Current progress of protein-based dressing for wound healing applications - A review.

Protein-based wound dressings have garnered increasing interest in recent years owing to their distinct physical, chemical, and biological characteristics. The intricate molecular composition of proteins gives rise to unique characteristics, such as exceptional biocompatibility, biodegradability, and responsiveness, which contribute to the promotion of wound healing. Wound healing is an intricate and ongoing process influenced by multiple causes, and it consists of four distinct phases. Various treatments have been developed to repair different types of skin wounds, thanks to advancements in medical technology and the recognition of the diverse nature of wounds. This review has literature reviewed within the last 3-5 years-the recent progress and development of protein in wound dressings and the fundamental properties of an ideal wound dressing. Herein, the recent strides in protein-based state-of-the-art wound dressing emphasize the significant challenges and summarize future perspectives for wound healing applications.

Metal-organic frameworks: synthesis, properties, wound dressing, challenges and scopes in advanced wound dressing.

Wound healing is a critical but complex biological process of skin tissue repair and regeneration resulting from various systems working together at the cellular and molecular levels. Quick wound healing and the problems associated with traditional wound repair techniques are being overcome with multifunctional materials. Over time, this research area has drawn significant attention. Metal-organic frameworks (MOFs), owning to their peculiar physicochemical characteristics, are now considered a promising class of well-suited porous materials for wound healing in addition to their other biological applications. This detailed literature review provides an overview of the latest developments in MOFs for wound healing applications. We have discussed the synthesis, essential biomedical properties, wound-healing mechanism, MOF-based dressing materials, and their wound-healing applications. The possible major challenges and limitations of MOFs have been discussed, along with conclusions and future perspectives. This overview of the literature review addresses MOFs-based wound healing from several angles and covers the most current developments in the subject. The readers may discover how the MOFs advanced this discipline by producing more inventive, useful, and successful dressings. It influences the development of future generations of biomaterials for the healing and regeneration of skin wounds.

Current Perspectives of Protein in Bone Tissue Engineering: Bone Structure, Ideal Scaffolds, Fabrication Techniques, Applications, Scopes, and Future Advances.

In view of their exceptional approach, excellent inherent biocompatibility and biodegradability properties, and interaction with the local extracellular matrix, protein-based polymers have received attention in bone tissue engineering, which is a multidisciplinary field that repairs and regenerates fractured bones. Bone is a multihierarchical complex structure, and it performs several essential biofunctions, including maintaining mineral balance and structural support and protecting soft organs. Protein-based polymers have gained interest in developing ideal scaffolds as emerging biomaterials for bone fractured healing and regeneration, and it is challenging to design ideal bone substitutes as perfect biomaterials. Several protein-based polymers, including collagen, keratin, gelatin, serum albumin, etc., are potential materials due to their inherent cytocompatibility, controlled biodegradability, high biofunctionalization, and tunable mechanical characteristics. While numerous studies have indicated the encouraging possibilities of proteins in BTE, there are still major challenges concerning their biodegradability, stability in physiological conditions, and continuous release of growth factors and bioactive molecules. Robust scaffolds derived from proteins can be used to replace broken or diseased bone with a biocompatible substitute; proteins, being biopolymers, provide excellent scaffolds for bone tissue engineering. Herein, recent developments in protein polymers for cutting-edge bone tissue engineering are addressed in this review within 3-5 years, with a focus on the significant challenges and future perspectives. The first section discusses the structural fundamentals of bone anatomy and ideal scaffolds, and the second section describes the fabrication techniques of scaffolds. The third section highlights the importance of proteins and their applications in BTE. Hence, the recent development of protein polymers for state-of-the-art bone tissue engineering has been discussed, highlighting the significant challenges and future perspectives.

Genomic characterization of Escherichia coli harbor a polyketide synthase ( pks ) island associated with colorectal cancer (CRC) development.

