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Pineapple aroma is one of the most important sensory quality traits that influences consumer purchasing patterns. Reported in this paper is a high throughput method to quantify in a single analysis the key volatile organic compounds that contribute to the aroma of pineapple cultivars grown in Australia. The method constituted stable isotope dilution analysis in conjunction with headspace solid-phase microextraction coupled with gas-chromatography mass spectrometry. Deuterium labelled analogues of the target analytes purchased commercially were used as internal standards. Twenty-six volatile organic compounds were targeted for quantification and the resulting calibration functions of the matrix -matched validated method had determination coefficients (R 2 ) ranging from 0.9772 to 0.9999. The method was applied to identify the key aroma volatile compounds produced by popular pineapple cultivars such as 'Aus Carnival', 'Aus Festival', 'Aus Jubilee', 'Aus Smooth (Smooth Cayenne)' and 'Aussie Gold (73-50)', grown in Queensland, Australia. Pineapple cultivars varied in its content and composition of free volatile components, which were predominantly comprised of esters, followed by terpenes, alcohols, aldehydes, and ketones., 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 Elsevier Ltd. All rights reserved.)

Biosurfactants are amphiphilic biomolecules with promising tensoative and emulsifying properties that find application in the most varied industrial sectors: environment, food, agriculture, petroleum, cosmetics, and hygiene. In the current work, a 2 3 full-factorial design was performed to evaluate the effect and interactions of pineapple peel and corncob as substrates for biosurfactant production by Bacillus subtilis LMA-ICF-PC 001. In a previous stage, an alkaline pretreatment was applied to corncob samples to extract the xylose-rich hydrolysate. The results indicated that pineapple peel extract and xylose-rich hydrolysate acted as partial glucose substitutes, minimizing production costs with exogenous substrates. Biosurfactant I (obtained at 8.11% pineapple peel extract, 8.11% xylose-rich hydrolysate from corncob, and 2.8109 g/L glucose) exhibited a significant surface tension reduction (52.37%) and a promising bioremediation potential (87.36%). On the other hand, biosurfactant III (obtained at 8.11% pineapple peel extract, 31.89% xylose-rich hydrolysate from corncob, and 2.8109 g/L glucose) exhibited the maximum emulsification index in engine oil (69.60%), the lowest critical micellar concentration (68 mg/L), and the highest biosurfactant production (5.59 g/L). These findings demonstrated that using pineapple peel extract and xylose-rich hydrolysate from corncob effectively supports biosurfactant synthesis by B. subtilis, reinforcing how agro-industrial wastes can substitute traditional carbon sources, contributing to cost reduction and environmental sustainability., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pineapple Fruitlet Core Rot (FCR) is a fungal disease characterized by a multi-pathogen pathosystem. Recently, Fusarium proliferatum , Fusarium oxysporum , and Talaromyces stollii joined the set of FCR pathogens until then exclusively attributed to Fusarium ananatum . The particularity of FCR relies on the presence of healthy and diseased fruitlets within the same infructescence. The mycobiomes associated with these two types of tissues suggested that disease occurrence might be triggered by or linked to an ecological chemical communication-promoting pathogen(s) development within the fungal community. Interactions between the four recently identified pathogens were deciphered by in vitro pairwise co-culture bioassays. Both fungal growth and mycotoxin production patterns were monitored for 10 days. Results evidenced that Talaromyces stollii was the main fungal antagonist of Fusarium species, reducing by 22% the growth of Fusarium proliferatum . A collapse of beauvericin content was observed when FCR pathogens were cross-challenged while fumonisin concentrations were increased by up to 7-fold. Antagonism between Fusarium species and Talaromyces stollii was supported by the diffusion of a red pigmentation and droplets of red exudate at the mycelium surface. This study revealed that secondary metabolites could shape the fungal pathogenic community of a pineapple fruitlet and contribute to virulence promoting FCR establishment.

The cultivation of pineapple ( Ananas comosus ) is threatened worldwide by mealybug wilt disease of pineapple (MWP), whose etiology is not yet fully elucidated. In this study, we characterized pineapple mealybug wilt-associated ampeloviruses (PMWaVs, family Closteroviridae ) from a diseased pineapple plant collected from Reunion Island, using a high-throughput sequencing approach combining Illumina short reads and Nanopore long reads. Reads co-assembly resulted in complete or near-complete genomes for six distinct ampeloviruses, including the first complete genome of pineapple mealybug wilt-associated virus 5 (PMWaV5) and that of a new species tentatively named pineapple mealybug wilt-associated virus 7 (PMWaV7). Short reads data provided high genome coverage and sequencing depths for all six viral genomes, contrary to long reads data. The 5' and 3' ends of the genome for most of the six ampeloviruses could be recovered from long reads, providing an alternative to RACE-PCRs. Phylogenetic analyses did not unveil any geographic structuring of the diversity of PMWaV1, PMWaV2 and PMWaV3 isolates, supporting the current hypothesis that PMWaVs were mainly spread by human activity and vegetative propagation., Competing Interests: The authors declare no conflicts of interest.

Ethnopharmacological Relevance: Infections caused by parasitic worms or helminth continue to pose a great burden on human and animal health, particularly in underdeveloped tropical and subtropical countries where they are endemic. Current anthelmintic drugs present serious limitations and the emergence of drug resistance has made it increasingly challenging to combat such infections (helminthiases). In Bangladesh, medicinal plants are often used by indigenous communities for the treatment of helminthiases. Knowledge on such plants along with screening for their anthelmintic activity has the potential to lead to the discovery of phytochemicals that could serve as novel molecular scaffolds for the development of new anthelminthic drugs., Aim of the Study: The purpose of this study was i) to conduct an ethnobotanical survey to gather data on Bangladeshi medicinal plants used in the treatment of helminthiases, ii) to test plants with the highest use values for their in vitro anthelmintic activity, and iii) to carry out in silico screening on phytochemicals present in the most active plant extract to investigate their ability to disrupt β-tubulin function in helminths., Methods: The ethnobotanical survey was conducted across three sub-districts of Bangladesh, namely Mathbaria, Phultala and Khan Jahan Ali. The in vitro screening for anthelmintic activity was performed in a motility test using adult Haemonchus contortus worms. Virtual screening using PyRx was performed on the phytochemicals reported from the most active plant, exploring their interactions with the colchicine binding site of the β-tubulin protein target (PDB ID: 1SA0)., Results: The survey respondents reported a total of 32 plants for treating helminthiases. Based on their use values, the most popular choices were Ananas comosus (L.) Merr., Azadirachta indica A.Juss., Carica papaya L., Citrus maxima (Burm.) Merr., Curcuma longa L., Momordica charantia L., Nigella sativa L. and Syzygium cumini (L.) Skeels. In vitro anthelmintic testing revealed that A. indica leaves and bark had the highest activity with LC 50 values of 16 mg/mL in both cases. Other plant extracts also exhibited good anthelmintic activity with LC 50 values ranging from 16 to 52 mg/mL, while the value for albendazole (positive control) was 8.39 mg/mL. The limonoids nimbolide and 28-deoxonimbolide showed a binding affinity of -8.9 kcal/mol, and satisfied all drug-likeness parameters. The control ligand N-deacetyl-N-(2-mercaptoacetyl)colchicine had a binding affinity of -6.9 kcal/mol., Conclusion: Further in silico and in vitro studies are warranted on the identified limonoids to confirm the potential of these derivatives as novel drug templates for helminthiases. The current study supports the need for an ethnobotanical survey-based approach to discover novel drug templates for helminthiases., 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. Jamil Ahmad Shilpi reports financial support was provided by Government of the People's Republic of Bangladesh Ministry of Science and Technology., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The recent increase in the harvesting and industrial processing of tropical fruits such as pineapple and papaya is leading to unavoidable amounts of byproducts rich in valuable compounds. Given the significance of the chemical composition of these byproducts, new research avenues are opening up to exploit them in the food industry. In this sense, the revalorization of pineapple and papaya byproducts is an emerging trend that is encouraging the full harnessing of these tropical fruits, offering the opportunity for developing innovative value-added products. Therefore, the main aim of this review is to provide an overview of the state of the art of the current valorization applications of pineapple and papaya byproducts in the field of food industry. For that proposal, comprehensive research of valorization applications developed in the last years has been conducted using scientific databases, databases, digital libraries, and scientific search engines. The latest valorization applications of pineapple and papaya byproducts in the food industry have been systematically revised and gathered with the objective of synthesizing and critically analyzing existing scientific literature in order to contribute to the advancement of knowledge in the field of tropical byproduct revalorization providing a solid foundation for further research and highlighting scientific gaps and new challenges that should be addressed in the future., (© 2024 The Authors. Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Bromelain is a complex natural mixture of sulfhydryl-containing proteolytic enzymes that can be extracted from the stem or fruit of the pineapple. This compound is considered a safe nutraceutical, has been used to treat various health problems, and is also popular as a health-promoting dietary supplement. There is continued interest in bromelain due to its remarkable therapeutic properties. The mechanism of action of bromelain appears to extend beyond its proteolytic activity as a digestive enzyme, encompassing a range of effects (mucolytic, anti-inflammatory, anticoagulant, and antiedematous effects). Little is known about the clinical use of bromelain in pediatrics, as most of the available data come from in vitro and animal studies, as well as a few RCTs in adults. This narrative review was aimed at highlighting the main aspects of the use of bromelain in children, which still appears to be limited compared to its potential. Relevant articles were identified through searches in MEDLINE, PubMed, and EMBASE. There is no conclusive evidence to support the use of bromelain in children, but the limited literature data suggest that its addition to standard therapy may be beneficial in treating conditions such as upper respiratory tract infections, specific dental conditions, and burns. Further studies, including RCTs in pediatric settings, are needed to better elucidate the mechanism of action and properties of bromelain in various therapeutic areas.

