported for relative tail length, which decreased with elevation, but not for ear and hindfoot length, with the former remaining constant and the latter increasing with elevation. This evidence together with changes in presumed diet-related traits (rostrum length, zygomatic breadth and upper tooth row length) along elevation suggest that selective pressures other than temperature, are playing a more important role shaping the morphological variation across the distribution of the Mountain Treeshrew. Diet, food acquisition, predation pressure, and/or intra- and inter-specific competition, are some of the potential factors driving the phenotypic variation of this study system. The lack of variation in body condition might suggest local adaptation of this species across its elevational range, perhaps due to generalist foraging strategies. Finally, a highly significant temporal effect was detected in several traits but not in others, representing the first phenotypic variation temporal trends described on treeshrews.

New distribution record of the rare bat Hypsugo cf. vordermanni (Chiroptera, Vespertilionidae) from the Crocker Range Park, Sabah, Malaysian Borneo.

A female Hypsugocf.vordermanni was caught at a stream near a village road in Mantailang, Crocker Range Park, Sabah, on 12 November 2018. This bat is a new record for the national park and the second record for Sabah. It was first recorded from Banggi Island, Kudat, in 1991. The species' few records throughout its range is most likely due to sampling effort bias towards forest interior and cave-dwelling species, as this species is more likely an edge-space aerial forager. It is morphologically similar to Peninsular Malaysia's Hypsugomacrotis, but the latter has never been reported from Borneo. Therefore, additional specimen collection and molecular data for H.cf.vordermanni are needed for further species confirmation. More information on H.vordermanni ecology is also crucial in aiding the management plan for this species as it is currently classified as Data Deficient by the IUCN.

Bat species diversity trend along an elevation gradient: A study in Crocker Range Park, Sabah, Borneo.

Bats (Order: Chiroptera) is a recognised group of bioindicators due to their sensitivity towards alterations in their immediate surroundings. With the threats of climate change becoming more severe on a daily basis, it is reasonable to collect data on how bat diversity is influenced by elevation. This will be useful to predict and monitor possible upslope shifting of bat species due to increase in surrounding temperature or anthropogenic pressure. Hence, this study aims to uncover the bat diversity trend at different elevations in Crocker Range Park (CRP), Sabah, Malaysia. Bat trappings were conducted in four substations within this park, covering an elevation spectrum from 450 to 1900 m a.s.l. The overall sampling managed to capture 133 individuals of bats, predominantly Pteropodidae, with the addition of two new species locality records for CRP, Murinapeninsularis and Hypsugovondermanni. Simple linear regression analyses revealed that both bat diversity and richness have an inverse linear relationship with elevation. Likewise, the Pearson's correlation value, associating bat diversity with elevation, also shows that they have a negative relationship at r = -0.852. Heterogeneity of habitats explain this trend, as in the lower counterpart of CRP, lowland forests, which are richer in fruit and insect resources persist. Besides, lower land forests have better niche assortment, due to the distinctive layers stratification, allowing bats utilising different guilds to thrive in the same vegetation profile. This study further emphasises the role of CRP to protect most of the bat species found in Borneo, as well as serving as the baseline data for the future studies that look into the impact of temperature increment towards the upslope shifting of the bat population in CRP.

Effectiveness of strip footing with geogrid reinforcement for different types of soils in Mosul, Iraq.

The main cause of problematic soil failure under a certain load is due to low bearing capacity and excessive settlement. With a growing interest in employing shallow foundation to support heavy structures, it is important to study the soil improvement techniques. The technique of using geosynthetic reinforcement is commonly applied over the last few decades. This paper aims to determine the effect of using geogrid Tensar BX1500 on the bearing capacity and settlement of strip footing for different types of soils, namely Al-Hamedat, Ba'shiqah, and Al-Rashidia in Mosul, Iraq. The analysis of reinforced and unreinforced soil foundations was conducted numerically and analytically. A series of conditions were tested by varying the number (N) and the width (b) of the geogrid layers. The results showed that the geogrid could improve the footing's bearing capacity and reduce settlement. The soil of the Al-Rashidia site was sandy and indicated better improvement than the other two sites' soils (clayey soils). The optimum geogrid width (b) was five times the footing width (B), while no optimum geogrid number (N) was obtained. Finally, the numerical results of the ultimate bearing capacity were compared with the analytical results, and the comparison showed good agreement between both the analyses and the optimum range published in the literature. The significant findings reveal that the geogrid reinforcement may induce improvement to the soil foundation, however, not directly subject to the width and number of the geogrid alone. The varying soil properties and footing size also contribute to both BCR and SRR values supported by the improvement factor calculations. Hence, the output complemented the benefit of applying reinforced soil foundations effectively.