The E. coli strain harboring the polyketide synthase ( Pks) island encodes the genotoxin colibactin, a secondary metabolite reported to have severe implications for human health and for the progression of colorectal cancer. The present study involved whole-genome-wide comparison and phylogenetic analysis of pks harboring E. coli isolates to gain insight into the distribution and evolution of these organism. Fifteen E. coli strains isolated from patients with ulcerative colitis were sequenced, 13 of which harbored pks islands. In addition, 2,654 genomes from the public database were also screened for pks harboring E. coli genomes, 158 of which were pks -positive isolates. Whole-genome-wide comparison and phylogenetic analysis revealed that 171 (158+13) pks -positive isolates belonged to phylogroup B2, and most of the isolates associated to sequence types ST73 and ST95. One isolate from an ulcerative colitis (UC) patient was of the sequence type ST8303. The maximum likelihood tree based on the core genome of pks -positive isolates revealed horizontal gene transfer across sequence types and serotypes. Virulome and resistome analyses revealed the preponderance of virulence genes and a reduced number of antimicrobial genes in Pks -positive isolates. This study strongly contributes to understanding the evolution of pks islands in E. coli .

Emotion brain network topology in healthy subjects following passive listening to different auditory stimuli.

A large body of research establishes the efficacy of musical intervention in many aspects of physical, cognitive, communication, social, and emotional rehabilitation. However, the underlying neural mechanisms for musical therapy remain elusive. This study aimed to investigate the potential neural correlates of musical therapy, focusing on the changes in the topology of emotion brain network. To this end, a Bayesian statistical approach and a cross-over experimental design were employed together with two resting-state magnetoencephalography (MEG) as controls. MEG recordings of 30 healthy subjects were acquired while listening to five auditory stimuli in random order. Two resting-state MEG recordings of each subject were obtained, one prior to the first stimulus (pre) and one after the final stimulus (post). Time series at the level of brain regions were estimated using depth-weighted minimum norm estimation (wMNE) source reconstruction method and the functional connectivity between these regions were computed. The resultant connectivity matrices were used to derive two topological network measures: transitivity and global efficiency which are important in gauging the functional segregation and integration of brain network respectively. The differences in these measures between pre- and post-stimuli resting MEG were set as the equivalence regions. We found that the network measures under all auditory stimuli were equivalent to the resting state network measures in all frequency bands, indicating that the topology of the functional brain network associated with emotional regulation in healthy subjects remains unchanged following these auditory stimuli. This suggests that changes in the emotion network topology may not be the underlying neural mechanism of musical therapy. Nonetheless, further studies are required to explore the neural mechanisms of musical interventions especially in the populations with neuropsychiatric disorders.

From port to planet: Assessing NO2 pollution and climate change effects with Sentinel-5p satellite imagery in maritime zones.

The growing effects of climate change on Malaysia's coastal ecology heighten worries about air pollution, specifically caused by urbanization and industrial activity in the maritime sector. Trucks and vessels are particularly noteworthy for their substantial contribution to gas emissions, including nitrogen dioxide (NO2), which is the primary gas released in port areas. The application of advanced analysis techniques was spurred by the air pollution resulting from the combustion of fossil fuels such as fuel oil, natural gas and gasoline in vessels. The study utilized satellite photos captured by the Tropospheric Monitoring Instrument (TROPOMI) on the Sentinel-5P satellite to evaluate the levels of NO2 gas pollution in Malaysia's port areas and exclusive economic zone. Before the COVID-19 pandemic, unrestricted gas emissions led to persistently high levels of NO2 in the analyzed areas. The temporary cessation of marine industry operations caused by the pandemic, along with the halting of vessels to prevent the spread of COVID-19, resulted in a noticeable decrease in NO2 gas pollution. In light of these favourable advancements, it is imperative to emphasize the need for continuous investigation and collaborative endeavours to further alleviate air contamination in Malaysian port regions, while simultaneously acknowledging the wider consequences of climate change on the coastal ecology. The study underscores the interdependence of air pollution, maritime activities and climate change. It emphasizes the need for comprehensive strategies that tackle both immediate environmental issues and the long-term sustainability and resilience of coastal ecosystems in the context of global climate challenges.

