Development and Characterization of Monoclonal Antibodies for the Detection of Fish Protein

This study developed and characterized anti-fish monoclonal antibodies (mAbs) capable of detecting fish, a major allergenic food, in processed food products to protect fish sensitized individuals. Of the three mAbs raised against crude protein extract of cooked fish muscle, mAb 8F5 exhibited a positive reaction to all 50 common food fish species tested with no cross-reactions to shellfish, land animals, or food additives. Although the ELISA results were negative against swordfish and yellowfin tuna, western blot clearly detected both after cooking. The ~36 kDa antigenic protein of mAb 8F5, which was found in all fish species, was detectable by mAb 8F5 in all of the fish samples even after prolonged heat treatment (100 °C, up to 60 min). These findings suggest that mAb 8F5 has great potential utility as a probe for the immunochemical detection of fish tissue in cooked food.

Spray-Dried Plasma Improves Body Weight, Intestinal Barrier Function, and Tibia Strength during Experimental Constant Heat Stress Conditions

The aim of this study was to see how spray-dried plasma (SDP) supplementation affected broiler chicken performance, intestinal permeability, and bone strength during persistent heat stress. One-day-old chicks (n = 480) were randomly assigned into twelve environmental corrals; four thermoneutral (TN-negative control, maintained at 24 °C from d 21–42); four heat stress (HS, exposed to 35 °C from d 21–42); and four heat stress treated with 2% SDP in the feed until d 28 followed by 1% SDP until d 42 (HS-SDP). The performance and serum levels of fluorescein isothiocyanate-dextran (FITC-d) were evaluated at d 21, 28, 35, and 42. The tibias strength was evaluated on d 21 and 42. The increment in chicken temperature (p < 0.05) was observed two h following the increase in environmental temperature in both HS groups and was associated with decreased performance parameters compared with the TN group. At d 42 of age, the chickens exposed to HS had an impaired gut permeability and decreased tibia strength compared to the TN group (p < 0.05). However, partially feeding SDP mitigated these adverse effects significantly. These findings imply that using SDP strategically during stressful times, such as prolonged heat stress, may help mitigate its negative consequences.

The role of glycation end products in the development and progression of diabetic neuroarthropathy

Diabetic neuroarthropathy (DNOAP, Charcot’s foot) is a serious complication of diabetes mellitus, the genesis of which is not fully understood. In most cases, this pathology is diagnosed late, which leads to the development of severe deformities of the foot, up to the loss of support ability of the limb. There is no single hypothesis for the formation of Charcot’s foot, but there are factors predisposing to its development, as well as a few likely provoking events. Excessive formation and accumulation of end products of glycation may play an important role in the pathogenesis of this complication of diabetes. End products of glycation (AGE) are a variety of compounds formed as a result of a non-enzymatic reaction between carbohydrates and free amino groups of proteins, lipids and nucleic acids. There are various factors that lead to the accumulation of AGE in the human body. Allocate endogenous and exogenous factors. The former include certain diseases, such as diabetes mellitus, renal failure, which accelerate glycation processes. Exogenous factors leading to the formation of lipo-oxidation and glyco-oxidation products include tobacco smoke and prolonged heat treatment of food.This review provides information on the role of glycation end products in the development and progression of complications in patients with diabetes mellitus.

