5 research outputs found
SÜRDÜRÜLEBİLİR BİR PROTEİN KAYNAĞI OLARAK DOMATES POSASI: GELENEKSEL ALKALİ, ULTRASON VE MİKRODALGA DESTEKLİ EKSTRAKSİYONUN PROTEİN İÇERİĞİ, VERİMİ VE FONKSİYONEL ÖZELLİKLERİ ÜZERİNE ETKİSİ
No full textTomato pomace, an abundant by-product of the tomato processing industry, is rich in protein and other valuable compounds. However, it is currently mostly utilized as animal feed, with considerable potential remaining for higher-value applications in human nutrition. Valorizing this agro-industrial waste as a sustainable protein source aligns with efforts to promote circular food systems and plant-based alternatives. In this study, protein extraction from tomato pomace was carried out using three methods: conventional alkaline extraction (CAE), microwave-assisted alkaline extraction (MWAE), and ultrasound-assisted alkaline extraction (USAE). The objective was to evaluate and compare these methods in terms of protein content, yield, and functional properties.
Protein content was determined using the Kjeldahl method, while solubility was evaluated using the Bradford assay. Functional properties, including foaming capacity and stability, emulsifying activity and stability, water-holding capacity, oil-holding capacity, and color parameters (L, a, b*), were assessed. Structural differences between protein extracts were analyzed using FTIR spectroscopy, and antioxidant activity was determined by DPPH⸱ radical scavenging assay.
CAE and MWAE yielded similar protein contents (44.46% and 45.12%, respectively). However, MWAE significantly reduced the extraction time from 1 h to 3 min. USAE yielded the lowest protein content (42.61%). MWAE showed the highest protein recovery yield (36.31%), followed by USAE (31.80%) and CAE (28.20%). Protein solubility was comparable across all methods. MWAE and USAE showed better functional properties than CAE, especially in foaming and emulsifying capacity. FTIR analysis revealed structural differences, and the highest antioxidant activity was observed in MWAE extracts.
These findings support MWAE and USAE as effective, sustainable alternatives for protein recovery from tomato pomace.Domates posası, domates işleme endüstrisinin bol miktarda üretilen bir yan ürünüdür ve yüksek oranda protein ile diğer değerli bileşenleri içermesine rağmen yeterince değerlendirilmemektedir. Bu tarımsal sanayi atığının sürdürülebilir bir protein kaynağı olarak değerlendirilmesi, döngüsel gıda sistemlerini ve bitki bazlı alternatifleri teşvik etmeye yönelik çalışmaları desteklemektedir. Bu çalışmada, domates posasından protein ekstraksiyonu üç yöntemle gerçekleştirilmiştir: konvansiyonel alkali ekstraksiyonu (CAE), mikrodalga destekli alkali ekstraksiyonu (MWAE) ve ultrason destekli alkali ekstraksiyonu (USAE). Amaç, bu yöntemleri protein içeriği, verimi ve fonksiyonel özellikler açısından karşılaştırmaktır.
Protein içeriği Kjeldahl yöntemiyle, çözünürlük ise Bradford yöntemi kullanılarak belirlenmiştir. Fonksiyonel özellikler arasında köpürme kapasitesi ve stabilitesi, emülsifiye etme aktivitesi ve stabilitesi değerlendirilmiş, ayrıca renk parametreleri (L*, a*, b*) ölçülmüştür. FTIR spektroskopisiyle yapısal farklılıklar analiz edilmiştir. DPPH⸱ yöntemiyle antioksidan aktivite belirlenmiştir.
CAE ve MWAE özütlerinin protein içerikleri benzer (44.46% ve 45.12%) bulunmuştur. MWAE süreci ekstraksiyon süresini 1 saatten 3 dakikaya düşürmüştür. En düşük protein içeriği USAE'de (42.61%), en yüksek geri kazanım ise MWAE’de (36.31%) elde edilmiştir. MWAE ve USAE, CAE’ye kıyasla daha iyi fonksiyonel özellikler göstermiştir. FTIR sonuçları yapısal farklılıkları, MWAE ise en yüksek antioksidan aktiviteyi ortaya koymuştur. Sonuçlar, mikrodalga ve ultrason destekli yöntemlerin etkili ve sürdürülebilir alternatifler olduğunu göstermektedir.M.S. - Master of Scienc
Tomato pomace as a sustainable protein source: impact of conventional alkaline, ultrasound, and microwave-assisted extraction on protein content, yield, and functional properties
No full textAssociation between tocilizumab and emerging multidrug-resistant organisms in critically ill patients with COVID-19: A multicenter, retrospective cohort study
No full textBackground: Tocilizumab is an IgG1 class recombinant humanized monoclonal antibody that directly inhibits the IL-6 receptor. Several randomized clinical trials have evaluated its safety and efficacy in patients with coronavirus disease 2019 (COVID-19), and these studies demonstrate conflicting results. Our study aimed to determine the association between tocilizumab treatment and microbial isolation and emergence of multidrug-resistant bacteria in critically ill patients with COVID-19. Methods: A multicenter retrospective cohort study was conducted at two tertiary government hospitals in Saudi Arabia. All critically ill patients admitted to intensive care units with a positive COVID-19 PCR test between March 1 and December 31, 2020, who met study criteria were included. Patients who received tocilizumab were compared to those who did not receive it. Results: A total of 738 patients who met our inclusion criteria were included in the analysis. Of these, 262 (35.5%) received tocilizumab, and 476 (64.5%) were included in the control group. Patients who received tocilizumab had higher odds for microbial isolation (OR 1.34; 95% CI 0.91–1.94, p = 0.13); however, the difference was not statistically significant. Development of resistant organisms (OR 1.00; 95% CI 0.51–1.98, p = 0.99) or detection of carbapenem-resistant Enterobacteriaceae (CRE) (OR 0.67; 95% CI 0.29–1.54, p = 0.34) was not statistically significant between the two groups. Conclusions: Tocilizumab use in critically ill patients with COVID-19 is not associated with higher microbial isolation, the emergence of resistant organisms, or the detection of CRE organisms. © 2021, The Author(s).Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]
The role of emerin and LEM domain proteins in nuclear envelope assembly and cytoskeleton organisation
