Analysis Of Haptoglobin Research Articles

Evaluation of Zonulin Expression and Its Potential Clinical Significance in Glioblastoma.

Glioblastoma, the deadliest adult brain tumor, poses a significant therapeutic challenge with a dismal prognosis despite current treatments. Zonulin, a protein influencing tight junctions and barrier functions, has gained attention for its diverse roles in various diseases. This study aimed to preliminarily analyze the circulating and tumor zonulin levels, evaluating their impact on disease prognosis and clinical-radiological factors. Additionally, we investigated in vitro zonulin expression in different glioblastoma cell lines under two different conditions. The study comprised 34 newly diagnosed glioblastoma patients, with blood samples collected before treatment for zonulin and haptoglobin analysis. Tumor tissue samples from 21 patients were obtained for zonulin expression. Clinical, molecular, and radiological data were collected, and zonulin protein levels were assessed using ELISA and Western blot techniques. Furthermore, zonulin expression was analyzed in vitro in three glioblastoma cell lines cultured under standard and glioma-stem-cell (GSC)-specific conditions. High zonulin expression in glioblastoma tumors correlated with larger preoperative contrast enhancement and edema volumes. Patients with high zonulin levels showed a poorer prognosis (progression-free survival [PFS]). Similarly, elevated serum levels of zonulin were associated with a trend of shorter PFS. Higher haptoglobin levels correlated with MGMT methylation and longer PFS. In vitro, glioblastoma cell lines expressed zonulin under standard cell culture conditions, with increased expression in tumorsphere-specific conditions. Elevated zonulin levels in both the tumor and serum of glioblastoma patients were linked to a poorer prognosis and radiological signs of increased disruption of the blood-brain barrier. In vitro, zonulin expression exhibited a significant increase in tumorspheres.

38 Effect of Breed on Dairy Calf Vigor from Birth to Preweaning

Abstract There is a stigma in the dairy industry that Brown Swiss calves are not as quick to stand and nurse following birth compared to other dairy breeds. The objective of this study is to determine if Brown Swiss calves are less vigorous at birth and if this affects their health status and early calf growth compared with the dairy industry’s most prominent breed, the Holstein. All calves (n = 25) were scored shortly after birth using a modified Apgar scoring system used for infants. Thirteen parameters divided into 5 categories were evaluated to determine overall vigor of each calf. These included visual scores for meconium staining and swelling of the head; initial mobility scores for time to lift head, time to stand, and ear position; general responsiveness scores included strength of the suckling reflex, head shake in response to straw in nasal cavity, tongue withdrawal when pinched, and eye reflex; oxygenation score for mucus membrane color; rates scores included heart rate, respiration rate, and temperature. In addition to the vigor scoring, a weight was taken at birth and 2 weeks post calving. Jugular blood samples were collected at birth, 24 hours later, and at 2 weeks of age, and serum was stored for later analysis of haptoglobin and IgG concentration. Vigor scores and weight data were analyzed by breed using the mixed procedure in SAS. The vigor score for suckling reflex was lower (P = 0.04) for Brown Swiss calves (1.69, n = 12) than Holstein calves (2.39, n = 13). A weakened suckling reflex can lead to more challenges when consuming colostrum. Early colostrum intake is vital to obtaining passive immunity and maintaining health status in young calves. All other vigor scores collected for the Brown Swiss and Holstein calves did not differ (P > 0.05).

