Averrhoa carambola Leaf Extract Induces Apoptosis-Like Death with Increased ROS Generation in Leishmania donovani.

PURPOSE: The parasitic disease leishmaniasis is responsible for high mortality and morbidity rates worldwide. The visceral form is the most severe form of leishmaniasis (or leishmaniosis), which is caused predominantly by Leishmania donovani. Currently, clinically recommended antileishmanial drugs are not convenient because of several medical complications and resistance issues. Phytocompounds are the best candidates in this regard. The present study aimed to evaluate the antileishmanial activity of Averrhoa carambola leaf extract. METHODS: The antipromastigote activity and cytotoxicity were assessed using the MTT assay. Morphological distortions were determined using phase contrast microscopy and scanning electron microscopy (SEM). Reactive oxygen species (ROS) production, nonprotein thiol depletion and apoptotic death in promastigotes were determined via flow cytometry. UV-visible spectroscopy and energy dispersive X-ray (EDX) spectroscopy was performed for elemental analysis. Fourier-transform infrared spectroscopy (FTIR) and liquid chromatography‒mass spectrometry (LCMS) were used to characterize the phytocomponent(s) present in the extract. RESULTS: The chloroform extract of Averrhoa carambola leaf (ACCEX) (IC50 = 50.76 ± 1.7 µg/mL) exhibited the highest activity, followed by the ethyl acetate, hexane, and methanol extracts. ACCEX has also exhibited lower toxicity towards host macrophages. ACCEX also induced morphological distortions in promastigotes, with significant generation of ROS and the concomitant apoptosis initiation followed by a decrease in the nonprotein thiol level. The major phytometabolites present in ACCEX were identified from the National Institute of Standards and Technology (NIST) database and from a literature review. CONCLUSIONS: This study suggested that Averrhoa carambola leaf extracts are rich in some classes of biologically active phytocompounds and exhibit good antileishmanial activity.

Effect of edible coating of gelatin-sodium alginate with the addition of green tea (Camellia sinensis) extract on the characteristics of star fruit (Averrhoa carambola L.) during storage.

Star fruit has a good nutritional value but was very easy to damage. Edible coating can be used to extend the shelf life of star fruit. Green tea had been added to improve the mechanical properties and functional value of edible coating. This study aimed to evaluate the application of edible coating gelatin-sodium alginate with the addition of green tea to the physicochemical and microbiological characteristics of star fruit. This research was conducted by making edible coating solutions from gelatin, sodium alginate, glycerol, and green tea of various concentrations (0%, 5%, 10%, and 15%). The coating solution was applied to star fruit and stored for 1, 6, and 13 days to determine the effect of coating on the physicochemical and microbiological properties of star fruit. The results showed that adding green tea was not significantly different from the color change of the coating solution. However, there was a change in viscosity and pH along with the concentration of green tea extract (p < 0.05). FTIR analyses indicated that an interaction existed between gelatin-sodium alginate and green tea extract. The addition of green tea to star fruit with an edible coating of gelatin and sodium alginate could prevent weight loss (25.84%), reduce respiration rate (11.035 mg CO2/kg/h), maintain fruit anatomy, protect against color change, inhibit pH changes (4.22), total titrated acid (0.22%), increase vitamin C (244.55 mg/g), and even reduce damage for up to 13 days of storage. This study indicates that edible coating with the addition of green tea might be effective to retain the quality and extend the storage life star fruit.

Interpretation and explanation of computer vision classification of carambola (Averrhoa carambola L.) according to maturity stage.