Manganese Benefits Heat-Stressed Corals at the Cellular Level

The intensity and frequency of coral bleaching events have increased worldwide especially due to thermal stress and seawater pollution. Although it has been observed that metal concentration in seawater can affect the coral’s ability to adopt cellular defensive mechanisms to counteract bleaching, more investigations are needed to better understand the role of metals in coral physiology. In this study, we analyzed the individual and combined effects of prolonged heat stress and manganese (Mn) and iron (Fe) enrichments at the cellular level on the coral Stylophora pistillata. Thermal stress caused an up-regulation in the expression of the host Heat shock proteins (Hsps) 60 and 70, which showed a parallel pattern of modulation in all treatments, as well as an increase of lipid peroxidation (LPO) in the holobiont. Fe enrichment did not induce any change in Hsp expression or in the oxidative status of the corals both at the ambient temperature of 26°C or at increased temperature, suggesting that Fe didn’t seem to play a role in mitigating the cellular damages and the coral bleaching. Mn or MnFe enrichment at 26°C seemed to increase the oxidative stress in zooxanthellae, since high LPO and glutathione reductase (GR) levels were recorded, but it did not cause any effect on polyp Hsp expression, probably due to the antioxidant action of GR. With the temperature increase, Mn enrichments prevented any increase in Hsp levels and caused a significant decrease of LPO and GR activity, strengthening a previous hypothesis suggesting that Mn could mitigate the negative cellular effects produced by the thermal stress.

Heat Stress Pre-Exposure May Differentially Modulate Plant Defense to Powdery Mildew in a Resistant and Susceptible Barley Genotype

Heat stress negatively affects barley production and under elevated temperatures defense responses to powdery mildew (Blumeria graminis f. sp. hordei, Bgh) are altered. Previous research has analyzed the effects of short-term (30 s to 2 h) heat stress, however, few data are available on the influence of long-term exposure to heat on powdery mildew infections. We simultaneously assessed the effects of short and long term heat pre-exposure on resistance/susceptibility of barley to Bgh, evaluating powdery mildew infection by analyzing symptoms and Bgh biomass with RT-qPCR in barley plants pre-exposed to high temperatures (28 and 35 °C from 30 s to 5 days). Plant defense gene expression after heat stress pre-exposure and inoculation was also monitored. Our results show that prolonged heat stress (24, 48 and 120 h) further enhanced Bgh susceptibility in a susceptible barley line (MvHV118-17), while a resistant line (MvHV07-17) retained its pathogen resistance. Furthermore, prolonged heat stress significantly repressed the expression of several defense-related genes (BAX inhibitor-1, Pathogenesis related-1b and Respiratory burst oxidase homologue F2) in both resistant and susceptible barley lines. Remarkably, heat-suppressed defense gene expression returned to normal levels only in MvHV07-17, a possible reason why this barley line retains Bgh resistance even at high temperatures.

Relative contribution of Trend and Interannually-varying SST Anomalies to the 2018 Heat Waves in the Extratropical Northern Hemisphere

The northern extratropics—including regions in northern Europe, northeast Asia, and North America—experienced extremely prolonged heat waves during May–August 2018. Record-breaking surface temperatures, which caused numerous deaths, were observed in several cities. The 2018 heat waves exhibited a circumglobal characteristic owing to a circumpolar perturbation (CCP) in the middle–upper troposphere of the Northern Hemisphere (NH). The CCP had two parts: a wave-like perturbation and a hemispheric perturbation that was almost zonally symmetric. Singular value decomposition analysis revealed that the zonally symmetric perturbation was coupled to the SST warming trend, whereas the wave-like perturbation was primarily coupled to the interannually-varying SST anomaly (SSTA), particularly in the tropical North Pacific, which reached an extreme in 2018. Numerical experiments confirmed that the zonally symmetric component was primarily resulted from the SSTA associated with the warming trend, whereas the interannually-varying SSTAs in the NH contributed mostly to the wave-like perturbation. The warming trend component of SSTA, especially that in the tropics, compounded by the unusually large SSTAs in 2018, was hypothesized to have contributed to inducing the circumpolar circulation anomaly that caused the record-breaking heat waves in the extratropical NH in 2018.

