Α-helix Protein Research Articles

Study on effects of neoadjuvant chemotherapy on ovarian cancer tissues using Raman spectroscopy

Abstract Ovarian cancer is one of the most prevalent malignancies in women, with over 70% of patients diagnosed at advanced stages (III–IV) due to a lack of early symptoms. Neoadjuvant chemotherapy (NACT) is frequently employed to alleviate systemic symptoms and prepare patients for surgery. However, no objective method currently exists to accurately assess its efficacy. Raman spectroscopy (RS), an analytical technique that detects molecular vibrations, offers a rapid, non-invasive, and cost-effective means of evaluation. This study aims to determine whether RS can serve as a novel approach for assessing the impact of NACT on fresh ovarian tissues. Fresh ovarian tissues were collected from patients treated at our hospital between September 2021 and November 2023; additionally, fresh normal ovarian tissues were obtained from six patients undergoing unrelated surgeries (the control group). The Raman spectra of different groups were analyzed using a partial least squares discriminant analysis model for interpretation. We observed that characteristic peaks at 673 and 1223 cm−1 appeared exclusively in IDS + NACT and normal control tissue maps, while the peak at 758 cm−1 was unique to the primary debulking surgery map. Following NACT, significant reductions in tissue intensity were noted at key characteristic peaks: glycogen at 477 cm−1 (P= 0.028), phenylalanine at 1004 cm−1 (P= 0.039), nucleic acids at 1342 cm−1 (P< 0.001), and lipids at 1446 cm−1 (P < 0.001). Conversely, a significant increase in amide III (α-helix) protein peak intensity was observed (P < 0.001). The results demonstrate that RS can effectively differentiate benign from malignant ovarian tissue characteristics while sensitively revealing microscopic changes in early-stage ovarian cancer before and after NACT. As an innovative method for evaluating NACT efficacy, it provides objective insights into molecular morphology that enhance our understanding of disease progression and treatment outcomes.

The effect of ionic regulation on the structure of whey protein during ultrafiltration process and hydraulic reverse cleaning efficiency.

The effects of ionic regulation on the structure of membrane surface proteins and backwashing efficiency during ultrafiltration were investigated to reveal the mechanism of ionic mitigating membrane fouling. The repulsion between proteins and membrane was enhanced after ion regulation. With the extension of ultrafiltration time, the α-helix and random coil of membrane surface proteins were decreased, while the β-turn structure increased which was subjected to continuous regulation by Na+, Zn2+ and K+ at 4min, 8min and 12min, respectively. Fourier transform infrared spectroscopy and Raman spectroscopy of membrane surface proteins showed that intra- and intermolecular hydrogen bonds of proteins were reduced on membrane surface. The exposed hydrophobic amino acid groups were reduced. The experimental group which was regulated by Na+ Zn2+ and K+ was favourable for the reduction of cake resistance (Rc) and had a lower square roughness (Rq) on membrane surface during filtration process. However, this could lead to the increased pore blockage (Rp), which was not conducive to hydraulic reverse cleaning efficiency compared to the experimental group regulated by K+, Zn2+ and Mg2+. Therefore, regulation of the whey ultrafiltration process by different ions at specific time is helpful in alleviating membrane fouling.

Textural enhancement and glycemic potency reduction of sugarcane fiber-incorporated white bread with ascorbic acid and xanthan gum

In this study, ascorbic acid (0.02 % w/w), xanthan (0.75 % or 1.5 % w/w), and their combination have been added into sugarcane fiber (SCF) incorporated (5 % or 10 % w/w) wheat flour-based white breads. The effects of different additives on the physical characteristics, and the in-vitro and in-vivo glycemic potency of breads were evaluated. Addition of xanthan alone and the combination of additives reduced hardness and increased specific volume. SEM images showed that xanthan caused larger, more uneven holes in breadcrumbs due to xanthan's high elasticity and viscosity. FTIR spectrum indicated that the combination of SCF, xanthan, and ascorbic acid resulted in higher β-turn, lower α-helix protein structures, and lower ratios of α-helix/β-sheet, indicating a more flexible gluten structure formed. In-vitro digestibility results suggested that all SCF-incorporated breads had a lower glycemic index (GI) value than reference. Samples with 0.02 % (w/w) ascorbic acid and 1.5 % (w/w) xanthan reported the lowest in-vitro and in-vivo GI values.

