A smartphone aptasensor for fipronil detection in honey samples

In this study, an electrochemical smartphone-based aptasensor for the determination of fipronil was developed by modifying a screen-printed carbon electrode (SPCE). Fipronil is a broad-spectrum insecticide that has been widely used in various applications such as agriculture, veterinary, and household pest control. Recently, its use has raised concerns over the potential impact on the environment and human health. The absence of effective methods for this purpose poses a significant obstacle. To tackle this problem, we have developed a cutting-edge aptamer-based portable sensor capable of rapidly and conveniently detecting fipronil in situ. Considering that the detection of small molecules, such as fipronil, can be a challenging task, a competitive replacement assay was set up based on the aptamer’s preference for the free form of fipronil over the immobilized one on the electrode. The analytical performance provided by the sensor on standard solutions of a known fipronil content made it possible to estimate a limit of detection (LOD) equal to 1.07 μg kg−1 and a limit of quantification (LOQ) of 3.21 μg kg−1. Selectivity tests were conducted using atrazine as a possible interferent. The use and performance of the developed portable aptasensor was assessed on honey samples, which were simultaneously analyzed using an HPLC–MS method. This aptasensor could be an affordable and effective tool for accurately quantifying fipronil not only in honey samples but also in other food products. Graphical abstract: [Figure not available: see fulltext.

Effect of pasteurization on in vitro \u3b1-glucosidase inhibitory activity of apple juice

The in vitro alpha-glucosidase inhibitory activity of raw, mildly (F-71.7(5) = 0.4 min, 5 Log reductions of Cryptosporidium parvum) and intensively (F-90(12) = 14.8 min, 2 Log reductions of Alicyclobacillus acidoterrestris) pasteurized apple juice was studied. Raw apple juice (23 mg(dw)/mL) caused 90% alpha-glucosidase inhibition. Analogous results were obtained for the mildly treated sample. The most intense treatment reduced by 35% the alpha-glucosidase inhibition. However, such a decrease was associated with an increase in the phenolic content, suggesting that alpha-glucosidase inhibition might not rely on these compounds, but depend on more complex mechanisms. Apple juice was combined with acarbose to investigate their interaction towards alpha-glucosidase inhibition. A synergistic behavior was observed for concentrations < 2 mg/mL. Increasing the concentration of the combined system (up to 9 mg/mL) produced an antagonistic effect, while a further increase (< 9 mg/mL) allowed approaching an addictive behavior

Effects of manipulating slowpoke calcium-dependent potassium channel expression on rhythmic locomotor activity in Drosophila larvae

Rhythmic motor behaviors are generated by networks of neurons. The sequence and timing of muscle contractions depends on both synaptic connections between neurons and the neurons’ intrinsic properties. In particular, motor neuron ion currents may contribute significantly to motor output. Large conductance Ca2+-dependent K+ (BK) currents play a role in action potential repolarization, interspike interval, repetitive and burst firing, burst termination and interburst interval in neurons. Mutations in slowpoke (slo) genes encoding BK channels result in motor disturbances. This study examined the effects of manipulating slo channel expression on rhythmic motor activity using Drosophila larva as a model system. Dual intracellular recordings from adjacent body wall muscles were made during spontaneous crawling-related activity in larvae expressing a slo mutation or a slo RNA interference construct. The incidence and duration of rhythmic activity in slo mutants were similar to wild-type control animals, while the timing of the motor pattern was altered. slo mutants showed decreased burst durations, cycle durations, and quiescence intervals, and increased duty cycles, relative to wild-type. Expressing slo RNAi in identified motor neurons phenocopied many of the effects observed in the mutant, including decreases in quiescence interval and cycle duration. Overall, these results show that altering slo expression in the whole larva, and specifically in motor neurons, changes the frequency of crawling activity. These results suggest an important role for motor neuron intrinsic properties in shaping the timing of motor output

Reformulation and food combination as strategies to modulate glycaemia: the case of apple pomace containing biscuits administered with apple juice to healthy rats

Conventional (CB) and apple-pomace-reformulated (RB) biscuits were administered to healthy rats. Although the areas under curve (AUC) of glucose concentration were comparable between samples, differences in the glycaemic profile of CB and RB were observed. RB caused an initial steeper increase in glycaemia but a shift in the glycaemic peak from 45 to 60 min, as compared to CB. When CB or RB was ingested with apple juice (AJ) no differences were observed as compared to their ingestion with a soft drink (SD) simulating AJ sugar content, indicating that reformulation, more than the presence of AJ, was crucial in affecting the glycaemic response. Consumer acceptability towards reformulation was assessed through conjoint analysis, by simulating labels reporting information on reformulation. Consumers preferred information generally referring to the health-promoting effect (i.e. \u201clow sugar\u201d and "high fibre\u201d contents), despite directly relating to a specific disease (i.e. \u201csuitable for diabetics\u201d and \u201clow glycaemic index\u201d)

Phenolic content and potential bioactivity of apple juice as affected by thermal and ultrasound pasteurization

