Levels Of Control Samples Research Articles

Key genes and biological pathways in pulmonary arterial hypertension related to endoplasmic reticulum stress identified by bioinformatics

Pulmonary arterial hypertension (PAH) is a cardiopulmonary vascular condition with an unclear pathogenesis. Targeting endoplasmic reticulum (ER) stress has been suggested as a novel treatment approach for PAH, but the mechanisms involving ER stress-related genes in PAH are not well understood. Microarray data for PAH and ER stress-related genes were analyzed. Differential and Venn analyses identified 17 differentially expressed ER stress-related genes in PAH. Candidate drugs targeting these genes were predicted using the CMap database. A protein-protein interaction (PPI) network was constructed, and hub genes (LCN2, IGF1, VCAM1, EDN1, HMOX1, TLR4) with complex interplays were identified using the STRING database and Cytoscape plugins. The clinical diagnostic performance of the hub genes was evaluated using ROC curves. The GeneMANIA website was utilized to predict enriched pathways associated with the hub genes and their functionally similar genes. MiRNAs and transcription factors (TFs) targeting the hub genes were predicted using the Networkanalyst website. The immune levels in control samples and PAH samples were assessed using various algorithms. Nine drug candidates were found to potentially target the identified ER stress-related genes. The hub genes and their correlated genes were significantly enriched in immune-related pathways. The PAH group showed increased immune cell infiltration, indicating a heightened immune response. This study sheds light on the role of ER stress-associated hub genes in PAH and proposes potential drugs targeting these genes. These findings provide valuable insights into PAH mechanisms and support the exploration of ER stress as a therapeutic target.

Comprehensive assessment of markers of apoptosis and cell proliferation during progression of atherosclerosis after surgery in patients with peripheral arterial disease.

Determination of predictors that can affect development of atherosclerosis progression in the postoperative period is an urgent problem in vascular surgery. Integrated assessment of markers of apoptosis and cell proliferation in atherosclerotic lesions and their progression after surgery in patients with peripheral arterial diseases. The investigation included 30 patients with stage IIB-III peripheral arterial disease. All patients have undergone open surgical interventions on the arteries of the aorto-iliac and femoral-popliteal segments. During these interventions, intraoperative specimens were obtained from the vascular wall with atherosclerotic lesions. The following values were evaluated: VEGF А165, PDGF BB, and sFas. Samples of normal vascular wall were obtained from post-mortem donors and used as a control group. The levels of Bax and p53 were increased (p<0.001) in samples from arterial wall with atherosclerotic plaque, while sFas values were reduced (p<0.001), compared to their levels in control samples. Values of PDGF BB and VEGF A165 were 1.9 and 1.7 times higher in atherosclerotic lesion samples (p=0.001), in comparison with the control group. The levels of p53 and Bax were increased against a background of reduced sFas levels in samples with progression of atherosclerosis compared to their baseline values in samples with atherosclerotic plaque (p<0.05). Initially increased values of the Bax marker against a background of reduced sFas values in vascular wall samples from patients with peripheral arterial disease is associated with risk of atherosclerosis progression in the postoperative period.

Inhibitory effect of mulberry leaf (Morus alba L.) extract on the formation of free and bound heterocyclic amines in pan-fried muscovy duck (Cairina moschata) patties

The effect of different levels (0.01%, 0.03%, 0.05% and 0.10%) of mulberry leaf (Morus alba L.) extract (MLE) on the formation of free and bound heterocyclic amines (HCAs) in pan-fried muscovy duck (Cairina moschata) patties were investigated for the first time. Precursors of HCAs, oxidant index of patties and phenolic composition of MLE were measured. The total free and bound HCAs levels in control samples were 28.37 ng/g and 1935.65 ng/g, and the inhibition rates were 6.71–68.92% and 6.71–14.05% in MLE treatment groups, respectively, and 0.03% showed the strongest inhibitory effect. OPLS-DA result indicated that MeIQ, 8-MeIQx and Norharman were crucial differential compounds in total HCAs. Furthermore, the addition of MLE significantly inhibited the lipid and protein oxidation of patties (P < 0.05). Fifteen phenolic compounds, mainly flavonoids, were identified by LC-MS. The mitigation of HCAs and oxidation may be attributed to them. Correlation analysis indicated that the total HCAs content was negatively correlated with precursors but positively correlated with oxidation. In conclusion, MLE helped mitigate HCAs formation during muscovy duck processing and appeared to be a viable natural additive for producing healthier processed meat products.