Diabetes affects millions globally and poses treatment challenges. Targeting the enzyme fructose-1,6-bisphosphatase (FBPase) in gluconeogenesis and exploring plant-based therapies offer potential solutions for improving diabetes management while supporting sustainability and medicinal advancements. Utilizing pineapple ( Ananas comosus L. Merr.) waste as a source of drug precursors could be valuable for health and environmental care due to its medicinal benefits and abundant yearly biomass production. Therefore, this study conducted a virtual screening to identify potential natural compounds from pineapple that could inhibit FBPase activity. A total of 112 compounds were screened for drug-likeness and ADMET properties, and molecular docking simulations were performed on 20 selected compounds using blind docking. The lead compound, butane-2,3-diyl diacetate, was subjected to 100 ns MD simulations, revealing a binding energy of -5.4 kcal/mol comparable to metformin (-5.6 kcal/mol). The MD simulation also confirmed stable complexes with crucial hydrogen bonds. Glu20, Ala24, Thr27, Gly28, Glu29, Leu30, Val160, Met177, Asp178, and Cys179 were identified as key amino acids that stabilized the human liver FBPase-butane-2,3-diyl diacetate complex, while Tyr215 and Asp218 played a crucial role in the human liver FBPase-Metformin complex. Our study indicates that the lead compound has high intestinal solubility. Therefore, it would show rapid bloodstream distribution and effective action on the target protein, making butane-2,3-diyl diacetate a potential antidiabetic drug candidate. However, further investigations in vitro , preclinical, and clinical trials are required to thoroughly assess its efficacy and safety.Communicated by Ramaswamy H. Sarma.

Lysinibacillus fusiformis PwPw_T2 isolated from deteriorating Ananas comosus sample collected from Lagos State, Nigeria putatively possesses genomic features like potential enzymes catalyzing acetic acid production and xenobiotic compounds degradation via various pathways as indicated by its genome sequences. These could make the organism relevant in food waste valorization and micro-biotechnology., Competing Interests: The authors declare no conflict of interest.

This research aimed to optimize the processing conditions to obtain ready-to-eat extruded snacks with a high fiber content from mixtures of pineapple byproduct powder (PBP) and nixtamalized maize flour (PBP-NMF) or maize flour (PBP-MF). The effects of barrel temperature, feed moisture content, and PBP were evaluated. The increase in barrel temperature has a negative effect on the bulk density, the water absorption index, and the texture in both mixtures (PBP-MF and PBP-NMF) and increases the expansion index and the water solubility index in the mixture with MF. The increase in the feed moisture content increased the bulk density and water absorption index in both mixtures and the texture in the mixtures with MF. The increasing PBP decreases the expansion index and increases the water solubility index in both mixtures. The increase in PBP in the mixtures with MF decreases the water absorption index, texture, and bulk density. From the optimization, four products were obtained, two for the NMF mixture and two for the MF mixtures. The optimal formulations can be considered a good source of total fiber (12.46-12.78 g/100 g) and protein (8.27-8.85 g/100 g) with good acceptance by consumers. PRACTICAL APPLICATION: Pineapple byproducts in combination with nixtamalized and nonnixtamalized maize flour are viable raw materials for the development of ready-to-eat extruded snacks with a high content of dietary fiber and good acceptance by consumers. Due to their characteristic nutritional properties, the consumption of this ready-to-eat snack could present potential benefits for human health., (© 2023 Institute of Food Technologists.)

Background: Pineapple has an important role in ethnopharmacology and its enzyme, bromelain, has been extensively investigated for its medicinal properties. Aim: This systematic review and meta-analysis aimed to assess clinical evidence concerning the efficacy and safety of bromelain. Methods: A systematic search was conducted from conception to August 2022 using CINAHL Complete, MEDLINE, ScienceDirect, Scopus, and Thai Journal Online (TJO). The risk of bias was assessed using Risk of Bias 2 or ROBIN-I. A random-effect model with inverse variance weighting and DerSimonian and Laird method was used for meta-analysis. The heterogeneity was evaluated by I 2 statistics. Results: We included 54 articles for qualitative summary and 39 articles for meta-analysis. The systematic review found that bromelain presented in serum with retained proteolytic activity after oral absorption. Bromelain may be effective against sinusitis but was not effective for cardiovascular diseases. Pain reduction from oral bromelain was slightly but significantly better than controls (mean difference in pain score = -0.27; 95% CI: -0.45, -0.08; n  = 9; I 2  = 29%). Adverse events included flatulence, nausea, and headache. Topical bromelain significantly reduced the time to complete debridement (mean difference in time = -6.89 days; 95% CI: -7.94, -5.83; n  = 4; I 2  = 2%). Adverse events may be irrelevant and include burning sensation, pain, fever, and sepsis. Conclusions: Moderate-quality studies demonstrated the potential of oral bromelain in pain control and topical bromelain in wound care. Major health risks were not reported during the treatment with bromelain.

Pineapple [ Ananas comosus (L.) Merr.] is the most economically important crop possessing crassulacean acid metabolism (CAM) photosynthesis which has a higher water use efficiency by control of nocturnal opening and diurnal closure of stomata. To provide novel insights into the diel regulatory landscape in pineapple leaves, we performed genome-wide mapping of DNase I hypersensitive sites (DHSs) in pineapple leaves at day (2a.m.) and night (10a.m.) using a simplified DNase-seq method. As a result, totally 33340 and 28753 DHSs were found in green-tip tissue, and 29597 and 40068 were identified in white-base tissue at 2a.m. and 10a.m., respectively. We observed that majority of the pineapple genes occupied less than two DHSs with length shorter than 1 kb, and the promotor DHSs showed a proximal trend to the transcription start site (>77% promotor DHSs within 1 kb). In addition, more intergenic DHSs were identified around transcription factors or transcription co-regulators (TFs/TCs) than other functional genes, indicating complex regulatory contexts around TFs/TCs. Through combined analysis of tissue preferential DHSs and genes, we respectively found 839 and 888 coordinately changed genes in green-tip at 2a.m. and 10a.m. (AcG2 and AcG10). Furthermore, AcG2-specific, AcG10-specific and common accessible DHSs were dissected from the total photosynthetic preferential DHSs, and the regulatory networks indicated dynamic regulations with multiple cis -regulatory elements occurred to genes preferentially expressed in photosynthetic tissues. Interestingly, binding motifs of several cycling TFs were identified in the DHSs of key CAM genes, revealing a circadian regulation to CAM coordinately diurnal expression. Our results provide a chromatin regulatory landscape in pineapple leaves during the day and night. This will provide important information to assist with deciphering the circadian regulation of CAM photosynthesis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ouyang, Liang, Miao, Zhang and Li.)

Pineapple ( Ananas comosus ), one of the most flavorful and popular tropical fruits consumed worldwide, is known to contain many volatile organic compounds (VOCs) at varying concentrations. Much attention has been paid to understand which VOC plays a significant role in the sensory aroma notes of the fruit. Though, nearly 480 VOCs have been identified to date using different analytical techniques, only 40 compounds are reported to contribute to the unique flavor of pineapple. A consolidated database of the reported VOCs and key aroma compounds of pineapple is currently not available. This review discusses the available published data regarding the analytical methodologies, volatile profile of different varieties of pineapple at different maturities, and their characteristic aroma compounds. The output of this review is a subset of key pineapple aroma volatiles that can be targeted in analytical method development for utilization in varietal improvement or other research of pineapple.

