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Purpose: This work investigated carboxymethyl hydroxypropyl guar (CMHPG) as a potential base polymer for a fracturing fluid used in deep tight gas wells, synonym to high-temperature reservoirs. Fracturing is a common method to improve oil production by creating fractures in potentially productive reservoirs using fracturing fluids. Fracturing fluids are typically composed of water, sand, proppant, and many different additives to alter the capabilities of the fluid. Design/methodology/approach: A stable high-density fracturing fluid (HDFF) was developed to cope with scorching good temperatures up to 350°F (177°C)as well as to reduce the surface treating pressure. The HDFF consists of NaBr heavy brine, CMHPG, clay stabilizers, and zirconium and borate crosslinkers. A linear gel fluid was prepared by combining NaBr brine with CMHPG before adding the additives. The resulting fluid was then tested using a high-pressure, high-temperature (HPHT) rheometer. Findings: The results showed that the performance of the HDFF was optimum in an alkaline environment between pH 10 to 12. The efficiency of zirconium and borate crosslinkers were optimum at 0.7 ppm and 1.5 ppt, respectively. Concentrations higher than these values are not only uneconomical but will cause the fluid to be overly crosslinked, consequently reducing efficiency. In actual field operation, this is disastrous when the fluid does not flow to the fracturing column but instead swirls around the drill string as the drill string rotates. On the other hand, a reduction of concentration below the optimum values can cause fluid instability at high temperature reservoirs. Research limitations/implications: The study was limited to using two different kinds of crosslinkers and various concentrations. Future studies can be conducted using other kinds of crosslinkers, as well as an investigation into the effects of varying temperatures, pressures, and pH on the HDFF using CMHPG. Practical implications: It can be concluded that HDFF using CMHPG as the base polymer can be a potential use in formulating fracturing fluids. [ABSTRACT FROM AUTHOR]

Fibroma of the Tendon Sheath (FTS) is a rare soft tissue tumor commonly found in upper extremities. Usually, the lesions occur in wrists and hands. There are only a few cases of fibroma of the tendon sheath arising from the lower extremities have been reported. This case report describes a rare presentation of painful FTS that arises from flexor hallucis longus at the plantar aspect of the forefoot in a middle age lady. To our knowledge there is only one similar case reported previously. [ABSTRACT FROM AUTHOR]

In this study, the experiment on the modification of the photoanode with organic polymer material as copolymer template for dye-sensitized solar cell (DSSC) applications has been conducted. The two organic copolymer templates are polystyrene sphere (PS) and poly[2- methoxy-5(2-ethylhexyloxy)-1,4-phenylenevinylene] (MEH-PPV). The modification photoanodes were made using Dr. Blade’s method. These organic copolymer templates were added to improve the surface of the mesoporous titanium dioxide (TiO2) layer, which is used as the main component in DSSC photoanode. The unmodified TiO2 photoanode has poor aggregation and porosity of TiO2. The addition of either MEH-PPV or PS sphere to the photoanode layer was found to affect the surface of mesoporous TiO2 in terms of porosity, particle size distribution and shape. The analysis of the TiO2 modification was conducted using an image analysis processing method via a 2D scanning electron microscope (SEM) image. The image analysis processing method used was the ImageJ program. The DSSC of modified photoanode is fabricated using metal complex dye, Ruthenium (N719) dye. The data collected from the ImageJ program showed that by adding organic copolymer templates into TiO2, the porosity of TiO2 decreased from 45 % to 42 %. From the photovoltaic analysis obtained, the J-V characteristic is recorded with the photoanode of TiO2 mixed with 1.00 wt% MEH-PPV gave the highest efficiency, which is 0.01 % with the following parameters – Voc = 0.43 V, Jsc = 0.17 mA/cm2 and FF = 0.20. Meanwhile, the photoanode of TiO2 mixed with 0.50 wt% PS sphere gave the highest efficiency which is 0.08 % with the following parameters – Voc = 0.39 V, Jsc = 0.86 mA/cm2 and FF = 0.25. [ABSTRACT FROM AUTHOR]