39 Effect of Breed on Dairy Calf Vigor from Birth to Preweaning

Abstract There is a stigma in the dairy industry that Brown Swiss calves are not as quick to stand and nurse following birth compared to other dairy breeds. The objective of this study is to determine if Brown Swiss calves are less vigorous at birth, and does this impact their health status and early calf growth compared with the dairy industry’s most prominent breed, the Holstein. All calves (n = 25) were scored shortly after birth using a modified Apgar scoring system used for infants. Thirteen parameters divided into 5 categories were evaluated to determine overall vigor of each calf, these included visual scores for meconium staining and swelling of the head; initial mobility scores for time to lift head, time to stand, and ear position; general responsiveness scores included strength of the suckling reflex, head shake in response to straw in nasal cavity, tongue withdrawal when pinched, and eye reflex; oxygenation score for mucus membrane color; rates scores included heart rate, respiration rate, and temperature. In addition to the vigor scoring a weight was taken at birth and 2 weeks post calving. Jugular blood samples were collected at birth, 24 hours later, and at 2 weeks of age and serum was stored for later analysis of haptoglobin and IgG concentration. Vigor scores and weight data were analyzed by breed using the mixed procedure in SAS. The vigor score for suckling reflex was lower (P = 0.04) for Brown Swiss calves (1.69, n = 12) than Holstein calves (2.39, n = 13). A weakened suckling reflex can lead to more challenges when consuming colostrum. Early colostrum intake is vital to obtaining passive immunity and maintaining health status in young calves. All other vigor scores collected for the Brown Swiss and Holstein calves did not differ (P > 0.05).

Serum haptoglobin response in red deer naturally infected with tuberculosis

The analysis of haptoglobin (Hp) serum concentration is a very sensitive, but non-specific, indicator of inflammation or infection. Methods to accurately diagnose infection in vivo in wildlife are usually constrained by low sensitivity due to the effects of stress on individual immune response and the challenging logistics of performing tests in the wild. Firstly, we sought to determine serum Hp concentration in red deer (Cervus elaphus) naturally infected with bovine tuberculosis (TB). Secondly, we assessed the complementary diagnostic value of serum Hp levels in conjunction with the cervical comparative skin test (CCT) performed in a subsample (n = 33). Serum Hp concentrations were significantly higher in TB-infected individuals (based on the presence of macroscopic lesions confirmed by culture) compared to those uninfected. In addition, serum Hp significantly changed with the type of animal handling, with captured and handled animals showing higher levels of Hp than hunted animals. Four out of 6 TB positive individuals that tested negative to the CCT (false negatives) showed Hp levels higher than the 95th percentile of healthy animals. These findings indicate that an acute phase response develops in animals with TB. In this paper, we demonstrate for the first time that an acute phase protein can provide a complementary assessment for specific diagnosis tests in wild species.

Analysis of haptoglobin N‐glycome alterations in inflammatory and malignant lung diseases by capillary electrophoresis

A CE-based method was introduced to compare the N-glycosylation profile of haptoglobin in normal and pathologic conditions. To assess the biomarker potential of glycosylation changes in various lung diseases, haptoglobin was isolated from plasma samples of healthy, pneumonia, chronic obstructive pulmonary disease, and lung cancer patients by means of two haptoglobin-specific monoclonal antibodies. Haptoglobin N-glycans were then enzymatically released, fluorescently labeled, and profiled by CE. Disease-associated changes of core and antennary fucosylation were identified by targeted exoglycosidase digestions and their levels were compared in the different patient groups. Terms such as core- and arm-fucosylation degree, as well as branching degree, were introduced for easier characterization of the changes and statistical analysis was used to examine which structures were responsible for the observed differences. Increased level of α1-6 fucosylated tri-antennary glycans was found in all disease groups compared to the control. Elevated amounts of core- and arm-fucosylation on tetra-antennary glycans were detected in the lung cancer group compared to the chronic obstructive pulmonary disease group. A larger scale study is necessary to confirm and validate these preliminary findings in the glycosylation changes of haptoglobin, so could then be used as biomarkers in the diagnosis of malignant and inflammatory lung diseases.

Proteomic Identification Network Analysis of Haptoglobin as a Key Regulator Associated with Liver Fibrosis

Liver fibrosis (LF) is the final stage of liver dysfunction, characterized by diffuse fibrosis which is the main response to the liver injury. Haptoglobin (HP) protein, produced as an acute phase reactant during LF, preventing liver damage, may be potential molecular targets for early LF diagnostics and therapeutic applications. However, protein networks associated with the HP are largely unknown. To address this issue, we used a pathological mouse model of LF that was induced by treatment with carbon tetrachloride for 8days. HP protein was separated and identified by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. HP protein was subjected to functional pathway analysis using STRING and Cytoscape software for better understanding of the protein-protein interaction (PPI) networks in biological context. Bioinformatics analyses revealed that HP expression associated with fibrosis was upregulated, and suggested that HP responsible for fibrosis may precede the onset and progression of LF. Using the web-based database, functional pathway analysis suggested the modulation of multiple vital physiological pathways, including antioxidation immunity, signal transduction, metabolic process, energy production, cell apoptosis, oxidation reduction, DNA repair process, cell communication, and regulation of cellular process. The generation of protein interaction networks clearly enhances the interpretation and understanding of the molecular mechanisms of HP. HP protein represents targets for further experimental investigation that will provide biological insight and potentially could be exploited for novel therapeutic approaches to combat LF.