Synthesis, In vivo, and In silico Evaluation of New Pyrazoline-Benzothiazole Conjugates as Antiepileptic Agents.

New 2-(4-benzothiazol-2-yl-phenoxy)-1-(3,5-diphenyl-4,5-dihydro-pyrazol-1-yl)-ethanones (9a-o) have been designed and synthesized. All the synthesized compounds were characterized by thin layer chromatography and spectral analysis. The antiepileptic potential of the synthesized compounds has been tested by following standard animal screening models, including maximal electroshock (MES) and subcutaneous pentylenetetrazole (scPTZ) models. The neurotoxic and antidepression effects of the synthesized compounds were checked by utilizing rotarod apparatus, and motor impairment test (by actophotometer) respectively. The study concluded that compounds 9c, 9d, 9f, 9i, 9n, and 9o possessed good antiepileptic potential compared to standard drugs like carbamazepine and phenytoin. The results of the rotarod performance test also established them without any neurotoxicity. The motor impairment test revealed that the synthesized compounds are also good antidepressants. In-silico studies have been performed for calculation of pharmacophore pattern, prediction of pharmacokinetic properties which determine the eligibility of synthesized compounds as orally administered molecules and interactions with the target proteins. The result of in-silico studies reinforced results obtained by in vivo study of the synthesized compounds and their possible mechanism of antiepileptic action i. e. via inhibiting voltage-gated sodium channels (VGSCs) and gamma-aminobutyric acid-A receptor.

Angiomyolipoma during pregnancy: A forgotten risk factor for rupture and massive haemorrhage - A case report.

Benign renal lesions are relatively rare. Angiomyolipoma (AML) is the most commonly encountered benign renal lesion. One of the complications of AML is rupture, which results in retroperitoneal haemorrhage with a mortality rate of up to 20%. Pregnancy poses a major risk for the rupture of AML. This is attributed to its hormonal effect, which causes the tumour to grow rapidly during pregnancy. The possibility of AML rupture should be considered when encountering pregnant patients with hypovolemic shock but with normal initial obstetric ultrasound findings. We present a case of a pregnant patient who was admitted with hypovolemic shock and CT scan confirmed rupture of AML.

Surgical Management of Huge Ranula Resembling Double Tongue in Pediatric Patients: A Successful Treatment of Two Cases.

Ranula is a fluid collection in a pseudo cystic wall secondary to the damage of the sublingual salivary gland causing blockage of salivary flow, leading to the extravasation phenomena. The growth rate of ranula varies depending on its severity. Due to its tendency to recur, the gold standard management of ranula has yet to be decided. The authors described two cases of young girls with huge ranulas on the floor of the mouth (FOM) resembling double tongue, which caused pain and discomfort during mastication. Following surgical excision of the ranulas along with the affected sublingual glands, both cases demonstrated successful treatment outcomes with no recurrence observed during post-operative follow-up. These cases highlight the importance of surgical excision of ranulas and removal of affected sublingual glands to prevent recurrence.

Real-time fluorescence loop-mediated isothermal amplification assays for detection of zoonotic malaria Plasmodium parasites.

