Dietary enzymatically-treated Artemisia annua L. supplementation could alleviate oxidative injury and improve reproductive performance of sows reared under high ambient temperature

The medicinal plant Artemisia annua L. is well known for its antimalarial compound artemisinin and the antioxidant capacity of its active ingredients. However, low bioavailability of Artemisia annua L. limits its therapeutic potential, fermentation of Artemisia annua L. can improve its bioavailability. This study was aimed to investigate the effects of dietary supplementation of enzymatically-treated Artemisia annua L. (EA) on reproductive performance, antioxidant status, milk composition of heat-stressed sows and intestinal barrier integrity of their preweaning offspring. 135 multiparous sows of average parity 4.65 (Landrace × large white) at day 85 of pregnancy were randomly distributed into 3 treatments. Sows in the control group were housed at control rooms (temperature: 27.12 ± 0.18 °C, temperature-humidity index (THI): 70.90 ± 0.80) and fed the basal diet. Sows in the HS, HS + EA groups were fed the basal diet supplemented with 0 or 1.0 g/kg EA respectively, and reared at heat stress rooms (temperature: 30.11 ± 0.16 °C, THI: 72.70 ± 0.60). Heat stress increased the malondialdehyde (MDA) content, reduced the activities of total antioxidant capacity (T-AOC) and total superoxide dismutase (T-SOD) of sows and piglets, and seriously compromised the antioxidant capacity of the sows and the intestinal integrity of their offspring. However, dietary supplementation of 1.0 g/kg EA reduced the MDA content, increased the activities of T-SOD and T-AOC in serum, colostrum, and milk of heat-stressed sows, and increased colostrum yield and 14-d milk fat content. EA supplementation also increased piglet weaning weight and the activities of T-SOD and T-AOC in serum. In addition, the abundances of intestinal tight junction proteins claudin-1 and occludin were up-regulated in piglets in EA-supplemented group. In conclusion, dietary EA supplementation at 1.0 g/kg can alleviate the oxidative stress in heat-stressed sows, improve the antioxidant capacity in both sows and their offspring, and promote the intestinal barrier integrity in their offspring. EA may be a potent dietary supplement that ameliorates oxidative stress in livestock production by improving the antioxidant capacity.     

Protein oxidation marker, α-amino adipic acid,impairs proteome of differentiated human enterocytes: Underlying toxicological mechanisms

Protein oxidation and oxidative stress are involved in a variety of health disorders such as colorectal adenomas, inflammatory bowel's disease, neurological disorders and aging, among others. In particular, the specific final oxidation product from lysine, the α-amino adipic acid (α-AA), has been found in processed meat products and emphasized as a reliable marker of type II diabetes and obesity. Currently, the underlying mechanisms of the biological impairments caused by α-AA are unknown. To elucidate the molecular basis of the toxicological effect of α-AA, differentiated human enterocytes were exposed to dietary concentrations of α-AA (200 μM) and analyzed by flow cytometry, protein oxidation and proteomics using a Nanoliquid Chromatography-Orbitrap MS/MS. Cell viability was significantly affected by α-AA (p < 0.05). The proteomic study revealed that α-AA was able to alter cell homeostasis through impairment of the Na⁺/K⁺-ATPase pump, energetic metabolism, and antioxidant response, among other biological processes. These results show the importance of dietary oxidized amino acids in intestinal cell physiology and open the door to further studies to reveal the impact of protein oxidation products in pathological conditions.     

Plant sterol oxidation products – Analogs to cholesterol oxidation products from plant origin?