The paradigm of choice practice represents the psychological theory of learning in the development of moral judgment. It is concerned with evaluating the implications of several choices and selecting one of them to implement. The goal of this work is to provide a generic functional equation to observe the behavior of animals in such circumstances. Our suggested functional equation can be employed to describe several well-known psychology and learning theories. The fixed point theorem proposed by Banach is utilized to show that the solution of a given functional problem exists and is unique. In addition, the stability of the given functional equation’s solution is discussed in terms of the Hyers-Ulam and Hyers-Ulam-Rassias results. Furthermore, two examples are provided to highlight the relevance of the significant outcomes in the context of the literature.

Primary villous adenoma originating from the urinary tract is an infrequent entity. We present a rare case of villous adenoma arising from a prostatic urethra with no sign of malignant transformation. Villous adenoma should be considered as one of the differential diagnoses of urethral lesions, especially if it has similar magnetic resonance imaging features as its colonic counterpart. Due to its potential for malignant transformation, its complete resection is mandatory. [ABSTRACT FROM AUTHOR]

In this paper, a collocation method which based on polynomial approximation of Taylor's series is proposed to approximate the solution of fractional pantograph differential equations (FPDE). The collocation method with truncated Taylor's polynomial is shown to be an applicable technique in solving FDDE. Some examples of the non-linear fractional pantograph differential equations are solved and compared with the exact solution to confirm the accuracy and applicability of the collocation method with Taylor's polynomial. [ABSTRACT FROM AUTHOR]

Sustainable urban farming is a strategy to improve food availability and food access, and to support food security for the urban population in Malaysia. However, the development of these activities has been affected by several constraints. This article aims to identify the challenges faced by urban farmers in Kuala Lumpur. The challenges of practicing urban gardening were categorised into five groups (technical, resource-related, economic, social and environmental factors). Data were collected via a questionnaire survey distributed to 106 urban farming practitioners from 17 urban gardens in Kuala Lumpur and were analysed using descriptive analysis by tabulating the frequency and percentage. The result showed that highly fluctuating weather, problems with access to available land and financial problems were the main challenges faced by urban farmers in Kuala Lumpur. Furthermore, difficulty in access to a financial institution, lack of commitment and the increased number of pests were also the problems faced by the urban garden. Availability of technical factors is the least issue in this study. Correlation analysis was used to determine the relationship between the challenges of urban gardens and socio-demographics. The result showed that there was a weak correlation between technical factors of educational level (r = 0.225) and race (r = 0.210), respectively, as well as between race and social factor (r = 0.201), while there was a moderate correlation between age and environment factor (r = −0.410). There is a need for further work, and comprehensive research should be conducted to capture what actions can be taken to create a policy-making space for urban farmers.

Heat treatment by heating furnace has limited the capability of heat treatment processes to control the desired heating volume and products' sizes. This study introduced a heat treatment process using light heat sourced from a spot-type halogen lamp, with the aim of clarifying the thermal phenomenon of aluminium (Al) foam. Here, the temperature and deformation of Al foam were observed experimentally. In addition, thermal analysis was conducted numerically using a uniform heating model by neglecting the energy consumption of the blowing agent and deformation and thermal conductivity of Al. The experimental results revealed that the precursor initially formed at almost the same time as the phase change. The numerical results almost corresponded with the experimental results until the melting point of Al, after which a slight disagreement was observed. Moreover, the phase transformation appeared slightly earlier as a result of neglecting the deformation of Al foam and energy consumption of the blowing agent. Although the formation of Al foam cannot be expressed perfectly using the proposed uniform heating model, a qualitative phenomenon of such formation was successfully explained. In the future, the effects of deformation and energy consumption of the blowing agent and the thermal conductivity of Al can be considered in the proposed uniform heating model.. [ABSTRACT FROM AUTHOR]