Characteristics of organic matter driven by Eichhornia crassipes during co-contamination with per(poly)fluoroalkyl substances (PFASs) and microplastics (MPs)

Co-contamination with MPs and PFASs has been recorded, particularly in surface-water environments. Floating macrophyte microcosms are an important part of the surface water ecosystem, and dissolved organic matter (DOM) driven by floating macrophytes (FMDDOM) is critical for maintaining material circulation. However, knowledge gaps remain regarding the impact of MPs and PFASs co-pollution on FMDDOM. An greenhouse simulation experiment was conducted in this study to investigate the effects of four PFASs, perfluorooctanoic acid (PFOA), perfluoro-octane-sulfonic acid (PFOS), perfluoro-2-methyl-3-oxahexanoic acid (Gen X), and potassium 9-chlorohexadecafluoro-3-oxanonane-1-sulfonate (F-53B), on FMDDOM sourced from Eichhornia crassipes (E. crassipes), a typical floating macrophyte, in the presence and absence of polystyrene (PS) MPs. Four PFASs increased FMDDOM release from E. crassipes, leading to a 32.52–77.49 % increase in dissolved organic carbon (DOC) levels. PS MPs further increased this, with results ranging from −21.28 % to 26.49 %. Based on the parallel factor analysis (PARAFAC), FMDDOM was classified into three types of fluorescent components: tryptophan-like, humic-like, and tyrosine-like compounds. Contaminants of MPs and PFASs modified the relative abundance of these three components. Protein secondary structure analysis showed that fluorocarbon bonds tended to accumulate on the α-helix of proteins in FMDDOM. The relative abundance of fluorescent and chromophorous FMDDOMs varied from 0.648 ± 0.044 to 0.964 ± 0.173, indicating that the photochemical structures of the FMDDOM were modified. FMDDOM exhibits decreased humification and increased aromaticity when contaminated with MPs and PFASs, which may be detrimental to the geochemical cycling of carbon. This study offers a theoretical basis for assessing the combined ecological risks of MPs and PFASs in floating macrophyte ecosystems.

Mixed Stereochemistry Macrocycle Acts as a Helix-Stabilizing Peptide N-Cap.

Interactions between proteins and α-helical peptides have been the focus of drug discovery campaigns. However, the large interfaces formed between multiple turns of an α-helix and a binding protein represent a significant challenge to inhibitor discovery. Modified peptides featuring helix-stabilizing macrocycles have shown promise as inhibitors of these interactions. Here, we tested the ability of N-terminal to side-chain thioether-cyclized peptides to inhibit the α-helix binding protein Mcl-1, by screening a trillion-scale library. The enriched peptides were lariats featuring a small, four-amino-acid N-terminal macrocycle followed by a short linear sequence that resembled the natural α-helical Mcl-1 ligands. These "Heliats" (helical lariats) bound Mcl-1 with tens of nM affinity, and inhibited the interaction between Mcl-1 and a natural peptide ligand. Macrocyclization was found to stabilize α-helical structures and significantly contribute to affinity and potency. Yet, the 2nd and 3rd positions within the macrocycle were permissible to sequence variation, so that a minimal macrocyclic motif, of an N-acetylated d-phenylalanine at the 1st position thioether connected to a cysteine at the 4th, could be grafted into a range of peptides and stabilize helical conformations. We found that d-stereochemistry is more helix-stabilizing than l- at the 1st position in the motif, as the d-amino acid can utilize polyproline II torsional angles that allow for more optimal intrachain hydrogen bonding. This mixed stereochemistry macrocyclic N-cap is synthetically accessible, requiring only minor modifications to standard solid-phase peptide synthesis, and its compatibility with peptide screening can provide ready access to helix-focused peptide libraries for de novo inhibitor discovery.

