1-acid Glycoprotein Research Articles

Association of Anti-Rotavirus IgA Seroconversion with Growth, Environmental Enteric Dysfunction and Enteropathogens in Rural Pakistani Infants

BackgroundThe underperformance of oral vaccines in children of low- and middle-income countries is partly attributable to underlying environmental enteric dysfunction (EED). MethodologyWe conducted a longitudinal, community-based study to evaluate the association of oral rotavirus vaccine (Rotarix®) seroconversion with growth anthropometrics, EED biomarkers and intestinal enteropathogens in Pakistani infants. Children were enrolled between three to six months of their age based on their nutritional status. We measured serum anti-rotavirus immunoglobulin A (IgA) at enrollment and nine months of age with EED biomarkers and intestinal enteropathogens. ResultsA total of 391 infants received two doses of rotavirus (RV) vaccine. 331/391 provided paired blood samples. Of these 331 children, 45% seroconverted at 9 months of age, 35% did not seroconvert and 20% were seropositive at baseline. Non-seroconverted children were more likely to be stunted, wasted and underweight at enrollment. In univariate analysis, insulin-like growth factor (IGF) concentration at 6 months were higher in seroconverters, median (25th, 75th percentile): 26.3 (16.5, 43.5) ng/ml vs. 22.5 (13.6, 36.3) ng/ml for non-seroconverters, p-value = 0.024. At nine months, fecal myeloperoxidase (MPO) concentrations were significantly lower in seroconverters, 3050(1250, 7587) ng/ml vs. 4623.3 (2189, 11650) ng/ml in non-seroconverted children, p-value = 0.017. In multivariable logistic regression analysis, alpha-1 acid glycoprotein (AGP) and IGF-1 concentrations were positively associated with seroconversion at six months. The presence of sapovirus and rotavirus in fecal samples at the time of rotavirus administration, was associated with non-seroconversion and seroconversion, respectively. ConclusionWe detected high baseline RV seropositivity and impaired RV vaccine immunogenicity in this high-risk group of children. Healthy growth, serum IGF-1 and AGP, and fecal shedding of rotavirus were positively associated with RV IgA seroconversion following immunization, whereas the presence of sapovirus was more common in non-seroconverters.Trial registration: Clinical Trials ID: NCT03588013.

Parkinson's Disease

Parkinson’s disease (PD) is a neurodegenerative disease that involves a progressive loss of dopaminergic neurons within the substantia nigra. Patients with PD experience a loss of motor function over time. In normal patients, muscle contractions are driven by action potentials through the binding of the neurotransmitter dopamine. Disruptions to dopaminergic activity would thus result in less efficient contractions. Patients with Parkinson’s can experience akinesia, tremors and other motor dysfunction.Glucocerebrosidase (GCase), specifically β‐GCase, is a lysosomal enzyme involved in sphingolipid metabolism. β‐GCase falls under the hydrolase classification, allowing it to catabolize glucosylceramide (GlcCer) into glucose and ceramide. β‐GCase consists of 497 amino acid glycoproteins within its four domains. The glucocerebrosidase gene (GBA) mutations are often associated with Gaucher disease. However, β‐GCase synthesized from mutated GBA1 is often studied as a biomarker for PD susceptibility. In a healthy cell with the non‐mutated GBA gene, the GBA gene would be transcribed into mRNA and then transported out to the rough Endoplasmic Reticulum, where GCase is then synthesized. Lysosomal integral membrane protein‐2 (LIMP2) is a protein that transports GCase through the Golgi Apparatus and transfers the β‐GCase into a late endosome. When the late endosome fuses with a lysosome, where β‐GCase can hydrolyze its substrates. Mutated β‐GCase may affect this autophagy pathway.This research will explore how mutated β‐GCase may disturb lysosomal functions, which would consequently lead to the aggregation of alpha‐synuclein (ASN), a protein that regulates synaptic vesicle trafficking and subsequently releases dopamine. Therefore, the detection and accumulation of mutated lysosomal enzymes, such as β‐GCase, in the cerebrospinal fluid might serve as biomarkers for Parkinson’s Disease.