In the evolution of the complement system, a major humoral innate immune factor, the existence of multiple isotypes of the complement components is considered as a key strategy to enhance innate immune defense. Complement C4 is also diversified in a wide range of vertebrate species including teleost fish, possibly supporting the robust activation mechanism of the complement. To better understand the functional diversity of C4 isotypes in the teleost complement system, two C4 isotypes, C4-1 and C4-2, sharing only 32% amino acid sequence identity, were examined for binding specificities towards model target molecules representing microbe antigens and towards Gram-positive and -negative bacteria. The results suggest that C4-1 and C4-2 behave similarly in binding to the tested targets, despite the predicted difference in binding specificity based on the thioester catalytic site. The participation of C4-1 in the classical and lectin pathways of complement activation was also explored using pathway-specific activating enzyme complexes, C1r/s and MBL-MASP2. As a result, C4-1 can be activated in both the classical and the lectin pathways, at higher efficiency in the classical pathway. Taken together, the present results imply that both C4-1 and C4-2 isotypes are fully functional in the complement activation cascades, probably playing comparable roles in innate immunity.

Knowledge of species richness and distribution is decisive for the composition of conservation areas. Plants typically contain many bioactive compounds are used for medicinal purposes for several disease treatment. This study aimed to identify the plant species distribution in area of UiTM Kuala Pilah, providing research scientific data and to contribute to knowledge of the use of the plants as therapeutic resources. Three quadrat frames (1x1 m), which was labeled as Set 1, 2 and 3 was developed, in each set consists of 4 plots (A, B, C and D). Characteristics of plant species were recorded, identified and classified into their respective groups. A total of 28 plant species belonging to 18 families were identified in all sets with the largest family of Rubiaceae. The most distribution species are Desmodium triflorum, Dactyloctenium aegyptium, Flavoparmelia caperata, Xanthoria elegans and Phlyctis argena. Most of the plant possesses their potential to treat skin diseases, fever, ulcers, diabetes as well as digestive problems with their antimicrobial, anti-inflammatory and antioxidant properties. This study suggests that study site and plant species can be delineated as an important area to preserve these therapeutic sources.

Internship is critical as it allows hands-on experience for what they have learned in classroom in real settings. It meant to test their career interests and acts as a personal development opportunity. However, little is known about the work environment they were in, as well as their internship experience had on their personal development. In this paper, we present a descriptive analysis from a survey where the responses were collected from 104 students who had underwent a 16 weeks paid internship. The purpose is to assess the work environment factors especially the amount work stress and workplace bullying they have had experience. Later we conducted an inferential analysis to determine the effect of work stress and workplace bullying have on their personal development. The outcomes from this study have the potential to bridge the expectation-reality gap on the career that they will be joining, to strengthen the bonds between university-industry and to fill the internship literature gap on personal development.

The aims of this study are to report on the extraction and characterization of Asian swamp eel (Monopterus albus) skin gelatin. The characterization conducted were includes chemical composition, pH, gel strength, viscosity, thermal property, color and structure determination of extracted eel skin gelatin. Eel skin contains 70.28% moisture, 11.07% protein, 4.21% fat, and 5.01% ash. The chemical composition of eel skin gelatin (yield of 12.75%) was 18.8% moisture, 67.64% protein, 0.34% fat and 1.08% ash, with a pH of 4.62 and gel strength of 215.96 g (± 9.62 g). Although viscosity (2.8 cPa/min) profile of eel skin gelatin showed lower than that of bovine gelatin, the higher melting temperature (35 °C) of eel skin gelatin indicating its higher stability than bovine gelatin with FTIR spectrum similar to that of typical bovine gelatin. Eel skin gelatin has a 71.4 (± 1.14), a +3.2 (± 0.29), and a +7.52 (± 0.29) for L*, a* and b* value respectively, indicate a darker and less yellow colour. These findings show promising potential for the application of eel skin gelatin as an alternative to commercial gelatin. [ABSTRACT FROM AUTHOR]