ASSESSMENT OF ACCURACY AND PRECISION STATISTICS IN ROUTINE BIOCHEMISTRY AUTOANALYZER USING INTERNAL QUALITY CONTROL SPECIMENS IN A TERTIARY CARE HOSPITAL LABORATORY

Background: Accuracy and precision are two important yardsticks of a reliable analytical system in the clinical laboratory. The study was designed to determine the accuracy and precision statistics of a routine biochemistry auto-analyzer using two levels of quality control materials and to compare these statistics with the company provided values. The study also aimed to compare the month-wise variations in these statistics. Methods: It was a cross-sectional study conducted in the Department of Biochemistry at Birat Medical College Teaching Hospital, Nepal. Laboratory data for the months of May–July, 2021 were retrieved from the laboratory information system (LIS). The retrieved data comprised of the results of two levels of quality control specimens run routinely on Beckman Coulter AU480 biochemistry autoanalyzer for most of the biochemical parameters. Accuracy and precision statistics were calculated as mean and coefficient of variation, respectively. Results: In both levels of control samples, the laboratory determined accuracy statistics were greater in magnitude than the company provided ones for albumin, alkaline phosphatase, aspartate transaminase, creatinine, unsaturated iron binding capacity, urea, direct bilirubin, and amylase; the precision statistics were similarly greater in magnitude for total protein and magnesium. In month-wise comparison of laboratory determined accuracy statistics, the overall mean differences were statistically significant (p&lt;0.05) for all parameters except lactate dehydrogenase and magnesium (both levels of control). Conclusions: The laboratory determined accuracy and precision statistics showed variations from the company provided ones apart from the month-wise variation. Therefore, continuous monitoring of these values is mandatory for ensuring reliable test reports.

RARE-32. Phase 0 and feasibility single-institution clinical trial of intravenous tocilizumab for adamantinomatous craniopharyngioma

BACKGROUND: Adamantinomatous craniopharyngioma (ACP) is a devastating skull-base tumor believed to derive from epithelial remnants of the primordial craniopharyngeal duct (Rathke’s pouch), which gives rise to the anterior pituitary gland. ACP lacks medical antitumor therapies. Current standard therapy with surgery and radiation is associated with poor quality of life. Clinical and preclinical data indicate that IL-6 blockade may contribute to ACP tumor control. METHODS: Children aged 2–21 years with newly diagnosed or previously treated ACP with measurable disease are eligible for the Phase 0/feasibility single-institution clinical trial (NCT03970226) of intravenous (IV) tocilizumab at Children’s Hospital Colorado. The phase I stratum involves IV tocilizumab prior to a standard-of-care surgical resection. The feasibility portion of the trial involves IV tocilizumab every two weeks for up to 13 28-day cycles. Tocilizumab is administered at the established weight-based pediatric dosage of 8 mg/kg for patients who weigh ≥30kg or 12 mg/kg for patients who weigh <30kg. RESULTS: To date, three patients have been enrolled on the Phase 0 component of the trial. These patients demonstrated clinically relevant levels of tocilizumab (≥ 4µg/mL) in serum, cyst fluid, and/or tumor tissue, compared to undetectable levels in control samples. Two patients (1 male and 1 female; median age 10.5 years) have enrolled on the feasibility stratum; one patient had best response of minor response but met definition of progressive disease at cycle 11. One patient with extensive disease required dose reduction for myelosuppression. CONCLUSION: Systemic delivery of tocilizumab at the established pediatric dosage is promising for treatment of ACP based on preclinical work and its demonstrated penetration into cystic and solid portions of ACP tumors. The therapy to date has been well tolerated overall. Further study is planned through a CONNECT consortium international Phase II trial.