Pineapple is one of the most economically important fruits in tropical countries, particularly in Thailand. Canned pineapple is currently Thailand's main exported commodity to many countries, including the United States, Russia, Germany, Poland, and Japan. Fungal diseases are considered a permanent threat to fruits in the pre- and post-harvest stages, leading to considerable economic losses. Fungal disease is one of the primary causes of massive yield losses in pineapples around the world. Colletotrichum species are the most common fungal pathogens affecting different tropical fruits. Although there are many reports regarding Colletotrichum species associated with pineapple, they do not have molecular data to confirm species identification. However, the occurrence of Colletotrichum species on pineapple has not been reported in Thailand so far. In this study, we isolated and identified Colletotrichum fructicola on pineapple in northern Thailand and have proven its pathogenicity to the host. This is the first report of the occurrence of Colletotrichum in pineapple, based on morpho-molecular approaches.

Pineapple ( Ananas comosus L.) is one of the most valuable subtropical fruit crop in the world. The sweet-acidic taste of the pineapple fruits is a major contributor to the characteristic of fruit quality, but its formation mechanism remains elusive. Here, targeted metabolomic and transcriptomic analyses were performed during the fruit developmental stages in two pineapple cultivars ("Comte de Paris" and "MD-2") to gain a global view of the metabolism and transport pathways involved in sugar and organic acid accumulation. Assessment of the levels of different sugar and acid components during fruit development revealed that the predominant sugar and organic acid in mature fruits of both cultivars was sucrose and citric acid, respectively. Weighted gene coexpression network analysis of metabolic phenotypes and gene expression profiling enabled the identification of 21 genes associated with sucrose accumulation and 19 genes associated with citric acid accumulation. The coordinated interaction of the 21 genes correlated with sucrose irreversible hydrolysis, resynthesis, and transport could be responsible for sucrose accumulation in pineapple fruit. In addition, citric acid accumulation might be controlled by the coordinated interaction of the pyruvate-to-acetyl-CoA-to-citrate pathway, gamma-aminobutyric acid pathway, and tonoplast proton pumps in pineapple. These results provide deep insights into the metabolic regulation of sweetness and acidity in pineapple., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gao, Yao, Chen, Wu, Wu, Liu, Guo and Zhang.)

Research Background: Morinda citrifolia L. (noni), Ananas comosus L. cv. Sarawak (pineapple) and Mangifera indica L. cv. Apple (mango) represent fruits capable of coagulating milk and forming a curd. Plant-derived milk coagulants have antidiabetic phytochemicals that enrich the curd. Hence this work evaluates the dual benefits of the fruits in coagulating milk and the antidiabetic activities found in the obtained curd., Experimental Approach: The three fruits were mixed to form a supercoagulant (a milk coagulant mixture of the extracts at a ratio of 1:1:1), and the milk coagulation time was measured. The milk was coagulated by the supercoagulant, and thus fortified curd was tested for its ability to inhibit α-glucosidase and α-amylase activities. Then, the fortified curd was fed daily to streptozotocin-induced diabetic rats and their biochemical markers such as blood glucose level, aspartate aminotransferase, alanine transaminase, etc. as well as histopathology of their liver and kidney tissues were compared with the untreated diabetic rats and normal rats., Results and Conclusion: The supercoagulant had a milk coagulation time of (28±3) s at a 50 mg/mL concentration. Its fortified curd inhibited α-glucosidase and α-amylase activities, with IC 50 values of (4.04±0.03) and (3.42±0.02) mg/mL, respectively. The average mass of the streptozotocin-induced diabetic rats fed daily with curd formed by the supercoagulant was (201±10) g on day 20 compared to diabetic control rats with (149±16) g. The blood glucose concentration for rats treated with the supercoagulant after fasting was (15±1) mmol/L compared to the diabetic control rats ((26±2) mmol/L). Blood tests on the treated rats showed aspartate aminotransferase, alanine transaminase, γ-glutamyl transferase and alkaline phosphatase (liver function tests) amounts of (214±78), (91±13), 3 and (510±138) U/L, respectively, while the total protein and renal function tests showed the concentrations of albumin, globulin, urea and creatinine of (37±2) g/L, (30±2) g/L, (11±1) mmol/L and (42±3) µmol/L, respectively. These concentrations were found to be similar to those of the normal rats on day 20. Furthermore, a histopathological study performed on the liver and kidney of the rats found no apparent damage., Novelty and Scientific Contribution: This supercoagulant derived from a mixture of fruits is able to coagulate milk rapidly, and its curd is fortified with safe antidiabetic agents. The supercoagulant is potentially useful in producing functional dairy food to prevent diabetes or as a supplement for diabetics to control their blood sugar. Such products are capable of replacing dairy products derived from animal enzymes and provide consumers with additional functional dairy products., Competing Interests: CONFLICT OF INTEREST The authors declare no conflict of interest.

Excessive production of reactive oxygen species (ROS) leads to potential toxicity in an organism. Class III peroxidases (PRXs) play an important role in maintaining ROS homeostasis in plants. Internal browning (IB) limits industrial development of pineapple, which is the third most important fruit trade in the world. IB is mainly caused by ROS, and the mechanism underlying IB is still unknown from the perspective of ROS. Here, we soaked pineapples in ascorbic acid after harvest and before storage to decrease excessive ROS and polyphenol oxidase (PPO) activity, ultimately restraining the spread and deterioration of IB. Using phylogenetic analysis; we identified 78 pineapple PRX genes ( AcPRXs ) and divided them into five subgroups. Gene structure analysis indicated that the exon numbers ranged from 2 to 14, and conserved motif analysis verified that all of the AcPRXs identified here have standard peroxidase domains. Analysis of duplication events suggested that tandem and segmental duplication events may have played equal and important roles in expanding the AcPRX family. Comprehensive transcriptomic analysis uncovered that AcPRXs may play an important role in negatively regulating the occurrence of IB. In summary, we found that ROS scavenging delayed IB occurrence. The results of characterized AcPRX family revealed that AcPRXs family responded to growth and development, and negatively regulated to IB occurrence in storage stage. This research provides potential target genes for future in-depth analysis of the molecular mechanisms underlying IB and contributes to develop IB-resistant pineapple varieties., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hou, Lu, Hong, Song, Gu, Hu and Yao.)

Pineapple core is considered a processing by-product. This study proposed and evaluated an ohmic heating extraction-based valorization platform to obtain value-added bioactive compounds from pineapple core and studied the effects of four important processing parameters. In this sense, a Taguchi design (L16(4)4) was used to assess the effects of temperature (70, 80, 90, and 100 °C), time (15, 30, 45, and 60 min), voltage (110, 160, 210, and 260 V), and frequency (60, 340, 620, and 900 Hz) on heating rate, come-up time, energy consumption, system performance efficiency, total phenolic compounds (TPC), DPPH, and ABTS. Finally, a side-by-side comparison of optimized ohmic heating (OOH) and conventional extraction was performed, and chemical composition was compared by ultra-performance liquid chromatography equipped with photodiode array detection-mass (UPLC-DAD-ESI-MS-MS). According to the results, increasing temperatures enhanced system performance efficiency but negatively affected TPC and antioxidant values above 90 ℃. Similarly, prolonging the extraction (>30 min) decreased TPC. Further, increasing voltage (from 110 to 260 V) shortened the come-up time (from 35.75 to 5.16 min) and increased the heating rate (from 2.71 to 18.80 °C/min−1). The optimal conditions were 30 min of extraction at 80 °C, 160 V, and 900 Hz. Verification of the optimal conditions revealed that OOH yielded an extract with valuable bioactive compounds and saved 50% of the time and 80% of energy compared to the conventional treatment. The UPLC-DAD-ESI-MS-MS showed that there were similarities between the chemical profiles of the extracts obtained by conventional and OOH methods, while the concentration of major compounds varied depending on the extraction method. This information can help achieve sustainable development goals (SDGs) by maximizing the yield and minimizing energy and time consumption.