This study is an attempt to investigate the adsorption of petroleum hydrocarbon (toluene) from aqueous solutions using granular activated carbon (GAC) synthesized from oil palm shell (OPS) (referred as OPSbased GAC). This study involved a series of batch experiments to determine the adsorption equilibrium and kinetics. The batch experiments were conducted by shaking 200 mL toluene solution containing 0.4 g GAC (initial concentrations of 5, 15, 25 and 30 mg/L) at 180 rpm at 30°C. The OPS-based GAC achieved more than 80% toluene removal in all the experiments. The adsorption capacity of the OPSbased GAC estimated using Freundlich isotherm was 6.039 mg/g (L/mg)1/n. The adsorption kinetic study showed that the adsorption of toluene was of chemisorption as the experimental data fitted better to the pseudo-second-order kinetic model than the pseudo-first-order kinetic model. [ABSTRACT FROM AUTHOR]

Misconduct often happens in the construction industry because of its scale, complex contract structure, competitive and fragmented nature. It leads to an unethical issue. The unethical issues and the impacts hindered the development of the construction industry in Malaysia and affected the national growth development. As the construction industry is now moving forward to the 4th industrial revolution, it is worth investigating whether the adoption of digital transformation can make some changes to the unethical issues in the Malaysian construction industry. This research paper aims to examine the construction players' perception of the usage of Building Information Modelling (BIM) in reducing the unethical issues in the Malaysian construction industry. A quantitative research method is adopted and the data is collected by a questionnaire survey. There were a total of 101 responses received from the construction industry practitioners in Malaysia. Data collected was analysed by reliability test and descriptive test for further evaluation and analysis. The finding shows the usage of BIM through its function of 3D project visualization, clash detection, better communication flow, and centralised single repository data storage are potentially reduced unethical issues. This research provides a better understanding to the construction industry practitioners on the possibility of BIM implementation as a potential tool in reducing unethical issues in the construction industry.

In construction industry, almost every construction project has to deal with construction contract, due to the project complexity and high financial risk. The contracts are developed and published by various associations and professional bodies and are widely used in the construction industry. Contract conditions are intended to specify the duties and rights of the contractual parties and must be properly drafted. Contractual parties under a particular contract must be aware of the details of the contract conditions. In Malaysian construction industry, there are several standard forms of main contracts and nominated subcontracts. However, for domestic subcontract, main contractors prefer to use bespoke contract, which the parties modify the standard contracts by omitting several clauses or adding new clauses; thus, potentially compromise the completeness and accuracy of the contract. Hence, the aim of this study is to examine the crucial conditions in the domestic subcontract commonly approached by main contractor in construction projects. A quantitative research method is adopted and the data have been collected by survey questionnaires. There are a total of 100 valid responses received from the construction industry practitioners in Malaysia. The survey objective is to rank the conditions in domestic subcontract from the most to the less important. As the result, the top 5 of relatively important domestic subcontract conditions, out of 27 conditions, are the terms of payment and final account, extension of time, variation, contractor’s and subcontractor’s obligations, and performance bond. The results of these surveys will enhance the contractual knowledge of main contractor and domestic subcontractor and improve awareness of crucial conditions that are prerequisite for the development of domestic subcontract.

Selected endophytic fungi have been report to be inulin degraders to produce fructose or other oligosaccharides. In this study, the Aspergillus sp. producing inulinase were isolated from selected plant species at Serdang area in Malaysia. Fungal isolates were screened solely based on inulin degrading enzymes production and two isolates named Asf1 and Onf1 were selected as the best inulinase enzyme producers. Genomic DNA of these two isolates were extracted and amplified by polymerase chain reaction (PCR). A 1,341 bp DNA fragment containing inulinase gene was successfully amplified from Asf1 fungal isolate and was named as inu2 gene in this study. Based on the morphological characteristics, rDNA and neighbour-joining phylogenetic analysis, Asf1 fungal isolate could display closely-related to the genus of Aspergillus. The complete sequence designated Asf1 Inu2 gene was successfully obtained via rapid-amplification of cDNA ends-polymerase chain reaction (RACE-PCR). A 2.3 kb DNA fragment encoding endoinulinase, inu2, from Asf1 fungal isolate includes an open reading frame of 1,552 bp with calculated molecular weight of 55,954.1 Da and signal peptide sequence of 23 amino acids. The deduced amino acid sequence of the Asf1 inu2 displayed 97, 96, 69 and 22% identities to that of A. ficuum inu2, A. niger inuB, P. purpurogenum and K. marxianus, respectively. Phylogenetic analysis showed that fungal endo- and exo-inulinases have indepently evolved with the respective hydrolytic activities toward terminal and internal β-(2 → 1)-fructofuranosidic linkages in inulin. [ABSTRACT FROM AUTHOR]