Methods for restoration of ki67 antigenicity in aged paraffin tissue blocks

Pathology archives are a treasure trove of paraffin embedded tissue spanning many years and covering a wide variety of tissues and diseases. The possibility of using old archival formalin fixed paraffin embedded (FFPE) tissues for diagnostic updates and research projects is a widespread need and it requires archives of stable, well-preserved samples. Immunohistochemistry performed on old archival paraffin blocks may give unreliable results, in particular for some antigens, such as Ki67. In consideration of this phenomenon, our aim is to comprehensively test and identify methods which may be used to obtain Ki67 immunohistochemical reactions of good quality from old archival FFPE blocks. Various methods were tested in order to evaluate their possible efficacy in increasing Ki67 immunointensity in a collection of 40-year-old, archival blocks including re-embedding, with deeper sectioning of tissue from the block and increasing heat-based pretreatment times (20 cases) and re-processing (20 cases). All reactions were performed using an automated immunostainer and Ki67 stained immunosections compared using a visual colour-based scale (the first immunostained section was considered as baseline). The combination of deep sectioning (1000 µM) and prolonged heat-based pretreatment (64 min) markedly increased immunoreactivity for Ki67. Re-embedding and reprocessing did not have a significant effect. Large tissue samples showed heterogeneity of Ki67 immunoexpression between the periphery of the sample and the central area. In conclusion, the study defines a useful protocol to increase antigen retrieval applicable to dated archival tissues.

Gene and Metabolite Integration Analysis through Transcriptome and Metabolome Brings New Insight into Heat Stress Tolerance in Potato (Solanum tuberosum L.)

Potatoes are particularly vulnerable to elevated temperatures, with short heat stress (6 h) inducing stomatal opening and reducing membrane stability and prolonged heat stress (3-day) decreasing the photosynthetic capacity of potato leaves. The integration of transcriptomics and metabolomics methods demonstrated that 448 heat upregulated and 918 heat downregulated genes and 325 and 219 compounds in the positive and negative ionization modes, respectively, were up- or downregulated in leaves in response to short and prolonged heat stress. Differentially expressed genes enriched in photosynthesis, cell wall degradation, heat response, RNA processing, and protein degradation were highly induced during heat exposure, and differentially expressed metabolites involved in amino acid biosynthesis and secondary metabolism were mostly induced during heat exposure, suggesting a possible role of these genes and metabolites in the heat tolerance of the potato. Metabolite and transcript abundances for the upregulation of flavone and flavonol biosynthesis under prolonged heat stress were closely correlated. Heat-induced gene expression in Arabidopsisthaliana shoots and potato leaves overlapped, and heat stress-responsive genes overlapped with drought stress-related genes in potato. The transient expression of four heat-induced genes in Nicotiana benthamiana exhibited increased heat tolerance. This study provides a new transcriptome and metabolic profile of the potato’s response to heat.

Quality Markers for Astragali Radix and Its Products Based on Process Analysis

Due to the complex nature of traditional medicines, quality control methods need to cover two aspects: compliance of raw materials with quality standards and process control. Astragali radix (AR), the roots of Astragalus mongholicus Bunge, was selected in this study as an example of a widely used traditional medicine in various formulations. Astragaloside IV (AG IV) and calycosin 7-O-β-D-glucoside (CG) are used as the markers for the quality control of AR and its products in the Chinese Pharmacopoeia. However, in the raw materials, malic acid esters of the CG and acetate esters of the astragaloside are easily decomposed into CG and AG IV during storage and processing of AR to make extracts for various preparations. The thermal stability of the isoflavonoids and astragalosides in decoction was studied. The level of CG and astragalosides (AG I/AG II/AG IV) was strongly affected by prolonged heat during processing, while calycosin was stable in the conditions. Also the major astragalosides in AR could fully converted into AG IV which eventually reaches a stable level under certain conditions. With calycosin and AG IV as marker components, practical, reproducible, and precise methods were established and applied to the quality analysis of AR from its raw materials to its intermediates and products. This study demonstrates that a full chemical profiles analysis of the whole manufacturing process (from “raw materials—intermediates/extracts—final product”) is important to identify quality markers (Q-markers) and even to establish proper analysis methods for traditional Chinese medicine products.