Systemic and ovarian inflammation in women with polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is a common endocrine/reproductive/metabolic disorder. The etiology of PCOS is complex and has been linked to low-grade chronic inflammation. Local inflammation of the ovary affects ovulation and induces or aggravates systemic inflammation. PCOS patients demonstrated significantly higher concentrations of circulating inflammatory cells, such as lymphocytes, neutrophils, eosinophilic granulocytes, monocytes and Th17 cells than women without PCOS, while the percentage of Treg cells was lower. Inflammatory factors, such as serum CRP, hs-CRP, IL-1Ra, IL-6, IL-17A, IL-17F, IL-18, IL-23, TNF-α, α-1 acid glycoprotein,monocyte chemoattractant protein-1 (MCP-1) and adipokines and their paralogs, including chemerin, C1q and TNF-related 6 (C1QTNF6), were also found to be significantly increased in the peripheral blood of PCOS patients. Levels of anti-inflammatory cytokines, such as IL-10, IL-17E, IL-27, IL-35 and IL-37, TGF-β, omentin-1, Secreted frizzled-related protein5 (SFRP5) were significantly lower. An analogous situation occurs locally in the ovary. Some vital inflammatory cells and cytokines may initially be released from the ovary and then enter the circulation. The systemic inflammation underlying PCOS is thought to interact with obesity, insulin resistance (IR) and hyperandrogenism. Traditional Chinese medicine, multitargeted treatment, anti-inflammatory and antioxidant medicine, and lifestyle modification can benefit PCOS women by alleviating inflammatory responses.

Role of Medicinal Mushroom Lentinula edodes in Nutrition, Nutraceutics and Ethnopharmacology

Lentinula edodes (shiitake mushroom) is the medicinal macro-fungus popular in Japan and China. It is the second most well- known edible fungus in the global market after Agaricus bisporus Because of presence of dietary fibers, vitamins, proteins, minerals and carbohydrates, shiitake mushroom have significant nutritional worth. It is commonly cultivated in winter season and known as “elixir of life”. Bioactive compounds such as polysaccharides, amino acid derivatives, sterols, glycoproteins, chitin and sulfurous compounds are all extracted from shiitake mushrooms. Some of these are lentinans, eritadenine and KS-2-α- mannan peptide etc. Shiitake mushroom is used therapeutically to cure cancers, cardiovascular disorders, viral and fungal infections, hepatitis, hypertension and hepatic disorders. Shiitake mushroom have also grown in popularity as a result of their unusual and well-liked flavor.

Rhamnose Is Superior to Mannitol as a Monosaccharide in the Dual Sugar Absorption Test: A Prospective Randomized Study in Children With Treatment-Naïve Celiac Disease

Background and AimWe sought to correlate two different measures of gut permeability [lactulose:mannitol (L:M) and lactulose:rhamnose (L:R)] to the severity of duodenal histopathology in children with and without elevated antibodies to tissue transglutaminase (tTG). A secondary objective was to correlate gut permeability with celiac disease (CD) serology and indices of inflammation and bacterial product translocation.MethodsWe prospectively randomized children undergoing endoscopy with abnormal (n = 54) and normal (n = 10) concentrations of circulating antibodies to tTG, to either L:M or L:R. Biopsies underwent modified Marsh scoring to measure mucosal injury. Circulating anticore Escherichia coli lipopolysaccharide (LPS) IgG, α-1 acid glycoprotein, LPS-binding protein, and C-reactive protein concentrations were measured by enzyme immunoassays.ResultsOf the 54 cases with positive celiac serology, 31 and 69% had modified Marsh 0/1 scores or ≥3a, respectively. Circulating tTG IgA correlated with the modified Marsh score (p = 0.03). L:R, but not L:M or percent L excreted, differed according to modified Marsh scores (p = 0.01). There was no significant association between any systemic marker of inflammation or gut injury, and modified Marsh scores. Concerningly, most participants had evidence of urinary M before the challenge sugar was administered.ConclusionsL:R, but not L:M, is associated with modified Marsh scores in children undergoing small bowel biopsy for suspected CD. Despite increased intestinal permeability, we see scant evidence of systemic exposure to gut microbes in these children. Gut permeability testing with L:R may predict which patients with abnormal celiac serology will have biopsy evidence for celiac disease and reduce the proportion of such patients undergoing endoscopy whose Marsh scores are ≤1. M should not be used as a monosaccharide for permeability testing in children.

Circular RNA MTCL1 promotes advanced laryngeal squamous cell carcinoma progression by inhibiting C1QBP ubiquitin degradation and mediating beta-catenin activation