The classification of carambola, also known as starfruit, according to quality parameters is usually conducted by trained human evaluators through visual inspections. This is a costly and subjective method that can generate high variability in results. As an alternative, computer vision systems (CVS) combined with deep learning (DCVS) techniques have been introduced in the industry as a powerful and an innovative tool for the rapid and non-invasive classification of fruits. However, validating the learning capability and trustworthiness of a DL model, aka black box, to obtain insights can be challenging. To reduce this gap, we propose an integrated eXplainable Artificial Intelligence (XAI) method for the classification of carambolas at different maturity stages. We compared two Residual Neural Networks (ResNet) and Visual Transformers (ViT) to identify the image regions that are enhanced by a Random Forest (RF) model, with the aim of providing more detailed information at the feature level for classifying the maturity stage. Changes in fruit colour and physicochemical data throughout the maturity stages were analysed, and the influence of these parameters on the maturity stages was evaluated using the Gradient-weighted Class Activation Mapping (Grad-CAM), the Attention Maps using RF importance. The proposed approach provides a visualization and description of the most important regions that led to the model decision, in wide visualization follows the models an importance features from RF. Our approach has promising potential for standardized and rapid carambolas classification, achieving 91 % accuracy with ResNet and 95 % with ViT, with potential application for other fruits.

Infraspecific Chemical Variability and Metabolomic Profiling by Paper Spray Ionization (PSI-MS) of Averrhoa carambola from Different Brazilian Biomes.

This research focused on the molecular diversity of A. carambola collected from three Brazilian biomes (Cerrado, Amazônia, and Mata Atlântica), whose results revealed significant differences in metabolite profiles among these biomes through PSI-MS analysis. Chemometric analysis provided valuable insights into the clustering patterns and metabolic distinctions. Cerrado and Mata Atlântica biomes exhibited a 70 % similarity, indicating a notable degree of resemblance. In Cerrado, carambolaside A was notably abundant, while carambolaside M was low in Amazônia and moderate in Cerrado samples. Carambolaside B was abundant in Amazônia but relatively low in the Cerrado and Mata Atlântica. In contrast, the Amazônia biome samples appeared to be more dissimilar. In Cerrado, epicatechin, kaempferol, and procyanidin B showed lower abundance, while apigenin, quercetin, myricetin, and rutin displayed moderate levels. Mata Atlântica showed relatively higher levels of kaempferol, quercetin, and rutin. This study indicated the environmental influence on secondary metabolites production in A. carambola fruits.

Bioactive aerogels based on native and phosphorylated potato (Solanum tuberosum L.) starches incorporated with star fruit extract (Averrhoa carambola L.).

The aim of this study was to develop a star fruit extract (SFE) and incorporate it into aerogels based on native and phosphorylated potato starches. The phosphorylation of starch enhances its properties by incorporating phosphate groups that increase the spaces between starch molecules, resulting in a more resilient, intact aerogel with enhanced water absorption. The bioactive aerogels based on potato starch and 10, 15, and 20 % (w/w) of SFE were characterized by their morphological and thermogravimetric properties, infrared spectra, water absorption capacity, loading capacity, and antioxidant activity. Epicatechin was the major compound present in SFE. The thermal stability of SFE increased when incorporated into phosphorylated starch aerogels at a concentration of 20 %. The water absorption capacity was higher in phosphorylated starch aerogels (reaching 1577 %) than in their native counterparts (reaching 1100 %). Native starch aerogels with 15 and 20 % SFE exhibited higher antioxidant activity against hydroxyl free radicals compared to phosphorylated starch aerogels, achieving 79.9 % and 86.4 % inhibition for the hydroxyl and nitric oxide radicals, respectively. The ideal choice of freeze-dried aerogel depends on the desired effect, either to act as an antioxidant agent by releasing bioactive compounds from SFE or as a water-absorbent agent in food products.

Cytotoxic/antiproliferative and nutraceutical activity of aqueous and ethanolic extracts of green and mature Averrhoa carambola.