BACKGROUND: Natural human infections by Plasmodium cynomolgi and P. inui have been reported recently and gain the substantial attention from Southeast Asian countries. Zoonotic transmission of non-human malaria parasites to humans from macaque monkeys occurred through the bites of the infected mosquitoes. The objective of this study is to establish real-time fluorescence loop-mediated isothermal amplification (LAMP) assays for the detection of zoonotic malaria parasites by combining real-time fluorescent technology with the isothermal amplification technique. METHODS: By using 18S rRNA as the target gene, the primers for P. cynomolgi, P. coatneyi and P. inui were newly designed in the present study. Four novel real-time fluorescence LAMP assays were developed for the detection of P. cynomolgi, P. coatneyi, P. inui and P. knowlesi. The entire amplification process was completed in 60 min, with the assays performed at 65 °C. By using SYTO-9 as the nucleic acid intercalating dye, the reaction was monitored via real-time fluorescence signal. RESULTS: There was no observed cross-reactivity among the primers from different species. All 70 field-collected monkey samples were successfully amplified by real-time fluorescence LAMP assays. The detection limit for P. cynomolgi, P. coatneyi and P. knowlesi was 5 × 109 copies/µL. Meanwhile, the detection limit of P. inui was 5 × 1010 copies/µL. CONCLUSION: This is the first report of the detection of four zoonotic malaria parasites by real-time fluorescence LAMP approaches. It is an effective, rapid and simple-to-use technique. This presented platform exhibits considerable potential as an alternative detection for zoonotic malaria parasites.

Relational aggression in romantic relationship: empirical evidence among young female adults in Malaysia.

BACKGROUND: Aggressive behaviour in romantic relationship is a social problem of great concern. Studies related to the influence of psychosocial factors on relational aggression are still limited. Furthermore, these factors have not been widely studied in the local context, resulting in the issue of relational aggression among young female adults still not being addressed. This study aims to explore whether psychosocial factors such as big five personality traits, adult attachment style and loneliness could predict relational aggression in romantic relationships among young female adults in Malaysia. In addition, this study aims to identify the moderating effect of social support in the relationship between psychosocial factors and relational aggression in romantic relationship. METHODS: A quantitative research approach was used with 424 young female adults in Malaysia aged between 18 and 30 years old (mean age = 24.18) were recruited through multistage sampling design by completing a questionnaire consisting of the Big Five Inventory (BFI), Experiences in Close Relationships Scale II (ECRS-II), Revised UCLA Loneliness Scale, Measure of Relational Aggression and Victimization (MRAV) and Multidimensional Scale of Perceived Social Support (MSPSS). RESULTS: Multiple regression analysis predicted significant relationship between agreeableness personality, loneliness, avoidant attachment style and anxious attachment style with relational aggression in romantic relationships. Hierarchical regression analysis found a significant effect of social support as a moderator between loneliness with relational aggression in romantic relationships. CONCLUSIONS: Thus, the results show that young female adults with low level of agreeableness, high level of loneliness, avoidant attachment style and anxious attachment style are at a higher risk of engaging in relational aggression in romantic relationships. The implication of this study can help in understanding the psychosocial factors that form the basis of relational aggression in romantic relationships. Hence, the gap in knowledge warrants further research.

Anorectal malformation and Hirschsprung's disease: a cross-sectional multicentre comparison of quality of life and bowel function to a healthy population.

PURPOSE: Children with anorectal malformation (ARM) and Hirschsprung's disease (HD) often experience bowel symptoms into adulthood, despite definitive surgery. This study evaluates the quality of life (QOL) and bowel functional outcome of children treated for ARM and HD in comparison to healthy controls. METHODS: Between December 2020 and February 2023, we recruited patients with ARM and HD aged 3-17 years at four tertiary referral centres, who had primary corrective surgery done >12 months prior. Healthy controls were age-matched and sex-matched. All participants completed the Pediatric Quality of Life Inventory Generic Core Scales 4.0, General Well-Being (GWB) Scale 3.0 and Family Impact (FI) Module 2.0 Questionnaires. Bowel Function Score (BFS) Questionnaires were also administered. We also performed subgroup analysis according to age categories. Appropriate statistical analysis was performed with p<0.05 significance. Ethical approval was obtained. RESULTS: There were 306 participants: 101 ARM, 87 HD, 118 controls. Patients with ARM and HD had significantly worse Core and FI Scores compared with controls overall and in all age categories. In the GWB Scale, only ARM and HD adolescents (13-17 years) had worse scores than controls. ARM and HD had significantly worse BFSs compared with controls overall and in all age categories. There was significant positive correlation between BFS and Core Scores, GWB Scores and FI Scores. CONCLUSION: Patients with ARM and HD had worse QOL than controls. Lower GWB Scores in adolescents suggests targeted interventions are necessary. Bowel function influences QOL, indicating the need for continuous support into adulthood.