BackgroundCircular RNAs (circRNAs) are involved in regulatory processes of ubiquitination and deubiquitination in various tumors at post-transcriptional epigenetic modification level. However, the underlying mechanism and its biological functions of circRNAs in the advanced laryngeal squamous cell carcinoma (LSCC) remain obscure.MethodsRNA sequencing and quantitative real-time PCR (qRT-PCR) assays were applied to screen for circRNAs differentially expressed in LSCC tissues and cell lines. The candidate RNA-binding proteins and target signalling pathway were detected by RNA pull-down and mass spectrometry, in situ hybridization (ISH), immunohistochemistry (IHC), qRT-PCR assays, and bioinformatics analysis. The functional roles of these molecules were investigated using in vitro and in vivo experiments including EdU, transwell, wound healing, western blot assays, and the xenograft mice models. The molecular mechanisms were identified using RNA pull-down assays, RNA immunoprecipitation (RIP), Co-IP, ISH, Ubiquitination assay, bioinformatics analysis, and the rescue experiments.ResultsHere, we unveil that microtubule cross-linking factor 1 circRNA (circMTCL1, circ0000825) exerts its critical oncogenic functions by promoting complement C1q-binding protein (C1QBP)-dependent ubiquitin degradation and subsequently activating Wnt/β-catenin signalling in laryngeal carcinoma initiation and development. Specifically, circMTCL1 was remarkably up-regulated in the paired tissues of patients with LSCC (n = 67), which predicted a worse clinical outcome. Functionally, circMTCL1 exerted oncogenic biological charactersistics by promoting cell proliferative capability and invasive and migrative abilities. Ectopic circMTCL1 augumented cell proliferation, migration, and invasion of LSCC cells, and this effect could be reversed by C1QBP knocking down in vitro and in vivo. Mechanistically, circMTCL1 directly recruited C1QBP protein by harboring the specific recognized sequence (+ 159 − + 210), thereby accelerating the translation of C1QBP expression by inhibiting its ubiquitin–proteasome-mediated degradation. Importantly, the direct interaction of C1QBP with β-catenin protein was enhanced via suppressing the β-catenin phosphorylation and accelerating its accumulation in cytoplasm and nucleus.ConclusionOur findings manifested a novel circMTCL1-C1QBP-β-catenin signaling axis involving in LSCC tumorigenesis and progression, which shed new light on circRNAs-ubiquitous acidic glycoprotein mediated ubiquitin degradation and provided strategies and targets in the therapeutic intervention of LSCC.

Alpha-1 Acid Glycoprotein Reduction Differentiated Recovery from Remission in a Small Cohort of Cats Treated for Feline Infectious Peritonitis.

Feline infectious peritonitis (FIP) is a systemic immune-mediated inflammatory perivasculitis that occurs in a minority of cats infected with feline coronavirus (FCoV). Various therapies have been employed to treat this condition, which was previously usually fatal, though no parameters for differentiating FIP recovery from remission have been defined to enable clinicians to decide when it is safe to discontinue treatment. This retrospective observational study shows that a consistent reduction of the acute phase protein alpha-1 acid glycoprotein (AGP) to within normal limits (WNL, i.e., 500 μg/mL or below), as opposed to duration of survival, distinguishes recovery from remission. Forty-two cats were diagnosed with FIP: 75% (12/16) of effusive and 54% (14/26) of non-effusive FIP cases recovered. Presenting with the effusive or non-effusive form did not affect whether or not a cat fully recovered (p = 0.2). AGP consistently reduced to WNL in 26 recovered cats but remained elevated in 16 cats in remission, dipping to normal once in two of the latter. Anaemia was present in 77% (23/30) of the cats and resolved more quickly than AGP in six recovered cats. The presence of anaemia did not affect the cat’s chances of recovery (p = 0.1). Lymphopenia was observed in 43% (16/37) of the cats and reversed in nine recovered cats but did not reverse in seven lymphopenic cats in the remission group. Fewer recovered cats (9/24: 37%) than remission cats (7/13: 54%) were lymphopenic, but the difference was not statistically different (p = 0.5). Hyperglobulinaemia was slower than AGP to return to WNL in the recovered cats. FCoV antibody titre was high in all 42 cats at the outset. It decreased significantly in 7 recovered cats but too slowly to be a useful parameter to determine discontinuation of antiviral treatments. Conclusion: a sustained return to normal levels of AGP was the most rapid and consistent indicator for differentiating recovery from remission following treatment for FIP. This study provides a useful model for differentiating recovery from chronic coronavirus disease using acute phase protein monitoring.

Genomic Confirmation of the P-IIIe Subclass of Snake Venom Metalloproteinases and Characterisation of Its First Member, a Disintegrin-Like/Cysteine-Rich Protein.

Disintegrin-like/cysteine-rich (DC) proteins have long been regarded just as products of proteolysis of P-III snake venom metalloproteinases (SVMPs). However, here we demonstrate that a DC protein from the venom of Vipera ammodytes (Vaa; nose-horned viper), VaaMPIII-3, is encoded per se by a P-III SVMP-like gene that has a deletion in the region of the catalytic metalloproteinase domain and in part of the non-catalytic disintegrin-like domain. In this way, we justify the proposal of the introduction of a new subclass P-IIIe of SVMP-derived DC proteins. We purified VaaMPIII-3 from the venom of Vaa in a series of chromatographic steps. A covalent chromatography step based on thiol-disulphide exchange revealed that VaaMPIII-3 contains an unpaired Cys residue. This was demonstrated to be Cys6 in about 90% and Cys19 in about 10% of the VaaMPIII-3 molecules. We further constructed a three-dimensional homology model of VaaMPIII-3. From this model, it is evident that both Cys6 and Cys19 can pair with Cys26, which suggests that the intramolecular thiol-disulphide exchange has a regulatory function. VaaMPIII-3 is an acidic 21-kDa monomeric glycoprotein that exists in at least six N-glycoforms, with isoelectric points ranging from pH 4.5 to 5.1. Consistent with the presence of an integrin-binding motif in its sequence, SECD, VaaMPIII-3 inhibited collagen-induced platelet aggregation. It also inhibited ADP- and arachidonic-acid-induced platelet aggregation, but not ristocetin-induced platelet agglutination and the blood coagulation cascade.