Patient Heterogeneity in Acute Myeloid Leukemia: Leukemic Cell Communication by Release of Soluble Mediators and Its Effects on Mesenchymal Stem Cells.

Acute myeloid leukemia (AML) is an aggressive bone marrow malignancy, and non-leukemic stromal cells (including mesenchymal stem cells, MSCs) are involved in leukemogenesis and show AML-supporting effects. We investigated how constitutive extracellular mediator release by primary human AML cells alters proteomic profiles of normal bone marrow MSCs. An average of 6814 proteins (range 6493−6918 proteins) were quantified for 41 MSC cultures supplemented with AML-cell conditioned medium, whereas an average of 6715 proteins (range 6703−6722) were quantified for untreated control MSCs. The AML effect on global MSC proteomic profiles varied between patients. Hierarchical clustering analysis identified 10 patients (5/10 secondary AML) showing more extensive AML-effects on the MSC proteome, whereas the other 31 patients clustered together with the untreated control MSCs and showed less extensive AML-induced effects. These two patient subsets differed especially with regard to MSC levels of extracellular matrix and mitochondrial/metabolic regulatory proteins. Less than 10% of MSC proteins were significantly altered by the exposure to AML-conditioned media; 301 proteins could only be quantified after exposure to conditioned medium and 201 additional proteins were significantly altered compared with the levels in control samples (153 increased, 48 decreased). The AML-modulated MSC proteins formed several interacting networks mainly reflecting intracellular organellar structure/trafficking but also extracellular matrix/cytokine signaling, and a single small network reflecting altered DNA replication. Our results suggest that targeting of intracellular trafficking and/or intercellular communication is a possible therapeutic strategy in AML.

Effects of active batter coatings enriched by quince seed gum and carvacrol microcapsules on oil uptake and quality loss of nugget during frying.

The effects of active batter coatings containing quince seed gum (QSG) and carvacrol microcapsules (CM) on oil uptake, moisture loss, lipid oxidation, texture, color and organoleptic properties of chicken nuggets during the frying process were investigated. Active coatings on the surface of nuggets reduced oil uptake and moisture loss of fried samples decreased by 33.21% and 29.64%, respectively. Antioxidant activity tests showed that oxygen radical absorbance capacity (ORAC) and DPPH radicals scavenging activity of carvacrol microcapsules were 152.23 ± 4.11μmol TE/g, and 51.09 ± 3.32%, respectively. Investigation of primary and secondary oxidation products in the fried nugget samples showed that the peroxide value and thiobarbituric acid levels in control samples were 7.43meq peroxide/kg and 1.35mg MDA/kg, respectively. The results of this study showed that the highest reduction in PV and TBA were 41.85 and 37.04% for the QSG-coated samples containing 1% carvacrol microcapsules. The color of QSG-coated samples did not change significantly compared to control samples, although their hardness was reduced compared to the control samples (p < 0.05). The results showed that the use of active edible coatings made from quince seed gum and containing carvacrol microcapsules did not show any negative effects on the sensory properties of nuggets.