First description of male worms of Enterobius (Colobenterobius) serratus (Nematoda: Oxyuridae), the pinworm parasite of proboscis monkeys.

Males of Enterobius (Colobenterobius) serratus Hasegawa et al., 2003 (Nematoda: Oxyuridae) are described for the first time based on six individuals collected from the feces of proboscis monkeys, Nasalis larvatus, in the Lower Kinabatangan Wildlife Sanctuary, Sabah, Malaysian Borneo. The males show identical cephalic morphology to females, being readily distinguishable from their congeners by the serrated inner margins of the lips. The bicolored esophageal corpus, long thin spicule and developed spicular pouch with paired muscular bands are also remarkable characteristics, presumably shared by other Asian members of the subgenus.

Phytochemical Modulation of Apoptosis and Autophagy: Strategies to Overcome Chemoresistance in Leukemic Stem Cells in the Bone Marrow Microenvironment.

Advances in scientific research and targeted treatment regimes have improved survival rates for many cancers over the past few decades. However, for some types of leukemia, including acute lymphoblastic and acute myeloid leukemia, mortality rates have continued to rise, with chemoresistance in leukemic stem cells (LSCs) being a major contributing factor. Most cancer drug therapies act by inducing apoptosis in dividing cells but are ineffective in targeting quiescent LSCs. Niches in the bone marrow, known as leukemic niches, behave as "sanctuaries" where LSCs acquire drug resistance. This review explores the role of the bone marrow environment in the maintenance of LSCs and its contribution to chemoresistance and considers current research on the potential use of phytochemicals to overcome chemoresistance through the modulation of signaling pathways involved in the survival and death of leukemic clonal cells and/or leukemic stem cells. Phytochemicals from traditional Chinese medicine, namely baicalein, chrysin, wogonin (constituents of Scutellaria baicalensis; huáng qín; ), curcumin (a constituent of Curcuma longa, jiang huáng, ), and resveratrol (a constituent of Polygonum cuspidatum; hu zhàng, ) have been shown to induce apoptosis in leukemic cell lines, with curcumin and resveratrol also causing cell death via the induction of autophagy (a nonapoptotic pathway). In order to be effective in eliminating LSCs, it is important to target signaling pathways (such as Wnt/ß-catenin, Notch, and Hedgehog). Resveratrol has been reported to induce apoptosis in leukemic cells through the inhibition of the Notch and Sonic hedgehog signaling pathways, therefore showing potential to affect LSCs. While these findings are of interest, there is a lack of reported research on the modulatory effect of phytochemicals on the autophagic cell death pathway in leukemia, and on the signaling pathways involved in the maintenance of LSCs, highlighting the need for further work in these areas.

A Novel Recombinant Canine Adenovirus Type 1 Detected from Acute Lethal Cases of Infectious Canine Hepatitis.

In this study, canine adenoviruses (CAdVs) from two acute fatal cases of infectious canine hepatitis (ICH) were analyzed using molecular detection and sequencing of the pVIII, E3, and fiber protein genes. Pathological findings in affected dogs were typical for CAdV-1 associated disease, characterized by severe centrilobular to panlobular necrohemorrhagic hepatitis and the development of disseminated intravascular coagulation in the terminal stages of disease. Comparison of partial genome sequences revealed that although these newly detected viruses mainly had CAdV-1 genome characteristics, their pVIII gene was more similar to that of CAdV-2. This likely suggests that a recombination has occurred between CAdV-1 and CAdV-2, which possibly explains the cause of vaccine failure or increased virulence of the virus in the observed ICH cases.