Physicochemical property and antioxidant activity of polysaccharide from the seed cakes of Camellia oleifera Abel.

Seed cake refers to the food by‐product of Camellia oleifera Abel, and its insufficient utilization can cause serious environment pollution and resource waste. This study aimed to investigate antioxidant activities of the polysaccharide from the seed cakes of Camellia oleifera Abel (COCP) in vitro and in vivo. The physicochemical property of COCP was also determined. COCP was characterized to be an acidic glycoprotein and mainly consisted of rhamnose (Rha), arabinose (Ara), galactose (Gal), glucose (Glc), xylose (Xyl), mannose (Man), and galacturonic acid (Gal‐UA). COCP exhibited the polysaccharide's characteristic absorption in the Fourier transform infrared (FT‐IR) spectroscopy and showed as sheet‐like structures with a smooth surface under the scanning electron microscope (SEM). COCP exerted good scavenging activities on ABTS, DPPH, and OH radicals, with IC50 values of 2.94, 2.24, and 5.09 mg/ml, respectively. COCP treatment improved learning and memory abilities of D‐galactose‐induced aging mice. Significant decreases were found in the levels of alanine transaminase (ALT), aspartate aminotransferase (AST), creatinine (CRE), blood urea nitrogen (BUN), creatine kinase (CK), and lactate dehydrogenase (LDH) in serum, as aging mice were supplemented with COCP. Aging mice showed obviously higher malondialdehyde (MDA) contents and lower superoxide dismutase (SOD) and glutathione peroxidase (GSH‐Px) activities in serum, brain, liver, kidney, and heart. The phenomena were noticeably reversed when mice were treated with COCP. Results indicated that COCP exerted excellent antioxidant activities in vitro and in vivo, which support its potential application as a natural antioxidant in food and medicine industries.

HISTOLOGICAL AND HISTOCHEMICAL STUDY OF BRUNNER'S GLANDS IN LOCAL ADULT COWS

The study was conducted to add information to the database for concentrate on theBrunner's glands of the duodenum in adult local cows. The histological study showedthat Brunner's glands present in the tunica submucosa of the duodenum in local adultcows, the glands were branched tubuloalveolar secreting mucous, extended from theend of the pylorus to the first and middle third of duodenum, and absent at the lastthird of the duodenum. The excretory ducts of Brunner's glands opened at the cryptsof lieberkuhn at the base of intestinal villi penetrating the muscularis mucosa layer.The statistical analysis showed there was a significant variation in the diameter andlumen of Brunner's glands between the first and middle third of duodenum at (P≤0.05). The diameter and lumen of Brunner's glands at the first third of duodenum werelower than that of the middle third of duodenum. The statistical study showed , thatthere was no significant variation of the lining epithelium of Brunner's glands betweenfirst and middle third of duodenum, and there was no significant variation in thethickness of Brunner's glands aggregation between first and middle third of theduodenum.The histochemical study revealed that there were different types of glycoprotein inBrunner's glands and excretory duct in local adult cows, at first third of duodenumBrunner's glands contain a large amount of neutral glycoprotein and little amount ofacidic glycoprotein, while at middle third of duodenum the Brunner's glands containthe same amount of neutral and acidic glycoprotein. At the first third of duodenum,the excretory ducts of Brunner's glands contain neutral glycoprotein more than acidicglycoprotein, while at the middle third of duodenum the excretory ducts of Brunner's glands contain a large amount of acidic glycoprotein and little amount of neutralglycoprotein.

Single-virus assay reveals membrane determinants and mechanistic features of Sendai virus binding

Sendai virus (SeV, formally murine respirovirus) is a membrane-enveloped, negative-sense RNA virus in the Paramyxoviridae family and is closely related to human parainfluenza viruses. SeV has long been utilized as a model paramyxovirus and has recently gained attention as a viral vector candidate for both laboratory and clinical applications. To infect host cells, SeV must first bind to sialic acid glycolipid or glycoprotein receptors on the host cell surface via its hemagglutinin-neuraminidase (HN) protein. Receptor binding induces a conformational change in HN, which allosterically triggers the viral fusion (F) protein to catalyze membrane fusion. While it is known that SeV binds to α2,3-linked sialic acid receptors, and there has been some study into the chemical requirements of those receptors, key mechanistic features of SeV binding remain unknown, in part because traditional approaches often convolve binding and fusion. Here, we develop and employ a fluorescence microscopy-based assay to observe SeV binding to supported lipid bilayers (SLBs) at the single-particle level, which easily disentangles binding from fusion. Using this assay, we investigate mechanistic questions of SeV binding. We identify chemical structural features of ganglioside receptors that influence viral binding and demonstrate that binding is cooperative with respect to receptor density. We measure the characteristic decay time of unbinding and provide evidence supporting a “rolling” mechanism of viral mobility following receptor binding. We also study the dependence of binding on target cholesterol concentration. Interestingly, we find that although SeV binding shows striking parallels in cooperative binding with a prior report of Influenza A virus, it does not demonstrate a similar sensitivity to cholesterol concentration and receptor nanocluster formation.