RARE-10. ADAMANTINOMATOUS CRANIOPHARYNGIOMA RESIDES OUTSIDE THE BLOOD BRAIN BARRIER

BACKGROUNDAdamantinomatous craniopharyngioma (ACP) is a devastating skull-base tumor believed to derive from epithelial remnants of the primordial craniopharyngeal duct (Rathke’s pouch), which gives rise to the anterior pituitary gland. Genetically engineered mouse models of ACP demonstrate that perturbation of the fetal anterior pituitary can generate tumors analogous to ACP. Clinical and preclinical data indicate that IL-6 blockade may contribute to ACP tumor control, with the most common agent being the humanized monoclonal antibody, tocilizumab. This agent demonstrated poor blood-brain barrier (BBB) penetration in primates. We present findings from two children enrolled on a phase 0 clinical trial (NCT03970226) of a single dose of preoperative intravenous tocilizumab prior to resection of newly diagnosed ACP.METHODSBlood samples were obtained at multiple timepoints. Serum was isolated via ficoll separation. Tumor tissue and cyst fluid were obtained 4–6 hours following the single IV dose of tocilizumab. Tissue was snap-frozen. Tumor was homogenized in RIPA buffer. Free tocilizumab in serum, cyst fluid, and tumor tissue was measured using enzyme-linked immunosorbent assay (ELISA) against a standard curve.RESULTSBoth patients in this trial demonstrated clinically relevant levels of tocilizumab (≥ 4µg/mL) in serum, cyst fluid, and tumor tissue, compared to undetectable levels in control samples.CONCLUSIONACP resides outside BBB protection. In addition to demonstrating the feasibility of systemic delivery of tocilizumab, these findings indicate that other large molecules, including those known to have poor BBB penetration, may be systemically delivered as part of an antitumor regimen in the treatment of ACP.

COMBINED EFFECT OF REACTIVE OXYGEN FORMS AND SHUNGITE ON THE GROWTH AND DEVELOPMENT OF CYANOBACTERIA

It has been shown that hydrogen peroxide, singlet oxygen and shungite affect the development of the population of cyanobacteria Anabaena variabilis and Synechocystis sp. PCC 6803 in different directions. The growth of cyanobacteria culture slows down in the presence of photosensitizer and hydrogen peroxide. In this case, the number of cells depends on the amount of shungite in the environment. Cyanobacteria Anabaena variabilis and Synechocystis sp. PCC 6803 grow better in the presence of 10 g/L shungite, but growth is suppressed at 100 g/L. Shungite at 10 g/L protectes cultures from the toxic effects of Bengal pink and hydrogen peroxide, which was determined by the level of photosynthesis efficiency and cell amounts. The growth of Anabaena variabilis (in terms of optical density at 680 nm) has been found to slow down in the presence of only hydrogen peroxide relative to the control. The growth of Anabaena variabilis in the presence of shungite and hydrogen peroxide is restored to the level of control samples. The number of cyanobacterium Synechocystis sp. PCC 6803 cells slows down significantly in the presence of singlet oxygen. Being combined with singlet oxygen, shungite partially inactivates the action of Bengal pink. At the end of the experiment, the growth of the culture is restored to 60% compared to the control. A similar relationship is observed when evaluating the effectiveness of photosynthesis of cyanobacteria Anabaena variabilis and Synechocystis sp. PCC 6803. Thus, singlet oxygen is more toxic than hydrogen peroxide. This may be due to different mechanisms of action of hydrogen peroxide and singlet oxygen on cyanobacteria growth. Depending on the content in the water, shungite can stimulate or inhibit the growth of cyanobacteria cells, inactivate the action of a toxicant.

Effect of polymeric biocide polyhexamethylene guanidine hydrochloride on morpho-physiological and biochemical parameters of wheat seedlings under copper stress

Aim. To study the effect of polymeric biocide polyhexamethylene guanidine hydrochloride (PHMG-Cl) on morphophysiological and biochemical parameters of wheat seedlings in the presence of toxic concentration of copper ions. Methods. The growth-stimulating activity of PHMG-Cl in pure state, as well as in the presence of elevated concentrations of copper ions was studied by phytotesting method using wheat seed as a test culture. Physico-chemical interaction between PHMG-Cl and CuSO4 in water solutions was studied by spectrophotometry. Results. Wheat seed treatment with water solutions of polymeric biocide PHMG-Cl in the concentration range of 0.01–0.1 % causes an increase in the length of roots and shoots by over 50 % after 7 days of incubation. The concentration of PHMG-Cl of 1 % was found to have a depressive effect on growth characteristics of the tested plants. This fact indicates that the phytotoxicity of PHMG-Cl is at least two orders of magnitude lower compared with a common pesticide copper sulfate. Thus, wheat seed soaking in 200 μM (0.003 %) water solution of copper sulfate causes reduced length of shoots and roots (by 65 and 34 %, respectively), as well as reduced weight of shoots (by 20 %) and roots (by 74 %) The analysis of green mass showed signifi cantly reduced content of carotenoids (by 26 %) and water soluble fl avonoids (by 37 %), as well as chlorophyll a/b ratio (by 18 %) that indicates decreased plant metabolic activity. However, the toxic effect of copper sulfate ions signifi cantly diminished in the presence of PHMG-Cl (0.01 %). Thus, the length of shoots and roots was found to increase by 24 and 64 %, respectively, compared to the samples treated with CuSO4. Moreover, the content of pigments and fl avonoids in the green mass of seedlings reached the level of control samples. The results of spectrophotometric studies indicate the absence of physico-chemical interaction between PHMG-Cl and copper sulfate in water solutions. Conclusions. The polymeric biocide PHMG-Cl effectively stimulates growth and development of wheat seedlings in the concentration range of its water solutions of 0.01–0.1 %. In the presence of toxic copper sulfate concentration, PHMG-Cl does not possess the growth stimulating activity. However, the polymeric biocide signifi cantly improves copper stress resistance of plants, and normalizes their morpho-physiological indicators, photosynthetic characteristics and secondary exchange.