Averrhoa carambola L. presents in its composition diversity of nutrients and vitamins. The present study aimed to extract water and fat-soluble compounds from this fruit at different stages of maturation (green and mature), perform the physical-chemical characterization as well as evaluate its cytotoxicity against hepatoma cells of Rattus norvegicus (HTC). The physicochemical results showed that the pH and molar acidity is influenced by the fruit maturation state. The fruit presented high percentage of moisture, while the percentage of total minerals (ash) increased according to its maturation stage. The results of the phytochemical screening showed that star fruits present phenolic compounds. The antioxidant activity showed greater potential for the ethanolic extracts of the green and mature star fruit. For HTC cells treated with ethanolic extract of green and mature star fruit the data show absence of cytotoxic effect. The tests with the aqueous extract showed cytotoxic/antiproliferative effect of green and mature star fruit extract, in 24, 48 and 72 hours. The presence of nutraceutical compounds and the cytotoxic/antiproliferative activity were more expressive in the aqueous extract, being an option of easily accessible solvent economic and not harmful to organisms.

Micrograph analysis of morphological alteration and cellular damage of fruit rot fungal pathogens treated with Averrhoa bilimbi fruit and Garcinia mangostana pericarp ethanolic extracts.

The aim of this study is to assess the antifungal action of Averrhoa bilimbi fruit and Garcinia mangostana pericarp ethanolic extracts in altering the morphology and causing cellular damage of Fusarium oxysporum, Fusarium proliferatum, Colletotrichum gloeosporioides and Lasiodiplodia theobromae. The pathogens were cultured on media containing both extracts individually and carbendazim as positive control, whereas media alone as negative control. All samples were processed for microscopy observations using scanning (SEM) and transmission electron (TEM) microscopes. Observation via SEM showed significant alterations in the hyphae of F. oxysporum, F. proliferatum and C. gloeosporioides compared to the control in which the hyphae were in normal form. However, no significant alteration in hyphae had been observed in the treated plate compared to the control for L. theobromae. The development of calcium carbonate crystals was also observed abundantly in control compared to treated pathogens for F. oxysporum and F. proliferatum only. This indicated that the plant extracts can inhibit some metabolic processes in the pathogens too. Observations via TEM had been conducted for F. proliferatum and C. gloeosporioides, respectively. The results showed disintegration of cytoplasmic organelles and cell wall, intense vacuolization and lyses part of fungal cells. The plant extracts have equivalent or even greater effects compared to commercial fungicide carbendazim.

Anti-Obesity Evaluation of Averrhoa carambola L. Leaves and Assessment of Its Polyphenols as Potential α-Glucosidase Inhibitors.

Averrhoa carambola L. is reported for its anti-obese and anti-diabetic activities. The present study aimed to investigate its aqueous methanol leaf extract (CLL) in vivo anti-obese activity along with the isolation and identification of bioactive compounds and their in vitro α-glucosidase inhibition assessment. CLL improved all obesity complications and exhibited significant activity in an obese rat model. Fourteen compounds, including four flavone glycosides (1-4) and ten dihydrochalcone glycosides (5-12), were isolated and identified using spectroscopic techniques. New compounds identified in planta included (1) apigenin 6-C-(2-deoxy-ß-D-galactopyranoside)-7-O-ß-D-quinovopyranoside, (8) phloretin 3'-C-(2-O-(E)-cinnamoyl-3-O-ß-D-fucopyranosyl-4-O-acetyl)-ß-D-fucopyranosyl-6'-O-ß-D fucopyranosyl-(1/2)-α-L arabinofuranoside, (11a) phloretin3'-C-(2-O-(E)-p-coumaroyl-3-O-ß-D-fucosyl-4-O-acetyl)-ß-D-fucosyl-6'-O-(2-O-ß-D-fucosyl)-α-L-arabinofuranoside, (11b) phloretin3'-C-(2-O-(Z)-p-coumaroyl-3-O-ß-D-fucosyl-4-O-acetyl)-ß-D-fucosyl-6'-O-(2-O-ß-D-fucosyl)-α-L-arabinofuranoside. Carambolaside M (5), carambolaside Ia (6), carambolaside J (7), carambolaside I (9), carambolaside P (10a), carambolaside O (10b), and carambolaside Q (12), which are reported for the first time from A. carambola L. leaves, whereas luteolin 6-C-α-L-rhamnopyranosyl-(1-2)-ß-D-fucopyranoside (2), apigenin 6-C-ß-D-galactopyranoside (3), and apigenin 6-C-α-L-rhamnopyranosyl-(1-2)-ß-L-fucopyranoside (4) are isolated for the first time from Family. Oxalidaceae. In vitro α-glucosidase inhibitory activity revealed the potential efficacy of flavone glycosides, viz., 1, 2, 3, and 4 as antidiabetic agents. In contrast, dihydrochalcone glycosides (5-11) showed weak activity, except for compound 12, which showed relatively strong activity.