Cholesterol and plant sterols are lipids which are abundantly present in a western type diet of animal and plant origin, respectively. The daily intake averages 300 mg/day each. Over the past decades, a steadily increasing consumption of plant sterol enriched dairy products (2–3 g/day) took place to lower circulating LDL cholesterol concentrations. Like all unsaturated components, plant sterols can be attacked by reactive oxygen species resulting in plant sterol oxidation products (POPs). The most widespread methods for POP determination are high-performance liquid chromatography and gas–liquid chromatography. Yet, based on the low plasma POP concentrations in normophytosterolemic subjects (POPs: ∼0.3–4.5 ng/mL), a reliable quantification yielding an appropriate limit of detection remains a challenge. While the more abundantly present cholesterol oxidation products (COPs) have elaborately been studied, research on the metabolism and biological effects of POPs is only emerging. In relation to atherogenity, biological effects including modulation of cholesterol homeostasis, membrane functioning, and inflammation are attributed to POPs. Although mostly supra-physiological concentrations are applied in in vitro assays, anti-tumor activity, cytotoxicity and estrogen-competition have been attributed to specific POPs. However, it is not obvious, if and how POPs may exert in vivo adverse or beneficial health effects similar to those attributed to COPs. In the field of nutritional science, standardized methods for the determination of POPs are required to perform relevant biological studies and to assess their presence in complex foods or biological tissues and fluids. The aim of this review is to provide an overview and evaluation of the published methods and an update on the biological effects attributed to POPs.     

Both genetic and dietary factors underlie individual differences in DNA damage levels and DNA repair capacity

The interplay between dietary habits and individual genetic make-up is assumed to influence risk of cancer, via modulation of DNA integrity. Our aim was to characterize internal and external factors that underlie inter-individual variability in DNA damage and repair and to identify dietary habits beneficial for maintaining DNA integrity.Habitual diet was estimated in 340 healthy individuals using a food frequency questionnaire and biomarkers of antioxidant status were quantified in fasting blood samples. Markers of DNA integrity were represented by DNA strand breaks, oxidized purines, oxidized pyrimidines and a sum of all three as total DNA damage. DNA repair was characterized by genetic variants and functional activities of base and nucleotide excision repair pathways.Sex, fruit-based food consumption and XPG genotype were factors significantly associated with the level of DNA damage. DNA damage was higher in women (p = 0.035). Fruit consumption was negatively associated with the number of all measured DNA lesions, and this effect was mediated mostly by β-cryptoxanthin and β-tocopherol (p < 0.05). XPG 1104His homozygotes appeared more vulnerable to DNA damage accumulation (p = 0.001). Sex and individual antioxidants were also associated with DNA repair capacity; both the base and nucleotide excision repairs were lower in women and the latter increased with higher plasma levels of ascorbic acid and α-carotene (p < 0.05).We have determined genetic and dietary factors that modulate DNA integrity. We propose that the positive health effect of fruit intake is partially mediated via DNA damage suppression and a simultaneous increase in DNA repair capacity.     

The antioxidant properties and microbial load of Moringa oleifera leaves dried using a prototype convective air-dryer

The recent COVID-19 pandemic resulted in major postharvest losses because most fresh produce could not be sold. Drying is an important thermal-based food preservation method which could have prolonged the shelf-life of these produce, but most drying technologies are costly, and cannot be afforded by small-time farmers. From this context, we were interested in evaluating the drying of Moringa oleifera leaves (MOL) using a low-cost self-built prototype convective-air dryer (CAD), alongside conventional drying methods for its antioxidant properties, microbial load and phytoconstituents. Results showed total polyphenol content was the highest (p < 0.05) in our CAD samples, and it retained among the highest total flavonoid content, total antioxidant capacity, total alkaloid content and DPPH radical scavenging activity. Furthermore, methanolic CAD extract presented lower coliform and yeast and mold count than the aqueous CAD extract. We also briefly explored MOL as a sanitizer where the microbial load of the methanolic extract was comparable (p > 0.05) with several commercial non-alcoholic sanitizers, indicating its commercialization potential as a bio-friendly sanitizer. Finally, using GC–MS, we are the first to report (best of our knowledge) on the presence of caprolactam, an important bio-medical field compound, in the CAD sample’s aqueous extract.     

