Pineapple Stem Extract Research Articles

Enhancing the Efficacy of HIPEC Through Bromelain: A Preclinical Investigation in Appendiceal Cancer

IntroductionAppendiceal cancer (AC) excessive mucin production is a barrier to heated intraperitoneal chemotherapy (HIPEC) drug delivery. Bromelain is a pineapple stem extract with mucolytic properties. We explored bromelain treatment effects against mucinous AC in a patient-derived tumor organoid (PTO) model and an AC cell line.Patients and MethodsPTOs were fabricated from tumor specimens obtained from patients with AC undergoing cytoreductive surgery with HIPEC. PTOs underwent HIPEC treatment with bromelain, cisplatin, and mitomycin C (MMC) at 37 °C and 42 °C with and without bromelain pretreatment.ResultsFrom October 2020 to May 2023, 16 specimens were collected from 13 patients with low-grade (12/16, 75%) and high-grade AC (4/16, 25%). The mucin-depleting effects of bromelain were most significant in combination with N-acetylcysteine (NAC) compared with bromelain (47% versus 10%, p = 0.0009) or NAC alone (47% versus 12.8%, p = 0.0027). Bromelain demonstrated > 31% organoid viability reduction at 60 min (p < 0.001) and > 66% in 48 h (p < 0.0001). Pretreatment with bromelain increased cytotoxicity of both cisplatin and MMC HIPEC conditions by 31.6% (p = 0.0001) and 35.5% (p = 0.0001), respectively. Ki67, CK20, and MUC2 expression decreased after bromelain treatment; while increased caspase 3/7 activity and decreased Bcl-2 (p = 0.009) and Bcl-xL (p = 0.01) suggest induction of apoptosis pathways. Furthermore, autophagy proteins LC3A/B I (p < 0.03) and II (p < 0.031) were increased; while ATG7 (p < 0.01), ATG 12 (p < 0.04), and Becline 1(p < 0.03), expression decreased in bromelain-treated PTOs.ConclusionsBromelain demonstrates cytotoxicity and mucolytic activity against appendiceal cancer organoids. As a pretreatment agent, it potentiates the cytotoxicity of multiple HIPEC regimens, potentially mediated through programmed cell death and autophagy.

Determination of the crystal structure and substrate specificity of ananain

Ananain (EC 3.4.22.31) accounts for less than 10% of the total enzyme in the crude pineapple stem extract known as bromelain, yet yields the majority of the proteolytic activity of bromelain. Despite a high degree of sequence identity between ananain and stem bromelain, the most abundant bromelain cysteine protease, ananain displays distinct chemical properties, substrate preference and inhibitory profile compared to stem bromelain. A tripeptidyl substrate library (REPLi) was used to further characterize the substrate specificity of ananain and identified an optimal substrate for cleavage by ananain. The optimal tripeptide, PLQ, yielded a high kcat/Km value of 1.7 x 106 M−1s−1, with cleavage confirmed to occur after the Gln residue. Crystal structures of unbound ananain and an inhibitory complex of ananain and E−64, solved at 1.73 and 1.98 Å, respectively, revealed a geometrically flat and open S1 subsite for ananain. This subsite accommodates diverse P1 substrate residues, while a narrow and deep hydrophobic pocket-like S2 subsite would accommodate a non-polar P2 residue, such as the preferred Leu residue observed in the specificity studies. A further illustration of the atomic interactions between E−64 and ananain explains the high inhibitory efficiency of E−64 toward ananain. These data reveal the first in depth structural and functional data for ananain and provide a basis for further study of the natural properties of the enzyme.

Produksi enzimatis Virgin Coconut Oil (VCO) dengan enzim bromelin serta pemurniannya menggunakan adsorben zeolit