No full textThe nuclear envelope (NE) plays a fundamental role in the cell by separating nuclear from cytoplasmic activities, and mutations in NE proteins have been associated with a diverse array of diseases. In the present study the Xenopus cell-free system was used to investigate the function of the inner nuclear membrane protein, emerin, which is associated with the Emery-Dreifuss muscular dystrophy (X-EDMD).Initially, the order and dynamics of NE assembly in Xenopus egg extracts have been investigated. Using a panel of antibodies it was shown that NE assembly proceeds by the ordered recruitment of two membrane populations, Nuclear Envelope Precursor vesicles -A and -B (NEP-A and NEP-B), to chromatin. As shown by immunofluorescence NEP-B vesicles, together with nucleoporins (Nups), appear first around chromatin at about ten minutes after initiation of NE assembly while NEP-A vesicles appear at a later stage, at about twenty minutes. To investigate the role of different emerin domains in this process, four human emerin peptides consisting of amino acids (aa) 1-70, 1-176, 1-220 and 73-180 were added individually to Xenopus nuclear assembly reactions at different concentrations and the effect on nuclear vesicle recruitment and NPC formation was monitored. Immunofluorescence analysis showed that peptides containing the LEM domain of emerin interfere with a correct NE assembly by inhibiting chromatin decondensation and recruitment of membranes to chromatin. This inhibitory effect was shown to be exerted mainly on NEP-A membranes and on Nup62 and Nupl53. By the use of two antibodies, raised against the LEM domain of human emerin and LAP2ß, two proteins of 30 and 36 kD, respectively, were identified in Xenopus. Both proteins were shown to reside in the NEP-A membrane population providing an explanation for the preferential inhibition of NEP-A recruitment to chromatin by exogenously added LEM domain containing emerin peptides. To further investigate whether the domain specific inhibitory effects of emerin on nuclear assembly correlate with specific interacting proteins, co-precipitation experiments were performed to identify emerin binding proteins in the Xenopus cytosol. From these experiments ß -tubulin was identified as a protein able to interact with emerin peptides 1-70 and 73-180. Staining of X-EDMD cells, which lack emerin, with a ß -tubulin antibody revealed no alterations in the organisation of the microtubule (MT) network. The most prominent effect of emerin mutations regarding MTs was the position of the Microtubule Organising Centre (MTOC) relative to the NE. Staining for the centrosomal protein pericentrin revealed a mis-localisation of the MTOC away from the NE in X-EDMD cell lines at distances at least double compared to control cells
The Transcriptomic and Genomic Analysis of Lamin A/C Expression in the Colon and in Colorectal Cancer
No full textLamins A and C, also known as A-type lamins, are type V nuclear intermediate filament proteins which form an interlacing meshwork of filaments subjacent to the inner nuclear membrane termed the nuclear lamina. A-type lamins have been implicated in DNA replication, gene transcription regulation, apoptosis, regulation of growth promoters and nuclear migration. Traditionally, expression of A-type lamins has been associated with differentiated cells. As such, mutations in A-type lamins have been associated with a diverse range of genetic diseases, including premature ageing syndromes and with increased proliferation, especially in tumours.
In colorectal cancer, expression of A-type lamins, have been shown to impart an adverse prognosis. In order to understand the underlying biological processes responsible for this adverse outcome in patients with colorectal cancer, I sought to clarify the expression profile of A-type lamins and their binding partners in normal colonic/rectal mucosa, prior to investigating the expression of A-type lamins in colorectal cancers. I used fresh tissue specimens obtained from patients with colorectal cancer for my experiments. A unique finding was the expression of lamin A in the putative stem cell niche of colonic / rectal mucosal crypts.
Further studies in the form of a microarray analysis, revealed a very complex picture of up regulation involving various signalling cascades in the cancer samples expressing A-type lamins. There was no evidence to suggest a direct involvement of A-type lamins influencing the Wnt signalling cascade, however, direct involvement of other signalling cascades, such as the IGF signalling cascade, Shh signalling cascade and TGF-β signalling cascades were noted. These signalling cascades were known to influence the Wnt signalling cascades and hence could play a crucial role in the pathogenesis of colorectal cancers expressing A-type lamins.
In addition to these important signalling cascades, other key genes involved in apoptosis, growth promoters, cell adhesion, stem cell regulation, oncogenes and tumour suppression, were noted to have a unique expression profile in the cancer sample expressing A-type lamins, not observed in the cancer sample lacking A-type lamin expression. These observations were suggestive of A-type lamins having a diverse range of actions via, as yet, undefined pathways. It would appear that A-type lamins were imparting a more motile, less adherent phenotype with stem cell like features in colorectal cancers expressing A-type lamins. This could explain the observed poor prognosis of patients with colorectal cancers expressing A-type lamins.
Creatine kinase brain (CKB), was also identified as an additional, potential, prognostic indicator in the Duke’s B group of patients with colorectal cancer expressing A-type lamins. This potential marker, in conjunction with A-type lamin expression could be used to identify a sub group of Duke’s B patients at high risk. Whether adjuvant therapy in this group would help improve their long term survival is unknown since no study has been done to assess this