Diet and trophic structure in assemblages of montane frugivorous phyllostomid bats

Neotropical frugivorous bats display a trophic structure composed of bat species with dietary preferences of core plant taxa (Artibeus-Ficus  +  Cecropia, Carollia-Piper, Sturnira- Solanum  +  Piper). This structure is hypothesized to be an ancestral trait, suggesting that similar diets would be observed throughout a species' range. However, most evidence comes from lowlands where data from montane habitats are scarce. In high mountain environments both diversity of bats and plants decreases with altitude; such decline in plant diversity produces less plants to feed from, which should ultimately affect the trophic structure of frugivorous bats in mountain environments. Here, we present a comprehensive review of the diet of frugivorous bats in Neotropical montane environments and evaluate their trophic structure in middle and higher elevations by combining a literature database with field data. We use the concept of modularity to test whether frugivorous montane bats have dietary preferences on core plant taxa. Our database revealed 47 species of montane bats feeding on 211 plant species. We find that the networks are modular, reflecting the trophic structure previously reported. We also found that in highlands the tribe Ectophyllini are Cecropia  +  Cavendishia-specialists rather than Ficus-specialists, and we describe new interactions reflecting 14 species of plants, including three botanical families previously not reported to be consumed by bats.     

Rhizophagus irregularis and nitrogen fixing azotobacter enhances greater yam (Dioscorea alata) biochemical profile and upholds yield under reduced fertilization

Greater yam (Dioscorea alata L.) is a tropical plant with a large food reserve in its underground tubers. Cultivating the greater yam is considered an essential food security crop. Yam tuber yield and quality is decreased by poor soil fertility, heavy use of fertilizers and attack of insect pest. The heavy use of fertilizers impaired the soil structure polluted the environment, and adversely impacted human beings. We employed Rhizophagus irregularis (Arbuscular Mycorrhiza Fungus) and nitrogen fixing Azotobacter to help reduce the adverse effects of fertilisers on the plants. In this study, we applied five treatments such as (1) CF: normal with conventional package and practices, (2) 70%CF: 70% chemical fertilizer, (3) 70 %CF + RI: 70% CF + AMF (R. irregularis), (4) 70%CF + AC: 70% CF + PGPB (Azotobacter chroococum), and (5) 70%CF + RI + AC: 70% CF + R. irregularis + Azotobacter chroococum, as donated as T1, T2, T3, T4 and T5, obtained that 70%CF + RI + AC was found to be the most efficient treatment under reduce chemical fertilization for improving morphological traits and biochemical content of greater yam. Although some other treatments such as 70%CF + AC, 70%CF + RI, 70% CF and CF demonstrated considerable effects in yam compared with 70%CF: 70% chemical fertilizer.     

Yield,size, nutritional value,and antioxidant activity of oyster mushrooms grown on perilla stalks

Perilla is an edible medical plant with rapidly increasing acreage in China. In this study, we investigated the potential of perilla stalks (PSs) as an alternative substrate for the cultivation of oyster mushrooms (Pleurotus ostreatus). P. ostreatus was cultivated on cottonseed hulls (CSH) alone or mixed with PSs in different ratios. The production parameters, physical characteristics, nutritional values, and antioxidant activity of mushrooms cultivated on different substrate mixtures were determined. The addition of PSs to CSH significantly improved the growth rate, yield, biological efficiency, and proximate composition and shortened the cultivation cycle. Cultivation on PSs alone increased the amino acid content in P. ostreatus fruiting bodies and the antioxidant activity of mushroom extracts. The PS75 (25% CSH + 75% PS) substrate was deduced to be the most effective substrate on the basis of yield and biological efficiency obtained in a large area where perilla had been planted. The results demonstrate that mixtures of PS with CSHs could be used as novel, practical, and easily accessible alternative substrates for P. ostreatus cultivation.     

Purslane (Portulacae oleracea L.) attenuates cadmium-induced hepatorenal and colonic damage in mice: Role of chelation,antioxidant and intestinal microecological regulation