Epitope Mapping of Avian Influenza M2e Protein: Different Species Recognise Various Epitopes.

A common approach for developing diagnostic tests for influenza virus detection is the use of mouse or rabbit monoclonal and/or polyclonal antibodies against a target antigen of the virus. However, comparative mapping of the target antigen using antibodies from different animal sources has not been evaluated before. This is important because identification of antigenic determinants of the target antigen in different species plays a central role to ensure the efficiency of a diagnostic test, such as competitive ELISA or immunohistochemistry-based tests. Interest in the matrix 2 ectodomain (M2e) protein of avian influenza virus (AIV) as a candidate for a universal vaccine and also as a marker for detection of virus infection in vaccinated animals (DIVA) is the rationale for the selection of this protein for comparative mapping evaluation. This study aimed to map the epitopes of the M2e protein of avian influenza virus H5N1 using chicken, mouse and rabbit monoclonal or monospecific antibodies. Our findings revealed that rabbit antibodies (rAbs) recognized epitope 6EVETPTRN13 of the M2e, located at the N-terminal of the protein, while mouse (mAb) and chicken antibodies (cAbs) recognized epitope 10PTRNEWECK18, located at the centre region of the protein. The findings highlighted the difference between the M2e antigenic determinants recognized by different species that emphasized the importance of comparative mapping of antibody reactivity from different animals to the same antigen, especially in the case of multi-host infectious agents such as influenza. The findings are of importance for antigenic mapping, as well as diagnostic test and vaccine development.

Avian Influenza Virus and DIVA Strategies.

Vaccination is becoming a more acceptable option in the effort to eradicate avian influenza viruses (AIV) from commercial poultry, especially in countries where AIV is endemic. The main concern surrounding this option has been the inability of the conventional serological tests to differentiate antibodies produced due to vaccination from antibodies produced in response to virus infection. In attempts to address this issue, at least six strategies have been formulated, aiming to differentiate infected from vaccinated animals (DIVA), namely (i) sentinel birds, (ii) subunit vaccine, (iii) heterologous neuraminidase (NA), (iv) nonstructural 1 (NS1) protein, (v) matrix 2 ectodomain (M2e) protein, and (vi) haemagglutinin subunit 2 (HA2) glycoprotein. This short review briefly discusses the strengths and limitations of these DIVA strategies, together with the feasibility and practicality of the options as a part of the surveillance program directed toward the eventual eradication of AIV from poultry in countries where highly pathogenic avian influenza is endemic.

The promise and challenge of ovarian cancer models.

The complexity and heterogeneity of ovarian cancer cases are difficult to reproduce in in vitro studies, which cannot adequately elucidate the molecular events involved in tumor initiation and disease metastasis. It has now become clear that, although the multiple histological subtypes of ovarian cancer are being treated with similar surgical and therapeutic approaches, they are in fact characterized by distinct phenotypes, cell of origin, and underlying key genetic and genomic alterations. Consequently, the development of more personalized treatment methodologies, which are aimed at improving patient care and prognosis, will greatly benefit from a better understanding of the key differences between various subtypes. To accomplish this, animal models of all histotypes need to be generated in order to provide accurate in vivo platforms for research and the testing of targeted treatments and immune therapies. Both genetically engineered mouse models (GEMMs) and xenograft models have the ability to further our understanding of key mechanisms facilitating tumorigenesis, and at the same time offer insight into enhanced imaging and treatment modalities. While genetic models may be better suited to examine oncogenic functions and interactions during tumorigenesis, patient-derived xenografts (PDXs) are likely a superior model to assess drug efficacy, especially in concurrent clinical trials, due to their similarity to the tumors from which they are derived. Genetic and avatar models possess great clinical utility and have both benefits and limitations. Additionally, the laying hen model, which spontaneously develops ovarian tumors, has inherent advantages for the study of epithelial ovarian cancer (EOC) and recent work champions this model especially when assessing chemoprevention strategies. While high-grade ovarian serous tumors are the most prevalent form of EOC, rarer ovarian cancer variants, such as small cell ovarian carcinoma of the hypercalcemic type and transitional cell carcinoma, or non-epithelial tumors, including germ cell tumors, will also benefit from the generation of improved models to advance our understanding of tumorigenic mechanisms and the development of selective therapeutic options.