Hippocampal LMNA Gene Expression is Increased in Late-Stage Alzheimer's Disease.

Lamins are fibrillary proteins that are crucial in maintaining nuclear shape and function. Recently, B-type lamin dysfunction has been linked to tauopathies. However, the role of A-type lamin in neurodegeneration is still obscure. Here, we examined A-type and B-type lamin expression levels by RT-qPCR in Alzheimer’s disease (AD) patients and controls in the hippocampus, the core of tau pathology in the brain. LMNA, LMNB1, and LMNB2 genes showed moderate mRNA levels in the human hippocampus with highest expression for the LMNA gene. Moreover, LMNA mRNA levels were increased at the late stage of AD (1.8-fold increase; p-value < 0.05). In addition, a moderate positive correlation was found between age and LMNA mRNA levels (Pearson’s r = 0.581, p-value = 0.018) within the control hippocampal samples that was not present in the hippocampal samples affected by AD. A-type and B-type lamin genes are expressed in the human hippocampus at the transcript level. LMNA mRNA levels are up-regulated in the hippocampal tissue in late stages of AD. The effect of age on increasing LMNA expression levels in control samples seems to be disrupted by the development of AD pathology.

The expression profile of integrin receptors and osteopontin in thyroid malignancies varies depending on the tumor progression rate and presence of BRAF V600E mutation

Thyroid cancer (TC) is one of the most common malignancy of the human endocrine system. BRAF V600E mutation is the most frequent genetic alteration of papillary carcinoma, the most frequent TC, which effects RAS-RAF-MEK intracellular signaling pathway. These alterations in RAS-RAF-MEK pathway lead to changes in expression levels of cell membrane integrin receptors and their ligand - extracellular matrix protein osteopontin, which in turn increases the metastatic potential of tumor cells. Thus, integrins and their ligand osteopontin can be considered as potential biomarkers of tumor progression and aggressive tumor phenotypes.The aim of the study was to evaluate the expression levels of integrin receptors ITGA2, ITGA3, ITGAV, ITGA6, ITGA9, ITGB1, ITGB3 and their ligands OPNa, OPNb in the thyroid cancer with different BRAF V600E mutation status. MethodsThyroid tumor samples of 70 patients obtained during surgical treatment were analyzed. Expression levels of the investigated genes were evaluated by real time RT-PCR. Fluorescent immunohistochemistry (IHC) was used to confirm the PCR results and to estimate the amount of protein levels. For IHC frozen sections were used. BRAF V600E mutation was determined using allele-specific amplification. Nonparametric criteria (Kruskal Wallis, Wilcoxon and Mann-Whitney tests) were used to evaluate group differences. P values of less than 0.05 were considered as statistically significant. ResultsA higher gene expression level of ITGA2 (1.9-fold, p = 0.037), ITGA3 (21.1-fold, p = 0.041) and ITGA5 (2.08-fold, p = 0.048) was observed in papillary thyroid cancer (PTC) tissue in comparison with median expression level in control samples (conventionally normal tissue of thyroid gland). These changes were confirmed by IHC (significant changes for α2 integrin). ITGAV expression level was statistically significantly higher in follicular thyroid cancer (FTC) (2.0-fold, p = 0.040). Next, high gene expression levels in tissue samples of lymph node metastases were observed for ITGA5 (2.92-fold, p = 0.015), OPNb (4.36-fold, p = 0.037). For genes ITGA3 (37.48-fold, p = 0.017790), ITGA6 (18.76-fold, p = 0.028921) and ITGA9 (12.52-fold, p = 0.026710) higher expression level was detected in T3-4 tumors (TNM) compared to tumors classified as T1-2. Presence of BRAF V600E mutation was identified in 20 samples of PTC of 40 (50%). A significant increase of the expression level only of ITGA3 (3.1-fold, p = 0. 0422) was observed in BRAF V600E positive samples. Further, changes in expression levels of integrins and osteopontin were assessed in benign and malignant neoplasms. In PTC samples higher expression of ITGA2 (2.8-fold, p = 0.005), ITGA6 (2.11, p = 0.03) and ITGB1 (2.32-fold, p = 0.02) was detected. In FTC expression level of ITGA6 (2.67, p = 0.007) was higher than in benign thyroid nodules. ConclusionIdentified changes in expression levels of the studied genes indicate that they could play an important role in tumor progression, and their expression could be affected by the product of mutant BRAF gene. Integrins and their ligand osteopontin might be considered as potential markers in determining prognosis and treatment of TC.