Plant-specific calcineurin B-like proteins (CBLs) and their interacting kinases, CBL-interacting protein kinases (CIPKs) module, are essential for dealing with various biotic and abiotic stress. The kinases (CIPKs) of this module have been well studied in several plants; however, the information about pineapple CIPKs remains limited. To understand how CIPKs function against environmental cues in pineapple, the CIPK5 gene of pineapple was cloned and characterized. The phylogenetic analyses revealed that AcCIPK5 is homologous to the CIPK12 of Arabidopsis and other plant species. Quantitative real-time PCR (qRT-PCR) analysis revealed that AcCIPK5 responds to multiple stresses, including osmotic, salt stress, heat and cold. Under optimal conditions, AcCIPK5 gets localized to the cytoplasm and cell membrane. The ectopic expression of AcCIPK5 in Arabidopsis improved the germination under osmotic and salt stress. Furthermore, AcCIPK5 positively regulated osmotic, drought, salt and cold tolerance and negatively regulated heat and fungal stress in Arabidopsis. Besides, the expression of AcCIPK impacted ABA-related genes and ROS homeostasis. Overall, the present study demonstrates that AcCIPK5 contributes to multiple stress tolerance and has the potential to be utilized in the development of stress-tolerant crops., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The pineapple ( Ananas comosus ) is an important tropical fruit in the world market. Its pulp has significant nutritional value while the peel and the core, in spite of being high in dietary fibre and nutrients, are generally considered to be agro-industrial waste. The aim of this research was to evaluate the effect that the integrated enzymatic and shear homogenization processes have on the physicochemical stability of pineapple base suspensions (pulp, core, and peel extract). Initially, an enzymatic hydrolysis process was evaluated with a completely randomized factorial design. Independent variables: incubation time (t inc ) (1-4 h) and [enzyme] (0-200 ppm). Dependent variables: viscosity (μ) and particle sizes (D [3;2] and D [4;3] ). The results showed a reduction of (μ) (70.7%), D [3;2] (54.2%), and D [4;3] (61.8%) for the optimized treatment (t inc  = 3.2 h and [enzyme] = 200 ppm) compared to the control (t = 0, without enzyme). The effect of the integrated enzymatic treatment with a serial homogenization process was subsequently evaluated. Independent variables: high-speed homogenization time (t 1 ) (15-20 min), recirculation time in high pressure homogenizer (t 2 ) (3-7 min), and arabic gum (AG) (0.6-1.0%). Dependent variables: total suspension solids (TS S ), zeta potential (ζ), μ, spectral stability index (R), D [3;2] , and D [4;3] . The application of the integrated processes of enzymatic treatment and serial homogenization was more effective to be able to obtain a stable pineapple-based suspension. The experimental optimization of multiple responses defined t 1  = 16.4 min, t 2  = 7 min, AG = 0.98%, and TSs = 15.7 ± 0.5%, ζ = - 23.1 ± 0.4 mV, µ = 221 ± 11 cP, D [3;2]  = 56.8 ± 2 µm and D [4;3]  = 120.6 ± 4 µm and R = 0.58 ± 0.02 were obtained., Competing Interests: Conflicts of interestThe authors declare that they have no competing interests., (© Association of Food Scientists & Technologists (India) 2021.)

<b>Background and Objective:</b> Pineapple (<i>Ananas comosus</i>) is a popular fruit worldwide with natural antioxidant properties. This study examined how pineapple modified the expression of drug-metabolizing enzymes (CYP1A2, CYP2C9, CYP3A4, UGT1A6, NAT2 and SULT1A1) and a drug transporter (OATP1B1) in human hepatocarcinoma (HepG2) cells. <b>Materials and Methods:</b> HepG2 cells (2.5×10<sup>5</sup> cells/well in a 24-well plate) were incubated with pineapple juice extract (125-1,000 μg mL<sup>1</sup>) for 48 hrs in phenol red-free medium. Resazurin reduction, ROS, AST and ALT assays were performed. The mRNA expression of target genes was determined by RT/qPCR. <b>Results:</b> Pineapple juice slightly reduced HepG2 cell viability to 80% of the control, while ROS, AST and ALT levels were not changed. Pineapple juice did not alter the expression of CYP1A2, CYP2C9 and UGT1A6 mRNA. All tested concentrations of pineapple juice suppressed CYP3A4, NAT2 and OATP1B1 expression, while SULT1A1 expression was induced. <b>Conclusion:</b> Though pineapple juice slightly decreased the viability of HepG2 cells, cell morphology and cell function remained normal. Pineapple juice disturbed the expression of phase I (CYP3A4) and phase II (NAT2 and SULT1A1) metabolizing genes and the drug transporter OATP1B1. Therefore, the consumption of excessive amounts of pineapple juice poses a risk for drug interactions.

Pineapple ( Ananas comosus (L.) Merr.) is the second most important tropical fruit crop globally, and 'MD2' is the most important cultivated variety. A high-quality genome is important for molecular-based breeding, but available pineapple genomes still have some quality limitations. Here, PacBio and Hi-C data were used to develop a new high-quality MD2 assembly and gene prediction. Compared to the previous MD2 assembly, major improvements included a 26.6-fold increase in contig N50 length, phased chromosomes, and >6000 new genes. The new MD2 assembly also included 161.6 Mb additional sequences and >3000 extra genes compared to the F153 genome. Over 48% of the predicted genes harbored potential deleterious mutations, indicating that the high level of heterozygosity in this species contributes to maintaining functional alleles. The genome was used to characterize the FAR1-RELATED SEQUENCE (FRS) genes that were expanded in pineapple and rice. Transposed and dispersed duplications contributed to expanding the numbers of these genes in the pineapple lineage. Several AcFRS genes were differentially expressed among tissue-types and stages of flower development, suggesting that their expansion contributed to evolving specialized functions in reproductive tissues. The new MD2 assembly will serve as a new reference for genetic and genomic studies in pineapple.

The large pineapple's consumption and processing have generated a massive amount of waste yearly, which requires adequate treatment measures to avoid damages to the environment. Pineapple peel is one of the main residues obtained from this fruit and a promising strategy to take advantage of its potential is using it for biosurfactant production due to the peel's rich composition in fermentable sugars and nutrients, such as potassium and magnesium that favor the Bacillus subtilis growth and biosurfactant excretion as well. The current research performed a central composite design (CCD) with four independent variables (glucose, pineapple peel, potassium, and magnesium), evaluating substrates' influence on the surface tension reduction rate (STRR) and the emulsification index (EI 24 ). The results indicated that pineapple peel has the necessary potential to act as a partial substitute for glucose and salt nutrients, minimizing the costs of supplementing with exogenous minerals. The highest surface tension reduction rate (57.744%) was obtained at 2.18% glucose (w/v); 14.67% pineapple peel (v/v); 2.38 g/L KH 2 PO 4 ; and 0.15 g/L MgSO 4 .7H 2 O; whereas to obtain the maximum predicted value for EI 24 (61.92%) the medium was composed by 2.24% glucose (w/v); 12.63% pineapple peel (v/v); 2.53 g/L KH 2 PO 4 ; and 0.29 g/L MgSO 4 .7H 2 O., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Bromelain is a complex combination of multiple endopeptidases of thiol and other compounds derived from the pineapple fruit, stem and/or root. Fruit bromelain and stem bromelain are produced completely distinctly and comprise unique compounds of enzymes, and the descriptor "Bromelain" originally referred in actuality to stem bromelain. Due to the efficacy of oral administration in the body, as a safe phytotherapeutic medication, bromelain was commonly suited for patients due to lack of compromise in its peptidase efficacy and the absence of undesired side effects. Various in vivo and in vitro studies have shown that they are anti-edematous, anti-inflammatory, anti-cancerous, anti-thrombotic, fibrinolytic, and facilitate the death of apoptotic cells. The pharmacological properties of bromelain are, in part, related to its arachidonate cascade modulation, inhibition of platelet aggregation, such as interference with malignant cell growth; anti-inflammatory action; fibrinolytic activity; skin debridement properties, and reduction of the severe effects of SARS-Cov-2. In this paper, we concentrated primarily on the potential of bromelain's important characteristics and meditative and therapeutic effects, along with the possible mechanism of action.

Agroindustrial wastes contain macronutrients and micronutrients essential for the reproduction of lactic acid bacteria. In this research, the reproduction of Weissella cibaria was experimentally optimized in a supplemented fermentation substrate (SFS) formulated from pineapple and sacha inchi wastes. Response surface methodology was used to evaluate the influence of the following independent variables: temperature (32-40 °C), pH (5.0-6.0), and stirring speed (SS) (100-150 rpm) on the following dependent variables: viability (Log 10 CFU mL -1 ), biomass production (B Wc ), lactic acid production (LA), biomass yield (Y Bwc/S ), biomass volumetric productivity (VP Wc ), LA volumetric productivity (VP LA ), carbon source consumption (CSC), N 2 consumption (N 2 C), and specific growth rate (µ). The experimental optimization of multiple responses presented a desirability of 76.8%, thus defining the independent variables of the process: temperature = 35.1 °C, pH = 5.0, and SS = 139.3 rpm; and the dependent variables: viability = 10.01 Log 10 CFU mL -1 , B Wc  = 2.9 g L -1 , LA = 19.4 g mL -1 , Y Bwc/ S  = 43.9 g biomass/g CSC, VP Wc = 0.49 g L -1 h   -   1 , VP LA = 3.2 g L -1 h -1 , CSC = 17.2%, N 2 C = 63.6% and µ = 0.28 h -1 . From these, viability, Y Bwc/S , CSC, N 2 C, and LA presented significant statistical differences, while the independent variable with the least important effect on the process was pH. Under optimal conditions of temperature, pH and SS; SFS favors the reproduction and viability of W. cibaria . This provides evidence of a sustainable alternative for the production of probiotics in the context of circular economy., Competing Interests: 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., (© 2021 The Authors. Published by Elsevier B.V.)