Risk reporting is essential in notifying investors on how companies manage the risks they engage in. Even though companies nowadays release information through press releases, corporate websites, and other forms of communications, they still have to disclose more information in their annual reports. The objective of this study is to examine the determinants of voluntary risk disclosures of Shariah compliant companies (ShCCs) in Malaysia, focusing on the religiosity factor and the ownership structure. A quantitative analysis using secondary data was employed as a method to assess the annual reports of 116 Shariah compliant companies in Malaysia for the financial years of 2012 and 2013. The findings of this study reveal that the voluntary risk disclosures in ShCCs on average are not more than sixty per cent. This study indicates no relationship between the religiosity of board members and directors’ ownership to the level of voluntary risk disclosure. More importantly, this study finds the relationship between the existence of government ownership and voluntary risk disclosure in ShCCs. The study adds value to the current body of knowledge in voluntary risk disclosures and suggests the importance of risk management information in the annual reports of ShCCs as Malaysia grows as a leading country in the Islamic economy.

A review of methods development in lipase based biosensor for triglyceride determination was briefly discussed. This review focuses on the basic principle of triglyceride biosensor that includes performances of triglyceride biosensor such as limit of detection, response time, and optimization.

Motorcycle is one of private transportation which has been widely used in many countries including Malaysia. However, motorcycles are the most dangerous form of motorized transport. Royal Malaysian Police (PDRM) statistics recorded that motorcycle is the highest vehicle (45.9%) involved in traffic accident compared to other vehicles. The potential cause of the death to the motorcyclist was due to the head injury. One of strategy to mitigate this problem is through proper usage of safety helmet. Therefore, this paper was introduce a new approach on motorcyclist safety by using the Technology Acceptance Model (TAM) with additional determinants that contribute to behavioral intention and to increase the proper usage of safety helmets among Malaysian motorcyclists. The Structural Equation Modelling (SEM) was used to test the structural TAM proposed. The evaluation for structural model showed the goodness of fit indices are excellent fit. This study found that perceived ease of use, perceived usefulness and social norm are significant towards behavioral intention to use Safety Helmet Reminder System (SHR).

Understanding breast cancer genetic risk relies on identifying causal variants and candidate target genes in risk loci identified by genome-wide association studies (GWAS), which remains challenging. Since most loci fall in active gene regulatory regions, we developed a novel approach facilitated by pinpointing the variants with greater regulatory potential in the disease's tissue of origin. Through genome-wide differential allelic expression (DAE) analysis, using microarray data from 64 normal breast tissue samples, we mapped the variants associated with DAE (daeQTLs). Then, we intersected these with GWAS data to reveal candidate risk regulatory variants and analysed their cis-acting regulatory potential. Finally, we validated our approach by extensive functional analysis of the 5q14.1 breast cancer risk locus. We observed widespread gene expression regulation by cis-acting variants in breast tissue, with 65% of coding and noncoding expressed genes displaying DAE (daeGenes). We identified over 54 K daeQTLs for 6761 (26%) daeGenes, including 385 daeGenes harbouring variants previously associated with BC risk. We found 1431 daeQTLs mapped to 93 different loci in strong linkage disequilibrium with risk-associated variants (risk-daeQTLs), suggesting a link between risk-causing variants and cis-regulation. There were 122 risk-daeQTL with stronger cis-acting potential in active regulatory regions with protein binding evidence. These variants mapped to 41 risk loci, of which 29 had no previous report of target genes and were candidates for regulating the expression levels of 65 genes. As validation, we identified and functionally characterised five candidate causal variants at the 5q14.1 risk locus targeting the ATG10 and ATP6AP1L genes, likely acting via modulation of alternative transcription and transcription factor binding. Our study demonstrates the power of DAE analysis and daeQTL mapping to identify causal regulatory variants and target genes at breast cancer risk loci, including those with complex regulatory landscapes. It additionally provides a genome-wide resource of variants associated with DAE for future functional studies., (© 2024. The Author(s).)