BackgroundCadmium (Cd) is a representative pernicious metal, which has high biological toxicity. Its precaution through dietary administration is considered an important strategy. Considering that Portulaca oleracea L. (Por.L) has antioxidant, anti-inflammatory and other high medicinal value, and purslane insoluble dietary fiber (PIDF) has good binding property to metal ions, they could be good methods for Cd-induced biotoxicity therapy.PurposeTo investigate the beneficial effects of Por.L or PIDF against Cd-induced subchronic toxicity and identify its underlying mechanisms.Study design and MethodsC57BL/6 male mice (n = 12) were received 100 mg l⁻¹ CdCl₂ in water for 8 weeks. Mice were divided into four groups: Control, Cd-treated, 8% Por.L + Cd, and 8% PIDF + Cd. Histological evaluation, inductively coupled plasma-mass spectrometry, western blotting analysis, quantitative real time-PCR, gas chromatography-mass spectrometry and 16S rDNA analysis were used in the study.Results: Por.L treatment was able to inhibit inflammation and accumulation of Cd, enhance the activity of antioxidant enzymes, increase beneficial bacterial species of Akkermansia and Faecalibaculum and suppress the production of inflammatory cytokines in the colon, such as TNF-α, IL-6, IL-1β and IFN-γ. PIDF mainly relieved the toxicity of Cd by increasing the production of short chain fatty acids with anti-inflammatory functions and repressing the liver and kidney inflammation mediated by the TLR4/ MyD88/NF-κB pathway.Conclusion: Our study has demonstrated that the antagonistic-Cd effects of Por.L might be mediated via chelation, antioxidation, regulation of intestinal microecology. Thus, our study provides a novel insight into Por.L as a promising function food for the anti-Cd biotoxicity. Por.L supplement could be considered as a potential coping strategy to alleviate hazardous effects in Cd-exposed humans.     

Effects of maturity status on biochemical content,polyphenol composition and antioxidant capacity of pomegranate fruit arils (cv. ‘Bhagwa’)

Pomegranate fruit (Punica granatum L.) has gained commercial importance in recent years in the food and health industries due to increasing scientific evidence linking its consumption to better health outcomes. In the present study, the evolution of some chemical contents, individual organic acids and sugars, phenolic composition and antioxidant capacity of pomegranate (cv. ‘Bhagwa’) during maturation was investigated. The results showed significant (P < 0.05) increases in sugar content, ascorbic acid and total anthocyanins during fruit maturation, while significant decreases occurred in titratable acidity (TA), organic acids and total phenolic contents (TPCs). The significant increase in TSS/TA ratio, which plays a significant role in juice flavor, peaked at 140 DAFB, while the highest accumulation of anthocyanin content occurred at the full-ripe stage (165 DAFB). Total antioxidant capacity (both DPPH and FRAP) declined during fruit maturation, suggesting a decrease in antioxidant power of fruit juice. Strong correlations between TPC and antioxidant capacity measured by the DPPH (r² = 0.99) and FRAP (r² = 0.96) methods were observed. Fructose and glucose were established to be the major sugars in the fruit cultivar while tartaric acid was the predominant organic acid. Principal component analysis (PCA) showed that harvest maturity of ‘Bhagwa’ pomegranate fruit is dependent on time from full bloom hence could be fixed around 165 DAFB, where fruits were characterized by intense fruit and aril pigmentation as well as high juice quality parameters. This information provided could help pomegranate juice producers to assess and optimize juice quality and antioxidant value of ‘Bhagwa’ pomegranate cultivar through maturity.     

Amomum tsaoko fruit extract exerts anticonvulsant effects through suppression of oxidative stress and neuroinflammation in a pentylenetetrazol kindling model of epilepsy in mice

