Citrus Waste as Source of Bioactive Compounds: Extraction and Utilization in Health and Food Industry.

The current research was conducted to extract the bioactive compounds from citrus waste and assess their role in the development of functional foods to treat different disorders. The scientific name of citrus is Citrus L. and it belongs to the Rutaceae family. It is one of the most important fruit crops that is grown throughout the world. During processing, a large amount of waste is produced from citrus fruits in the form of peel, seeds, and pomace. Every year, the citrus processing industry creates a large amount of waste. The citrus waste is composed of highly bioactive substances and phytochemicals, including essential oils (EOs), ascorbic acid, sugars, carotenoids, flavonoids, dietary fiber, polyphenols, and a range of trace elements. These valuable compounds are used to develop functional foods, including baked products, beverages, meat products, and dairy products. Moreover, these functional foods play an important role in treating various disorders, including anti-aging, anti-mutagenic, antidiabetic, anti-carcinogenic, anti-allergenic, anti-oxidative, anti-inflammatory, neuroprotective, and cardiovascular-protective activity. EOs are complex and contain several naturally occurring bioactive compounds that are frequently used as the best substitutes in the food industry. Citrus essential oils have many uses in the packaging and food safety industries. They can also be used as an alternative preservative to extend the shelf lives of different food products.

Malformations in the Murine Kidney Caused by Loss of CENP-F Function.

Centromere-binding protein F (CENP-F) is a large and complex protein shown to play critical roles in mitosis and various other interphase functions. Previous studies have shown that the disruption of CENP-F function leads to detrimental effects on human development. Still, it is important to note the lack of studies focusing on the effects that the loss of this essential protein may have on specific adult organs. In the current study, we used a novel global knockout murine model to analyze the potential consequences deletion of CENP-F has on adult kidney structure and function. We discovered several structural abnormalities including loss of ciliary structure, tubule dilation, and disruption of the glomerulus. Along with these structural irregularities, renal dysfunction was also detected suggesting hydronephrosis and acute kidney injury in these knockout organs. Importantly, this is the first study linking CENP-F to kidney disease and hopefully these data will serve as a platform to further investigate the molecular mechanisms disrupted in the kidney by the loss of CENP-F. Anat Rec, 302:163-170, 2019. © 2018 Wiley Periodicals, Inc.

Thermal stability and degradation of chitosan modified by benzophenone.

N-(biphenylmethylidenyl) chitosan polymer was prepared, characterized and thermal stability was compared with chitosan. Thermal degradation products of the modified polymer were identified by GC-MS technique. It seems that the mechanism of degradation of the prepared polymer is characterized by formation of low molecular weight radicals, followed by random scission mechanism along the backbond chain.

Bves: ten years after.

Bves was discovered in 1999 by two independent laboratories using screens to identify novel genes that were highly expressed in the developing heart (Reese et al., 1999; Andree et al., 2000). As an evolutionarily conserved transmembrane protein, Bves is postulated to play a role in cell adhesion and cell motility. In studies of Bves protein disruption, there have been multiple phenotypes, but few molecular mechanisms have been advanced to explain the underlying cause of these phenotypes. As the molecular function of Bves protein begins to be uncovered, it is now time to review the literature to examine the significance of this work and future directions of study. This review summarizes the literature on this unique protein and explores new and exciting data that support emerging themes on its molecular function.

Bves directly interacts with GEFT, and controls cell shape and movement through regulation of Rac1/Cdc42 activity.

Bves is an integral membrane protein with no determined function and no homology to proteins outside of the Popdc family. It is widely expressed throughout development in myriad organisms. Here, we demonstrate an interaction between Bves and guanine nucleotide exchange factor T (GEFT), a GEF for Rho-family GTPases. This interaction represents the first identification of any protein that has a direct physical interaction with any member of the Popdc family. Bves and GEFT are shown to colocalize in adult skeletal muscle. We also demonstrate that exogenous expression of Bves reduces Rac1 and Cdc42 activity levels while not affecting levels of active RhoA. Consistent with a repression of Rac1 and Cdc42 activity, we show changes in speed of cell locomotion and cell roundness also result from exogenous expression of Bves. Modulation of Rho-family GTPase signaling by Bves would be highly consistent with previously described phenotypes occurring upon disruption of Bves function in a wide variety of model systems. Therefore, we propose Bves as a novel regulator of the Rac1 and Cdc42 signaling cascades.