Bromelain, a member of cysteine proteases, is found abundantly in pineapple (Ananas comosus), and it has a myriad of versatile applications. However, attempts to produce recombinant bromelain for commercialization purposes are challenging due to its expressibility and solubility. This study aims to express recombinant fruit bromelain from MD2 pineapple (MD2Bro; accession no: OAY85858.1) in soluble and active forms using Escherichia coli host cell. The gene encoding MD2Bro was codon-optimized, synthesized, and subsequently ligated into pET-32b( +) for further transformation into Escherichia coli BL21-CodonPlus(DE3). Under this strategy, the expressed MD2Bro was in a fusion form with thioredoxin (Trx) tag at its N-terminal (Trx-MD2Bro). The result showed that Trx-MD2Bro was successfully expressed in fully soluble form. The protein was successfully purified using single-step Ni 2+ -NTA chromatography and confirmed to be in proper folds based on the circular dichroism spectroscopy analysis. The purified Trx-MD2Bro was confirmed to be catalytically active against N-carbobenzoxyglycine p-nitrophenyl ester (N-CBZ-Gly-pNP) with a specific activity of 6.13 ± 0.01 U mg -1 and inhibited by a cysteine protease inhibitor, E-64 (IC 50 of 74.38 ± 1.65 nM). Furthermore, the catalytic efficiency (k cat /K M ) Trx-MD2Bro was calculated to be at 5.64 ± 0.02 × 10 -2  µM -1  s -1 while the optimum temperature and pH were at 50 °C and pH 6.0, respectively. Furthermore, the catalytic activity of Trx-MD2Bro was also affected by ethylenediaminetetraacetic acid (EDTA) or metal ions. Altogether it is proposed that the combination of codon optimization and the use of an appropriate vector are important in the production of a soluble and actively stable recombinant bromelain.

Bromelain is an effective chemoresponsive proteolytic enzyme derived from pineapple stems. It contains several thiol endopeptidases and is extracted and purified via several methods. It is most commonly used as an anti-inflammatory agent, though scientists have also discovered its potential as an anticancer and antimicrobial agent. It has been reported as having positive effects on the respiratory, digestive, and circulatory systems, and potentially on the immune system. It is a natural remedy for easing arthritis symptoms, including joint pain and stiffness. This review details bromelain's varied uses in healthcare, its low toxicity, and its relationship to nanoparticles. The door of infinite possibilities will be opened up if further extensive research is carried out on this pineapple-derived enzyme.

In the present study, protective effects of Ananas comosus i.e., pineapple pulp (PA) against cobalt chloride (CoCl 2 )‑induced hypoxia in Caco-2 cells were evaluated. PA reduces levels of lipid peroxidation, reactive oxygen and nitrogen species. It was proved to be cytoprotective and increased anti-oxidant activity against CoCl 2 -induced hypoxia. The inference drawn from this experiment was CoCl 2 -induced hypoxia that regulates hypoxia-inducible factor-1 (HIF-1), a transcription factor. It was also confirmed that PA pre-treatment inhibited the expression of HIF-1α, thereby downregulating the hypoxia-associated gene/protein expressions such as GLUT-1, VEGF, ANG and FGF. Finally, supplementation of PA could help in snow-balling the digestive hormones like leptin and CCK in hypoxic conditions. Therefore, this report provides substantial proof of reducing the hypoxia-induced loss of appetite at high-altitude environments. Graphical Abstract.

Background: Crassulacean acid metabolism (CAM) photosynthesis is an important carbon fixation pathway especially in arid environments because it leads to higher water-use efficiency compared to C3 and C4 plants. However, the role of DNA methylation in regulation CAM photosynthesis is not fully understood., Results: Here, we performed temporal DNA methylome and transcriptome analysis of non-photosynthetic (white base) and photosynthetic (green tip) tissues of pineapple leaf. The DNA methylation patterns and levels in these two tissues were generally similar for the CG and CHG cytosine sequence contexts. However, CHH methylation was reduced in white base leaf tissue compared with green tip tissue across diel time course in both gene and transposon regions. We identified thousands of local differentially methylated regions (DMRs) between green tip and white base at different diel periods. We also showed that thousands of genes that overlapped with DMRs were differentially expressed between white base and green tip leaf tissue across diel time course, including several important CAM pathway-related genes, such as beta-CA, PEPC, PPCK, and MDH., Conclusions: Together, these detailed DNA methylome and transcriptome maps provide insight into DNA methylation changes and enhance our understanding of the relationships between DNA methylation and CAM photosynthesis.

<b>Background and Objective:</b> Pineapple (<i>Ananas comosus</i> L.) has antioxidant and other pharmacological properties. This study examined how pineapple modified mitochondrial permeability transition and expression of drug-metabolizing enzymes, i.e., CYP1A2, CYP2C9, CYP3A4, UGT1A6, NAT2 and the drug transporter OATP1B1 in human colorectal adenocarcinoma (Caco-2) cells. <b>Materials and Methods:</b> Caco-2 cells (2.5×10<sup>5</sup> cells well<sup>1</sup> in 24-well plates) were incubated with pineapple (125 to 1,000 μg mL<sup>1</sup>) for 48 hrs in a phenol red-free medium. Mitochondrial permeability transition, resazurin cell viability and AST and ALT levels were investigated. The mRNA expression of target genes was determined by RT/qPCR. <b>Results:</b> Pineapple significantly reduced depolarized mitochondria, slightly decreased cell viability and did not change AST and ALT levels. Pineapple did not modify the mRNA expressions of CYP1A2, CYP2C9 and CYP3A4 but markedly induced UGT1A6 expression. The highest tested concentration of pineapple (1,000 μg mL<sup>1</sup>) significantly suppressed NAT2 and OATP1B1 expression. <b>Conclusion:</b> Although pineapple slightly decreased cell viability to ~80% of control, the morphology and functions of the cells were unaffected. Pineapple showed a beneficial effect to reduce depolarized mitochondria, which consequently decreased reactive oxygen species production. Pineapple did not modify the expression of CYPs, whilst it altered the expression of phase 2 metabolizing genes UGT1A6 and NAT2 and the transporter OATP1B1. Therefore, the consumption of large amounts of pineapple is of concern for the risk of drug interaction via alteration of UGT1A6, NAT2 and OATP1B1 expression.

Introduction: The potential efficacy of selected plant extracts to counteract the dermal toxicity of jellyfish envenomation was investigated using an in vitro cell culture model., Methods: We studied plant extracts from Carica papaya, Ananas comosus, and Bouvardia ternifolia, known for their antivenom properties, in pairwise combinations with tissue homogenates of the jellyfish Pelagia noctiluca, Phyllorhiza punctata, and Cassiopea andromeda, to evaluate modulations of jellyfish cytotoxic effects. L929 mouse fibroblasts were incubated with pairwise jellyfish/plant extract combinations and examined by MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide)., Results: C papaya and A comosus significantly lowered the cytotoxicity of P noctiluca and P punctata but induced a slight worsening of C andromeda cytotoxicity. Conversely, B ternifolia was protective against P punctata, ineffective against P noctiluca, and worsened C andromeda cytotoxicity., Conclusions: Data showed species-specific and contrasting effects of plant extracts, suggesting that those containing protease activities, namely A comosus and C papaya, are more effective in lowering the cytotoxicity of jellyfish venom containing toxic peptidic factors such as phospholipase A. However, all examined plants require further investigation in vivo to evaluate their ability to counteract jellyfish injury to the skin., (Copyright © 2020 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.)

Simple sequence repeat (SSR) markers provide a reliable tool for the identification of accessions and the construction of genetic linkage maps because of their co-dominant inheritance. In the present study, we developed new SSR markers with next-generation sequencing using the Roche 454 GS FLX+ platform. Five hundred SSR primer sets were tested for the genetic identification of pineapple, including 100 each for the di-, tri-, tetra-, penta-, and hexa-nucleotide motif SSRs. In total, 160 SSR markers successfully amplified fragments and exhibited polymorphism among accessions. The SSR markers revealed the number of alleles per locus (ranging from 2 to 13), the expected heterozygosity (ranging from 0.041 to 0.823), and the observed heterozygosity (ranging from 0 to 0.875). A total of 117 SSR markers with tri- or greater nucleotide motifs were shown to be effective at facilitating accurate genotyping. Using the SSR markers, 25 accessions were distinguished genetically, with the exception of accessions 'MD-2' and 'Yonekura'. The developed SSR markers could facilitate the establishment of efficient and accurate genetic identification systems and the construction of genetic linkage maps in the future., (Copyright © 2020 by JAPANESE SOCIETY OF BREEDING.)

