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Abstract Several drug screening campaigns identified Calpeptin as a drug candidate against SARS-CoV-2. Initially reported to target the viral main protease (Mpro), its moderate activity in Mpro inhibition assays hints at a second target. Indeed, we show that Calpeptin is an extremely potent cysteine cathepsin inhibitor, a finding additionally supported by X-ray crystallography. Cell infection assays proved Calpeptin’s efficacy against SARS-CoV-2. Treatment of SARS-CoV-2-infected Golden Syrian hamsters with sulfonated Calpeptin at a dose of 1 mg/kg body weight reduces the viral load in the trachea. Despite a higher risk of side effects, an intrinsic advantage in targeting host proteins is their mutational stability in contrast to highly mutable viral targets. Here we show that the inhibition of cathepsins, a protein family of the host organism, by calpeptin is a promising approach for the treatment of SARS-CoV-2 and potentially other viral infections.

Sour cherry is one of the most important horticultural crops in the Hungarian market. Its flavour combination makes it ideal for fresh consumption as well as canned products. The Hungarian and European markets have requested for new varieties to be introduced, making the evaluation of breeding and prebreeding material a crucial point. A total of 30 sour cherry accessions from the genebank collection were investigated for their potential inclusion into the breeding program. The main aim of the study was to identify candidates for future breeding programs, selecting their colour profiling and total polyphenolic content (TPC). This study follows the antioxidant activity of cherry species by determining the total content in polyphenols. Polyphenols are found in higher concentration in cherries when compared to other plants and have been identified as free radical scavengers, which are useful to prevent the occurrence of several diseases. Furthermore, TPC has been observed as a contributor of bitterness, acidity, colour, flavour, odour, and oxidative stability. The accessions were evaluated for their colour, fruit weight, flavour profile, firmness, and TPC. Colorimetric data were compared among four methods to illustrate to the Hungarian breeders which of the available approaches is the most accurate for sour cherry breeding. Results suggested that several accessions appear relevant for the breeding program, such as ‘Pipacs 1′, ‘Bosnyák’, ‘Hortenzia Királynője’, and ‘Mogyoródi Kései’. The total polyphenolic content was highest in ‘Pipacs1′ and lowest in ‘Kántorjánosi 3′. ‘Pipacs 1′ and ‘Hortenzia Királynője’ had interestingly high acidity content and light to very light fruit colours. ‘Bosnyák’ had a deep and dark colour with high soluble sugar content. ‘Mogyoródi Kései’ appears to have the biggest fruit. All mentioned varieties will be included in future breeding programs.

Abstract The aim of the present study was to evaluate the effect of post-flowering chilling of sweet cherry (Prunus avium L.) on the content of biochemical parameters in the leaf (chloroplast pigments, sugars and phenolics). The effect of chilling was investigated in two experiments. Potted 2-year-old trees of cv. 'Grace Star' and 'Schneiders' were exposed to one, two or three consecutive overnight chillings at an average air temperature of 4.7 °C (Experiment I), but in the following year only trees of 'Grace Star' were chilled at 2.2 °C (Experiment II), 3 to 7 weeks after flowering. The analysis of the biochemical parameters was performed by high performance liquid chromatography combined with electrospray ionization mass spectrometry. Chilling at 4.7 °C caused little or no stress, while 2.2 °C induced more intense stress with increased zeaxanthin, sugar and phenolic content in leaves, while exposure of trees to higher temperatures and closer to flowering showed no changes. Two or three consecutive overnight chilling periods increased the phenolic content and enhanced the accumulation of zeaxanthin in the leaves. Sucrose, sorbitol, fructose, total sugar, and total flavonoid content in leaves increased within 48 h after chilling. Zeaxanthin epoxidized within 24 h after one and 48 h after one and two consecutive overnight chillings.

Trees of the sweet cherry cultivar ‘Grace Star’ (Prunus avium L.) were exposed to low temperatures without frost for two consecutive nights under natural conditions 36 d after flowering, to study the effects on the physiological properties and metabolic status of leaves. The response was studied by measuring chlorophyll fluorescence and gas exchange parameters and by analyzing chloroplast pigments (i) immediately after exposure, (ii) 24 h and (iii) 48 h later. The first exposure at 2.4 (±0.2) °C and a minimum of 0.8 °C elicited more changes than the second exposure at 4.9 (±0.3) °C and a minimum of 2.4 °C. After the first exposure, the maximum quantum yield of PS II (Fv/Fm), effective quantum efficiency of PS II, net photosynthesis (PN), stomatal conductance (gs), transpiration, and intercellular CO2 concentration were significantly lower, and after the second exposure, the content of chlorophyll b, total chlorophyll, β-carotene, and lutein were lower. The content of antheraxanthin and zeaxanthin was higher immediately after both exposures, and that of antheraxanthin was also higher 24 h later. Recovery took longer in trees that were exposed twice. Fv/Fm recovered within 48 h, but the de-epoxidation state of the xanthophyll cycle pool, PN, and gs did not reach the level of controls, indicating that the stress effect lasted several days which is probably sufficient to cause fruit drop and reduce yield.