Acute Kidney Injury Following Star Fruit Ingestion: A Case Series.

Star fruit (Averrhoa carambola) is a popular fruit in many tropical countries, including Sri Lanka. It is rich in oxalic acid, which is nephrotoxic in higher concentrations. The development of both acute (AKI) and chronic kidney injury after oxalate nephropathy is often underrecognized. Here we discuss the risk factors, clinical features, treatment, and outcomes of 4 patients who developed AKI after star fruit ingestion. Baseline clinical characteristics, the amount of star fruit ingested, clinical presentation, investigation, and outcome of the patients (ages 28, 50, 54, and 55 y; all male) were traced. More common symptoms of acute star fruit intoxication were nausea, vomiting, and abdominal and back pain, followed by low urine output and high serum creatinine over hours to days. Urinary analysis of all patients demonstrated oxalate crystals. Histopathologic examination of renal tissues of all 4 patients revealed acute tubular damage with calcium oxalate crystals, interstitial edema, and inflammatory cellular infiltration. The presence of calcium oxalate crystals was further confirmed with the brilliant birefringence seen under polarized light. Two patients needed intermittent hemodialysis over a week owing to oliguria and uremia. The other 2 patients did not require hemodialysis and had improvement of renal function with supportive treatment. All had high renal function on discharge but were back to normal within a month. This study highlights AKI as a serious complication of star fruit ingestion. The type and quantity of star fruit ingested and some patient factors may play a role in the pathogenesis of AKI. Public education about this serious uncommon complication is important.

Protective effect of 2-dodecyl-6-methoxycyclohexa-2, 5-diene-1, 4-dione, isolated from Averrhoa carambola L., against Aß1-42-induced apoptosis in SH-SY5Y cells by reversing Bcl-2/Bax ratio.

BACKGROUND AND PURPOSE: Aß1-42-induced neurotoxicity has been considered as a possible mechanism to aggravate the onset and progression of Alzheimer's disease (AD). In this study, we aim to determine the protective effect of DMDD on the apoptosis of SH-SY5Y cells induced by Aß1-42 and elucidate potential mechanism of DMDD's protective function in apoptosis. EXPERIMENTAL APPROACH: CCK-8, AnnexinV-FITC/PI flow cytometry, and transmission electron microscopy analysis were used to determine the protection of DMDD on Aß1-42-evoked apoptosis of SH-SY5Y cells. Cytochrome c release, JC-1 staining, and measuring the protein of Bcl-2 family by Western blot were applied to elucidate the mechanism of DMDD's protective function in apoptosis. KEY RESULTS: Three concentration of DMDD (5 µmol/L, 10 µmol/L, and 20 µmol/L) rescues the cell viability loss and apoptosis of SH-SY5Y cells cultivated in Aß1-42. The expressions of cleaved Caspase-3, -8, -9, the cytochrome c release, and mitochondrial membrane potential loss were inhibited by DMDD in Aß1-42-insulted SH-SY5Y cells. The Western blot analysis showed that DMDD pretreatment clearly downregulated the protein of Bax and upregulated Bcl-2. Moreover, the Bcl-2/Bax ratio was obviously decreased in cells only exposed to Aß1-42, but, which was suppressed by treated with DMDD. CONCLUSION AND IMPLICATIONS: DMDD attenuated the apoptosis of SH-SY5Y cells induced by Aß1-42 through reversing the Bcl-2/Bax ratio.

Rapid Structure-Based Annotation and Profiling of Dihydrochalcones in Star Fruit (Averrhoa carambola) Using UHPLC/Q-Orbitrap-MS and Molecular Networking.