BackgroundChronic epilepsy is a multifaceted common brain disorder with manifold underlying factors. Epilepsy affects around 70 million peoples worldwide. Amomum tsaoko is a perennial herbaceous plant that is extensively cultivated in many provinces of China reported to exert immense biological activities.ObjectiveThis research work was aimed to reveal the therapeutic actions of ethanolic extract of A.tsaoko fruits (EE-ATF) against the pentylenetetrazol (PTZ)-provoked convulsive seizures in the mice.MethodologyThe convulsive seizures were provoked to the animals via administering 70 mg/kg of PTZ through intraperitoneally to trigger the convulsive seizures then treated with the EE-ATF at 50, 75, and 100 mg/kg orally 30 min prior to PTZ challenge. After the 30 min of PTZ challenge, animals closely monitored for signs of convulsion, generalized clonic and tonic convulsion durations, and mortality. A sub-convulsive dose 35 mg/kg of PTZ was used to provoke the kindling and seizure stages were examined using standard method. The levels of dopamine, GABA, glutamate, and Na + K + ATPase and Ca + ATPase activities in the brain tissues were studied using marker specific assay kits. The oxidative stress and antioxidant markers studied using standard methods. The mRNA expressions of COX-2, TNF-α, NF-κB, TLR-4, and IL-1β in the brain tissues were studied using RT-PCR analysis. The brain tissues were examined histologically.ResultsEE-ATF treatment remarkably decreased the onset and duration of convulsion and suppressed the seizure severity and mortality in the PTZ animals. EE-ATF treatment appreciably ameliorated the PTZ triggered modifications in the GABA, glutamate, dopamine levels and Ca + 2ATPase and Na + K + ATPase activities in the brain tissues. EE-ATF suppressed the mRNA expressions of NF-κB, IL-1β, TLR-4, TNF-α, and COX-2. The status of antioxidants were elevated by the EE-ATF. Histological findings also demonstrated the curative actions of EE-ATF.ConclusionOur findings evidenced that the EE-ATF substantially ameliorated the PTZ-provoked convulsive seizures in the mice.     

Plasma total antioxidant capacity is associated with dietary intake and plasma level of antioxidants in postmenopausal women

Increased plasma total antioxidant capacity (TAC) has been associated with a high consumption of fruits and vegetables. However, limited information is available on whether plasma TAC reflects the dietary intake of antioxidants and the levels of individual antioxidants in plasma. By using three different assays, the study aimed to determine if plasma TAC can effectively predict dietary intake of antioxidants and plasma antioxidant status. Forty overweight and apparently healthy postmenopausal women were recruited. Seven-day food records and 12-h fasting blood samples were collected for dietary and plasma antioxidant assessments. Plasma TAC was determined by vitamin C equivalent antioxidant capacity (VCEAC), ferric-reducing ability of plasma (FRAP) and oxygen radical absorbance capacity (ORAC) assays. TAC values determined by VCEAC were highly correlated with FRAP (r=0.79, P<.01) and moderately correlated with ORAC (r=0.34, P<.05). Pearson correlation analyses showed that plasma TAC values by VCEAC and ORAC had positive correlation with plasma uric acid (r=0.56 for VCEAC; r=0.49 for ORAC) and total phenolics (r=0.63 for VCEAC; r=0.36 for ORAC). However, TAC measured by FRAP was correlated only with uric acid (r=0.69). After multivariate adjustment, plasma TAC determined by VCEAC was positively associated with dietary intakes of γ-tocopherol (P<.001), β-carotene (P<.05), anthocyanidins (P<.05), flavones (P<.05), proanthocyanidins (P<.01) and TAC (P<.05), as well as with plasma total phenolics (P<.05), α-tocopherol (P<.001), β-cryptoxanthin (P<.05) and uric acid (P<.05). The findings indicate that plasma TAC measured by VCEAC reflects both dietary and plasma antioxidants and represents more closely the plasma antioxidant levels than ORAC and FRAP.     

Physiological relevance of dietary melanoidins

Melanoidins are the final products of the Maillard reaction. The main dietary sources of melanoidins are coffee, bread crust, bakery products, black beer and cocoa. Although the chemical structures of melanoidins are widely unknown, data from gravimetric techniques allow to roughly estimate a daily intake in the order of 10 g with a Western diet. Melanoidins contribute to the sensorial properties, modulating texture and flavour of foods. Growing evidence also suggests that melanoidins have health beneficial properties, such as chemopreventive, antioxidant and antimicrobial activities, and the ability to chelate different minerals. In the gastrointestinal tract, melanoidins behave not only as antioxidants, but also as dietary fibre by promoting the growth of bifidobacteria. This array of biological activities suggests the need for analytical techniques to identify the melanoidin structures and to control their formation during thermal food processing.     