Acute Phase Proteins as Early Predictors for Immunotherapy Response in Advanced NSCLC: An Explorative Study.

In the last decade, targeting the immune system became a promising therapy in advanced lung cancer stages. However, in a clinical follow-up, patient responses to immune checkpoint inhibitors widely differ. Peripheral blood is a minimally invasive source of potential biomarkers to explain these differences. We blindly analyzed serum samples from 139 patients with non-small cell lung cancer prior to anti-PD-1 or anti-PD-L1 therapies to assess whether baseline levels of albumin (ALB), alpha-1 acid glycoprotein (AGP), alpha1-antitrypsin (AAT), alpha2-macroglobulin (A2M), ceruloplasmin (CP), haptoglobin (HP), alpha1-antichymotrypsin (ACT), serum amyloid A (SAA), and high-sensitivity C-reactive protein (hs-CRP), have a predictive value for immunotherapy success. Disease progression-free survival (PFS) was calculated based on RECIST 1.1 criteria. A multivariate Cox regression analysis, including serum levels of acute-phase proteins and clinical parameters, revealed that higher pre-therapeutic levels of HP and CP are independent predictors of a worse PFS. Moreover, a combined panel of HP and CP stratified patients into subgroups. We propose to test this panel as a putative biomarker for assessing the success of immunotherapy in patients with NSCLC.

Wound Infusion of 0.35% Levobupivacaine Reduces Mechanical Secondary Hyperalgesia and Opioid Consumption After Cesarean Delivery: A Prospective, Randomized, Triple-Blind, Placebo-Controlled Trial.

Some patients still report moderate-to-severe postoperative pain after cesarean delivery. Local anesthetic wound infusion improves acute pain and might act on peripheral and central sensitization mechanisms; however, no studies have proved this hypothesis. We evaluated the potential benefits of continuous wound infusion of levobupivacaine after cesarean delivery on secondary hyperalgesia (primary end point) and primary hyperalgesia, pain relief, persistent pain, and inflammatory and metabolic stress response. Healthy women scheduled for elective cesarean delivery participated in this prospective, randomized, triple-blind, placebo-controlled trial (NCT01458431). All patients received spinal anesthesia with 0.5% hyperbaric bupivacaine with fentanyl and a multiholed wound catheter placed under the fascia. Women were randomized to receive continuous wound infusion (0.35% levobupivacaine 7 mL/h for 48 hours; group L) or an equal volume of saline (group S). Secondary hyperalgesia to punctate mechanical stimuli was evaluated using dynamic tests, and primary hyperalgesia was evaluated using an electronic von Frey anesthesiometer; both were assessed at 24, 48, and 72 hours. The following variables were collected: intensity of postoperative parietal and visceral pain at rest and on movement rated on a visual analog scale >72 hours, time to first bolus of patient-controlled analgesia (PCA), cumulative dose of rescue morphine (PCA) and acetaminophen, ability to sleep and sleep quality, and patient satisfaction. Persistent postoperative pain was evaluated during a telephone interview at 1, 3, 6, and 12 months after surgery. C-reactive protein, acid glycoprotein, preprandial glucose, insulin, cortisol, prolactin, growth hormone, and interleukin-6 were measured before cesarean delivery and at 8, 24, and 48 hours. Adverse events and patient outcomes were recorded. Seventy women were included. In group L, the area of secondary hyperalgesia was significantly reduced (43.4 [18.5-80] vs 68.4 [39.0-136] cm2 and 45.1 [0.9-89.8] vs 67.3 [31.3-175] cm2 at 24 and 48 hours, respectively; group:time interaction P value < .001), the mechanical pain threshold was significantly higher at 24 hours (633 [441-802] vs 417 [300-572] g.mm-2; P = .001), and morphine consumption was significantly lower at 24 hours (4 [2-11] vs 11[6-23]; P = .003) compared with group S. Levobupivacaine had no effect on persistent postoperative pain at 1, 3, 6, and 12 months. Plasma insulin levels in the immediate postoperative period and at 8, 24, and 48 hours were significantly lower in group L (P < .001). There were no significant differences in other biochemical parameters of inflammatory and endocrine-metabolic response. Levobupivacaine wound infusion provides adequate analgesia and might be an effective antihyperalgesic adjunct.