Dihydrochalcones are a subclass of flavonoids. There has been growing interest in dihydrochalcones for their health benefits and potential to modulate flavor, but their comprehensive profile in diverse plant species is lacking. Star fruit is a tropical fruit rich in dihydrochalcones. In this study, a systematic annotation using UHPLC/Q-Orbitrap-MS and molecular networking was established to rapidly identify dihydrochalcones in 12 star fruit cultivars. A total of 53 dihydrochalcones were characterized within a short retention time including one novel compound (phloretin-3'-C-(2-O-trans-p-coumaroyl)-ß-d-fucopyranoside) and 23 compounds identified from the Averrhoa genus for the first time. 3-Hydroxyphloretin was the most abundant dihydrochalcone in star fruit. All the identified dihydrochalcones had a higher abundance in leaves compared to fruits. This is the first report that systematically investigates dihydrochalcones in star fruit of multiple cultivars, and the results could provide a useful reference for the future development and utilization of plant genetic resources.

Anti-Breast Cancer Effect of 2-Dodecyl-6-Methoxycyclohexa-2,5-Diene-1,4-Dione in vivo and in vitro Through MAPK Signaling Pathway.

BACKGROUND: 2-Dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) has been reported to inhibit a variety of cancer cell lines. The purpose of this study was to investigate the effects of DMDD on 4T1 breast cancer cells and the effects of DMDD on 4T1 breast cancer in mice and its molecular mechanisms. METHODS: 4T1 breast cancer cells were treated with different concentrations of DMDD, and their proliferation, apoptosis, cell-cycle distribution, migration, and invasion were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT, Acridine orange and ethidium bromide dual staining analysis (AO/EB) dual staining, flow cytometry, scratch test, and the Transwell assay. Relative quantitative real-time qPCR analysis and Western blot were applied to examine the expression levels of related genes and proteins. In animal experiments, we established a xenograft model to assess the anti-breast cancer effects of DMDD by evaluating the inhibition rate. The apoptotic activity of DMDD was evaluated by hematoxylin-eosin (HE) staining, transmission electron microscope (TEM) analysis and TdT-mediated dUTP nick end labeling (TUNEL) assays. The mRNA expression levels of MAPK pathway components were detected by relative quantitative real-time qPCR. In addition, the protein expression levels of MAPK pathway components were assessed through immunohistochemical assays and Western blotting. RESULTS: Experiments showed that DMDD could inhibit the proliferation, migration, invasion of 4T1 cells and induce cellular apoptosis and G1 cell cycle arrest. Moreover, DMDD down-regulated the mRNA expressions of raf1, mek1, mek2, erk1, erk2, bcl2, and up-regulated the mRNA expression of bax. DMDD reduced the protein expressions of p-raf1, p-mek, p-erk, p-p38, Bcl2, MMP2, MMP9 and increased the protein expressions of Bax and p-JNK. The results showed that DMDD can effectively reduce the tumor volume and weight of breast cancer in vivo, up-regulate the expression of IL-2, down-regulate the expression of IL-4 and IL-10, induce the apoptosis of breast cancer cells in mice, and regulate the expression of genes and proteins of the MAPK pathway. CONCLUSION: Our study indicates that DMDD can inhibit proliferation, migration, and invasion and induces apoptosis and cell-cycle arrest of 4T1 breast cancer cells. Also, our findings indicate that DMDD induces the apoptosis of breast cancer cells and inhibits the growth in mice. Its mechanism may be related to the MAPK pathway.

Effect of Star Fruit (Averrhoa carambola L.)By-product on Oxidative Stability of Sesame (Sesamum indicum) Oil under Accelerated Oven Storage and during Frying.