The illegal trade in tigers (Panthera tigris) and their derivatives, such as bones, teeth and pelts, is a major threat to the species' long-term persistence. As wild tiger populations have dwindled, a large proportion of trafficked tiger products now derive from captive breeding facilities found throughout Asia. Moreover, wild tigers have been poached and laundered into captive facilities, then falsely designated as captive-bred. The establishment of a DNA registration system is recognized as a key tool to monitor compliance of captive facilities, support tiger trade investigations and improve prosecution outcomes. Here, we present a standardised wildlife forensic DNA profiling system for captive tigers called TigerBase. TigerBase has been developed in four South-East Asia countries with captive tiger facilities: Malaysia, Vietnam, Thailand and Lao PDR. TigerBase DNA profile data is based on 60 single nucleotide polymorphism (SNP) markers, genotyped using two different TaqMan®-based approaches: OpenArray® chip (capable of genotyping 60 SNPs for 48 samples in a single chip), and singleplex TaqMan® assays (capable of genotyping one SNP for one sample per reaction). Of the 60 SNPs, 53 are autosomal nuclear markers, suitable for individualisation and parentage applications, two are sex-linked markers, suitable for sexing, and five are mtDNA markers, suitable for maternal subspecies identification. We conducted a series of validation experiments to investigate the reliability and limitations of these SNP genotyping platforms. We found that the OpenArray® chip platform is more appropriate for generating reference data given its greater throughput, while the singleplex TaqMan® assays are more appropriate for genotyping lower quality casework samples, given their higher sensitivity and throughput flexibility. Only 19 autosomal nuclear markers were validated as singleplex TaqMan® assays, which generally provides ample power for individualisation analysis (probability of identity among siblings was <6.9 ×10 -4 ), but may lack power for specific parentage questions, such as determining parentage of an offspring when one of the parent's genotypes is missing. Further, we have developed pipelines to support standardised SNP calling and decrease the chance of genotyping errors through the use of analytical workflows and synthetic positive controls. We expect the implementation of TigerBase will enhance enforcement of tiger trafficking cases and encourage compliance among captive tiger facilities, together contributing to combatting the illegal tiger trade., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Dengue is a mosquito-borne disease caused by four distinct, closely related dengue viruses (DENV). Global dengue incidence has markedly increased in the past decades. The World Health Organization reported that cases increased from 505,430 in 2000 to 5.2 million in 2019. Similarly, the total dengue cases in Malaysia increased from 7,103 in 2000 to a peak of 130,101 in 2019. Knowledge, attitude, and practice (KAP) remain the most effective dengue prevention and control tools. Furthermore, school-based health education is key to enhancing knowledge and raising awareness of the seriousness of dengue among schoolchildren and transferring knowledge and practice from classrooms to homes. Thus, it is necessary to plan an integrated module for the primary prevention of dengue infection, specifically among schoolchildren., Aims: The present study intends to develop, implement, and evaluate the effectiveness of a theory-based integrated dengue education and learning (iDEAL) module in improving the KAP, environmental cleanliness index, and dengue index among schoolchildren in Selangor and Kuala Lumpur., Methods: This study is a single-blinded, cluster randomised controlled trial to be conducted from 1 September 2023 to 31 August 2025. The study will involve 20 primary and 20 secondary schools in Selangor and Kuala Lumpur. The 1600 participants will be randomly allocated to intervention and control groups based on selected clusters to avoid contamination. A cluster is a comparable school that fulfils the inclusion and exclusion criteria. The intervention group will receive the iDEAL module, while the control group will receive standard education. The iDEAL module will be developed following a systematic procedure and delivered in-person by trained researchers to the participants. The outcome will be measured using validated, self-administered questionnaires at baseline (T0), immediately (T1), one month (T2), and three months (T3) post-intervention