Deciphering retention effect of extracellular polymeric substances to typical heavy metals and their interaction with key inner enzymes of Candidatus Kuenenia

This study employed spectroscopy, metagenomics, and molecular simulation to investigate the inhibitory effects of Cd(II) and Cu(II) on the anammox system, examining both intracellular and extracellular effects. At concentrations of 5 mg/L, Cd(II) and Cu(II) significantly reduced nitrogen removal efficiency by 41.46 % and 62.03 %, respectively. Additionally, elevated metal concentrations were correlated with decreased extracellular polymeric substances (EPS), thereby reducing their capacity to absorb heavy metals, particularly Cu(II), which decreased from 76.47 % to 14.67 %. Spectral analysis revealed alterations in the secondary structures of EPS induced by Cd(II) and Cu(II), decreasing the ratio of extracellular protein α-helix to (β-sheet + random coil), which resulted in looser extracellular protein configurations. The results of the metagenomics study showed that the abundance of Candidatus Kuenenia and its genes encoding nitrogen removal-related enzymes was reduced. The abundance of hzs-γ was reduced by 35.09 % at a concentration of 5 mg/L Cu(II). Conversely, genes associated with metal efflux enzymes, like czcR, increased by 54.86 % at 2 mg/L Cd(II). Molecular docking revealed robust bindings of Cd(II) to HZS-α (−342.299 ± 218.165 kJ/mol) and Cu(II) to HZS-γ (−880.934 ± 55.526 kJ/mol). This study elucidated the inhibitory mechanisms of Cd(II) and Cu(II) on the anammox system, providing insights into the resistance of anammox bacteria to heavy metals.

Exploring the effect of high-pressure processing conditions on the deaggregation of natural major royal jelly proteins (MRJPs) fibrillar aggregates

High pressure processing is a safe and green novel non-thermal processing technique for modulating food protein aggregation behavior. However, the systematic relationship between high pressure processing conditions and protein deaggregation has not been sufficiently investigated. Major royal jelly proteins, which are naturally highly fibrillar aggregates, and it was found that the pressure level and exposure time could significantly promote protein deaggregation. The 100–200 MPa treatment favoured the deaggregation of proteins with a significant decrease in the sulfhydryl group content. Contrarily, at higher pressure levels (>400 MPa), the exposure time promoted the formation of disordered agglomerates. Notably, the inter-conversion of α-helix and β-strands in major royal jelly proteins after high pressure processing eliminates the solvent-free cavities inside the aggregates, which exerts a 'collapsing' effect on the fibrillar aggregates. Furthermore, the first machine learning model of the high pressure processing conditions and the protein deaggregation behaviour was developed, which provided digital guidance for protein aggregation regulation.

Effects of air frying on protein digestive properties in scallop (Patinopecten yessoensis) adductor muscles and the mechanisms involved

The effects of air frying at different temperatures (140 °C, 160 °C, 180 °C, and 200 °C) on the simulated digestive properties of proteins in scallop adductor muscle (SAM) and the mechanisms involved were investigated. The results of the simulated gastrointestinal digestion showed that the 160 °C-treated SAM group had significantly higher protein digestibility than the fresh SAM group, which was reflected in higher degree of hydrolysis, more low molecular weight (<0.5 kDa) protein digestive products, and less insoluble residues, while the 140 °C-, 180 °C- and 200 °C-treated SAM groups all had opposite results. Moreover, air frying altered the solubility, secondary structure percentage and surface hydrophobicity of SAM proteins, as well as the microstructure of insoluble residues in simulated gastric juice. Correlation analysis showed that the protein digestive level of the air fried SAMs was positively correlated with the ratio of soluble proteins before digestion and the surface hydrophobicity of insoluble residues, but negatively correlated with the content of α-helix of soluble proteins dissolved in simulated gastric juice. Therefore, the highest digestibility of the 160 °C-treated SAM group can be attributed to the lowest α-helix content of soluble proteins, as well as the highest surface hydrophobicity and the largest pores size of insoluble residues. The present study provides a good basis for understanding the effects of air frying temperature on the digestive properties of aquatic muscle foods and the mechanisms involved, which contributes to establish precise air frying method for aquatic muscle foods.