Aim of this study was to evaluate the effect of star fruit (Averrhoa carambola L.) by-products (peel and residue) on stability of sesame (Sesamum indicum) oil against oxidation. Antioxidant properties of extract of peel and residue at different time durations of extraction were determined and found that peel contains higher antioxidant potential than residue. Thus, extract of peel obtained after 24 h extraction was used to study its effectiveness on oxidative stability of sesame oil during accelerated oven storage and frying using the butylated hydroxytoluene (BHT) (200 ppm) as the reference antioxidant (positive control) and oil without added antioxidant as the negative control. The oxidative stability of the oil was determined by evaluating peroxide value, p-anisidine value, thiobarbituric acid reactive substances (TBARS) value, total oxidation (TOTOX) value, conjugated diene (CD) and conjugated triene (CT) values, and iodine value. Peel extract at different concentrations (200-1000 ppm) was tested. The oil added with peel extract exhibited higher stability against oxidation than the controls during oven storage test. Extract at 1000 ppm significantly increased the stability of sesame oil during frying as compared with controls. Thus, star fruit peel extract could be an alternative to synthetic antioxidants to suppress oxidation of edible oils.

Averrhoa bilimbi pectin-based edible films: Effects of the linearity and branching of the pectin on the physicochemical, mechanical, and barrier properties of the films.

The potential of Averrhoa bilimbi pectin (ABP) as a source of biopolymer for edible film (EF) production was explored, and deep eutectic solvent (DES) (1% w/w) containing choline chloride-citric acid monohydrate at a molar ratio of 1:1 was used as the plasticizer. The EF-ABP3:1, which was produced from ABP with large branch size, showed a higher value of melting temperature (175.30 °C), tensile stress (7.32 MPa) and modulus (33.64 MPa). The EF-ABP3:1 also showed better barrier properties by obtaining the lowest water vapor transmission rates (1.10-1.18 mg/m2.s) and moisture absorption values (2.61-32.13%) depending on the relative humidity compared to other EF-ABPs (1.39-1.83 mg/m2.s and 3.48-51.50%, respectively) that have linear structure with smaller branch size. From these results, it was suggested that the galacturonic acid content, molecular weight, degree of esterification and pectin structure of ABP significantly influenced the properties of EFs. The interaction of highly branched pectin chains was stronger than the linear chains, thus reduced the effect of plasticizer and produced a mechanically stronger EF with better barrier properties. Hence, it was suggested that these EFs could be used as alternative degradable packaging/coating materials.

Nutrient and Sensory Metabolites Profiling of Averrhoa Carambola L. (Starfruit) in the Context of Its Origin and Ripening Stage by GC/MS and Chemometric Analysis.

Averrhoa carambola L. is a tropical tree with edible fruit that grows at different climatic conditions. Despite its nutritive value and reported health benefits, it is a controversial fruit owing to its rich oxalate content. The present study aimed at investigating aroma and nutrient primary metabolites distribution in A. carambola fruits grown in Indonesia, Malaysia (its endemic origin) versus Egypt, and at different ripening stages. Two techniques were employed to assess volatile and non-volatile metabolites including headspace solid-phase micro-extraction (HS-SPME) joined with gas chromatography coupled with mass-spectrometry (GC-MS) and GC-MS post silylation, respectively. Twenty-four volatiles were detected, with esters amounting for the major class of volatiles in Egyptian fruit at ca. 66%, with methyl caproate as the major component, distinguishing it from other origins. In contrast, aldehydes predominated tropically grown fruits with the ether myristicin found exclusively in these. Primary metabolites profiling led to the identification of 117 metabolites viz. sugars, polyols and organic acids. Fructose (38-48%) and glucose (21-25%) predominated sugar compositions in ripe fruits, whereas sorbitol was the major sugar alcohol (2.4-10.5%) in ripe fruits as well. Oxalic acid, an anti-nutrient with potential health risks, was the major organic acid detected in all the studied fruits (1.7-2.7%), except the Malaysian one (0.07%). It increases upon fruit ripening, including considerable amounts of volatile oxalate esters detected via SPME, and which must not be omitted in total oxalate determinations for safety assessments.

2-Dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione, isolated from the root of Averrhoa carambola L., protects against diabetic kidney disease by inhibiting TLR4/TGFß signaling pathway.