Improved screening of microcystin genes and toxins in blue-green algal dietary supplements with PCR and a surface plasmon resonance biosensor

Microcystins (MCs) comprise a group of cyclic heptapeptide toxins that share a common backbone and have two variable l-amino acids that yield at least 21 known analogs of varying potency. These hepatotoxins and potential tumor promoters are produced by certain cyanobacteria, including Microcystis aeruginosa. The cyanobacterium M. aeruginosa blooms in freshwater lakes and can potentially co-occur with other species such as Aphanizomenon flos-aquae, which is targeted and harvested for the production of dietary supplements known as blue-green algae (BGA). BGA supplements are currently marketed in the U.S. and internationally as a product that may elevate mood, increase energy, and alleviate attention deficit hyperactivity disorder. However, the potential for BGA dietary supplements to be contaminated with MCs is of concern, and there are currently no validated methods for detection of MCs in these products. This research focused on establishing screening methods for toxic Microcystis and MCs in BGA supplements. A DNA-based method employing polymerase chain reaction (PCR) was used as a prescreening tool to evaluate the dietary supplements and to detect the presence of toxin genes (i.e., presence of toxic Microcystis). A rapid, sensitive surface plasmon resonance (SPR) biosensor, directed towards recognition of all MC forms, was also developed and validated. This improved SPR biosensor incorporates a commercial Adda-group antibody (Ab) that has the capacity for broader recognition of MCs than previously developed sensors for BGA supplements that rely solely on an arginine-reactive Ab and can quantitate MC levels down to 0.24 ng/mL (equivalent to 0.24 μg per gram of BGA supplement) in less than 10 min. Such a rapid, quantitative screening method may allow for further surveillance of BGA products to assist risk assessment efforts, establishment of regulatory guidance levels, and response to potential consumer complaints related to BGA products. The PCR technique and SPR biosensor may be used in concert as prescreening and screening tools, respectively or individually, thereby limiting the number of samples that must be evaluated with confirmatory methods.     

Antioxidant activity of blueberry fruit is impaired by association with milk

The antioxidant properties of dietary phenolics are believed to be reduced in vivo because of their affinity for proteins. In this study we assessed the bioavailability of phenolics and the in vivo plasma antioxidant capacity after the consumption of blueberries (Vaccinium corymbosum L.) with and without milk. In a crossover design, 11 healthy human volunteers consumed either (a) 200 g of blueberries plus 200 ml of water or (b) 200 g of blueberries plus 200 ml of whole milk. Venous samples were collected at baseline and at 1, 2, and 5 h postconsumption. Ingestion of blueberries increased plasma levels of reducing and chain-breaking potential (+ 6.1%, p < 0.001; + 11.1%, p < 0.05) and enhanced plasma concentrations of caffeic and ferulic acid. When blueberries and milk were ingested there was no increase in plasma antioxidant capacity. There was a reduction in the peak plasma concentrations of caffeic and ferulic acid (? 49.7%, p < 0.001, and ? 19.8%, p < 0.05, respectively) as well as the overall absorption (AUC) of caffeic acid (p < 0.001). The ingestion of blueberries in association with milk, thus, impairs the in vivo antioxidant properties of blueberries and reduces the absorption of caffeic acid.     

Pharmacological properties and preliminary phytochemical analysis of Pseudocaryopteris foetida (D.Don) P.D. Cantino leaves

Medicinal plants have significant contribution in pharmaceutical industries being producers of compounds utilized as precursors for drug development. A plant of Lamiaceae family; Pseudocaryopteris foetida had not been investigated for its biomedical potential. Current research was aimed to investigate phytochemical analysis, cytotoxic potential and antioxidant activity of crude methanolic extract and fractions of Pseudocaryopteris foetida (leaves). The preliminary phytochemical analysis of crude methanolic extracts and fractions of Pseudocaryopteris foetida revealed that plant is rich in phenolic and flavonoid classes of secondary metabolites while presence of tannin was observed only in crude methanolic extract. The cytotoxicity was determined using brine shrimp lethality test. Different concentrations (25, 50, 100, 150, 200 and 250 µg/mL) of crude methanolic extract and fractions exhibited dose dependent cytotoxicity. However, The LD₅₀ for all the extracts was more than 200 µg/mL indicating weak cytotoxic potential of Pseudocaryopteris foetida. The antioxidant capabilities of crude methanolic extract and fraction of Pseudocaryopteris foetida were analyzed by in vitro bio assays including DPPH, ABTS, Reducing power and phosphomolybdate antioxidant assays using ascorbic acid as standard. The crude methanolic extract showed IC₅₀ (256.38 ± 0.6 and 314.95 ± 1.1 µg/mL) for DPPH and ABTS respectively, while total antioxidant capacity was calculated as 55.79 ± 0.5 µg/mL for crude methanolic extract of Pseudocaryopteris foetida while ascorbic acid indicated total antioxidant capacity of 71.89 ± 2.3 µg/mL. Study concluded that leaves of Pseudocaryopteris foetida were the rich source of antioxidant phytochemicals. Based on preliminary investigations further research should be focused to isolate bioactive phytochemicals as leading source of clinical medicines in future.     