The synthesis of MgO/Zein nanocomposites to improve anti-bacteria characteristics of mouthwash

Mouthwash is an alternative to address problems in the oral cavity, such as caries, periodontal inflammation, and microbial abscesses generally caused by pathogens. However, chlorhexidine contained in the mouthwash was limited concentration, i.e., 0.2 %, due to the side effects caused by these ingredients. Even though chlorhexidine has bactericidal and bacteriostatic properties against various bacteria found in plaque, for this reason, to increase the antibacterial activity of these mouthwashes, it is necessary to add other ingredients to the mouthwash formula, namely MgO/Zein nanocomposites with a composition of 1:4 and 1:5. The method used in this study was the preparation of MgO nanoparticles from dolomite using a solid acid solvent, the results of which were then composited with Zein using alcohol and NaOH. Synthesized samples were characterized using XRD, FTIR, SEM, and TEM. The antibacterial activity of S. aureus and mushrooms C. albicans in mouthwash with a concentration of 0.5 to 2.5 % can be determined by conducting a diffusion test. The results showed that the MgO/Zein nanocomposite fabrication had been successfully carried out and had the following characteristics: having a MgO phase with a polycrystalline cubic structure, having a zein phase with α-helix and β-sheet structures of proteins shows that Zein has an amorphous structure. The results of the antibacterial activity test showed that mouthwash with adding MgO/Zein nanocomposites more effectively inhibited bacterial growth of S. aureus and mushrooms C. albicans. It is known that the MgO/Zein nanocomposite composition 1:4 has the largest inhibition zone in S. aureus bacteria with a concentration of 1.5 % mouthwash and C. albicans fungi with a concentration of 0.5 %, namely 40.17 mm and 8.42 mm

Label-Free, Noninvasive Bone Cell Classification by Hyperspectral Confocal Raman Microscopy.

Characterizing and identifying cells in multicellular in vitro models remain a substantial challenge. Here, we utilize hyperspectral confocal Raman microscopy and principal component analysis coupled with linear discriminant analysis to form a label-free, noninvasive approach for classifying bone cells and osteosarcoma cells. Through the development of a library of hyperspectral Raman images of the K7M2-wt osteosarcoma cell lines, 7F2 osteoblast cell lines, RAW 264.7 macrophage cell line, and osteoclasts induced from RAW 264.7 macrophages, we built a linear discriminant model capable of correctly identifying each of these cell types. The model was cross-validated using a k-fold cross validation scheme. The results show a minimum of 72% accuracy in predicting cell type. We also utilize the model to reconstruct the spectra of K7M2 and 7F2 to determine whether osteosarcoma cancer cells and normal osteoblasts have any prominent differences that can be captured by Raman. We find that the main differences between these two cell types are the prominence of the β-sheet protein secondary structure in K7M2 versus the α-helix protein secondary structure in 7F2. Additionally, differences in the CH2 deformation Raman feature highlight that the membrane lipid structure is different between these cells, which may affect the overall signaling and functional contrasts. Overall, we show that hyperspectral confocal Raman microscopy can serve as an effective tool for label-free, nondestructive cellular classification and that the spectral reconstructions can be used to gain deeper insight into the differences that drive different functional outcomes of different cells.

Non-crystallographic helices in polymers and close-packed metallic crystals determined by the four-dimensional counterpart of the icosahedron