OBJECTIVE: Diabetic kidney disease (DKD) is the leading cause of death and disability of diabetes mellitus. However, there is still a lack of specific drugs for the treatment of DKD. The chief aim of this research is to investigate the role and mechanism of 2-Dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) for DKD. METHODS: Wild type and TLR4 knockout mice were induced to diabetes. After 4-week treatment with DMDD, blood sugar, renal function, blood lipid and pathological changes were assessed. Real-time PCR, western blotting, and immunohistochemistry were employed to detect the expressions of TLR4, TGFß1 and Smad2/3 in the renal tissue. RESULTS: DMDD improved the serum lipid and decreased fasting blood glucose levels in diabetic mice. CysC and urinary albumin levels increased markedly in the diabetic group, and they were obviously decreased after 4 weeks of DMDD treatment. Compared with the WT diabetic mice, the urinary albumin and CysC in the TLR4-/- mice were expressed at lower levels. HE and Masson's staining revealed that DMDD clearly ameliorated pathological changes and renal fibrosis. When TLR4 gene was knock out, the pathological was improved. Mechanistically, TLR4, TGF-ß1 and Smad2/3 were obvious up-regulation in the renal tissues of diabetic mice. The expressions of these proteins were significantly down-regulated after DMDD treatment (p < 0.05). In the TLR4-/- mice, mRNA and protein levels of TGF-ß1 and Smad2/3 were obviously lower than those in the WT mice. In addition, IHC revealed that a strong in situ expressions of TLR4, TGF-ß1 and Smad2/3 were seen in the kidney tissues of diabetic mice, which were distinctly weakened in the DMDD-treated mice. In the TLR4-/- mice, however, expressions of TGF-ß1 and Smad2/3 were not remarkable increase in the diabetic mice compared with normal mice. CONCLUSIONS: These results strongly indicate that TLR4 is essential for DMDD protection against renal dysfunction in diabetic mice. Its hypoglycemic and anti-fibrosis effects were likely mediated by the TLR4/TGFß signaling pathway.

Flavan-3-ols and 2-diglycosyloxybenzoates from the leaves of Averrhoa carambola.

Averrhoa carambola L. (Oxalidaceae) was widely cultivated for fruits (star fruit), whereas the value of leaves remains to be discovered. Our study on the leaves yielded five flavan-3-ols (1-5) and two 2-diglycosyloxybenzoates. Their structures were determined by spectroscopic and chemical methods. Epicatechin-(5,6-bc)-4ß-(p-hydroxyphenyl)-dihydro-2(3H)-pyranone (1) and benzyl 2-ß-d-apiofuranosyl-(1 â†’ 6)-ß-d-glucopyranosyloxybenzoate (6) were new structures. 6-(S-2-Pyrrolidinone-5-yl)epicatechin (4) and 6-(R-2-pyrrolidinone-5-yl)epicatechin (5) were obtained as monomeric diastereomer for the first time and their absolute configurations were determined by electronic circular dichroism (ECD) computation. Epicatechin-(7,8-bc)-4α-(p-hydroxyphenyl)-dihydro-2(3H)-pyranone (2), epicatechin-(7,8-bc)-4ß-(p-hydroxyphenyl)-dihydro-2(3H)-pyranone (3), and methyl 2-ß-d-apiofuranosyl-(1 â†’ 6)-ß-d-glucopyranosyloxybenzoate (7) were not previously reported from the genus Averrhoa. Compounds 1-5 showed more potent 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical cation and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities and ferric reducing antioxidant power (FRAP) than l-ascorbic acid. Meanwhile 1 and 3 exhibited lipase and α-glucosidase inhibitory activities, respectively. The results clarified the structures of flavan-3-ols and 2-diglycosyloxybenzoates in the leaves and their antioxidant, lipase, and α-glucosidase inhibitory activities.

Chemo-treated 4T1 breast cancer cells radiation response measured by single and multiple cell ionization using infrared laser trap.