The identity of the fungi responsible for fruitlet core rot (FCR) disease in pineapple has been the subject of investigation for some time. This study describes the diversity and toxigenic potential of fungal species causing FCR in La Reunion, an island in the Indian Ocean. One-hundred-and-fifty fungal isolates were obtained from infected and healthy fruitlets on Reunion Island and exclusively correspond to two genera of fungi: Fusarium and Talaromyces. The genus Fusarium made up 79% of the isolates, including 108 F. ananatum , 10 F. oxysporum , and one F. proliferatum . The genus Talaromyces accounted for 21% of the isolated fungi, which were all Talaromyces stollii . As the isolated fungal strains are potentially mycotoxigenic, identification and quantification of mycotoxins were carried out on naturally or artificially infected diseased fruits and under in vitro cultures of potential toxigenic isolates. Fumonisins B 1 and B 2 (FB 1 -FB 2 ) and beauvericin (BEA) were found in infected fruitlets of pineapple and in the culture media of Fusarium species. Regarding the induction of mycotoxin in vitro, F. proliferatum produced 182 mg kg⁻ 1 of FB 1 and F. oxysporum produced 192 mg kg⁻ 1 of BEA. These results provide a better understanding of the causal agents of FCR and their potential risk to pineapple consumers.

Gold honey variety pineapple wastes and sacha inchi sub-products (SIS) were characterized in their elemental, physical, and chemical form in order to formulate a supplemented fermentation substrate (SFS) for the growth Weissella cibaria . The peels and fresh cores of the pineapple (FPP, FPC) were dried and ground (PPP, PPC) and then mixed (MCPP). The following procedures were then undertaken: a physicochemical characterization (moisture, a w , pH, acidity, and soluble solids) of the SIS, FPP, FPC, PPP, and PPC; a proximal characterization of he FPP, FPC, SIS, and SFS; and an elemental analysis (C-N 2 -H 2 -O 2 -S) of the MCPP, SIS, and W. cibaria , which allowed the stoichiometric equation to be defined and the SFS to be formulated. We then evaluated the effect that homogenization and heating to boiling point had on the concentration of reducing sugars in the SFS (g L -1 ). Finally, W. cibaria´s kinetic fermentation parameters were evaluated in the SFS and in a commercial substrate (control). The results showed FPP and FPC yields of 26.02 ± 0.58 and 14.69 ± 1.13%, respectively; a higher total sugar content in FPC (7.21%) than in FPP (6.65%); a high crude protein content in SIS (56.70%), and a C:N 2 ratio of 6.50:1.00. Moreover, the highest concentration of reducing sugars (4.44 ± 0.29 g L -1 ) in the SFS was obtained with 5 h of hydrolysis under homogenization pre-treatments and heating until boiling. The SFS allowed the adaptation of W. cibaria , and there was a biomass production of 2.93 g L -1 and a viability of 9.88 log CFU mL -1 . The formulation of an unconventional fermentation substrate from -Agro-industrial wastes of pineapple and sacha inchi to produce valuable products (such as lactic acid biomass through fermentation), is an excellent perspective for large-scale application., (© 2020 Published by Elsevier Ltd.)

Background: Antiplatelet, anticoagulant and fibrinolytic activities of stem bromelain (EC 3.4.22.4) are well described, but more studies are still required to clearly define its usefulness as an antithrombotic agent. Besides, although some effects of bromelain are linked to its proteolytic activity, few studies were performed taking into account this relationship., Objective: We aimed at comparing the effects of stem bromelain total extract (ET) and of its major proteolytic compounds on fibrinogen, fibrin, and blood coagulation considering the proteolytic activity., Methods: Proteolytic fractions chromatographically separated from ET (acidic bromelains, basic bromelains, and ananains) and their irreversibly inhibited counterparts were assayed. Effects on fibrinogen were electrophoretically and spectrophotometrically evaluated. Fibrinolytic activity was measured by the fibrin plate assay. The effect on blood coagulation was evaluated by the prothrombin time (PT) and activated partial thromboplastin time (APTT) tests. Effects were compared with those of thrombin and plasmin., Results: Acidic bromelains and ananains showed thrombin-type activity and low fibrinolytic activity, with acidic bromelains being the least effective as anticoagulants and fibrinolytics; while basic bromelains, without thrombin-like activity, were the best anticoagulant and fibrinolytic proteases present in ET. Procoagulant action was detected for ET and its proteolytic compounds by the APTT test at low concentrations. The measured effects were dependent on proteolytic activity., Conclusion: Two sub-populations of cysteine proteases exhibiting different effects on fibrin (ogen) and blood coagulation are present in ET. Using well characterized stem bromelain regarding its proteolytic system is a prerequisite for a better understanding of the mechanisms underlying the bromelain action., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Tropical fruits rich in polyphenols, ascorbic acid, and high antioxidant capacity can enhance the functional composition, flavor as well as the overall acceptance of their juices. The aim of this study was to evaluate the effects of pasteurization and storage time on the physical, chemical, and sensory quality parameters of two formulations of tropical fruit juice blends with high antioxidant capacity. Two formulations with different concentrations of acai, camu-camu, acerola, cashew apple, yellow mombin, and pineapple were pasteurized at 85 °C for 30 s, hot-filled in glass bottles, and stored in refrigerated conditions (5 °C). Analyses for color, total antioxidant activity, total polyphenols, carotenoids, ascorbic acid, and sensory quality were performed before and after pasteurization as well as at 90 and 180 day of storage. Pasteurization did not negatively affect the concentration of functional components and sensory properties. However, the storage time negatively influenced the sensory quality and reduced the ascorbic acid content of both the formulations. Further, the acceptability and the nutritional quality were still high in these products after 180-days of storage, thus, not preventing their consumption., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Association of Food Scientists & Technologists (India) 2019.)

Fruitlet core rot is one of the major postharvest disease of pineapple ( Ananas comosus var. comosus ). In the past, control strategies were designed to eliminate symptoms without addressing their causes or mechanisms, thus achieving only moderate success. In this study, (i) we focused on the anatomy of the fruitlets in the resistant "MD-2" and susceptible "Queen" pineapple cultivars; (ii) we identified the key role of the carpel margin in the infection process; (iii) we identified the key role of the sinuous layer of thick-walled cells in the inhibition of Fusarium ananatum colonization; and (iv) we linked the anatomy of the fruitlets with the phenolic content of cell walls. The fruitlet anatomy of the two cultivars was studied using X-ray, fluorescence, and multiphoton microscopy. Sepals and bracts were not perfectly fused with each other, allowing the pathogen to penetrate the fruit even after flowering. In fact, the fungi were found in the blossom cups of both cultivars but only became pathogenic in the flesh of the "Queen" pineapple fruit under natural conditions. The outer layer of the "MD-2" cavity was continuous with thick cell walls composed of ferulic and coumaric acids. The cell walls of the "Queen" blossom cup were less lignified at the extremities, and the outer layer was interspersed with cracks. The carpel margins were fused broadly in the "MD-2" pineapple, in contrast to the "Queen" pineapple. This blemish allows the fungus to penetrate deeper into the susceptible cultivar. In pineapple fruitlets, the hyphae of F. ananatum mainly progressed directly between cell walls into the parenchyma but never reached the vascular region. A layer of thick-walled cells, in the case of the resistant cultivar, stopped the colonization, which were probably the infralocular septal nectaries. Anatomical and histochemical observations coupled with spectral analysis of the hypodermis suggested the role of lignin deposition in the resistance to F. ananatum . The major phenolics bound to the cell walls were coumaric and ferulic acids and were found in higher amounts in the resistant cultivar postinoculation. The combination of fruitlet anatomy and lignification plays a role in the mechanism of host resistance to fruitlet core rot.

The cellulose synthase (Ces) and cellulose synthase-like (Csl) gene families belonging to the cellulose synthase gene superfamily, are responsible for the biosynthesis of cellulose and hemicellulose of the plant cell wall, and play critical roles in plant development, growth and evolution. However, the Ces/Csl gene family remains to be characterized in pineapple, a highly valued and delicious tropical fruit. Here, we carried out genome-wide study and identified a total of seven Ces genes and 25 Csl genes in pineapple. Genomic features and phylogeny analysis of Ces/Csl genes were carried out, including phylogenetic tree, chromosomal locations, gene structures, and conserved motifs identification. In addition, we identified 32 pineapple AcoCes/Csl genes with 31 Arabidopsis AtCes/Csl genes as orthologs by the syntenic and phylogenetic approaches. Furthermore, a RNA-seq investigation exhibited the expression profile of several AcoCes/Csl genes in various tissues and multiple developmental stages. Collectively, we provided comprehensive information of the evolution and function of pineapple Ces/Csl gene superfamily, which would be useful for screening out and characterization of the putative genes responsible for tissue development in pineapple. The present study laid the foundation for future functional characterization of Ces/Csl genes in pineapple., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Xyloglucan endotransglycosylase/hydrolase (XTH) is a cell-wall-modifying enzyme participating in diverse cell morphogenetic processes and adaptation to stress. In this study, 48 XTH genes were identified from two pineapple ( Ananas comosus ) cultivars ('F153' and 'MD2') and designated Ac(F153)XTH1 to -24 and Ac(MD2)XTH1 to -24 based on their orthology with Arabidopsis thaliana genes. Endoglucanase family 16 members were identified in addition to XTHs of glycoside hydrolase family 16. Phylogenetic analysis clustered the XTHs into three major groups (Group I/II, III and Ancestral Group) and Group III was subdivided into Group IIIA and Group IIIB. Similar gene structure and motif number were observed within a group. Two highly conserved domains, glycosyl hydrolase family 16 (GH16-XET) and xyloglucan endotransglycosylase C-terminus (C-XET), were detected by multiple sequences alignment of all XTHs. Segmental replication were detected in the two cultivars, with only the paralogous pair Ac(F153)XTH7-Ac(F153)XTH18 presented in 'F153' prior to genomic expansion. Transcriptomic analysis indicated that XTHs were involved in the regulation of fruit ripening and crassulacean acid metabolism with tissue specificity and quantitative real-time PCR analysis suggested that Ac(MD2)XTH18 was involved in root growth. The results enhance our understanding of XTHs in the plant kingdom and provide a basis for further studies of functional diversity in A. comosus .