The modulatory effect of carvacrol on viral shedding titer and acute phase response in broiler chickens experimentally infected with infectious bronchitis virus

Infectious bronchitis virus (IBV) is one of the major respiratory diseases of broiler causing huge economic losses. The inability to control IBV using different vaccination programs owing to the high mutation rate and recombination ability of the RNA genome generates IBV variants. This study was designed to give a specific perspective of carvacrol effect on early immune response, viral shedding titer, oxidative stress, serum biochemical parameters and clinical consequences in broilers experimentally infected by IBV. One hundred and twenty-one-day old commercial broiler chicks were equally divided into 4 groups. First group was considered as control. Second group was given carvacrol, third group was infected with IBV and fourth group was given carvacrol and infected with IBV. Infection with variant IBV induced significant upregulation of chicken interferon-inducible transmembrane protein 3 (chIFITM3) gene in trachea, elevations in serum levels of Alpha-1 acid glycoprotein (α1-AGP) and Interleukin 6 (IL-6), total leucocytic count (TLC), heterophil/lymphocyte (H/L) ratio and oxidative stress in lung and kidney tissues. Beside, histopathological changes in trachea, lung and kidney induced by IBV, elevation of kidney function tests was detected. The pretreatment with carvacrol significantly reduced viral shedding titer, chIFITM3 gene expression, IL-6 and α1-AGP levels, leucocytic response and H/L ratio with minimization of clinical signs intensity. Also, carvacrol relieved oxidative stress, ameliorated the increased uric acid level and histopathological alterations in kidney and lung caused by viral infection.

Shell water-soluble matrix protein from oyster shells promoted proliferation, differentiation and mineralization of osteoblasts in vitro and vivo

Matrix protein is secreted by the membrane of bivalve shellfish to and used to regulate shell biomineralization. In this study, we extracted water-soluble matrix protein (WSMP) from oyster shells to investigate its effects on osteogenic differentiation and mineralization of MC3T3-E1 cells and osteoporosis rats. Our results suggested that WSMP was an acidic glycoprotein by amino acid analysis and secondary structure analysis. In vitro, WSMP could promote osteoblastic proliferation. Moreover, alkaline phosphatase (ALP) and osteocalcin (OCN) were increased, mineralized nodules were increased, and BMP-2 expression was up-regulated. Additionally, in vivo, tartrate-resistant acid phosphatase (TRAP) and Bone alkaline phosphatase (BALP) expressions in the medium-dose and high-dose groups were significantly decreased compared with the model group, while OCN expression was significantly increased. Bone mineral density (BMD) and bone mineral content (BMC) of bone recovered significantly. In summary, WSMP can promote the proliferation, differentiation and mineralization of osteoblasts in vitro and in vivo.

Choice of assay affects serum albumin measurements in HIV patients: A comparison of bromocresol green and bromocresol purple dye binding methods

Background: Dye-binding assays – bromocresol green (BCG) and bromocresol purple (BCP) –are widely used for serum albumin measurement. Differences between the results reported by these assays can have important clinical implications. There are limited studies elucidating the differences between the results of two assays in HIV patients. We aimed to determine whether the choice of assay affects serum albumin measurements in HIV patients. Materials and Methods: Hundred newly diagnosed HIV patients were included in the study. The subjects were followed up for 6–12 months after initiating ART. Hundred HIV-negative healthy controls matched for age and body mass index were taken as controls. Serum albumin and three acute phase proteins (APPs) (Alpha-1 acid glycoprotein, C-reactive protein, ceruloplasmin) were measured. Serum albumin measurement was done by both BCG and BCP dye binding methods. Paired t-test was used to compare the groups. Results: The concentration of APPs reduces with ART indicating reduced inflammation, and is lowest in the control group. The difference in serum albumin concentrations measured by the two dye binding methods reduces with reduction in APPs. The study further indicates that the BCG method gives higher values of serum albumin as compared to BCP method in the presence of other proteins in the sample. Conclusion: The choice of albumin assay affects the serum albumin assessment of HIV patients. We recommend that the BCP assay be used to measure serum albumin in HIV patients, particularly in patients not on ART.