Effects of k-carrageenan supplementation or in combination with cholesterol-loaded cyclodextrin following freezing-thawing process of rooster spermatozoa

This experimental research purposely seeks to explore the effect of supplementing k-carrageenan (k-CRG) or CLC (cholesterol-loaded cyclodextrins) or the combined effect of k-CRG and CLC as supplements of antioxidants to an extender for rooster semen freezing. A total of 75 neat pooled ejaculates were collected twice a week from twenty-five (25) commercial line arbor acres broiler roosters (30 wks) during the experimental period. In each replicate, semen samples (n= 15, three ejaculates per rooster) were pooled and divided into nine equal aliquots, and each aliquot was diluted with one of the following extender supplemented with k-CRG, CLC, and k-CRG + CLC after which it was subjected to cryopreservation process using the “pellet” method. In study I, the supplementation of extenders with k-CRG was in five equal aliquots as follows; (0.2, 0.4, 0.6, 0.8) mg/mL and control group (k-CRG 0) mg/mL while in Study II, there was a combination of both k-CRG + CLC (0.4 mg/mL + 1.5 mg/mL, respectively), 0.4 mg/mL k-CRG, 1.5 mg/mL CLC and control group. Sperm quality parameters, endogenous antioxidant enzymes, lipid peroxidation (MDA) and ROS were all assessed after the freeze-thaw process. Our findings in study I indicated that at post-thaw, an optimum 0.4 mg/mL k-CRG supplementation in the extender improved semen quality parameters, endogenous enzymes, MDA and ROS in comparison to the control group. Interestingly prior to the freeze-thaw process, it was depicted in study II that combined k-CRG + CLC (0.4 mg/mL+1.5 mg/mL) inclusion in the extender provided maximum protection to sperm quality parameters, endogenous enzymes, MDA and ROS in comparison to 1.5 mg/mL CLC and control group at post-thaw. Besides, there was also a significant difference observed in the extenders supplemented with combined k-CRG + CLC (0.4 mg/mL +1.5 mg/mL) when compared to 0.4 mg/mL k-CRG for semen quality parameters and endogenous antioxidant enzymes (SOD, CAT, and GPx) but no significant difference was observed for MDA and ROS. Also, there was a significant difference observed in the extender supplemented with 1.5 mg/mL CLC when compared to the control group for semen quality parameters, SOD, CAT, and MDA but no significant difference for GPx and ROS at post-thaw. In conclusion, k-CRG at an optimal dosage of 0.4 mg/mL proved effective for improving post-thaw sperm quality but its combined addition k-CRG + CLC at an optimal concentration of (0.4 + 1.5) mg/mL in the extender provided greater protection to the rooster spermatozoa at post-thaw.     