Bves: prototype of a new class of cell adhesion molecules expressed during coronary artery development.

Bves is a protein expressed in cells of the developing coronary vascular system, specifically in the proepicardium, migrating epithelial epicardium, delaminated vasculogenic mesenchyme and vascular smooth muscle cells. Here, we show that Bves protein undergoes a dynamic subcellular redistribution during coronary vessel development. Bves is a membrane protein with three predicted transmembrane helices, an extracellular C terminus and an intracellular N terminus, and is confined to the lateral membrane compartment of epithelial cells. When epicardial cells are dissociated into single cells in vitro, Bves accumulates in a perinuclear region until cells make contact, at which time Bves is trafficked to the cell membrane. Bves accumulates at points of cell/cell contact, such as filopodia and cell borders, before the appearance of E-cadherin, suggesting an early role in cell adhesion. While Bves shares no homology with any known adhesion molecule, transfection of Bves into L-cells readily confers adhesive behavior to these cells. Finally, Bves antibodies inhibit epithelial migration of vasculogenic cells from the proepicardium. This study provides direct evidence that Bves is a novel cell adhesion molecule and suggests a role for Bves in coronary vasculogenesis.

Cloning and expression of hbves, a novel and highly conserved mRNA expressed in the developing and adult heart and skeletal muscle in the human.

bves is a novel mRNA expressed in the developing heart in chick and mouse. Here we describe hbves, the human homolog of chick and mouse bves. Northern and dot blot analyses reveal restricted expression in the heart and skeletal muscle in the embryo and adult. BLAST searches of the NCBI databases confirm that hbves is novel. Portions of the sequence are an exact match with genomic PAC 52202, which localizes to Chromosome (Chr) 6q21. Presumably, these matches and intervening sequences match the intron-exon borders of the gene. Computer conformation analysis of the derived amino acid predicts three transmembrane helices with an extracellular C-terminus that is conserved in chick, mouse, and human. bves is highly conserved among all three species at the amino acid level with 75% identity and 92% similarity.

Forensic science program: a community effort.

A forensic science program has been developed to assist the professional community population in using a scientific approach in the investigation of criminal offenses. The science of medicine and the principles of the law come together to form a multidisciplinary group of professionals to instruct in the collection of evidence after a crime has been committed. Specialists in these areas include forensic pathologists, forensic anthropologists, forensic odontologists, entomologists, and a radiologist with a specialty in forensics. No longer can educators ignore the necessity of community involvement in the apprehension and prosecution of the perpetrators of crime. This program is the first to offer basic forensic science courses to professionals in a variety of related fields.

Establishment of the mesodermal cell line QCE-6. A model system for cardiac cell differentiation.

The QCE-6 cell line was derived from precardiac mesoderm of the Japanese quail. As previously reported, these cells are able to differentiate into two distinct cardiac cell types with myocardial or endocardial endothelial cell properties. This present communication describes in detail the derivation of this cell line and further characterizes the nontreated and induced myocardial and endothelial phenotypes of these cells. The QCE-6 cells exhibit an epithelial morphology, as well as the pattern of protein expression, that is characteristic of precardiac mesoderm. Treatment with retinoic acid, basic fibroblast growth factor (bFGF), transforming growth factor (TGF)-beta 2, and TGF-beta 3 induces these cells to differentiate and produce mixed cultures of epithelial and mesenchymal cells. The epithelial cells express myosin, desmin, and cardiac troponin I in a punctate pattern throughout the cytoplasm. These sarcomeric proteins become organized in a premyofibrillar pattern when TGF-beta 1, platelet-derived growth factor (PDGF)-BB, and insulin-like growth factor (IGF) II are added in combination along with retinoic acid, bFGF, TGF-beta 2, and TGF-beta 3. Also, these treatments induce Na+,K(+)-ATPase expression. When the QCE-6 cells are cultured on collagen type I, the mesenchymal cells that are promoted by retinoic acid, bFGF, TGF-beta 2, and TGF-beta 3 will invade the gel. These mesenchymal cells are positive for QH1 and JB3, which are both markers for presumptive endocardial cells within the early cardiogenic mesoderm. The addition of both PDGF-BB and IGF II to QCE-6 cell cultures will inhibit the ability of retinoic acid, bFGF, TGF-beta 2, and TGF-beta 3 to induce both the mesenchymal morphology and QH1 and JB3 expression. Collectively, these results suggest that the proces of cardiac cell differentiation is regulated by multiple signals and that early cardiogenic mesoderm contains a bipotential stem cell that can give rise to both the myocardial and endocardial lineages. More important, since the QCE-6 cells are representative of early cardiogenic cells, this cell line offers a unique model system to study cardiac cell differentiation.