Non-crystallographic fractional screw axes are inherent to the constructions of n-dimensional crystallography, where 3 &lt; n ≤ 8. This fact allows one to consider experimentally obtained helices as periodic approximants of helices from the four-dimensional {3, 3, 5} polytope and its derivative constructions. For the tetrahedral Coxeter–Boerdijk helix (tetrahelix) with a 30/11 axis from the {3, 3, 5} polytope, approximants with 11/4 and 8/3 axes in three-dimensional Euclidean space {\bb E}^{3} are considered. These determine the structure of rods composed of deformed tetrahedra in close-packed crystals of α-Mn and β-Mn. In the {3, 3, 5} polytope, highlighted here for the first time, is a 40-vertex helix with a 20/9 axis composed of seven-vertex quadruples of tetrahedra (tetrablocks), whose 7/3 approximants determine in a crystal of an α-Mn rod of deformed tetrablocks with the same period as the 11/4 approximant of the tetrahelix. In the spaces of the three-dimensional sphere and {\bb E}^{3}, the parameters of 20/9, 40/9 and 40/11 helices, as well as of their 20- and 40-vertex approximants, are calculated. The parameters of the approximant of the 40/11 helix in {\bb E}^{3} correspond to experimentally determined parameters of the α-helix, which allows us to explain the versatility of the α-helix in proteins by the symmetry of the polytope. The set of fractional axes of all periodic approximants of helices with 30/11, 20/9, 40/9, 40/11 axes, as well as the powers of these axes, are combined into a tetrahedral-polytope class of 50 basic axes. The basic axes as well as composite (defined as a combination of basic ones) fractional axes of this class cover all fractional axes known to us according to literature data for polymers, biopolymers and close-packed metals.

Nanoscale structural characterization of transthyretin aggregates formed at different time points of protein aggregation using atomic force microscopy-infrared spectroscopy.

Transthyretin (TTR) amyloidosis is a progressive disease characterized by an abrupt aggregation of misfolded protein in multiple organs and tissues TTR is a tetrameric protein expressed in the liver and choroid plexus. Protein misfolding triggers monomerization of TTR tetramers. Next, monomers assemble forming oligomers and fibrils. Although the secondary structure of TTR fibrils is well understood, there is very little if anything is known about the structural organization of TTR oligomers. To end this, we used nano-infrared spectroscopy, also known as atomic force microscopy infrared (AFM-IR) spectroscopy. This emerging technique can be used to determine the secondary structure of individual amyloid oligomers and fibrils. Using AFM-IR, we examined the secondary structure of TTR oligomers formed at the early (3-6 h), middle (9-12 h), and late (28 h) of protein aggregation. We found that aggregating, TTR formed oligomers (Type 1) that were dominated by α-helix (40%) and β-sheet (~30%) together with unordered protein (30%). Our results showed that fibril formation was triggered by another type of TTR oligomers (Type 2) that appeared at 9 h. These new oligomers were primarily composed of parallel β-sheet (55%), with a small amount of antiparallel β-sheet, α-helix, and unordered protein. We also found that Type 1 oligomers were not toxic to cells, whereas TTR fibrils formed at the late stages of protein aggregation were highly cytotoxic. These results show the complexity of protein aggregation and highlight the drastic difference in the protein oligomers that can be formed during such processes.

Low-voltage electrostatic field enhances the frozen force of −12 ℃ to suppress oxidative denaturation of the lamb protein during the subsequent frozen storage process after finishing initial freezing

The effect of low-voltage electrostatic field (LVEF) assisted −9 °C (LVEF-9) and −12 °C (LVEF-12) frozen, non-LVEF-assisted −9 °C (NLVEF-9) and −12 °C (NLVEF-12) frozen, and conventional frozen (CF-18, −18 °C) storage on the muscle microstructure and the oxidative denaturation of the lamb protein during the subsequent frozen storage process after finishing initial freezing was investigated. Compared with NLVEF-9, LVEF-9, and NLVEF-12, LVEF-12 maintained the better integrity of muscle microstructure, demonstrated by smaller holes, more complete Z-line and M-line, and no significant difference with CF-18 (P > 0.05). Furthermore, LVEF-12 effectively inhibited protein oxidative denaturation as shown by the lower carbonyl content, surface hydrophobicity, and higher total/active sulfhydryl groups and Ca2+-ATPase activity. Moreover, LVEF-12 effectively maintained the integrity of the secondary and tertiary structure of proteins, reduced cross-linking aggregation of proteins, and sustained better functional properties, as shown by higher α-helix content, fluorescence intensity, protein solubility, and lower R-value, disulfide bonds.