Continuous production of extracellular antioxidants in suspension cells attenuates the oxidative burst detected in plant microbe interactions

Suspension cells of Solanacearum tuberosum and Nicotiana tabacum placed in fresh buffer rapidly produce and maintain significant pools of extracellular antioxidants. The extracellular antioxidant was detected by first adding a known amount of exogenous H₂O₂ to samples and then immediately measuring the remaining H₂O₂. The difference between the amount added and amount remaining was used to determine the antioxidant capacity of the sample. This extracellular antioxidant pool attenuates levels of hydrogen peroxide produced during plant–bacterial interactions. When tobacco cells were inoculated with an isolate Pseudomonas syringae pv. syringae that causes a hypersensitive response much of the antioxidant capacity had been expended neutralizing the oxidative burst characteristic of such plant–microbe interactions. After a brief delay, the levels of extracellular phenolics increased commensurate to antioxidative capacity in freshly transferred cells within 2–4 h. The strong UV absorbance of these extracellular phenolics within 250 and 350 nm was used to follow oxidation upon reaction with H₂O₂. This extracellular antioxidant pool is an important consideration in cell suspension studies of the plant–microbe oxidative burst. This study demonstrates that the true magnitude and timing of the oxidative burst in cell suspensions is masked by extracellular antioxidants.     

Analysis of the chemical composition of root essential oil from Indian sarsaparilla (Hemidesmus indicus) and its application as an ecofriendly insecticide and pharmacological agent

The Indian sarsaparilla (Hemidesmus indicus) is a commonly used plant in Indian traditional medicine of Ayurveda for the preparation of various non-alcoholic beverages. However, limited studies are available on the essential oil of H. indicus roots (HRO); therefore, the study evaluated the antioxidant, anti-inflammatory and antidiabetic activities of H. indicus root essential oil as well as insecticide potential against the common pests of stored food materials (Sitophilus oryzae, Callosobruchus maculatus and Tribolium castaneum). The repellant efficacy of HRO was found to be high against S. oryzae (8.21 ± 0.55 μg/mL). Likewise, the fumigant potential was also observed for HRO against these pests; the higher activities were observed against S. oryzae and C. maculatus (32.46 ± 1.42 and 35.18 ± 1.62 μg/L). Besides, the essential oil was also found to be active as a contact poison, however, against all the three pests, the toxicity was above 100 μg/mm³, being the highest against C. maculatus (122.8 ± 3.57 μg/mm³). To analyze the possible effect of the essential oil on grains, the different grains were allowed to germinate and compared to that of normal; thus, the non-toxic nature of HRO against the stored products is also confirmed. The essential oil shown to have DPPH hydrogen peroxide and ABTS radical scavenging, nitric oxide scavenging potential, and inhibition of lipoxgenase, alpha-amylase and alpha-glucosidase. Overall, the present study concludes that the H. indicus may be a suitable repellant and fumigant agent against different pests of stored products and a possible antioxidant, anti-inflammatory, and anti-diabetic agent.     

Optimization of growing conditions for pigments production from microalga Navicula incerta using response surface methodology and its antioxidant capacity

Navicula incerta is a marine microalga distributed in Baja California, México, commonly used in aquaculture nutrition, and has been extended to human food, biomedical, and pharmaceutical industries due to its high biological activity. Therefore, the study aimed to optimize culture conditions to produce antioxidant pigments. A central composite experimental design and response surface methodology (RSM) was employed to analyze the best culture conditions. The medium (nitrogen-deficient concentrations), salinity (PSU = Practical Salinity Unity [g/kg]), age of culture (days), and solvent extraction (ethanol, methanol, and acetone) were the factors used for the experiment. Chlorophyll a (Chl a) and total carotenoids (T-Car), determined spectroscopically, were used as the response variables. The antioxidant capacity was evaluated by DPPH^? and ABTS^?+ radical inhibition, FRAP, and anti-hemolytic activity. According to the overlay plots, the optimum growth conditions for Chl a and T-Car production were the following conditions: medium = 0.44 mol·L^-1 of NaNO₃, salinity = 40 PSU, age of culture: 3.5 days, and solvent = methanol. The pigment extracts obtained in these optimized conditions had high antioxidant activity in ABTS^?+ (86.2–92.1% of inhibition) and anti-hemolytic activity (81.8–96.7% of hemolysis inhibition). Low inhibition (33–35%) was observed in DPPH^?. The highest value of FRAP (766.03 ± 16.62 μmol TE/g) was observed in the acetonic extract. The results demonstrated that RSM could obtain an extract with high antioxidant capacity with potential applications in the biomedical and pharmaceutical industry, which encourages the use of natural resources for chemoprevention of chronic-degenerative pathologies.