We present a study that uses a laser trapping technique for measurement of radiation sensitivity of untreated and chemo-treated cancer cells. We used a human mammary tumor cell line (4T1) treated by an antitumor compound, 2-Dodecyl-6-methoxycyclohexa-2, 5-diene-1,4-dione (DMDD), which was extracted from the root of Averrhoa carambola L. The untreated control group, and both 2-hour and 24-hour treated groups of 4T1 cells were used in this study. The absorbed threshold ionization energy (TIE) and the threshold radiation dose (TRD) were determined using a high-power infrared laser (at 1064 nm) trap by single and multiple cells trapping and ionization. The results were analyzed using descriptive and t-statistics. The relation of the TIE and TRD to the mass of the individual cells were also analyzed for different hours of treatment in comparison with the control group. Both TIE and TRD decrease with increasing treatment periods. However, the TRD decreases with mass regardless of the treatment. Analyses of the TRD for single vs multiple cells ionizations within each group have also consistently showed this same behavior regardless of the treatment. The underlying factors for these observed relations are explained in terms of radiation, hyperthermia, and chemo effects.

Nitrogen-Doped Carbon Dots from Averrhoa carambola Fruit Extract as a Fluorescent Probe for Methyl Orange.

In this study, a simple and green hydrothermal treatment was performed to prepare nitrogen-doped carbon dots (NCDs) from Averrhoa carambola (AC) fruit extract as a carbon precursor and L-arginine (Arg) as a nitrogen dopant. The AC-NCDs were characterized by UV light, fluorescence spectroscopy, transmission electron microscopy, FTIR spectroscopy, Raman spectroscopy, UV-vis spectroscopy, and zeta potential analyzer. The AC-NCDs were spherical and the average diameter was estimated to be 6.67 nm. The AC-NCDs exhibited the maximum emission intensity at 446 nm with 360 nm excitation wavelength. The fluorescence quenching behavior of AC-NCDs after interacting with methyl orange (MO) dye was studied. The interaction of AC-NCDs and MO was achieved within 3 min and the fluorescence quenching was maintained to a fixed value even after 30 min. The linearity was obtained in the range of 1 to 25 µM MO with a 0.30 µM detection limit. Furthermore, the pH values affected the quenching behavior of the AC-NCDs/MO system where the interaction mechanisms were driven by the electrostatic interaction, π-π interaction, inner filter effect, and energy transfer. The pH 5 maintained higher quenching efficiency while other pH values slightly decreased the quenching efficiency. Incoming applications, the AC-NCDs can be used in various important fields, especially for environmental protection.

Novel discovery of Averrhoa bilimbi ethanolic leaf extract in the stimulation of brown fat differentiation program in combating diet-induced obesity.

BACKGROUND: Brown adipocytes are known to promote energy expenditure and limit weight gain to combat obesity. Averrhoa bilimbi, locally called belimbing buluh (DBB), is mainly used as an ethnomedicine in the treatment of metabolic disorders including diabetes mellitus, hypertension and obesity. The present study aims to investigate the browning activity on white adipocytes by A. bilimbi leaf extract and to evaluate the potential mechanisms. METHODS: Ethanolic leaf extract of A. bilimbi was exposed to Myf5 lineage precursor cells to stimulate adipocyte differentiation. Protein expressions of brown adipocyte markers were determined through high content screening analysis and validated through western blotting. Mito Stress Test assay was conducted to evaluate the cellular oxygen consumption rate upon A. bilimbi treatment. RESULTS: A. bilimbi ethanolic leaf extract exhibited an adipogenesis effect similar to a PPARgamma agonist. It also demonstrated brown adipocyte differentiation in myoblastic Myf5-positive precursor cells. Expression of UCP1 and PRDM16 were induced. The basal metabolic rate and respiratory capacity of mitochondria were increased upon A. bilimbi treatment. CONCLUSIONS: The findings suggest that Averrhoa bilimbi ethanolic leaf extract induces adipocyte browning through PRDM16 activation and enhances mitochondria activity due to UCP1 up-regulation.