Salah satu produk olahan buah kelapa yang bermanfaat bagi kesehatan dan telah banyak dijadikan sebagai bahan baku industri adalah VCO. Masyarakat tradisional pedesaan dan rumah tangga telah memproduksi sendiri VCO untuk konsumsi pribadi karena khasiatnya bagi kesehatan. Akan tetapi proses produksi VCO yang masih tradisional menghasilkan produk VCO dengan standar kualitas dan rendemen yang belum maksimal. Penelitian dilakukan dengan metode enzimatis, yaitu menambahkan enzim bromelain yang terkandung dalam ekstrak batang buah nanas ke dalam santan kelapa. Produk VCO yang diperoleh selanjutnya dimurnikan dengan adsorben zeolit, dan dilakukan pengujian kadar air dan kadar asam lemak bebas. Hasil penelitian menunjukkan rendemen VCO tertinggi terdapat pada konsentrasi 20%, yaitu sebesar 36% serta pemurnian VCO dengan adsorben zeolit terbukti dapat menurunkan kadar air dan kadar asam lemak bebas dengan persentasi penurunan kadar air tertinggi sebesar 66%, dan persentasi penurunan kadar asam lemak bebas tertinggi sebesar 63%. Dari hasil penelitian dapat disimpulkan bahwa penambahan ekstrak batang buah nanas yang mengandung enzim bromelain dalam produksi VCO dapat meningkatkan rendemen VCO maksimal sebesar 35,9 % dan pemurnian sampel VCO dengan adsorben zeolit terbukti dapat membantu menurunkan kadar air dan kadar asam lemak bebas.

Pineapple stem extract (Bromelain) as an environmental friendly novel corrosion inhibitor for low carbon steel in 1 M HCl

The anticorrosion behavior of bromelain on low carbon steel (LCS) in 1 M HCl solution was studied employing weight loss, potentiodynamic polarization measurement (PDP), electrochemical impedance spectroscopy (EIS), UV–visible spectrophotometry, and surface assessment techniques like scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDAX) at 308–338 K. The obtained results suggest that bromelain is an excellent corrosion inhibitor and its inhibition efficiency (%η) is both concentration and temperature dependent. %η is observed to increase with an increase in bromelain concentration and an increase in electrolyte temperature. The maximum %η of 97.6% is observed at bromelain concentration of 1000 ppm at 338 K. The inhibitor adsorption on the LCS surface is in accordance with the Langmuir adsorption isotherm. As evidenced by PDP measurements, bromelain behaves as a mixed-type inhibitor and controls both anodic and cathodic processes. Adsorption free energy of the bromelain on LCS surface together with an increase in %η with the rise in temperature is suggestive of chemical adsorption. SEM micrographs show a smoother surface for inhibited LCS specimen. Analysis of variance statistically compare the difference existing between inhibition efficiencies from gravimetric, PDP and EIS technique and suggests that they are not significantly different.

Uji Aktivitas Antidiabetes Ekstrak Bonggol Nanas (Ananas comusus L.) pada Tikus yang Di induksi Aloksan

This research aims to knowing about decrease glucose levels in rat induced alloxan with the provision of pineapple stem extract (Ananas comusus L.). Previous studies have shown that bromelin can significantly reduce CD4+ lymphocytes, which are included in inflammatory diseases. One of the inflammatory diseases is diabetes mellitus. Blood glucose levels can be lowered by pineapple extract as it contains bromelin, and bromelin most widely found on the stem. The test animals used white rat strain wistar (Rattus norvegicus L.) with weight is 250-300 gram acclimated for seven days. The research was divided six treatment groups: positive control, negative control, P1 (25% concentration), P2 (50% concentration), P3 (75% concentration), P4 (100% concentration). After treatment for rat in category or sampel was administered pineapple stem extract for 14 days after that experiment status blood glucose level. The results showed that rat had increased (hyperglicemic) after alloxan induced,and also in blood glucose levels in all treatment. the biggest decrease was in P4 is 44 mgdl. The statistic of Kruskal-Wallis test showed that the decrease of rat blood glucose concentration at various concentration of pineapple extract influence signifificantly. According to the result that conclusion is the difference of pineapple extract concentration (25%, 50%, 75%, 100%) influence for decrease blood glucose levels and decreasing blood glucose level after 14 days, but in this time blood glucose not returned as normal conditions.