Control of Histamine Level in Oreochromis Niloticus Fillets Using Essential Oils

It was studied that effect of different essential oils on the histamine level in examined Oreochromis niloticus fillet samples. Accordingly, the addition of essentials oils (cinnamon and Oregano) was applied by (1%) against control group. The obtained results revealed that histamine levels in control samples increased by 24.8%; 70.1% and 94.5% with mean values of 18.84; 25.67, and 29.35 after chilling for 3; 6 and 9 days, respectively; meanwhile, the histamine mean concentration at zero time was 15.09 mg/kg. Concerning addition of cinnamon oil 1%, levels of histamine decreased to 18.7%; 46.6% and 77.0 % with mean values of 17.92; 22.13, and 26.71 after 3; 6 and 9 days, respectively. In contrast the addition of oregano oil 1%, to the examined O. niloticus fillet samples lead to significant decrease in histamine levels by 8.2%, 23.9% and 32.3% with mean values 16.33, 18.7 and 19.96 after 3, 6 and 9 days in 4C storage temperature, respectively. Finally, it can have concluded that the addition of oregano oil was superior for control of undesirable histamine in Oreochromis niloticus fillets.

Paper-based enzymatic platform coupled to screen printed graphene-modified electrode for the fast neonatal screening of phenylketonuria

IntroductionThe PKU is an inborn error of amino acid metabolism, in which phenylalanine (Phe) accumulated in the blood causing alterations at the central nervous system. We report a novel paper-based enzymatic platform coupled to screen printed graphene-modified electrode for the neonatal screening of phenylketonuria (PKU0. MethodsThe paper-based analytical device coupled to electrochemical detection (EPAD) is based on the use of paper microzones modified with phenylalanine dehydrogenase enzyme (PheDH). The modified PADs were placed on the surface of an electrode modified with electrochemically reduced graphene (ERGO). PheDH in the presence of NAD+ catalyzes the reversible deamination of Phe to form phenylpyruvate, ammonia, and NADH. The electrochemical oxidation of NADH was monitored by differential pulse amperometry (DPA) at 0.6 V. The method was linear in the concentration range from 1 to 600 μmol/L of Phe with a LOQ of 1 μmol/L and LOD of 0.2 μmol/L. Within day precision was 5.7% across 3 levels of control samples. Between-day precision was 8.3%. The comparison with the standard Phe enzyme assay kit showed good agreement. The time required for the overall assay was <5 min. The non-sophisticated equipment required, the short assay time and the appropriate LOQ and LOD achieved by our EPAD make it an attractive and easy to use alternative compared to existing methods applied to the screening of PKU in neonatal samples.