The plasma membrane forms a permeable barrier that separates the cytoplasm from the external environment, defining the physical and chemical limits in each cell in all organisms. The movement of molecules and ions into and out of cells is controlled by the plasma membrane as a critical process for cell stability and survival, maintaining essential differences between the composition of the extracellular fluid and the cytosol. In this process aquaporins (AQPs) figure as important actors, comprising highly conserved membrane proteins that carry water, glycerol and other hydrophilic molecules through biomembranes, including the cell wall and membranes of cytoplasmic organelles. While mammals have 15 types of AQPs described so far (displaying 18 paralogs), a single plant species can present more than 120 isoforms, providing transport of different types of solutes. Such aquaporins may be present in the whole plant or can be associated with different tissues or situations, including biotic and especially abiotic stresses, such as drought, salinity or tolerance to soils rich in heavy metals, for instance. The present review addresses several aspects of plant aquaporins, from their structure, classification, and function, to in silico methodologies for their analysis and identification in transcriptomes and genomes. Aspects of evolution and diversification of AQPs (with a focus on plants) are approached for the first time with the aid of the LCA (Last Common Ancestor) analysis. Finally, the main practical applications involving the use of AQPs are discussed, including patents and future perspectives involving this important protein family., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Ananas comosus (L.) Merr. has been used as a traditional medicine in inducing abortion in many countries. Our previous in vitro experiments showed that the aqueous fraction (F4) of A. comosus extract stimulated the rat and human uterine contractions., Purpose: The aim of this study was to identify the bioactive compound and further investigate the molecular mechanism of F4 induced contraction and the in vivo uterotonic effect of F4., Materials and Methods: Organ bath studies were employed to compare the stimulatory effect of F4 in non and late pregnant uterine tissue followed by isolation of protein from late pregnant uterine tissue for the western blot analysis. The PhysioTel transmitter was implanted in pregnant SD rats to measure the changes in intrauterine pressure (IUP). Analyses of the crude extract and active principle in F4 was performed using LC-HRMS., Results: Ripe F4 in a similar manner as serotonin produced a greater stimulatory response in late pregnant than non-pregnant uterine tissue without significant change in potency; ripe F4 also increased ERK phosphorylation which eventually led to a significant increase of the final product, MMP-13. In pregnant rats (E18), oral ripe F4 (1.5 g.100 g -1 body weight) and ergometrine (1 mg) did not stimulate the uterine contraction probably due to the low level of estradiol and as a consequence low 5-HT receptors at the time of administration. In contrast, in postpartum rats, oral administration of F4 and ergometrine produced a significant increase in maximal IUP to 4.3 and 4.9 folds of basal IUP respectively. Contrary to the folklore use, unripe F4 did not stimulate the uterine activity during pregnancy and postpartum. Bioassay guided fractionation identified serotonin as a major bioactive compound in ripe F4., Conclusions: Our data clearly indicate that the uterotonic effect of ripe F4 is mediated via the serotonergic pathway and suggest that serotonin rich diet may increase the peripheral serotonin and implicate in diverse physiological functions, including uterine motility., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The aim of this study was to investigate the tocolytic properties of Ananas comosus extract in rat and human uterine tissue in vitro and in the rat in vivo. Organ bath technique was employed to perform functional studies in vitro. The PhysioTel transmitter was implanted in SD rats to measure the changes in intrauterine pressure (IUP) in vivo. Analyses of F2 was performed using LC-HRMS. F2 produced a non-selective inhibitory response on oxytocin, prostaglandin F2α, acetylcholine and KCl. The inhibitory activity of F2 on oxytocin-induced contraction was not attenuated by propranolol, TEA, glibenclamide and indomethacin. Nω-Nitro-L-arginine, a nitric oxide synthase inhibitor, suppressed the maximal tocolytic activity of F2 by 25%. DIDS, an inhibitor of chloride channels, appeared to suppress the relaxant effect of F2. F2 suppressed the oxytocin-induced contraction in Ca 2+ free solution. The in vivo tocolytic activity of F2 and ritodrine were observed in non-pregnant rats during the estrous stage by suppressing the frequency and amplitude of IUP peaks following intrauterine administration. Chemical analysis confirmed the involvement of citric acid in the tocolytic activity of F2. However, another less polar fraction is essential to accompany citric acid to produce such potent inhibitory response of F2. It is likely that F2 exerted tocolytic activity by multiple mechanisms, including antagonizing L-type Ca 2+ channels, interfering with the intracellular Ca 2+ release mechanism and releasing nitric oxide. F2 would be a promising candidate to develop as a tocolytic agent., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Ananas comosus (L.) Merr (Pineapple) is a tropical plant with an edible fruit. In the present study, the potential anti-inflammatory activity of A. comosus leaf extract (ALE) was studied. ALE prepared using soxhlet apparatus was subjected to preliminary qualitative phytochemical analysis and quantitative estimations of flavonoids and tannins. The components present in ALE were identified using liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS). Inhibitory effects of ALE on protein denaturation, and proteinase activity were assessed. Its effect on secretion of pro-inflammatory cytokines and inflammatory mediators by lipopolysaccharide-stimulated macrophages was also analyzed. Further, its anti-inflammatory activity in carrageenan-induced inflammatory rat model was examined. The preliminary qualitative phytochemical analysis revealed presence of flavonoids, phenols, tannins, carbohydrates, glycosides, and proteins in the extract. Total flavonoids and total tannins were 0.17 ± 0.006 mg equivalent of quercetin/g of ALE and 4.04 ± 0.56 mg equivalent of gallic acid/g of ALE. LC-MS analysis identified the presence of 4-hydroxy pelargonic acid, 3,4,5-trimethoxycinnamic and 4-methoxycinnamic acid, whereas GC-MS analysis identified the presence of campesterol and ethyl isoallocholate that have been previously reported for anti-inflammatory activity. ALE showed significant inhibition of protein denaturation and proteinase activity and also controlled secretion of tumour necrosis factor-α, interleukin-1β and prostaglandins, as well as the generation of reactive oxygen species by activated macrophages. ALE also significantly decreased carrageenan-induced acute paw edema. The study, therefore, identified the components present in ALE that may be responsible for its anti-inflammatory activity and thus demonstrated its potential use against acute inflammatory diseases.

Cryopreservation is a technique that allows the conservation of many species for long periods. Among the protocols used for cryopreservation, droplet vitrification has shown efficient results in preserving shoot tips of various wild and cultivated pineapple genotypes. The method consists of extraction of shoot tips from plants grown in vitro, dehydration for a period of 48 h in a preculture medium supplemented with a high concentration of sucrose, treatment in a plant vitrification solution (PVS2), and immersion in liquid nitrogen. The method described in this chapter has produced survival and regeneration indices of around 70%, depending on the genotype and physiological conditions of the initial explants. The objective of this chapter is to describe in detail a droplet vitrification protocol for shoot tips that is easy to perform for cryopreservation of pineapple germplasm.