Biotechnological valorization of pineapple stem for pectinase production by Bacillus subtilis BKDS1: Media formulation and statistical optimization for submerged fermentation

Abstract Agro-residues are mainly comprised of complex polysaccharides that might serve as nutrients for microbial growth and production of enzymes. India is one of the largest producers of pineapple and along with the expanding pineapple production, associated agro-wastes are also increasing proportionally. In this background, the present study has been endeavored to accomplish valorization of pineapple stem for the economical production of the enzyme, pectinase by Bacillus subtilis BKDS1 using pineapple stem extract (PSE) medium. The results revealed that, the effective concentration of the extract towards producing maximum pectinase enzyme was found to be 12.5% in distilled water. Enhancement of pectinase production in terms of media optimization has been attempted using Plackett-Burman design (PBD) and response surface methodology (RSM). Out of nine variables screened by PBD, only four variables; yeast extract, CaCl2, CaCO3 and inoculum have been selected to affect the pectinase production significantly. The single and interactive effects of all the selected variables have been analyzed using Central composite design (CCD) of RSM. The data revealed that, optimized medium produces a higher concentration of enzyme than unoptimized medium. Further, the efficiency of modified PSE medium for the production of pectinase by submerged fermentation (Smf) has confirmed in laboratory scale fermenter. It has also been found that, enzyme production is achieved at a faster rate in fermenter compared to shaker flasks. Thus, the study as whole highlights the fact that, pineapple stems can be successfully employed in the production of the industrially important enzyme, pectinase using B. subtilis BKDS1. This will in turn, can deliver a lot towards the proper management of agro-wastes and also ensure a better cleaner environment.

Nanoparticle Formulation of Pineapple Stem Extract (Ananas Comosus (L).(Merr) as Antimicrobial Agent

Pineapple stem contains bromelain enzyme with highest concentration compared to the other parts. The aim of the research was to formulate an anti dandruff creambath containing nanoparticles of pineapple stem which was physically and chemically stable and could inhibit the growth of Pityrosporum ovale. The pineapple stem extract was then formulated into nanoparticles prepared by cross-linking method using chitosan. The resulting nanoparticles were dried using freeze drying. The dried powder of extract was used to determine the minimum inhibitory concentration (MIC) against Pityrosporum ovale. The zone of inhibition of extract and nanoparticles were measured at the MIC. The extracts were then formulated into anti dandruff creambath. The creambath in the concentration of 3% (MIC) whereas the nanoparticles were formulated in the consentration of 1, 2, 3 fold MIC. Each formula was evaluated for its physical characteristics including organoleptics, homogenity, viscosity and flow properties, spread ability and for its chemical characteristic pH value. The stability was performed for one month at temperature of 25 oC and 40 oC. The results showed that the nanoparticles have particle size of 860.03 nm and zeta potensial of 18.63 mV. The best formula was formula IV with inhibitory zone of 17 mm, and had stable organoleptic properties, homogenity, viscosity and flow properties, and pH value, as well as capaability of spreading with diameter of 4817.53 mm. Accordingly the nanoparticlesin creambath could increase the antimicrobial activity.

A standardised challenge model with an enterotoxigenic F4+ Escherichia coli strain in piglets assessing clinical traits and faecal shedding of fae and est-II toxin genes

This study evaluated the effect of five feed additives on post weaning diarrhoea (PWD) in piglets challenged 3 d after weaning with an enterotoxigenic Escherichia coli strain (ETEC). In three experimental runs, a total of 84 piglets was weaned at 21 days of age and randomly assigned to seven treatments. As dietary treatment, piglets were fed a basal diet or diets with addition of bovine colostrum (0.2%), pineapple stem extract containing bromelain (0.2%), an autolysed yeast preparation (Saccharomyces cerevisiae) (0.1%), a combination of organic acids (0.7%) and a phytogenic product with thyme essential oil (0.015%). A porcine ETEC, serotype O149:K91:K88ac was given twice via oral infection on day 3 after weaning at 1010 colony forming units/animal. One group of piglets was fed the basal diet without ETEC challenge. Traits included clinical sores, body temperature, faecal scoring and determination of faecal dry matter and the shedding of fae and est-II ETEC toxin genes. After weaning, non-challenged control piglets did not show signs of diarrhoea or impaired health, while the majority of infected piglets had a drop in body temperature, signs of diarrhoea and impaired general health. Mortality, the decrease of faecal dry matter and shedding of the toxin genes fae and est-II were not affected by the different additives. In conclusion, the ETEC challenge model induced distinct clinical signs of PWD in piglets, but the tested feed additives had no preventive effect under these conditions.