HMG-CoA synthase isoenzymes 1 and 2 localize to satellite glial cells in dorsal root ganglia and are differentially regulated by peripheral nerve injury

In dorsal root ganglia (DRG), satellite glial cells (SGCs) tightly ensheathe the somata of primary sensory neurons to form functional sensory units. SGCs are identified by their flattened and irregular morphology and expression of a variety of specific marker proteins. In this report, we present evidence that the 3-hydroxy-3-methylglutaryl coenzyme A synthase isoenzymes 1 and 2 (HMGCS1 and HMGCS2) are abundantly expressed in SGCs. Immunolabeling with the validated antibodies revealed that both HMGCS1 and HMGCS2 are highly colabeled with a selection of SGC markers, including GS, GFAP, Kir4.1, GLAST1, GDNF, and S100 but not with microglial cell marker Iba1, myelin sheath marker MBP, and neuronal marker β3-tubulin or phosphorylated CaMKII. HMGCS1 but not HMGCS2 immunoreactivity in SGCs is reduced in the fifth lumbar (L5) DRGs that contain axotomized neurons following L5 spinal nerve ligation (SNL) in rats. Western blot showed that HMGCS1 protein level in axotomized L5 DRGs is reduced after SNL to 66±8% at 3 days (p<0.01, n=4 animals in each group) and 58±13% at 28 days (p<0.001, n=9 animals in each group) of its level in control samples, whereas HMGCS2 protein was comparable between injured and control DRGs. These results identify HMGCSs as the alternative markers for SGCs in DRGs. Downregulated HMGCS1 expression in DRGs after spinal nerve injury may reflect a potential role of abnormal sterol metabolism of SGCs in the nerve injured-induced neuropathic pain.

High pressure processing extend the shelf life of fresh salmon, cod and mackerel

High pressure processing (HPP) is used on several types of seafood, though its potential for extending the shelf life of fresh fish has not been fully exploited. This study compared the effect of HPP (200 and 500 MPa, 120 s) on salmon, cod and mackerel. Immediately after processing, and during storage up to 26 days different analysis were carried out to evaluate the microbiological shelf life, oxidation, acid phosphatase level and pH of the different fish species during the storage period. For cod and mackerel, HPP at 500 MPa restrained the bacterial flora and it did not reach spoilage level, opposite to the salmon samples exposed to 500 MPa. Analysis of TBARS is often used as a measure of lipid oxidation. The TBARS-level was differently affected by pressure in salmon and cod. HPP at 200 MPa did not have any effect on the oxidation level in salmon during the storage period while this was observed for cod. The TBARS level in mackerel was high, independent of pressure treatment or not. Phosphatase activity in fish has been suggested to be related to freshness. The acid phosphatase (ACP) levels in control samples showed significant differences between salmon and cod. For salmon both control and samples treated at 200 MPa showed a decrease in ACP level at day 11 compared with day 0, whereas the HPP500 values were even throughout the study. This latter observation was also the case for cod treated at 500 MPa. This study have shown that HPP induce different levels of changes in various fish species.

Proteomic analysis of Daphnia magna exposed to glyphosate and methidathion

Proteomic analysis was performed to identify proteins involved in the stress responses of Daphnia magna to glyphosate and methidathion. Lethal concentration of chemicals causing 50% mortality (LC50) determined by acute toxicity assays were used to expose to daphnias from 24 h to 21 days prior to total protein extraction. The proteomic profile of tested organisms was analyzed by the two-dimensional electrophoresis method. The average intensity of each spots was then analyzed to determine the differently expressed proteins (DEPs). The DEPs was considered as down- or up-regulations only if their expression level in control samples are linearly higher than LC50 or lower than LC50, respectively. The result showed that there are 22 and 27 DEPs responding to the exposure of glyphosate and methidathion, respectively. The expression pattern of established DEPs is helpful to understand the molecular response of D. magna to glyphosate and methidathion, and discover potential candidates as novel biomarkers.

