Pectins in Processed Fruits and Vegetables: Part II—Structure–Function Relationships

ABSTRACT: Pectin represents a very heterogeneous biopolymer whose functionality remains largely puzzling. The link between the pectin fine structure and functional properties with relevance to plant growth and development, as well as food processing, is continually being explored. This review describes the current knowledge of pectin structure–function relationships. Key mechanisms dictating pectin structure–function relationships are discussed, including the polymer biosynthesis, cross‐linking mechanisms, enzymatic and nonenzymatic conversion reactions, solubility properties, and more. Insight into the polymer structure–function relationships is highlighted by examining traditional and advanced methodologies used in pectin research. The role of pectin in modulating the quality characteristics of plant‐based foods is underlined while pin‐pointing some of the main challenges and perspectives. An integrated approach using the pectin structure–function relationship in the precision engineering of mechanical properties of tissue‐based systems is proposed.     

Extended study on the influence of z-value(s) of single and multicomponent time-temperature integrators on the accuracy of quantitative thermal process assessment

The possibilities and limitations of single- and multicomponent time-temperature integrators (TTIs) for evaluating the impact of thermal processes on a target food attribute with a Ztarget value different from the zTTI value(s) of the TTI is far from sufficiently documented. In this study, several thousand time-temperature profiles were generated by heat transfer simulations based on a wide range of product and process thermal parameters and considering a Ztarget value of 10 degrees C and a reference temperature of 121.1 degrees C, both currently used to assess the safety of food sterilization processes. These simulations included 15 different Ztarget=10 degrees CF121.1 degrees C values in the range 3 to 60 min. The integration of the time-temperature profiles with ZTTI values of 5.5 to 20.5 degrees C in steps of 1 degrees C allowed generation of a large database containing for each combination of product and process parameters the correction factor to apply to the process value FmultiTTI, which was derived from a single- or multicomponent TTI, to obtain the target process value 10 degrees CF121.1 degrees C. The table and the graph results clearly demonstrated that multicomponent TTIs with z-values close to 10 degrees C can be used as an extremely efficient approach when a single-component TTI with a z-value of 10 degrees C is not available. In particular, a two-component TTI with z1 and z2 values respectively above and below the Ztarget value (10 degrees C in this study) would be the best option for the development of a TTI to assess the safety of sterilized foods. Whatever process and product parameters are used, such a TTI allows proper evaluation of the process value 10 degrees CF121.1 degrees C.     

Mutations in the EXT1 and EXT2 genes in hereditary multiple exostoses

In an in-vitro preparation of gastric mucosae of Rana pipiens, the effect of adding melittin to a concentration of 5x10-6 M in the secretory solution on the transepithelial potential difference (PD), resistance (R) and short-circuit current (Isc) was studied. In 20 min, melittin decreased the PD by 9.3 mV and R by 148 ohm cm2. These changes can be explained by a decrease in the resistance, RP, of the paracellular pathway. To determine whether specific-ion pathways were responsible for the decrease in R, the effect of melittin on the partial conductances of Cl-, K+ and Na+ was also studied using the ion substitution method. Melittin decreased the PD response to changes in nutrient Na+, K+ and Cl- and the PD response to changes in secretory Cl-, but did not affect PD responses to changes in secretory Na+ or K+. Therefore, melittin decreased the nutrient membrane partial conductances of Cl-, K+ and Na+ and secretory membrane partial conductance of Cl-, without affecting the secretory partial conductances of Na+ or K+. Initially, melittin increased Isc in regular and Cl--free but not in Na+-free solutions. There was a delayed decrease in Isc. The results indicate that melittin decreases RP, increases the Na+ conductance of the secretory membrane and inhibits, eventually, the Na+/K+-ATPase pump.     