Cytotoxic effects of bromelain in human gastrointestinal carcinoma cell lines (MKN45, KATO-III, HT29-5F12, and HT29-5M21)

BackgroundBromelain is a pineapple stem extract with a variety of therapeutic benefits arising from interaction with a number of different biological processes. Several preclinical studies and anecdotal clinical observations have reported the anticancer properties of bromelain. In the present study, we investigated the cytotoxic effects of bromelain in four human cancer cell lines of gastrointestinal origin and the mechanisms involved.MethodsThe gastric carcinoma cell lines (KATO-III and MKN45) and two chemoresistant subpopulations of the HT29 colon adenocarcinoma cell line (HT29-5M21 and HT29-5F12) were treated with a range of concentrations of bromelain, as well as with cisplatin as a positive control. The effect of bromelain on the growth and proliferation of cancer cells was determined using a sulforhodamine B assay after 72 hours of treatment. Expression of apoptosis-associated proteins in MKN45 cells treated with bromelain was analyzed by Western blotting.ResultsData from our sulforhodamine B assay showed that bromelain inhibited proliferation of HT29-5F12, HT29-5M21, MKN45, and KATO-III cells, with respective half maximal inhibitory concentration values of 29, 34, 94, and 142 μg/mL. Analyzing the expression of proapoptotic and antiapoptotic proteins in bromelain-treated MKN45 cells, we observed activation of the caspase system, cleavage of PARP and p53, overexpression of cytochrome C, attenuation of phospho-Akt and Bcl2, and removal of MUC1. Apart from the caspase-dependent apoptosis observed, emergence of cleaved p53 supports a direct, extranuclear apoptotic function of p53. Moreover, interrupted Akt signaling and attenuation of Bcl2 and MUC1 oncoproteins suggest impaired survival of cancer cells.ConclusionOur findings collectively indicate that bromelain exerts cytotoxic effects in a panel of human gastric and colon carcinoma cells. Our study of MKN45 cells implicated different mechanisms in bromelain-induced cell death. While promoting apoptosis with involvement of the caspase system and extranuclear p53, bromelain also appears to impair cancer cell survival by blocking the Akt pathway and attenuating Bcl2 and MUC1 oncoproteins.

Bromelain Treatment Leads to Maturation of Monocyte‐derived Dendritic Cells but Cannot Replace PGE2 in a Cocktail of IL‐1β, IL‐6, TNF‐α and PGE2

Immunotherapy using dendritic cells (DC) has shown promising results. However, the use of an appropriate DC population is critical for the outcome of this treatment, and the search for an optimal DC subset is still ongoing. The DC used in immunotherapy today are usually matured with a cytokine cocktail consisting of TNF-α, IL-1β, IL-6 and PGE(2). These cells have deficits in their cytokine production, particularly IL-12p70, mainly because of the presence of PGE(2). Bromelain is a pineapple stem extract containing a mixture of proteases that has been used clinically in adjuvant cancer treatment. In this study, we analysed the effect of bromelain on human monocyte-derived DC. We added bromelain to the cytokine cocktail and modified cytokine cocktails with either no PGE(2) or reduced amounts of PGE(2), respectively. Combining bromelain with the cytokine cocktails containing PGE(2) resulted in an increased surface expression of CD83, CD80 and CD86. The chemokine receptor CCR7 was also considerably upregulated in these DC populations compared with DC treated with the cytokine cocktail alone. Removal or reduction of PGE(2) from the cytokine cocktail did not increase the IL-12p70 secretion from stimulated DC, and addition of bromelain to the different cytokine cocktails resulted in only a minor increase in IL-12p70 production. Moreover, combining bromelain with the cytokine cocktails did not improve the T cell stimulatory capacity of the generated DC populations. In conclusion, bromelain treatment of monocyte-derived DC does not improve the functional quality compared with the standard cytokine cocktail.

Utilization of pineapple stem juice to enhance enzyme-hydrolytic efficiency for sugarcane bagasse after an optimized pre-treatment with alkaline peroxide

The enzymatic hydrolysis of sugarcane bagasse was investigated by treating a peroxide–alkaline bagasse with a pineapple stem juice, xylanase and cellulase. Pre-treatment procedures of sugarcane bagasse with alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2 4 factorial designs, with pre-treatment time, temperature, magnesium sulfate and hydrogen peroxide concentration as factors. The responses evaluated were the yield of cellobiose and glucose released from pretreated bagasse after enzymatic hydrolysis. The results show that the highest enzymatic conversion was obtained for bagasse using 2% hydrogen peroxide at 60 °C for 16 h in the presence of 0.5% magnesium sulfate. Bagasse (5%) was treated with pineapple stem extract, which contains mixtures of protease and esterase, in combination with xylanase and cellulase. It was observed that the amount of glucose and cellobiose released from bagasse increased with the mixture of enzymes. It is believed that the enzymes present in pineapple extracts are capable of hydrolyze specific linkages that would facilitate the action of digesting plant cell walls enzymes. This increases the amount of glucose and other hexoses that are released during the enzymatic treatment and also reduces the amount of cellulase necessary in a typical hydrolysis.