to measure the intervention module effectiveness. The data will be analysed using IBM Statistical Package for Social Science (SPSS) version 28 and descriptive and inferential statistics. Within-group changes over time will be compared using one-way repeated measure analysis of variance for continuous and normally distributed variables. Within-group analysis of categorical data will use Cochran's Q test. The main effect and interaction between and within the intervention and control groups at T0, T1, T2, and T3 will be tested using the generalised linear mixed model (GLMM). Hypothetically, the KAP, environmental cleanliness index, and dengue index among the intervention group will be significantly improved compared to the control group. The hypothesis will be tested using a significance level with a p-value of 0.05 and a confidence interval of 95%., Conclusions: The study protocol outlines developing and testing an iDEAL module for schoolchildren in Selangor and Kuala Lumpur, with no socio-demographic differences expected. The intervention aims to improve KAP, environmental cleanliness index, and dengue index, potentially reducing dengue risk. Results could inform public health policies, emphasizing school-based interventions' importance in combating diseases like dengue., Competing Interests: The authors declare that they have no competing interests., (Copyright: © 2024 Dapari et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Coolant-assisted liquid nitrogen (LN) flash freezing of frozen tissues has been widely adopted to preserve tissue morphology for histopathological annotations in mass spectrometry-based spatial proteomics techniques. However, existing coolants pose health risks upon inhalation and are expensive. To overcome this challenge, we present our pilot study by introducing the EtOH-LN workflow, which demonstrates the feasibility of using 95 % ethanol as a safer and easily accessible alternative to existing coolants for LN-based cryoembedding of frozen tissues. Our study reveals that both the EtOH-LN and LN-only cryoembedding workflows exhibit significantly reduced freezing artifacts compared to cryoembedding in cryostat (p < 0.005), while EtOH-LN (SD = 0.56) generates more consistent results compared to LN-only (SD = 1.29). We have modified a previously reported morphology restoration method to incorporate the EtOH-LN workflow, which successfully restored the tissue architecture from freezing artifacts (p < 0.05). Additional studies are required to validate the impact of the EtOH-LN workflow on the molecular profiles of tissues., Competing Interests: Declaration of competing interest All authors declare no conflicts of interest to report., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Annona muricata is a common plant used in Africa and South America to manage various types of disease. However, there is insufficient toxicological information or published standard available regarding repeated dose animal toxicity data. As part of the safety assessment, we exposed Sprague Dawley rats to an acute oral toxicity of A. muricata . The intent of the current study was to use advanced proton nuclear magnetic resonance ( 1 H NMR) in serum and urinary metabolomics evaluation techniques to provide the in vivo acute toxicological profile of A. muricata leaf ethanol extract in accordance with the Organization for Economic Co-operation and Development's (OECD) 423 guidelines. A single 2000 mg/kg dose of A. muricata leaf ethanol extract was administered to Sprague Dawley rats over an observational period of 14 days. The toxicity evaluation (physical and behavior observation, body weight, renal function test, liver function test and 1 H NMR analysis) showed no abnormal toxicity. Histopathological analysis manifested mild changes, i.e., the treated kidney manifested mild hypercellularity of mesangial cells and mild red blood cell congestion. In addition, there was mild hemorrhage into tissue with scattered inflammatory cells and mild dilated central vein with fibrosis in the liver. However, the changes were very mild and not significant which correlate with other analyses conducted in this study (biochemical test and 1 H NMR metabolomic analysis). On the other hand, urinary 1 H NMR analysis collected on day 15 revealed high similarity on the metabolite variations for both untreated and treated groups. Importantly, the outcomes suggest that A. muricata leaf ethanol extract can be safely consumed at a dose of 2000 mg/kg and the LD 50 must be more than 2000 mg/kg.

Competing Interests: There are no conflicts of interest.