Correlation of Immunological and Histopathological Features with Gene Expression-Based Classifiers in Colon Cancer Patients

The purpose of this study was to evaluate the association between four distinct histopathological features: (1) tumor infiltrating lymphocytes, (2) mucinous differentiation, (3) tumor-stroma ratio, plus (4) tumor budding and two gene expression-based classifiers—(1) consensus molecular subtypes (CMS) plus (2) colorectal cancer intrinsic subtypes (CRIS). All four histopathological features were retrospectively scored on hematoxylin and eosin sections of the most invasive part of the primary tumor in 218 stage II and III colon cancer patients from two independent cohorts (AMC-AJCC-90 and AC-ICAM). RNA-based CMS and CRIS assignments were independently obtained for all patients. Contingency tables were constructed and a χ2 test was used to test for statistical significance. Odds ratios with 95% confidence intervals were calculated. The presence of tumor infiltrating lymphocytes and a mucinous phenotype (>50% mucinous surface area) were strongly correlated with CMS1 (p < 0.001 and p = 0.008) and CRIS-A (p = 0.006 and p < 0.001). The presence of mucus (≥ 10%) was associated with CMS3: mucus was present in 64.1% of all CMS3 tumors (p < 0.001). Although a clear association between tumor-stroma ratio and CMS4 was established in this study (p = 0.006), still 32 out of 61 (52.5%) CMS4 tumors were scored as stroma-low, indicating that CMS4 tumors cannot be identified solely based on stromal content. Higher budding counts were seen in CMS4 and CRIS-B tumors (p = 0.045 and p = 0.046). No other associations of the measured parameters were seen for any of the other CRIS subtypes. Our analysis revealed clear associations between histopathologic features and CMS or CRIS subtypes. However, identification of distinct molecular subtypes solely based on histopathology proved to be infeasible. Combining both molecular and morphologic features could potentially improve patient stratification.     

Asymmetric synthesis and conformational analysis by NMR spectroscopy and MD of Aba- and α-MeAba-containing dermorphin analogues

Dermorphin analogues, containing a (S)‐ and (R)‐4‐amino‐1,2,4,5‐tetrahydro‐2‐benzazepin‐3‐one scaffold (Aba) and the α‐methylated analogues as conformationally constrained phenylalanines, were prepared. Asymmetric phase‐transfer catalysis was unable to provide the (S)‐α‐Me‐o‐cyanophenylalanine precursor for (S)‐α‐MeAba in acceptable enantiomeric purity. However, by using a Schöllkopf chiral auxiliary, this intermediate was obtained in 88 % ee. [(S)‐Aba 3‐Gly 4]dermorphin retained μ‐opioid affinity but displayed an increased δ‐affinity. The corresponding R epimer was considerably less potent. In contrast, the [(R)‐α‐MeAba 3‐Gly 4]dermorphin isomer was more potent than its S epimer. Tar‐MD simulations of both non‐methylated [Aba 3‐Gly 4]dermorphin analogues showed a degree of folding at the C‐terminal residues toward the N terminus of the peptide, without however, adopting a stabilized β‐turn conformation. The α‐methylated analogues, on the other hand, exhibited a type I/I′ β‐turn conformation over the α‐MeAba 3 and Gly 4 residues, which was stabilized by a hydrogen bond involving Tyr 5‐H_N and D ‐Ala 2‐CO.     

The Protective Effects of the Autophagic and Lysosomal Machinery in Vascular and Valvular Calcification: A Systematic Review

Background: Autophagy is a highly conserved catabolic homeostatic process, crucial for cell survival. It has been shown that autophagy can modulate different cardiovascular pathologies, including vascular calcification (VCN). Objective: To assess how modulation of autophagy, either through induction or inhibition, affects vascular and valvular calcification and to determine the therapeutic applicability of inducing autophagy. Data sources: A systematic review of English language articles using MEDLINE/PubMed, Web of Science (WoS) and the Cochrane library. The search terms included autophagy, autolysosome, mitophagy, endoplasmic reticulum (ER)-phagy, lysosomal, calcification and calcinosis. Study characteristics: Thirty-seven articles were selected based on pre-defined eligibility criteria. Thirty-three studies (89%) studied vascular smooth muscle cell (VSMC) calcification of which 27 (82%) studies investigated autophagy and six (18%) studies lysosomal function in VCN. Four studies (11%) studied aortic valve calcification (AVCN). Thirty-four studies were published in the time period 2015–2020 (92%). Conclusion: There is compelling evidence that both autophagy and lysosomal function are critical regulators of VCN, which opens new perspectives for treatment strategies. However, there are still challenges to overcome, such as the development of more selective pharmacological agents and standardization of methods to measure autophagic flux.     