Interaction between Bovine leukemia virus (BLV) infection and age on telomerase misregulation.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). BLV can interact with telomerase and inhibits telomere shortening, contributing in leukemogenesis and tumour induction. The role of telomerase in BLV-induced lymphosarcoma and aging has been extensively studied. To date, the interaction of both BLV and aging on telomerase mis-regulation have, however, not been investigated. In the present study, telomerase activity in BLV positive and negative cows was compared over a wide range of ages (11-85 months). Lymphocyte counts were also measured in both BLV positive and negative groups. Telomerase activity was detected in all BLV infected animals with persistent lymphocytosis (PL), especially in older individuals. This study revealed that the cells undergo the natural telomerase shortening even in the presence of an existing viral infection. We also show that viral infection, especially during the PL phase of the disease, increases telomerase activity. A statistically significant interaction between age and viral infection was observed for telomere shortening during BLV infection. Older animals with BLV infection, especially those with persistent lymphocytosis or visible tumors, exhibited a sharp increase in telomerase activity. This study demonstrates that there is a significant interaction between BLV infection and telomerase up-regulation and lymphocytosis.

Advances in tumor screening, imaging, and avatar technologies for high-grade serous ovarian cancer.

The majority of high-grade serous ovarian carcinoma cases are detected in advanced stages when treatment options are limited. Surgery is less effective at eradicating the disease when it is widespread, resulting in high rates of disease relapse and chemoresistance. Current screening techniques are ineffective for early tumor detection and consequently, BRCA mutations carriers, with an increased risk for developing high-grade serous ovarian cancer, elect to undergo risk-reducing surgery. While prophylactic surgery is associated with a significant reduction in the risk of cancer development, it also results in surgical menopause and significant adverse side effects. The development of efficient early-stage screening protocols and imaging technologies is critical to improving the outcome and quality of life for current patients and women at increased risk. In addition, more accurate animal models are necessary in order to provide relevant in vivo testing systems and advance our understanding of the disease origin and progression. Moreover, both genetically engineered and tumor xenograft animal models enable the preclinical testing of novel imaging techniques and molecularly targeted therapies as they become available. Recent advances in xenograft technologies have made possible the creation of avatar mice, personalized tumorgrafts, which can be used as therapy testing surrogates for individual patients prior to or during treatment. High-grade serous ovarian cancer may be an ideal candidate for use with avatar models based on key characteristics of the tumorgraft platform. This review explores multiple strategies, including novel imaging and screening technologies in both patients and animal models, aimed at detecting cancer in the early-stages and improving the disease prognosis.

Multimeric recombinant M2e protein-based ELISA: a significant improvement in differentiating avian influenza infected chickens from vaccinated ones.

Killed avian influenza virus (AIV) vaccines have been used to control H5N1 infections in countries where the virus is endemic. Distinguishing vaccinated from naturally infected birds (DIVA) in such situations however, has become a major challenge. Recently, we introduced the recombinant ectodomain of the M2 protein (M2e) of H5N1 subtype as a novel tool for an ELISA based DIVA test. Despite being antigenic in natural infection the monomer form of the M2e used in ELISA had limited antigenicity and consequently poor diagnostic capability. To address this shortcoming, we evaluated the use of four tandem copies of M2e (tM2e) for increased efficiency of M2e antibody detection. The tM2e gene of H5N1 strain from Indonesia (A/Indonesia/CDC540/2006) was cloned into a pMAL- p4x expression vector and expressed in E.coli as a recombinant tM2e-MBP or M2e-MBP proteins. Both of these, M2e and tM2e antigens reacted with sera obtained from chickens following live H5N1 infection but not with sera from vaccinated birds. A significantly stronger M2e antibody reaction was observed with the tM2e compared to M2e antigen. Western blotting also supported the superiority of tM2e over M2e in detection of specific M2e antibodies against live H5N1 infection. Results from this study demonstrate that M2e tetramer is a better antigen than single M2e and could be more suitable for an ELISA based DIVA test.