Evaluating climate change impacts on reef environments via multibeam echosounder and Acoustic Doppler Current profiler technology.

Crucial to the Earth's oceans, ocean currents dynamically react to various factors, including rotation, wind patterns, temperature fluctuations, alterations in salinity and the gravitational pull of the moon. Climate change impacts coastal ecosystems, emphasizing the need for understanding these currents. This study explores multibeam echosounder (MBES), specifically R2-Sonic 2020 instrument, offering detailed seabed information. Investigating coral reefs, rocky reefs and artificial reefs aimed to map seafloor currents movement and their climate change responses. MBES data viz. Bathymetry and backscatter were classified and acoustic doppler current profiler (ADCP) ground data were validated using random forest regression. Results indicated high precision in currents speed measurement i.e. coral reefs with 0.96, artificial reefs with 0.94 and rocky reefs with 0.97. Currents direction accuracy was notable in coral reefs with 0.85, slightly lower in rocky reefs with 0.72 and artificial reefs with 0.60. Random forest identified sediment and backscatter as key for speed prediction while direction relies on bathymetry, slope and aspect. The study emphasizes integrating sediment size, backscatter, bathymetry and ADCP data for seafloor current analysis. This multibeam data on sediments and currents support better marine spatial planning and determine biodiversity patterns planning in the reef area.

Thermal Properties and Drying Shrinkage Performance of Palm Kernel Shell Ash and Rice Husk Ash-Based Geopolymer Concrete.

This study aims to develop suitable formulations of geopolymer concrete (GPC) by varying the percentages of the geopolymer with aggregates and evaluating the performances in thermal and mechanical properties of palm kernel shell ash (PKSA)-GPC compared to rice husk ash (RHA)-GPC and ordinary Portland cement concrete (OPCC). Preliminary tests were conducted to select the best mix design ratios before casting the specimens. Then, the performance of the PKSA-GPC, RHA-GPC and OPCC specimens was evaluated based on their thermal performance and drying shrinkage. The mix designs of PKSA-GPC 70:30, PKSA-GPC 60:40, PKSA-GPC 50:50 and PKSA-GPC 66.6:33.3 were found to produce an acceptable consistency, rheological and thixotropic behaviour for the development of the GPC. PKSA-GPC showed a better thermal performance than the RHA-GPC and OPCC due to their strong and dense intumescent layers and slow temperature increment upon exposure to a high flame temperature from ambient temperature to 169 °C. The low molar ratio of the Si/Al present in the PKSA-GPC created a thermally stable intumescent layer. In the drying shrinkage test, PKSA-GPC 60:40 and RHA-GPC 60:40 shared an equal drying shrinkage performance (5.040%) compared to the OPCC (8.996%). It was observed that microcrack formation could significantly contribute to the high shrinkage in the PKSA-GPC 50:50 and RHA-GPC 70:30 specimens. The findings of this study show that PKSA could be incorporated into GPC as a fire-retardant material due to its capability of prolonging the spread of fire upon ignition and acting as an alternative to the conventional OPCC.

The prevalence of simian malaria in wild long-tailed macaques throughout Peninsular Malaysia.