Effect of radio frequency explosion puffing on physicochemical, functional and crystalline properties, and in vitro digestibility of yam flour

Yam flour was modified by radio frequency explosion puffing at different moisture content, puffing temperature, and puffing pressure difference. After puffing, the protein content and lipid content increased by 0.56–1.28 % and 0.23–0.39 %, respectively. Puffing caused the flour granules to aggregate, increasing the thermal transition temperatures and reducing the pasting viscosities, enthalpy, 1047/1022 cm−1 ratio, and relative crystallinity. Puffing reduced the intensity of the infrared spectrum peak at 1641 cm−1 by breaking the hydrogen bonds without changing A-type crystalline structure. Puffing promoted the conversion of random-coil and α-helix protein structure to β-turn and β-sheet. Puffing retarded in vitro digestibility by reducing rapidly digestible starch content by 7.04–11.12 % and rising slowly digestible starch content and resistant starch content by 4.02–4.89 % and 2.77–3.10 %, respectively. Radio frequency explosion puffing altered flour's physicochemical, functional and digestibility properties by destroying the protein structure and promoting the interaction of starch and proteins/lipids.

Crotonaldehyde induced structural alterations in Low-Density Lipoprotein: Immunogenicity of the modified protein in experimental animals and auto-antibodies generation in various cancers

Crotonaldehyde (CA), a prominent component of cigarette smoke (CS) is a pervasive environmental pollutant that is a highly toxic, unsaturated aldehyde. Exposure to CA-rich pollutants has been linked to the emergence of many malignancies in humans. To better understand the role of CA in biomolecule modification, this study investigated the detailed structural alterations in low-density lipoprotein (LDL) modified by CA, as well as the immunogenicity of the modified protein in experimental animals and the search for autoantibodies in various cancers patients.In vitro, results indicated alterations in secondary and tertiary structures; examined using UV–visible, fluorescence, far-UV circular dichroism, and Fourier transform infrared spectroscopy techniques. Changes in the oxidation status of LDL were studied by carbonyl content assay and NBT assay. ThT binding assay, scanning, and transmission electron microscopy were used to study aggregate formation. The findings revealed significant structural damage in LDL modified by CA. The modification resulted in the unmasking of hydrophobic clusters, the loss of the protein α-helix, and the formation of β-pleated sheet structure. The amyloid aggregate formation was confirmed through ThT microscopy and electron spectroscopy. Rabbits immunized with crotonaldehyde; lead to structural changes in the LDL; that acted as extra antigenic determinants, eliciting strong antibody response. Immunoglobulin response is highly specific for modified LDL as demonstrated by the ELISA. The presence of antibodies against CA-modified LDL was confirmed by the immunoglobulin content of blood sera from human subjects with lung cancer, and competitive ELISA demonstrated the specificity of these antibodies. This study offers insights into the CA-mediated LDL modification and immunogenicity in lung cancer that will have diagnostic importance.

Changes in physicochemical and structural properties of pea protein during the high moisture extrusion process: Effects of carboxymethylcellulose sodium and different extrusion zones

This study investigated effects of carboxymethylcellulose sodium (CMC) on the conformational evolution of pea protein during the high moisture extrusion process. The morphological observation showed that the addition of CMC facilitated the formation of fibrous structure of pea protein. In comparison with the pea protein in the melting zone and extrudate, the combination of CMC increased the denaturation enthalpy of pea protein by 2.09 % and 2.34 %. Compared with the material in the mixing zone, the degree of grafting between CMC and pea protein in the die was enhanced by 98.95 %. In general, the supplementation of CMC depressed the exposure of hydrophobic groups in the pea protein. In the extrusion barrel, the CMC increased the unfolding of protein molecular chains while it promoted the refolding of protein chains in the die. For the extrudate, the addition of CMC decreased the contents of α-helix and β-sheet of pea protein by 9.67 % and 6.93 % while the contents of β-turn and random coil were increased, leading to changes in the molecular weight distribution of protein molecules. In conclusion, these results provided new strategies toward producing the high-quality pea protein-based meat analogues by adding CMC.