Improvement of fibrin clot structure after factor VIII injection in haemophilia A patients treated on demand

Patients with haemophilia A have seriously impaired thrombin generation due to an inherited deficiency of factor (F)VIII, making them form unstable fibrin clots that are unable to maintain haemostasis. Data on fibrin structure in haemophilia patients remain limited. Fibrin permeability, assessed by a flow measurement technique, was investigated in plasma from 20 patients with severe haemophilia A treated on demand, before and 30 minutes after FVIII injection. The results were correlated with concentrations of fibrinogen, FVIII and thrombin-activatable fibrinolysis inhibitor (TAFI), and global haemostatic markers: endogenous thrombin potential (ETP) and overall haemostatic potential (OHP). Fibrin structure was visualised using scanning electron microscopy (SEM). The permeability coefficient Ks decreased significantly after FVIII treatment. Ks correlated significantly with FVIII levels and dosage, and with ETP, OHP and levels of TAFI. SEM images revealed irregular, porous fibrin clots composed of thick and short fibers before FVIII treatment. The clots had recovered after FVIII replacement almost to levels in control samples, revealing compact fibrin with smaller intrinsic pores. To the best of our knowledge, this is the first description of fibrin porosity and structure before and after FVIII treatment of selected haemophilia patients. It seems that thrombin generation is the main determinant of fibrin structure in haemophilic plasma.

IL-17 and TNF Synergistically Modulate Cytokine Expression while Suppressing Melanogenesis: Potential Relevance to Psoriasis

Inflammation-associated pigmentation changes are extremely common, but the etiology behind this clinical observation remains elusive. Particularly, it is unclear how the myriad of cytokines known to be involved in inflammatory skin processes affect epidermal melanocytes. We sought to determine how IL-17 and TNF influence normal human melanocytes, as these two cytokines have been implicated in various skin diseases. IL-17 and TNF jointly stimulated broad inductions of cytokines , including melanoma mitogens CXCL1 and IL-8. Moreover, IL-17 and TNF synergistically inhibited pigmentation-related signaling and melanin production, and induced keratinocytes production of β-defensin 3, an antagonist for melanocortin-receptor 1. When analyzing psoriasis lesions that are known to over express IL-17 and TNF, we observed an increase in melanocyte number and a simultaneous decrease in pigmentation signaling. Furthermore, therapeutic neutralization of TNF and IL-17 with mAbs results in a rapid recovery of pigment gene expression in psoriasis lesions. These results demonstrate that IL-17 and TNF can impact both the growth and pigment production of melanocytes, which may contribute to the pigmentation changes associated with psoriasis. These findings may allow the development of novel therapeutics for pigmentary disorders and bring new insights into the immune milieu surrounding melanocytes and related neoplasms.

Identification of potential serum biomarkers for gastric cancer by a novel computational method, multiple normal tissues corrected differential analysis

Background Genes specifically expressed in one or a few tissues and upregulated in tumors are potentially good serum biomarkers. Methods By applying a recently developed computational method, called multiple normal tissues corrected differential analysis (MNTDA), we identified genes that are likely to be upregulated in the blood of gastric cancer patients as compared to normal controls. Results We identified four genes (MMP-1, MMP-3, MMP-12, and CXCL5) as potential serum biomarkers for gastric cancer. Of these four genes, only MMP-1 was significantly upregulated in the sera of 40 gastric cancer patients, as compared to 40 control sera. The same pattern was observed in the second cohort of 80 gastric cancer patients and 80 controls. In a combined analysis, the level of serum MMP-1 in gastric cancer patients was significantly higher than the level in control samples ( P < 0.0001). The use of MMP-1 was 62.5% sensitive and 62.5% specific in detecting gastric cancer patients. Patients with high serum levels of MMP-1 had a significantly worse outcome than patients with low serum MMP-1 levels. Finally, we determined that preoperative serum MMP-1 levels were prognostic, independent of tumor stage. Conclusions MMP-1 is a potential prognostic marker for gastric cancer patients after gastrectomy.