Behavior of mustard seed (<Emphasis Type="Italic">Sinapis alba</Emphasis> L.) myrosinase during temperature/pressure treatments: a case study on enzyme activity and stability

The activity of myrosinase, an enzyme found mainly in Brassicaceae, is influenced by some intrinsic (e.g. pH, ascorbic acid) and extrinsic (e.g. temperature, pressure) factors. In this study, the effect of intrinsic and extrinsic factors on the activity of mustard seed myrosinase (Sinapis alba L.) was determined in a buffer system and in broccoli juice. Ascorbic acid and to a much lesser extent MgCl₂ were found to enhance the myrosinase activity. In buffer solution, the optimal temperature for myrosinase activity at atmospheric pressure was 60 °C. At elevated pressure, the reaction rate increased until 200 MPa and the optimal temperature shifted to 40 °C in a buffer system. In broccoli juice, mustard seed myrosinase behaved somewhat different compared to the buffer system. The highest enzyme activity was found at 60 °C, both at atmospheric and elevated pressures. In broccoli juice, the enzymatic reaction rate also increased up to pressures of 200 MPa. Enzyme inactivation could be described by first order kinetics.     

Mild-Heat and High-Pressure Inactivation of Carrot Pectin Methylesterase: A Kinetic Study

ABSTRACT: Pectin methylesterase (PME) was extracted from carrots and purified by affinity chromatography. The thermal high-pressure inactivation of the PME was investigated in a model system. Under these conditions, the (thermo) stable fraction is not inactivated and the isobaric-isothermal inactivation followed a fractional-conversion model. At lower pressures (< 300 MPa) and higher temperatures (> 50°C), an antagonistic effect of pressure and heat was observed. A 2nd- and 3rd-degree polynomial model (derived from available thermodynamic model) was successfully used to describe the heat pressure dependence of the inactivation rate constants. From the purified carrot PME sample, the thermostable PME fraction was isolated. The thermal inactivation of this fraction followed first-order kinetics.     

Effect of high-pressure/high-temperature processing on chemical pectin conversions in relation to fruit and vegetable texture

Heat sterilization of plant derived food products entails considerable organoleptic and nutritional quality losses. For instance, texture loss of fruits and vegetables occurs, next to turgor pressure losses, mainly due to chemical changes in the cell-wall pectic polysaccharides. High-pressure sterilization, i.e. the combination of high temperature (?90 °C) with high pressure (?500 MPa), could present a positive alternative assuring safety while minimizing quality losses. In this study, the potential of high-pressure sterilization in preserving fruit and vegetable texture was evaluated by investigating the effect of combined high-pressure/high-temperature (HP/HT) treatments on two texture related chemical pectin conversions in model sytems. First, a protocol was developed to perform reproducible kinetic studies at HP/HT under constant processing conditions. Subsequently, apple pectin solutions at pH 6.5 were subjected to different HP/HT combinations (500, 600 and 700 MPa/90, 110 and 115 °C) and the extent of chemical demethoxylation and β-eliminative depolymerization was determined. At atmospheric pressure, both zero-order reaction rate constants increased with increasing temperature. At all temperatures, demethoxylation showed a higher rate constant than β-elimination. However, a temperature rise resulted in a stronger acceleration of β-elimination than of demethoxylation. When combining high temperature with high pressure, β-elimination was retarded or even stopped, whereas demethoxylation was stimulated. These results are very promising in the context of the texture preservation of high-pressure sterilized fruits and vegetables, as β-elimination is accepted to be one of the main causes of thermal softening and low methoxylated pectin can enhance tissue strength by forming cross-links with calcium ions present.