Retention of the Enzymatic Activity and Product Properties During Spray Drying of Pineapple Stem Extract in Presence of Maltodextrin

The aim of this work was to investigate the effects of drying parameters on the retention of the enzymatic activity and on the physical properties of spray-dried pineapple stem extract. A Box and Behnken experimental design was used to investigate the effects of the processing parameters on the product properties. The parameters studied were the inlet temperature of drying gas (Tgi), the feed flow rate of the pineapple extract relative to evaporative capacity of the system (Ws /Wmax), and the concentration of maltodextrin added to the extract (MD). Significant effects of the processing parameters on the retention of the proteolytic activity of the powdered extract were observed. High processing temperatures lead to a product with a smaller moisture content, particle size, and lower agglomerating tendency. A product with insignificant losses of the proteolytic activity (≈ 10%) and low moisture content (less than 6.5%) is obtained at selected conditions.

Removal of necrotic tissue with an ananain‐based enzyme‐debriding preparation

An enzymatic debriding preparation was formulated with purified enzyme derived from a crude pineapple stem extract. The primary component of this preparation was the sulfhydryl protease ananain which represented >/=85% of the proteolytic activity. The remaining proteolytic activity in the preparation was contributed by a co-purifying homologous cysteine protease comosain. Taken together these two proteases provided a protein purity of greater than 95% as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This ananain-based enzyme preparation exhibited both gelatinolytic and fibrinolytic activity in vitro. Ananain-based enzyme preparation was formulated in a hydrophilic cream vehicle at concentrations ranging from 115 to 260 U/gm vehicle. Ananain-based enzyme preparation formulated in this fashion is referred to as Vianain debriding agent. Vianain was applied to partial-thickness cutaneous burn wounds produced in the skin of domestic pigs. A maximum of two 4-hour applications of Vianain provided complete debridement of eschar from the partial-thickness burn wounds as judged by light and electron microscopic analyses of biopsy specimens harvested before and after debridement. Wounds debrided with Vianain exhibited more rapid reepithelialization as compared with wounds that were not debrided. Wounds on pigs that were hyperimmunized to ananain-based enzyme preparation before burning and debridement with Vianain exhibited a similar enhancement in reepithelialization as compared with wounds treated with vehicle alone. The capacity of Vianain to debride necrotic tissue was also evaluated in a guinea pig ischemic ulcer model. Full-thickness ischemic lesions were created on the back of guinea pigs. Vianain was applied to the hardened necrotic tissue for 6 hours per day for up to a maximum of 5 days. Complete debridement of these wounds was accomplished within 4 to 5 days. Treatment of ischemic cutaneous ulcerations in this animal model with two commercially available enzyme-debriding agents provided little or no debridement of the necrotic tissue. In vitro, Vianain treatment of surgically debrided human tissue samples, obtained from patients with burn injury or cutaneous ulcers, showed that the protease preparation was effective in rapidly digesting these necrotic tissues.

Sulphhydryl protease inhibitors from pineapple plant stem

1. 1. Pineapple stem extract, consisting of a mixture of the protease bromelain and sulphhydryl protease inhibitors, was fractionated by gel permeation chromatography. 2. 2. Inhibitor-containing fractions were further resolved by ion exchange chromatography on DEAE-cellulose, giving 12 chromatographically distinct inhibitory fractions. 3. 3. These 12 inhibitory fractions all show an inhibition specificity towards bromelain. 4. 4. Reduction, S-carboxymethylation and refractionation of each of these inhibitory fractions gave, for each fraction, two separated peptides of ca 13 and 40 amino acids in length, respectively. 5. 5. The amino acid compositions and the N-terminal sequences of these peptides show the inhibitors to be a closely homologous set. Both the constituent peptides of each fraction are microheterogeneous. Each DEAE-cellulose chromatogram peak contains a co-eluting set of iso-inhibitors. 6. 6. Structural microvariations within these isoinhibitors have a minor influence on inhibitor activity towards bromelain.