新生児ハプトグロビンの定量と型判別の問題点

新生児ハプトグロビンの定量と型判別の問題点

Accumulation of Golgi-processed secretory proteins in an organelle of high density upon reduction of ATP concentration in rat hepatocytes.

We have previously shown that when rat hepatocytes are incubated with 4 mM azide, which reduces the intracellular ATP concentration to about 30% of its normal level, secretory proteins are reversibly arrested within the cell. Analysis of haptoglobin after 150 min of azide incubation shows that its carbohydrates have been processed by Golgi enzymes (Persson, R., Ahlström, E., and Fries, E. (1988) J. Cell Biol. 107, 2503-2510). Here, we have further characterized the site of arrest. Subcellular fractionation by density gradient centrifugation showed that albumin and haptoglobin fractionated like a marker for the endoplasmic reticulum. Localization of albumin by immunoelectron microscopy showed, however, that it occurred in flattened cisternae and that the endoplasmic reticulum was devoid of the protein. A possible explanation for these results is that the azide treatment blocks transport through the Golgi complex, leading to an accumulation of secretory proteins in a pre- or early Golgi compartment of high density. This compartment could contain sufficient amounts of Golgi enzymes to carry out the observed carbohydrate processing upon prolonged incubation or possibly acquire them as an effect of the azide treatment.

Haptoglobin Phenotypes in Diabetes mellitus and Diabetic Retinopathy

An analysis of haptoglobin (HP) phenotypes in 81 cases of diabetes mellitus (DM) without retinopathy and 122 cases with diabetic retinopathy (DR) were studied in relation to 180 normal and healthy controls matched for age and sex. A significant decrease in HP 2-1 frequency was found, suggesting protection for heterozygotes in both DM and DR (with a relative risk of about 0.31). As an acute-phase reactant HP may be functionally involved in the etiology of DM and DR, which are associated with immunologic and inflammatory processes, respectively. No significant differences were found with respect to sex, age at onset, duration of DR, types of DM and DR, and family history.

Acute-phase reactants in experimental inhalation lung disease.

An investigation on the occurrence and role of acute-phase reactants in experimental inhalation lung disease (ILD) was undertaken. Using an experimental model of ILD in which rabbits are exposed to aerosols of appropriate fungal spores, haptoglobin analysis was compared with depressions in arterial oxygen tension (PaO2) with time following challenge. Haptoglobin values of rabbits exposed to single 30-min aerosol challenges of Aspergillus terreus, demonstrated a significant (two- to three fold) increase 24-48 hr after challenge. Haptoglobin elevation was found to be a more reliable and consistent indicator of pulmonary inflammation than depression of PaO2. In an effort to determine the role of haptoglobin in this response, acute phase reactant levels were elevated either actively or passively. When rabbits with augmented haptoglobin levels were exposed to aerosol challenges with A. terreus, the typical depressions in PaO2 at 1-4 hr postchallenge were not observed. This indicated the plasma containing elevated acute-phase reactants may be involved in limiting the pulmonary response that normally occurs following challenge. Haptoglobin was not found to bind to A. terreus spores in vitro.

An Efficient Method to Eliminate Streaking in the Electrophoretic Analysis of Haptoglobin in Bloodstains

A bloodstain extraction procedure that improves the analysis of haptoglobin in dried bloodstains has been developed. The streaking of electrophoresis gels caused by deteriorated hemoglobin can be eliminated by incorporating chloroform in the bloodstain extraction procedure. The method is easier to execute than previously published techniques for eliminating the adverse effects of deteriorated hemoglobin on the analysis of haptoglobin. Bloodstains up to two years old were correctly phenotyped in haptoglobin by this method.