Concentração de proteínas de fase aguda e vitamina D em cães com linfoma multicêntrico

Abstract This study aimed to evaluate the serum concentration of vitamin D (25-Hydroxyvitamin D) and acute phase proteins (APPs; alpha-1 acid glycoprotein, haptoglobin, transferrin, ceruloplasmin, albumin, IgA, IgG and alpha-1 - antitrypsin) as potential biomarkers for prognostic and therapy response in dogs with multicentric lymphoma submitted to the CHOP (Cyclophosphamide, Doxorubicin, Vincristine and Prednisone) chemotherapy protocol. Thirteen dogs with multicentric lymphoma classified as high grade by cytology were included in the treatment group (GL), while ten healthy dogs were included in the control group (GC). Serum was collected in the weeks T0, T5 and T10 of CHOP chemotherapy protocol, for the GL group, and in a single collection, for the GC group. All the collected samples were evaluated for the APPs and vitamin D concentrations through electrophoresis and chemiluminescence methods, respectively. Diagnostic and staging tests were performed for all the dogs in the GL group, and included cytopathology, histopathology and immunohistochemistry of the affected lymph node. Of these dogs, 9 achieved a complete response and 4 a partial response to the treatment. Data analysis was performed with the R software. The results demonstrated that serum concentrations of IgA, haptoglobin and α1-acid glycoprotein were significantly different between the groups and also between the different chemotherapy times analyzed (p&lt;0.05), indicating that these proteins can be considered as sensitive biomarkers for lymphoma in dogs. Furthermore, the α1-acid glycoprotein showed prognostic value for the disease, with 63% specificity. However, vitamin D concentration was not correlated with prognosis of the dogs with lymphoma.

CHARACTERISTICS OF THE MECHANISMS OF CYTOPROTECTIVE ACTIVITY OF CRYOPRESERVED PLACENTA EXTRACT ACCORDING TO MORPHOLOGICAL AND BIOCHEMICAL STUDIES OF THE GASTRIC MUCOSA IN THE EXPERIMENT

Relevance. Non-steroidal anti-inflammatory drugs (NSAIDs) are used as antipyretics and analgesics. In this case, NSAIDs have a side effect - ulcerogenic. Therefore, it is important to find means to correct the ulcerogenic action of NSAIDs. Potential antiulcerogenic action may be in cryopreserved human placenta extract "Cryocell-placental cryoextract".&#x0D; Objective: to investigate the effect of cryopreserved placenta (CEP) extract on the morphological state of the stomach and the processes of protein synthesis in the gastric mucosa with the introduction of NSAIDs diclofenac sodium in rats with experimental rheumatoid arthritis.&#x0D; Materials and methods. The study was performed on 28 nonlinear male rats weighing 200-220 g. The animals were divided into 4 groups: I (n = 7) – intact rats; II (n = 7) – rats with adjuvant arthritis (AA), this is a treatment without treatment; III (n = 7) – rats with AA, treated with diclofenac sodium (DN), 8.0 mg / kg, intragastrically (i/g); IV (n = 7) – rats with AA, treated with DN, 8.0 mg / kg, i/g and CEP, 0.16 ml / kg body weight, intramuscularly (i/m).&#x0D; AA was simulated by subplantar injection of complete Freund's adjuvant into the hind right limb at a rate of 0.1 ml per rat. The day of adjuvant administration was considered as the "0" day of the experiment. AA treatment was performed from 14 to 28 days. DN was applied daily, and CEP – with an interval of 3 days: for 14, 17, 20, 23 and 26 days. On day 28, the animals were removed from the experiment by cervical dislocation under inhalation anesthesia.&#x0D; To assess morphological changes, gastric fragments were fixed in 10.0% neutral formalin solution. Histological sections 5–7 μm thick were stained with hematoxylin and eosin, as well as Schiff's reagent (fuchsin sulfuric acid) with iodic acid. The concentration of total protein (TP) was determined spectrophotometrically by light absorption at a wavelength of 546 nm. The content of RNA and DNA in the homogenates of the gastric mucosa was determined spectrophotometrically by reaction with perchloric acid.&#x0D; Results. In rats with AA, the mucous membrane (MM) had a preserved structure in all animals. There were no signs of hyper- or hyposecretion of mucins when stained with a chic reaction with alcyan blue. There were foci of desquamation of the superficial epithelium and single lymphocytes. The introduction of DN in rats with AA led to signs of erosive changes in MM. Deep erosions and single ulcerative defects were found in 28.6% of rats, reaching the depth of the lesion to the muscle plate. There is a small amount of mucopolysaccharides with a predominance of acidic glycoproteins. Reduced content of neutral glycoproteins. Single desquamated epitheliocytes were detected. Focal lympho-plasmacytic infiltration with exudate with eosinophilic leukocytes was noted in the submucosal layer of MM. Against the background of the combined use of DN and CEP in rats with AA in any animal, no significant violations of the histoarchitectonics of the MM, except for isolated foci of desquamation of the superficial epithelium.&#x0D; In gastric MM homogenates in rats with AA there was a decrease (p &lt;0.01) in the level of TP by 14.1% and a decrease (p &lt;0.05) in the content of RNA and DNA by 27.5% and 14.3%, respectively, relative to indicators of intact animals.&#x0D; Against the background of DN monotherapy, there was an even greater decrease (p &lt;0.001) in the level of TP: by 26.2% (40.3 ± 1.43 μg / mg) relative to intact animals (54.6 ± 1.39 μg / mg). And the content of RNA and DNA was 0.59 ± 0.57 μg / ml and 2.21 ± 0.27 μg / ml, respectively.&#x0D; Against the background of the combined use of DN and CEP, comparable to the level of intact animals indicators of TP and DNA were noted. Thus, the level of TP was 53.0 ± 2.09 μg / mg, which was only 2.9% lower than intact animals (54.6 ± 1.39 μg / mg), and the DNA content by 5.1% (p = 0.3) was lower than intact animals (3.1 ± 0.12 μg / ml) and was respectively 2.94 ± 0.08 RNA level (1.72 ± 0.12 μg / ml) on 17.6% exceeded (p = 0.1) intact animals (1.46 ± 0.08 μg / ml).&#x0D; Conclusions. The combined use of placental cryoextract and diclofenac sodium eliminates the negative effect of the latter on protein metabolism in the gastric mucosa of rats with adjuvant arthritis. Thus, the level of total protein was 53.0 ± 2.09 μg / mg, which is only 2.9% lower than intact animals. Restoration of diclofenac sodium-induced mucin secretion by the integumentary epithelium of the gastric mucosa is one of the mechanisms of cytoprotective activity of cryopreserved placenta extract in experimental ulcerogenesis.