The parasite Plasmodium knowlesi has been the sole cause of malaria in Malaysia from 2018 to 2022. The persistence of this zoonotic species has hampered Malaysia's progress towards achieving the malaria-free status awarded by the World Health Organisation (WHO). Due to the zoonotic nature of P. knowlesi infections, it is important to study the prevalence of the parasite in the macaque host, the long-tailed macaque (Macaca fascicularis). Apart from P. knowlesi, the long-tailed macaque is also able to harbour Plasmodium cynomolgi, Plasmodium inui, Plasmodium caotneyi and Plasmodium fieldi. Here we report the prevalence of the 5 simian malaria parasites in the wild long-tailed macaque population in 12 out of the 13 states in Peninsular Malaysia using a nested PCR approach targeting the 18s ribosomal RNA (18s rRNA) gene. It was found that all five Plasmodium species were widely distributed throughout Peninsular Malaysia except for states with major cities such as Kuala Lumpur and Putrajaya. Of note, Pahang reported a malaria prevalence of 100% in the long-tailed macaque population, identifying it as a potential hotspot for zoonotic transmission. Overall, this study shows the distribution of the 5 simian malaria parasite species throughout Peninsular Malaysia, the data of which could be used to guide future malaria control interventions to target zoonotic malaria.

Bilateral Buccal Mucosa Graft for Urethroplasty: A Versatile Graft.

Male urethral stricture is scarring of the urethral tissue that narrows the urethral lumen causing reduced urinary flow. Urethral reconstruction or substitution urethroplasty using oral mucosa graft, especially from the buccal mucosa, is one of the most widely known techniques to manage urethral stricture. However, studies using bilateral buccal mucosa are still limited. Therefore, this study aims to report our experience and technique of bilateral buccal mucosa grafting for urethroplasty. The authors described a 66-year-old man with long-segment urethral stricture that was successfully treated with urethral reconstruction harvested from bilateral buccal mucosa.

The role of sheet-like TiO2 in polyamide reverse osmosis membrane for enhanced removal of endocrine disrupting chemicals.

Thin film composite (TFC) reverse osmosis (RO) membrane shows good promise for treating wastewater containing endocrine disrupting chemical (EDC) pollutants. The incorporation of functional materials with exceptional structural and physico-chemical properties offers opportunities for the membranes preparation with enhanced permselectivity and better antifouling properties. The present study aims to improve the EDC removal efficiency of TFC RO membrane using two-dimensional titania nanosheets (TNS). RO membrane was prepared by incorporating TNS in the dense layer of polyamide (PA) layer to form thin film nanocomposite (TFN) membrane. The TNS loading was varied and the influences on membrane morphology, surface hydrophilicity, surface charge, as well as water permeability and rejection of EDC were investigated. The results revealed that the inclusion of TNS in the membrane resulted in the increase of water permeability and EDC rejection. When treating the mixture of bisphenol A (BPA) and caffeine at 100 ppm feed concentration, the TFN membrane incorporated with 0.05% TNS achieved water permeability of 1.45 L/m2·h·bar, which was 38.6% higher than that of unmodified TFC membrane, while maintaining satisfactory rejection of >97%. The enhancement of water permeability for TFN membrane can be attributed to their hydrophilic surface and unique nanochannel structure created by the nanoscale interlayer spacing via staking of TiO2 nanosheets. Furthermore, the 0.05TFN membrane exhibited excellent fouling resistance towards BPA and caffeine pollutants with almost 100% flux recovery for three cycles of operations.

Unravelling trends, patterns and intellectual structure of research on bankruptcy in SMEs: A bibliometric assessment and visualisation.

Despite the burgeoning interest among academics in investigating the factors contributing to the high business failure rate among SMEs (small and medium-sized enterprises), the systematic synthesis of the literature on bankruptcy in SMEs is restricted. This article aims to significantly advance the understanding of the causes and repercussions of bankruptcy in SMEs and the preventative actions that may be taken to avoid it. This review assesses 282 articles from 175 outlets employing quantitative and statistics-based bibliometric tools. This bibliometric assessment helped delineate the citation and publication trends and the top contributors to the domain. The underlying thematic clusters of research on bankruptcy in SMEs were also identified, deciphered and elaborated, along with charting the future research vistas through the lens of theory, context, and methods framework. The authors believe this bibliometric variant of systematic literature review makes a significant contribution to bankruptcy and SME research by highlighting the development of the literature and some of the most active research fronts in the domain by offering insights that were not clasped thoroughly or assessed by prior literature assessments.