Identification of DNA-binding protein(s) in the developing heart.

An antiserum (anti-H2) directed at the second helix of the helix-loop-helix (HLH) protein MyoD1 reacts with a protein expressed during avian cardiac myocyte differentiation. Indirect immunohistochemical whole mount staining with anti-H2 detected a protein expressed in stage 11 hearts, but not in hearts of older embryos. At the cellular level, this staining is confined to the nucleus of cardiac cells suggesting that these proteins may have DNA-binding abilities. Several proteins were immunoprecipitated by anti-H2 from stage 11 heart tissue. Protein extracts from similarly staged hearts, when incubated with the muscle-specific enhancer sequence of muscle creatinine kinase (MCK), gave a stage-specific band shift in electromobility shift assays (EMSA), and these protein-DNA complexes were recognized and supershifted by anti-H2. Incubation with a MCK sequence containing a mutated E box did not produce a shift. The specific shift was present as early as stage 6, remained through stage 13, and disappeared by stage 17. These data suggest the presence of at least one protein that is transiently expressed in the differentiating cardiac myocyte, that is immunochemically reactive with an antiserum raised against the second helix of MyoD1, and that binds to a muscle-specific DNA enhancer sequence.

Expression of sarcomeric myosin in the presumptive myocardium of chicken embryos occurs within six hours of myocyte commitment.

The distribution of sarcomeric myosin heavy chain (MyHC) has been examined immunocytochemically in the presumptive myocardial cells of chicken embryos (stages 6-10) prior to the onset of the heart beat. Embryos were stained with monoclonal antibody MF20, a reagent which recognizes all chicken sarcomeric MyHCs (Bader et al., 1982), and then examined both in whole mount by immunofluorescence and in semithin, plastic-embedded sections following immunoperoxidase labeling. We observed that myosin could be detected as early as stage 7 (0-2 pairs of somites) in 29% of the 31 embryos examined, and by stage 8 (4 pairs of somites) more than 80% of the embryos were MF20+. Every embryo with 5 pairs of somites (stage 8+) labeled strongly with MF20. Labeling was first detected at stage 7 to 7+ as a diffuse fluorescent signal within pleomorphic cells of the splanchnic mesoderm located in two crescent-shaped regions bordering each side of the anterior intestinal portal (AIP). With progressive development, the two crescent-shaped regions merged at the apex of the AIP, and as the two heart tubes began fusion at stage 9, the MyHC+ regions extended cranially and medially. By somite stages 9-10, the myosin-positive cells completely encircled the heart tube. From stages 7 to 9 the myosin signal had no sarcomeric distribution; i.e., there were no MyHC striations nor periodic repeats evident in the presumptive myocytes until late stage 9 and stage 10. Semithin sections revealed that myosin was first distributed in apical regions of the myocytes, adjacent to the pericardial coelom. The implications of these findings for myocyte determination, differentiation and morphogenesis are discussed.

Myosin heavy chain expression in embryonic cardiac cell cultures.