Application of Fourier Transform InfraRed spectroscopy of machine learning with Support Vector Machine and principal components analysis to detect biochemical changes in dried serum of patients with primary myelofibrosis

Primary myelofibrosis (PM) is a myeloproliferative neoplasm characterized by stem cell-derived clonal neoplasms. Several factors are involved in diagnosing PM, including physical examination, peripheral blood findings, bone marrow morphology, cytogenetics, and molecular markers. Commonly gene mutations are used. Also, these gene mutations exist in other diseases, such as polycythemia vera and essential thrombocythemia. Hence, understanding the molecular mechanism and finding disease-related biomarker characteristics only for PM is crucial for the treatment and survival rate. For this purpose, blood samples of PM (n = 85) vs. healthy controls (n = 45) were collected for biochemical analysis, and, for the first time, Fourier Transform InfraRed (FTIR) spectroscopy measurement of dried PM and healthy patients' blood serum was analyzed. A Support Vector Machine (SVM) model with optimized hyperparameters was constructed using the grid search (GS) method. Then, the FTIR spectra of the biomolecular components of blood serum from PM patients were compared to those from healthy individuals using Principal Components Analysis (PCA). Also, an analysis of the rate of change of FTIR spectra absorption was studied. The results showed that PM patients have higher amounts of phospholipids and proteins and a lower amount of H-O=H vibrations which was visible. The PCA results indicated that it is possible to differentiate between dried blood serum samples collected from PM patients and healthy individuals. The Grid Search Support Vector Machine (GS-SVM) model showed that the prediction accuracy ranged from 0.923 to 1.00 depending on the FTIR range analyzed.Furthermore, it was shown that the ratio between α-helix and β-sheet structures in proteins is 1.5 times higher in PM than in control people. The vibrations associated with the CO bond and the amide III region of proteins showed the highest probability value, indicating that these spectral features were significantly altered in PM patients compared to healthy ones' spectra.The results indicate that the FTIR spectroscope may be used as a technique helpful in PM diagnostics. The study also presents preliminary results from the first prospective clinical validation study.

Insights into Intramuscular Connective Tissue Associated with Wooden Breast Myopathy in Fast-Growing Broiler Chickens.

Wooden breast myopathy (WBM) is a meat abnormality affecting pectoralis majors (PMs) of fast-growing broiler chickens. WBM-affected PMs exhibited varied meat qualities with increasing WBM severity. Normal PMs (NOR), mild WBM-affected PMs (MIL), moderate WBM-affected PMs (MOD), and severe WBM-affected PMs (SEV) were selected as raw materials. The structure and organization of connective tissue and fibrillar collagen were investigated through immersing with sodium hydroxide solution, Masson trichrome staining, and using an electron microscope. The mechanical strength of intramuscular connective tissue was analyzed via the shear force of samples treated with sodium hydroxide solution. The thermal property and secondary structure of connective tissue were analyzed by differential scanning calorimetry and Fourier transform infrared spectroscopy. The obtained connective tissue was dissolved in a sodium hydroxide solution for the evaluation of the physicochemical properties of proteins, including particle size, molecular weight, surface hydrophobicity, and intrinsic fluorescence. In particular, the particle size was measured using a zeta potential instrument. The molecular weight was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The surface hydrophobicity and intrinsic fluorescence were measured by spectroscopy technology. Histologically, macrophage infiltration, myodegeneration and necrosis, regeneration, fibrous connective tissue, and thickened perimysial connective tissue were observed in WBM-affected PMs, especially SEV with fibrosis, including blood vessels. Compared with NOR, WBM led to increased average diameter of the collagen fibrils in perimysial (36.61 nm of NOR to 69.73 nm of SEV) and endomysial (34.19 nm of NOR to 56.93 nm of SEV) layers. A significant increase (p < 0.05) was observed in the mechanical strength (2.05 N to 5.55 N) of fresh PMs and the thermal transition temperature (onset temperature (TO), 61.53 °C to 67.50 °C; maximum transition temperature (TM), 66.46 °C to 70.18 °C; termination temperature (TE), 77.20 °C to 80.88 °C) of connective tissue from NOR to SEV. Cooking decreased the mechanical strength, and MOD samples showed the highest mechanical strength (1.24 N, p < 0.05), followed by SEV (0.96 N), MIL (0.93 N), and NOR (0.72 N). For proteins in connective tissue, random coil (19.64% to 29.61%, p < 0.0001), particle size (p < 0.05), and surface hydrophobicity (p < 0.05) increased with the decrease in the α-helix (14.61% to 11.54%, p < 0.0001), β-sheet (45.71% to 32.80%, p < 0.0001), and intrinsic fluorescence of proteins from NOR to SEV. The molecular weights of intramuscular connective tissue proteins were in the ranges of >270 kDa, 180-270 kDa, 110-180 kDa, 95-100 kDa, and <15 kDa. Taken together, WBM resulted in thickened organization, tightly packed collagen fibrils, increased mechanical strength and thermal temperature, and increased particle size, surface hydrophobicity, and intrinsic fluorescence of proteins in connective tissue, as the WBM severity increased.