Thermal (T) and ultrasound (US) pasteurization processes were applied to apple juice and the phenolic compounds (TPC) were quantified before and after in vitro digestion by HPLC-DAD-ESI-MSn, with their bioaccessibility ascertained. Digested samples were analysed for their inhibitory capacity against α-glucosidase. Since some of the compounds exhibit fluorescence, both steady state and time-resolved fluorescence methods were used to investigate the binding to a blood transport protein, human serum albumin (HSA). It was found that processing induced an increase in the TPC content, which was more pronounced when US was applied. In contrast, digestion reduced the TPC content, evening out the overall effect. Still T and US pasteurized juices exhibited a higher quantity of TPC upon digestion as compared to the raw sample. No correlation was found between the TPC content and α-glucosidase inhibition, as the T and US pasteurized juices showed the highest and lowest inhibitory capacities against the enzyme, respectively. This is indicative that other compounds, such as those formed upon thermal treatment, may be involved in the antidiabetic effect of apple juice. The fluorescence study showed that binding occurred to HSA, at slightly different rates for different species present in the US treated extract. Considering energy consumption, US pasteurization is the most power consuming treatment despite its shorter duration. Overall, no univocal indication on the best pasteurization process can be gathered. Thus, it is necessary to define the desired target in order to drive technological interventions by a customized approach.</p

The role of processing on phenolic bioaccessibility and antioxidant capacity of apple derivatives

Fruit derivatives are commonly obtained by applying processing operations deemed responsible for the loss of phenol compounds, but very little information is available on the fate of phenols upon digestion of these products. The present study evaluated the effect of thermal and mechanical treatments, commonly applied to turn apple pulp into puree and homogenate, on phenolic bioaccessibility and antioxidant activity. Despite a 20 % decrease in polyphenols due to processing, their bioaccessibility was higher in apple derivatives (>20 %) compared to pulp (∼2 %). Polyphenol oxidase (PPO), inactivated by thermal treatments in apple derivatives but not in the pulp, was hypothesized to be responsible for this difference. Results acquired on an unprocessed PPO-free apple model, only featuring quercetin-3-glucoside and pectin, actually exhibited similar bioaccessibility as processed derivatives. The radical scavenging capacity was unaffected by the structural integrity of apples, indicating independence from the plant tissue's hierarchical arrangement. After digestion, radical scavenging capacity decreased in the real apple matrices, correlating with phenolic content, while it was retained in the apple model, further suggesting the pivotal food matrix role in modulating polyphenols bioaccessibility and subsequent biological activity. Translating these results to an industrial scale, processing conditions can be optimized not only to guarantee that the quality requirements are met, but also to achieve desired nutritional benefits

Cydonia oblonga Mill. Pulp Callus Inhibits Oxidative Stress and Inflammation in Injured Cells

The pharmacological activity of a callus extract from the pulp of Cydonia oblonga Mill., also known as quince, was investigated in murine macrophage (RAW 264.7) and human keratinocyte (HaCaT) cell lines. In particular, the anti-inflammatory activity of C. oblonga Mill. pulp callus extract was assessed in lipopolysaccharides (LPS)-treated RAW264.7 by the Griess test and in LPS-treated HaCaT human keratinocytes by examining the expression of genes involved in the inflammatory process, including nitric oxide synthase (iNOS), interleukin-6 (IL-6), interleukin-1 (IL-1 ), nuclear factor-kappa-B inhibitor alfa (ikB ), and intercellular adhesion molecule (ICAM). The antioxidant activity was evaluated by quantizing the reactive oxygen species (ROS) production in the hydrogen peroxide and tert-butyl hydroperoxide-injured HaCaT cell line. The obtained results indicate that C. oblonga callus from fruit pulp extract has anti-inflammatory and antioxidant activities, suggesting its possible application in delaying and preventing acute or chronic diseases associated with aging or in the treatment of wound dressing

Altered Motoneuron Properties Contribute to Motor Deficits in a Rabbit Hypoxia-Ischemia Model of Cerebral Palsy

Cerebral palsy (CP) is caused by a variety of factors attributed to early brain damage, resulting in permanently impaired motor control, marked by weakness and muscle stiffness. To find out if altered physiology of spinal motoneurons (MNs) could contribute to movement deficits, we performed whole-cell patch-clamp in neonatal rabbit spinal cord slices after developmental injury at 79% gestation. After preterm hypoxia-ischemia (HI), rabbits are born with motor deficits consistent with a spastic phenotype including hypertonia and hyperreflexia. There is a range in severity, thus kits are classified as severely affected, mildly affected, or unaffected based on modified Ashworth scores and other behavioral tests. At postnatal day (P)0–5, we recorded electrophysiological parameters of 40 MNs in transverse spinal cord slices using whole-cell patch-clamp. We found significant differences between groups (severe, mild, unaffected and sham control MNs). Severe HI MNs showed more sustained firing patterns, depolarized resting membrane potential, and fired action potentials at a higher frequency. These properties could contribute to muscle stiffness, a hallmark of spastic CP. Interestingly altered persistent inward currents (PICs) and morphology in severe HI MNs would dampen excitability (depolarized PIC onset and increased dendritic length). In summary, changes we observed in spinal MN physiology likely contribute to the severity of the phenotype, and therapeutic strategies for CP could target the excitability of spinal MNs