Chick embryonic heart cell isolates and monolayer cultures were prepared from atria and ventricles at selected stages of cardiac development. The cardiac myocytes were assayed for myosin heavy chain (MHC) content using monoclonal antibodies (McAbs) specific in the heart for atrial (B-1), ventricular (ALD-19), or conductive system (ALD-58) isoforms. Using immunofluorescence microscopy or radioimmunoassay, MHC accumulation was measured before plating and at 48 hr or 7 days in culture. Reproducible changes in MHC antigenicity were observed by 7 days in both atrial and ventricular cultures. The changes were stage dependent and tissue specific but generally resulted in a decreased reactivity with the tissue specific MHC McAbs. In addition, the isoform recognized by ALD-58, characteristic of the conductive system cells in vivo, was never present in cultured myocytes. These results indicate that MHC isoforms produced in vivo may be replaced in monolayer cultures by an isoform(s) not recognized by our tissue specific MHC McAbs. This suggests that the intrinsic program of cardiac myogenesis, within cardiac myocytes, may not be sufficient to establish and maintain differential expression of tissue specific MHC in monolayer cell culture.

Immunochemical analysis of C-protein isoform transitions during the development of chicken skeletal muscle.

Isoforms of C-protein in adult chickens which differ in fast (pectoralis major, PM) and slow (anterior latissimus dorsi, ALD) skeletal muscles can be distinguished immunochemically with monoclonal antibodies (McAbs) specific for the respective fast (MF-1) and slow (ALD-66) protein variants (Reinach et al., 1982 and 1983). The expression of these C-proteins during chick muscle development in vivo has been analyzed by immunoblot and immunofluorescence procedures. Neither MF-1 nor ALD-66 reacted with whole-cell lysates or myofibrils from PM of 12-day-old embryos. However, both McAbs bound to peptides of 145 kDa in PM from late embryonic and young posthatched chickens. All of the myofibers in these muscles reacted with both antibodies, but the binding of the anti-slow McAb (ALD-66) diminished progressively with age and was completely negative with PM by 2 weeks after hatching. In contrast, the ALD muscle from 17 days in ovo thru adulthood only reacted with ALD-66; no binding of MF-1 could be detected at these stages. Since both fast and slow myosin light chains (LC) coexist within embryonic pectoralis and ALD muscles (e.g., G. F. Gauthier, S. Lowey, P. A. Benfield, and A. W. Hobbs, 1982, J. Cell Biol. 92, 471-484) yet segregate to specific fast and slow muscle fibers at different stages of development, the temporal transitions of C-protein and myosin LC were compared during myogenesis. "Slow-type" C-protein appeared after the disappearance of slow myosin light chains, whereas the accumulation of the "fast-type" light chains occurred before the expression of "fast-type" C-protein. The pattern of isoform transitions appears to be far more complex than previously suspected.

Restoration of full mass in nerve-intact muscle grafts after delayed reinnervation.

A rat muscle freely grafted with the motor nerve intact becomes restored to full mass and contractile function, in contrast to the reduced weight of a standard free graft. By crushing the nerve to a nerve-intact graft and delaying reinnervation, full mass is still restored. One can conclude that earlier reinnervation is not the reason for the success of nerve-intact grafts, but that it is rather due to reinnervation along preserved Schwann cell channels.

Comparison between grafts with intact nerves and standard free grafts of the rat extensor digitorum longus muscle.

Standard grafts and nerve-intact grafts of the extensor digitorum longus muscle were compared in the rat. In standard grafts the muscle was completely removed from its bed and replaced; nerve-intact grafts were treated in an identical manner except that the muscle nerve was not severed. Nerve-intact grafts underwent the same sequence of skeletal muscle fibre degeneration and regeneration as standard grafts. In nerve-intact grafts the intramuscular portions of the nerve fibres initially degenerated, but within a week new nerve fibres had regenerated back to the original zone of motor end-plates. By 60 days the weight of nerve-intact grafts approached those of control muscles. Contractile tension in nerve-intact grafts was greater than that of standard grafts. In standard and nerve-intact grafts choline acetyltransferase activity rapidly decreased to low values and then increased along curves roughly paralleling the muscle weights. In nerve-intact grafts, neuromuscular transmission was established early in the second week whereas a considerably later return was seen in standard grafts. Either the early onset or the topographical pattern of reinnervation are potentially major factors in determining the success of free muscle grafts.