The binding of alkali metal cations with two tertiary-amide lower-rim calix[4]arenes was studied in methanol, N,N-dimethylformamide, and acetonitrile in order to explore the role of triazole and glucose functionalities in the coordination reactions. The standard thermodynamic complexation parameters were determined microcalorimetrically and spectrophotometrically. On the basis of receptor dissolution enthalpies and the literature data, the enthalpies for transfer of reactants and products between the solvents were calculated. The solvent inclusion within a calixarene hydrophobic basket was explored by means of 1H NMR spectroscopy. Classical molecular dynamics of the calixarene ligands and their complexes were carried out as well. The affinity of receptors for cations in methanol and N,N-dimethylformamide was quite similar, irrespective of whether they contained glucose subunits or not. This indicated that sugar moieties did not participate or influence the cation binding. All studied reactions were enthalpically controlled. The peak affinity of receptors for sodium cation was noticed in all complexation media. The complex stabilities were the highest in acetonitrile, followed by methanol and N,N-dimethylformamide. The solubilities of receptors were greatly affected by the presence of sugar subunits. The medium effect on the affinities of calixarene derivatives towards cations was thoroughly discussed regarding the structural properties and solvation abilities of the investigated solvents.

The influence of a water supply on the content of phytochemicals (sugars, organic acids, hydroxycinnamic acids, flavonols, flavanols and anthocyanins) in the bud, leaf and fruit of sweet cherry (Prunus avium L.) was studied in two growing seasons. In addition, the shoot length, yield efficiency and fruit weight were determined. The trees of the cultivar ‘Regina’ on Weiroot 72 or Gisela 5 rootstocks were either irrigated or non-irrigated. Irrigated trees received, in addition to rainfall, an amount of water equal to 100% of evapotranspiration, while non-irrigated trees received only rainwater (40% less). An analysis of phytochemicals was performed using high-performance liquid chromatography combined with electrospray ionization mass spectrometry. Irrigated trees had a higher content of total sugars in leaf and bud, higher content of total organic acids in the fruit, and lower content of total hydroxycinnamic acids, total flavonols and flavanols in the leaf and fruit. Irrigated trees also had higher shoot length, fruit weight and lower yield efficiency. The content of phytochemicals in bud and leaf was not affected by rootstock, but the fruit phytochemical composition, shoot length and yield efficiency were. The content of phytochemicals in the bud and leaf was influenced by the presence or absence of fruits. Our results show that irrigation, rootstock and the presence of fruits had an influence on the composition of phytochemicals in sweet cherry.

Plum pox virus (PPV), the causal agent of the sharka disease, is the most important viral disease in plums. To understand plant defense response against PPV-infection, changes in the composition of phenolics were studied in plum fruit (Prunus domestica L.). The phenolics were determined in visually undeformed and necrotic tissue during the last three ripening stages. The results indicated a significantly modified composition of anthocyanins, flavonols and hydroxycinnamic acids, in necrotic tissue the most. The phenolics differed significantly also between developmental stages and give insight into the phenolic profile of fallen unripe fruit. This study shows how PPV infection induces the biosynthesis of flavonoids in plum fruit.

Protein inhibitors of proteases are an important tool of nature to regulate and control proteolysis in living organisms under physiological and pathological conditions. In this review, we analyzed the mechanisms of inhibition of cysteine proteases on the basis of structural information and compiled kinetic data. The gathered structural data indicate that the protein fold is not a major obstacle for the evolution of a protease inhibitor. It appears that nature can convert almost any starting fold into an inhibitor of a protease. In addition, there appears to be no general rule governing the inhibitory mechanism. The structural data make it clear that the “lock and key” mechanism is a historical concept with limited validity. However, the analysis suggests that the shape of the active site cleft of proteases imposes some restraints. When the S1 binding site is shaped as a pocket buried in the structure of protease, inhibitors can apply substrate-like binding mechanisms. In contrast, when the S1 binding site is in part exposed to solvent, the substrate-like inhibition cannot be employed. It appears that all proteases, with the exception of papain-like proteases, belong to the first group of proteases. Finally, we show a number of examples and provide hints on how to engineer protein inhibitors.

Understanding the forces governing hydrophobically driven inclusion provides a path for aimed utilization of non-polar synthons and provides insights into the related hydration thermodynamics. To shed light on the factors that determine the stability of complexes with large, rigid guests, we studied the temperature and the solvent effect on the hosting of diamantane alcohols with heptameric and octameric cyclodextrins and cucurbiturils. The smaller cyclodextrin was a more efficient binder of the explored guests, while inclusion within γ-CD was observed solely in water. The higher stability of β-CD complexes in this solvent (298 K) was due to the strongly exothermic, entropically opposed inclusion, whereas endothermic hosting of alcohols by γ-CD was observed in all cases except for diamantan-1-ol. The entropically more demanding dehydration of the β-CD cavity hence masks the positive entropy changes accompanying the removal of guest-hydrating water. A strong decrease in ΔrH°(T) for all studied systems was noticed in water. In the case of cyclodextrins, the phenomenon shifts the driving force from completely or predominantly classical towards non-classical. Conversely, due to the particularly poor structuring of cucurbituril-confined water, the binding remained essentially non-classical over the explored temperature range. Unlike complexation in water, the complexation in formamide and ethylene glycol was entirely enthalpy-driven and weakly temperature-dependent. [ABSTRACT FROM AUTHOR]