Elucidation of the interaction mechanism of olmutinib with human α-1 acid glycoprotein: insights from spectroscopic and molecular modeling studies

Olmutinib, the third-generation tyrosine kinase inhibitor, is applied in treating non-small cell lung cancer (NSCLC). The aim of this study is to elucidate the interaction mechanism of olmutinib with human α-1 acid glycoprotein (HAG), an important carrier protein, by mean of multi-spectroscopic and molecular simulation techniques. Fluorescence spectral results confirmed that the fluorescence of this carrier protein can be quenched by olmutinib in the static quenching mode, and this anticancer drug possesses a moderate binding affinity on HAG. The evidence from thermodynamic analysis, replacement interaction with ANS and sucrose, and computational simulation results showed that hydrogen bonding, hydrophobic interactions, and van der Waals forces involved the olmutinib-HAG complexation process. The results from UV-vis, 3D fluorescence and synchronous fluorescence spectroscopy proved that binding anticancer drug olmutinib caused the alteration in the microenvironment around Trp residues. And, circular dichroism spectral results provided the support for the conformational alterations in the carrier protein. The data also proved that olmutinib preferably bound to the hydrophobic cavity of HAG and the binding distance between the two was 2.21 nm. In addition, it can be found that the presence of some metal ions such as Zn2+, Ca2+, Ni2+ and Cu2+ would exert a certain extent effect on the olmutinib-HAG complexation process. Communicated by Ramaswamy H. Sarma

Postexercise urinary alpha-1 acid glycoprotein is not dependent on hypoxia.

Proteinuria is a transient physiological phenomenon that occurs with a range of physical activities and during ascent to altitude. Exercise intensity appears to dictate the magnitude of postexercise proteinuria; however, evidence also indicates the possible contributions from exercise-induced hypoxemia or reoxygenation. Using an environmental hypoxic chamber, this crossover-designed study aimed to evaluate urinary alpha-1 acid glycoprotein (α1-AGP) excretion pre/postexercise performed in hypoxia (HYP) and normoxia (NOR). Sixteen individuals underwent experimental sessions in normoxia (NOR, 20.9% O2) and hypoxia (HYP, 12.0% O2). Sessions began with a 2-h priming period before completing a graded maximal exercise test (GXT) on a cycle ergometer, which was followed by continuation of exposure for an additional 2 h. Physiological responses (i.e., blood pressure, heart rate, and peripheral oxygenation), Lake Louise Scores (LLSs), and urine specimens (analyzed for albumin and α1-AGP) were collected pre- and postexercise (after 30, 60, and 120 min). Peak power output was significantly reduced in HYP (193 ± 45 W) compared with NOR (249 ± 59 W, P < 0.01). Postexercise urinary α1-AGP was greater in NOR (20.04 ± 14.84 µg·min-1) than in HYP (15.08 ± 13.46 µg·min-1), albeit the difference was not significant (P > 0.05). Changes in urinary α1-AGP from pre- to post-30 min were not related to physiological responses or performance outcomes observed during GXT in NOR or HYP. Despite profound systemic hypoxemia with maximal exercise in hypoxia, postexercise α1-AGP excretion was not elevated above the levels observed following normoxic exercise.NEW & NOTEWORTHY By superimposing hypoxic exposure and maximal exercise, we were able to investigate the impact of hypoxia on postexercise proteinuria. Urinalysis for α1-AGP (via particle-enhanced immunoturbidimetry) in specimens collected pre-/postexercise enabled the sensitive detection of altered glomerular permeability. Data indicated that exercise intensity, rather than the degree of exercise-induced hypoxemia, determines postexercise proteinuria.