Genetic polymorphism and clustering of the Plasmodium cynomolgi Duffy binding protein 1 region II of recent macaque isolates from Peninsular Malaysia.

Plasmodium cynomolgi is a simian malaria parasite that has been increasingly infecting humans. It is naturally present in the long-tailed and pig-tailed macaques in Southeast Asia. The P. cynomolgi Duffy binding protein 1 region II [PcDBP1(II)] plays an essential role in the invasion of the parasite into host erythrocytes. This study investigated the genetic polymorphism, natural selection and haplotype clustering of PcDBP1(II) from wild macaque isolates in Peninsular Malaysia. The genomic DNA of 50 P. cynomolgi isolates was extracted from the macaque blood samples. Their PcDBP1(II) gene was amplified using a semi-nested PCR, cloned into a plasmid vector and subsequently sequenced. The polymorphism, natural selection and haplotypes of PcDBP1(II) were analysed using MEGA X and DnaSP ver.6.12.03 programmes. The analyses revealed high genetic polymorphism of PcDBP1(II) (π = 0.026 ± 0.004; Hd = 0.996 ± 0.001), and it was under purifying (negative) selection. A total of 106 haplotypes of PcDBP1(II) were identified. Phylogenetic and haplotype analyses revealed two groups of PcDBP1(II). Amino acid length polymorphism was observed between the groups, which may lead to possible phenotypic difference between them.

Proteomic profiling of membrane vesicles from Mycobacterium avium subsp. paratuberculosis: Navigating towards an insilico design of a multi-epitope vaccine targeting membrane vesicle proteins.

Bacteria typically produce membrane vesicles (MVs) at varying levels depending on the surrounding environments. Gram-negative bacterial outer membrane vesicles (OMVs) have been extensively studied for over 30 years, but MVs from Gram-positive bacteria only recently have been a focus of research. In the present study, we isolated MVs from Mycobacterium avium subsp. paratuberculosis (MAP) and analyzed their protein composition using LC-MS/MS. A total of 316 overlapping proteins from two independent preparations were identified in our study, and topology prediction showed these cargo proteins have different subcellular localization patterns. When MVs were administered to bovine-derived macrophages, significant up-regulation of pro-inflammatory cytokines was observed via qRT-PCR. Proteome functional annotation revealed that many of these proteins are involved in the cellular protein metabolic process, tRNA aminoacylation, and ATP synthesis. Secretory proteins with high antigenicity and adhesion capability were mapped for B-cell and T-cell epitopes. Antigenic, Immunogenic and IFN-γ inducing B-cell, MHC-I, and MHC-II epitopes were stitched together through linkers to form multi-epitope vaccine (MEV) construct against MAP. Strong binding energy was observed during the docking of the 3D structure of the MEV with the bovine TLR2, suggesting that the putative MEV may be a promising vaccine candidate against MAP. However, in vitro and in vivo analysis is required to prove the immunogenic concept of the MEV which we will follow in our future studies. SIGNIFICANCE: Johne's disease is a chronic infection caused by Mycobacterium avium subsp. paratuberculosis that has a potential link to Crohn's disease in humans. The disease is characterized by persistent diarrhea and enteritis, resulting in significant economic losses due to reduced milk yield and premature culling of infected animals. The dairy industry in the United States alone experiences losses of approximately USD 250 million due to Johne's disease. The current vaccine against Johne's disease is limited by several factors, including variable efficacy, limited duration of protection, interference with diagnostic tests, inability to prevent infection, and logistical and cost-related challenges. Nevertheless, a multiepitope vaccine design approach targeting M. avium subsp. paratuberculosis has the potential to overcome these challenges and offer improved protection against Johne's disease.