Enhancing milk quality assessment with watermelon (Citrullus lanatus) urease immobilized on VS2-chitosan nanocomposite beads using response surface methodology

Article

作者: Divya ; Kayastha, Arvind M. ; Patel, Amit Kumar ; Srivastava, Anchal ; Kumar, Prince

An eco-friendly hydrothermal method synthesized VS₂ nanosheets. Several spectroscopic and microscopic approaches (TEM) were used to characterize the produced VS₂ nanosheet microstructure. VS₂, Chitosan, and nanocomposite were used to immobilize watermelon (Citrullus lanatus) urease. Optimization using the Response Surface Methodology and the Box-Behnken design yielded immobilization efficiencies of 65.23 %, 72.52 %, and 87.68 % for chitosan, VS₂, and nanocomposite, respectively. The analysis of variance confirmed the mathematical model's validity, enabling additional research. AFM, SEM, FTIR, Fluorescence microscopy, and Cary Eclipse Fluorescence Spectrometer showed urease conjugation to the matrix. During and after immobilization, FTIR spectra showed a dynamic connectivity of chemical processes and bonding. The nanocomposite outperformed VS₂ and chitosan in pH and temperature. Chitosan and VS₂-immobilized urease were more thermally stable than soluble urease, but the nanocomposite-urease system was even more resilient. The nanocomposite retained 60 % of its residual activity after three months of storage. It retains 91.8 % of its initial activity after 12 reuse cycles. Nanocomposite-immobilized urease measured milk urea at 23.62 mg/dl. This result was compared favorably to the gold standard p-dimethylaminobenzaldehyde spectrophotometric result of 20 mg/dl. The linear range is 5 to 70 mg/dl, with a LOD of 1.07 (±0.05) mg/dl and SD of less than 5 %. The nanocomposite's k_sel coefficient for interferents was exceptionally low (k_sel < 0.07), indicating urea detection sensitivity. Watermelon urease is suitable for dairy sector applications due to its availability, immobilization on nanocomposite, and reuse.

2022-06-01·Journal of Endocrinological Investigation

Role of chromogranin A-derived fragments after resection of nonfunctioning pancreatic neuroendocrine tumors

Article

作者: Manzoni, M F ; Partelli, S ; Crippa, S ; Andreasi, V ; Corti, A ; Di Filippo, L ; Muffatti, F ; Falconi, M

PURPOSENo single reliable biomarker is available for nonfunctioning pancreatic neuroendocrine tumors (NF-PanNETs). Vasostatin-1 (VS-1), the N-terminal fragment of chromogranin A (CgA), seems to be a more accurate biomarker compared to its precursor. Primary aim was to investigate the ability of VS-1, compared to total-CgA, to assess the effectiveness of surgical resection performed for NF-PanNETs. Secondary aim was to evaluate two additional CgA-derived fragments, pancreastatin (PST) and vasostatin-2 (VS-2), as possible biomarkers for NF-PanNETs.METHODSConsecutive patients who underwent surgery for NF-PanNETs at San Raffaele Scientific Institute were included (n = 35). Plasma levels of CgA and CgA-derived fragments were measured by Enzyme-Linked ImmunoSorbent Assay (ELISA), preoperatively and postoperatively.RESULTSPreoperative VS-1 was significantly higher compared to VS-1 measured on postoperative day 5 (POD5) (pre: 0.338 nM versus POD5: 0.147 nM, P < 0.001), whereas total-CgA significantly increased after surgery (pre: 1.123 nM versus POD5: 1.949 nM, P = 0.006). Overall, 24 patients showed ≥ 1 feature of tumor aggressiveness (T3-T4, nodal/distant metastases, Ki67 > 5%, microvascular/perineural invasion, necrosis). The median percentage decrease in VS-1 plasma levels was 63% (IQR 28-88%) among patients with aggressive tumors, compared to 13% (IQR 0-57%) in the remaining population (P = 0.033). No significant differences in terms of PST (P = 0.870) and VS-2 (P = 0.909) were observed between preoperative and postoperative time.CONCLUSIONVS-1 provides an early assessment of surgical efficacy in patients who undergo resection for NF-PanNETs, especially in those with aggressive neoplasms. Total-CgA, PST and VS-2 have no clinical utility in this setting.

2017-01-01·Thrombosis and Haemostasis2区 · 医学

The anti-inflammatory vasostatin-2 attenuates atherosclerosis in ApoE-/- mice and inhibits monocyte/macrophage recruitment

2区 · 医学

Article

作者: Xiong, Weixin ; Wang, Xiaoqun ; Zhang, Bao ; Dai, Daopeng ; Lu, Lin ; Tao, Rong

SummaryWe showed previously that reduced level of vasostatin-2 (VS-2) correlates to the presence and severity of coronary artery disease. In this study, we aimed to figure out the role of chromogranin A (CGA) derived VS-2 in the development of atherosclerosis and monocyte/macrophage recruitment. Apolipoprotein E-deficient (ApoE-/-) mice fed a high-fat diet exhibited attenuated lesion size by 65 % and 41 % in En face and aortic root Oil red O staining, MOMA-2 positive area by 64 %, respectively, in VS-2 treatment group compared with PBS group. Proinflammatory cytokines tumour necrosis factor-alpha (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and vascular cell adhesion molecule-1 (VCAM-1) were all remarkably reduced in aortic tissues after VS-2 treatment. Mechanistically, in adhesion assay using intravital microscopy in vivo, VS-2 suppressed the number of leukocytes adhering to the wall of apoE-/- mice mesenteric arteries. In chemotactic assay, flow cytometry analysis of peritoneal lavage exudate from C57BL/6 mice showed VS-2 significantly decreased the recruiment number of inflammatory monocytes/macrophages in a thioglycollate-induced peritonitis model. Furthermore, fewer fluorescent latex beads labelled Ly-6Chi monocytes accumulated in aortic sinus lesions of apoE-/- mice after VS-2 treatment. In addition, according to the microarray of human monocyte/macrophage, we found VS-2 stimulation caused a dose-dependent decrease of Rac1 expression and inactivation of Pak1 in mice primary monocytes as well as THP-1 cells and inhibited MCP-1/CCL-5 induced transmigration in vitro. In conclusion, the Chromogranin A-derived VS-2 attenuates atherosclerosis in apoE-/- mice and, in addition to its anti-inflammatory property, also acts as an inhibitor in monocyte/macrophage recruitment.Supplementary Material to this article is available online at www.thrombosis-online.com.

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