Fusarium ananatum causes fruitlet core rot (FCR) in pineapple (Ananas comosus var. comosus) when the fruit reaches maturity. Hidden symptoms make it difficult to assess the disease, regardless of its stage, and basic questions concerning the involvement of the phenolic compounds in response to infection remain unknown. A direct inoculation method of F. ananatum in pineapple fruitlets was developed to monitor the growth of black spots and the changes in phenolic acids and ascorbic acid concentration under controlled conditions. After inoculation, infection began with a flesh discolouration at the inoculation point and then spread in a darker shade to form a black spot. Coumaroyl-isocitric and caffeoyl-isocitric acids levels respectively showed a 150- and 200-fold increase in infected fruitlet when compared to healthy fruitlet. These hydroxycinnamic acids increased minimally in the adjacent fruitlet and remained stable in the other parts of the fruit. By contrast, sinapic acid and hydroxybenzoic acid isomers (HBA) decreased after F. ananatum inoculation in the infected fruitlet, whereas they remained stable in the adjacent and healthy fruitlets. Ascorbic acid decreased to zero in the infected fruitlet. The antifungal activity of phenolic compounds and ascorbic acid was evaluated against the mycelial growth of F. ananatum. p-Coumaric acid exhibited a total inhibition of the mycelial growth at 1000 μg g -1 . Ferulic acid inhibited 64 % of mycelial growth at a concentration of 1000 μg g -1 . Caffeoylquinic acid, sinapic acid, and ascorbic acid also showed significant antifungal activity, but to a lesser extent. Finally, coinoculation of the hydroxycinnamic acids with the pathogen restrains its development in the fruit. This is the first study to highlight the involvement of phenolic compounds in the pineapple FCR disease., (Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Terpene synthases (TPSs) are vital for the biosynthesis of active terpenoids, which have important physiological, ecological and medicinal value. Although terpenoids have been reported in pineapple (Ananas comosus), genome-wide investigations of the TPS genes responsible for pineapple terpenoid synthesis are still lacking. By integrating pineapple genome and proteome data, twenty-one putative terpene synthase genes were found in pineapple and divided into five subfamilies. Tandem duplication is the cause of TPS gene family duplication. Furthermore, functional differentiation between each TPS subfamily may have occurred for several reasons. Sixty-two key amino acid sites were identified as being type-II functionally divergence between TPS-a and TPS-c subfamily. Finally, coevolution analysis indicated that multiple amino acid residues are involved in coevolutionary processes. In addition, the enzyme activity of two TPSs were tested. This genome-wide identification, functional and evolutionary analysis of pineapple TPS genes provide a new insight into understanding the roles of TPS family and lay the basis for further characterizing the function and evolution of TPS gene family., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The altered carbon assimilation pathway of crassulacean acid metabolism (CAM) photosynthesis results in an up to 80% higher water-use efficiency than C 3 photosynthesis in plants making it a potentially useful pathway for engineering crop plants with improved drought tolerance. Here we surveyed detailed temporal (diel time course) and spatial (across a leaf gradient) gene and microRNA (miRNA) expression patterns in the obligate CAM plant pineapple [Ananas comosus (L.) Merr.]. The high-resolution transcriptome atlas allowed us to distinguish between CAM-related and non-CAM gene copies. A differential gene co-expression network across green and white leaf diel datasets identified genes with circadian oscillation, CAM-related functions, and source-sink relations. Gene co-expression clusters containing CAM pathway genes are enriched with clock-associated cis-elements, suggesting circadian regulation of CAM. About 20% of pineapple microRNAs have diel expression patterns, with several that target key CAM-related genes. Expression and physiology data provide a model for CAM-specific carbohydrate flux and long-distance hexose transport. Together these resources provide a list of candidate genes for targeted engineering of CAM into C 3 photosynthesis crop species., (© 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.)

Circadian clock provides fitness advantage by coordinating internal metabolic and physiological processes to external cyclic environments. Core clock components exhibit daily rhythmic changes in gene expression, and the majority of them are transcription factors (TFs) and transcription coregulators (TCs). We annotated 1,398 TFs from 67 TF families and 80 TCs from 20 TC families in pineapple, and analyzed their tissue-specific and diurnal expression patterns. Approximately 42% of TFs and 45% of TCs displayed diel rhythmic expression, including 177 TF/TCs cycling only in the nonphotosynthetic leaf tissue, 247 cycling only in the photosynthetic leaf tissue, and 201 cycling in both. We identified 68 TF/TCs whose cycling expression was tightly coupled between the photosynthetic and nonphotosynthetic leaf tissues. These TF/TCs likely coordinate key biological processes in pineapple as we demonstrated that this group is enriched in homologous genes that form the core circadian clock in Arabidopsis and includes a STOP1 homolog. Two lines of evidence support the important role of the STOP1 homolog in regulating CAM photosynthesis in pineapple. First, STOP1 responds to acidic pH and regulates a malate channel in multiple plant species. Second, the cycling expression pattern of the pineapple STOP1 and the diurnal pattern of malate accumulation in pineapple leaf are correlated. We further examined duplicate-gene retention and loss in major known circadian genes and refined their evolutionary relationships between pineapple and other plants. Significant variations in duplicate-gene retention and loss were observed for most clock genes in both monocots and dicots., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Crude pineapple proteases extract (aka stem bromelain; EC 3.4.22.4) is an important proteolytic mixture that contains enzymes belonging to the cysteine proteases of the papain family. Numerous studies have been reported aiming at the fractionation and characterization of the many molecular species present in the extract, but more efforts are still required to obtain sufficient quantities of the various purified protease forms for detailed physicochemical, enzymatic and structural characterization. In this work, we describe an efficient strategy towards the purification of at least eight enzymatic forms. Thus, following rapid fractionation on a SP-Sepharose FF column, two sub-populations with proteolytic activity were obtained: the unbound (termed acidic) and bound (termed basic) bromelain fractions. Following reversible modification with monomethoxypolyethylene glycol (mPEG), both fractions were further separated on Q-Sepharose FF and SP-Sepharose FF, respectively. This procedure yielded highly purified molecular species, all titrating ca. 1 mol of thiol group per mole of enzyme, with distinct biochemical properties. N-terminal sequencing allowed identifying at least eight forms with proteolytic activity. The basic fraction contained previously identified species, i.e. basic bromelain forms 1 and 2, ananain forms 1 and 2, and comosain (MEROPS identifier: C01.027). Furthermore, a new proteolytic species, showing similarities with basic bomelain forms 1 and 2, was discovered and termed bromelain form 3. The two remaining species were found in the acidic bromelain fraction and were arbitrarily named acidic bromelain forms 1 and 2. Both, acidic bromelain forms 1, 2 and basic bromelain forms 1, 2 and 3 are glycosylated, while ananain forms 1 and 2, and comosain are not. The eight protease forms display different amidase activities against the various substrates tested, namely small synthetic chromogenic compounds (DL-BAPNA and Boc-Ala-Ala-Gly-pNA), fluorogenic compounds (like Boc-Gln-Ala-Arg-AMC, Z-Arg-Arg-AMC and Z-Phe-Arg-AMC), and proteins (azocasein and azoalbumin), suggesting a specific organization of their catalytic residues. All forms are completely inhibited by specific cysteine and cysteine/serine protease inhibitors, but not by specific serine and aspartic protease inhibitors, with the sole exception of pepstatin A that significantly affects acidic bromelain forms 1 and 2. For all eight protease forms, inhibition is also observed with 1,10-phenanthrolin, a metalloprotease inhibitor. Metal ions (i.e. Mn 2+ , Mg 2+ and Ca 2+ ) showed various effects depending on the protease under consideration, but all of them are totally inhibited in the presence of Zn 2+ . Mass spectrometry analyses revealed that all forms have a molecular mass of ca. 24 kDa, which is characteristic of enzymes belonging to the papain-like proteases family. Far-UV CD spectra analysis further supported this analysis. Interestingly, secondary structure calculation proves to be highly reproducible for all cysteine proteases of the papain family tested so far (this work; see also Azarkan et al., 2011; Baeyens-Volant et al., 2015) and thus can be used as a test for rapid identification of the classical papain fold., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Ethnopharmacological Relevance: In folklore medicine Ananas comosus (pineapple) is reputed to act as an abortifacient and in expectant women as a means of inducing labor. Several reports have claimed abortifacient property of A. comosus fruit (ripe or unripe). Ripe fruit has been used orally as traditional medicine in inducing abortion in Kerala state of India while the juice of unripe fruit was used for abortion in Bangladesh. However, scientific evidence supporting the efficacy of pineapple extracts in inducing uterine contractions is clearly lacking., Aim of the Study: This study investigated the pharmacological effects of different fractions of pineapple extract with a range of maturities to identify the most potent uterotonic fraction., Materials and Methods: The ethanolic crude extracts of pineapple (edible part) were prepared and fractionated through a series of liquid-liquid partitions. Fractions were separately tested on isolated uterine muscle from pregnant SD rats and human pregnant myometrium, which were cut into strips along the longitudinal axis of uterus. The strips were mounted vertically in organ baths (37°C) and exposed to cumulative addition of fractions (0.1-10mgml -1 ), serotonin (0.05-5µM) and different inhibitors to delineate the mechanism of action of the active ingredients of the extract., Results: Aqueous fraction (F4) possesses uterine stimulant property which was blocked by verapamil but unaffected by indomethacin, prazosin and atosiban. Notably, ketanserin (10µM) diminished the maximal contractile response induced by both F4 and 5HT by 74.3% and 92.1% respectively., Conclusions: These results may indicate the presence of 5HT or 5HT-like compound(s) and serotonergic pathways may contribute to the uterotonic activity of pineapple extract., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)