Cytotoxicity, structural, conformational, and techno-functional properties of α-lactalbumin nanostructured aggregates.

Protein nanostructures can be used in food applications to improve the techno-functional properties of a food formulation. This study aims to find the best conditions for the production and conformational change of α-lactalbumin nanostructured aggregates. The criteria to determine the best operating conditions to produce α-lactalbumin nanostructured aggregates were intensification of foaming and emulsification, techno-functional proprieties, cytotoxic, and antibacterial activity of nanostructures compared with native α-lactalbumin. Conformational alterations occurred in the α-helix and sheet-β protein structures. The size obtained by dynamic light scattering was 163.84nm with a polydispersity index of 0.29. The nano protein improved the techno-functional property compared to the native protein. Additionally, nanostructures had no cytotoxic effect and were innocuous to bacterial activity. Thus, this study presents the best conditions to produce α-lactalbumin nanostructured aggregates with improved properties that allow new food industry applications.

Exploration of clinical and genetic findings in Ataxia-Telangiectasia (AT) patients from the Indian subcontinent

BackgroundAtaxia-Telangiectasia (AT) is a rare autosomal recessive neurodegenerative disorder. It is caused by mutations in the Ataxia-Telangiectasia mutated (ATM) gene, which codes for protein ATM serine/threonine kinase. ObjectiveWe aim to describe the clinical and radiological findings in children and adolescents of 20 molecularly confirmed cases of AT. We aim to correlate these findings with the genotype identified among them. MethodsThis retrospective study included 20 patients diagnosed clinically and genetically with AT over 10 years. The clinical, radiological and laboratory data were extracted from the hospital's electronic medical records. Molecular testing was done using next generation sequencing and Sanger sequencing. In silico predictions were performed for the variants identified by applying Cryp-Skip, Splice site prediction by Neural Network, Mutation Taster and Hope prediction tool. ResultsConsanguinity was documented in nearly half of the patients. Telangiectasia was absent in 10%. Microcephaly was seen in 40% cases. The incidence of malignancy in our study population was low. Molecular testing done in the 18 families (20 patients) identified 23 variants of which ten were novel. Biallelic homozygous variants were noted in 13 families and compound heterozygous in 5 families. Out of the 13 families who were homozygous, 8 families (61.5%) (9 patients) have history of consanguinity. In silico prediction of novel missense variants, NM_000051.4 (ATM_v201): c.2702T > C showed disruption of the α-helix of ATM protein and NM_000051.4 (ATM_v201): c.6679C > G is expected to disturb the rigidity of protein structure in the FAT domain. The four novel splice site variants and two intronic variants result in exon skipping as predicted by Cryp-Skip. ConclusionsAT should be confirmed by molecular testing in young-onset cerebellar ataxia, even without telangiectasia. Awareness of this rare disease will facilitate study of larger cohorts from Indian population to characterize variants and determine its prevalence in this population.