Blood Plasma Markers Research Articles

Blood Plasma Markers in Depressed Mice under Chronic Social Defeat Stress.

It has previously been shown that, in mice, chronic social defeat stress in daily agonistic interactions leads to a depression-like state similar to that in depressive patients. With this model, it has become obvious that it is possible to study peripheral markers of the depression-like state in an experiment. This paper was aimed at searching for protein markers in the blood plasma of depressed mice in the chronic social conflict model, which allows for us to obtain male mice with repeated experiences of defeat. Proteomic analysis of blood plasma samples was conducted to identify proteins differentially expressed in this state. There were changes in the expression levels of the amyloid proteins SAA1, SAA4, and SAMP and apolipoproteins APOC3, APOD, and ADIPO in the blood plasma of depressed mice compared with controls (unstressed mice). Changes in the expression of serine protease inhibitors and/or proteins associated with lipid metabolism, inflammation, or immune function [ITIH4, SPA3, A1AT5, HTP (HP), CO9, and A2MG] were also found. Here, we showed that chronic social stress is accompanied by increased levels of amyloid proteins and apolipoproteins in blood plasma. A similarity was noted between the marker protein expression changes in the depressed mice and those in patients with Alzheimer's disease. These data indicate a psychopathogenic role of chronic social stress, which can form a predisposition to neurodegenerative and/or psychoemotional disorders.

Thiazolidine-2,4-dione hybrids as dual alpha-amylase and alpha-glucosidase inhibitors: design, synthesis, in vitro and in vivo anti-diabetic evaluation.

Twelve 3,5-disubstituted-thiazolidine-2,4-dione (TZD) hybrids were synthesized using solution phase chemistry. Continuing our previous work, nine O-modified ethyl vanillin (8a-i) derivatives were synthesized and reacted with the TZD core via Knoevenagel condensation under primary reaction conditions to obtain final derivatives 9a-i. Additionally, three isatin-TZD hybrids (11a-c) were synthesized. The intermediates and final derivatives were characterized using 1H and 13C NMR spectroscopy, and the observed chemical shifts agreed with the proposed structures. The in vitro alpha-amylase and alpha-glucosidase inhibitory evaluation of newly synthesized derivatives revealed compounds 9F and 9G as the best dual inhibitors, with IC50 values of 9.8 ± 0.047 μM for alpha-glucosidase (9F) and 5.15 ± 0.0017 μM for alpha-glucosidase (9G), 17.10 ± 0.015 μM for alpha-amylase (9F), and 9.2 ± 0.092 μM for alpha-amylase (9G). The docking analysis of synthesized compounds indicated that compounds have a higher binding affinity for alpha-glucosidase as compared to alpha-amylase, as seen from docking scores ranging from -1.202 to -5.467 (for alpha-amylase) and -4.373 to -7.300 (for alpha-glucosidase). Further, the molecules possess a high LD50 value, typically ranging from 1000 to 1600 mg kg-1 of body weight, and exhibit non-toxic properties. The in vitro cytotoxicity assay results on PANC-1 and INS-1 cells demonstrated that the compounds were devoid of significant toxicity against the tested cells. Compounds 9F and 9G showed high oral absorption, i.e., oral absorption >96%, and their molecular dynamics simulation yielded results closely aligned with the observed docking outcomes. Finally, compounds 9F and 9G were evaluated for in vivo antidiabetic assessment by the induction of diabetes in Wistar rats using streptozotocin. Molecule 9G has been identified as the most effective anti-diabetic molecule due to its ability to modulate several biochemical markers in blood plasma and tissue homogenates. The results were further confirmed by histology investigations conducted on isolated pancreas, liver, and kidney.

Protein profiling in plasma for biomarkers of seizure

PurposeA biochemical way to measure seizures would greatly benefit epilepsy research and clinical follow-up. Short-term biomarkers like lactate exist, and interest in biomarkers representative of longer-term seizure burden is growing. In this exploratory study, we aimed to identify markers in blood plasma that differentiate persons with recent seizures from persons with epilepsy and long-standing seizure freedom. MethodsA proteomic analysis was performed on plasma samples of 120 persons with seizures using the Olink Neuro-exploratory panel. Participants were selected from a regional biobank study in Västra Götaland (Sweden) and categorized into two groups: recent seizure and seizure-free. The panel contained 92 proteins linked to neurological diseases and processes, and levels of these proteins were compared between the patient groups to identify potential markers of seizure activity. ResultsWe identified significant differences in protein levels between the recent seizure and seizure-free patient groups for Cadherin-15 [(CDH15; p = 0.008)], Latent transforming growth factor beta-binding protein 3 [(LTBP3; p = 0.002)], Phosphoethanolamine/phosphocholine phosphatase 1 [(PHOSPHO1; p = 0.011)], and Progestagen associated endometrial protein [(PAEP; p = 0.0005)]. ConclusionThe findings in this study present CDH15, LTBP3, PHOSPHO1 and PAEP as candidate markers of seizure activity. Further confirmatory studies are needed.

Abstract B049: Breast cancer-derived angiopoietin-like 7 regulates necrotic core formation and metastasis from the tumor interior

Abstract Necrosis in the tumor interior is a common feature of aggressive cancers that is associated with poor clinical prognosis and the development of metastasis. However, how the necrotic core promotes tumor cell dissemination and metastasis remains unclear. Foundational models for cancer metastasis research include mouse, zebrafish, and chick embryo, but identifying tumor cells in transit is painstaking. Here we used a rat model of breast cancer metastasis to increase detection of dissemination events. Owing to the large size and higher blood volume of rats, we collected 10 times more circulating tumor cells (CTCs) than from mice. In the rat model, tumor dissemination was temporally correlated with the emergence of necrosis in the tumor interior. These findings were corroborated by longitudinal study of CTC abundance and tissue necrosis markers in blood plasma from patients with metastatic breast cancer. Further, we observed that dilated blood vessels were located next to necrotic regions of the tumor and that their increase was concurrent with the initiation of intratumoral necrosis and increase in CTC abundance. Bulk RNA sequencing of mouse-to-rat xenograft tumor necrotic core compared to the non-necrotic rim revealed distinct tumor-specific and host-specific transcripts in the necrotic interior. We identified angiopoietin-like 7 (Angptl7) as a tumor-specific factor localized to the peri-necrotic zone. Functional studies showed that Angptl7 loss normalizes central necrosis, peri-necrotic dilated vessels, metastasis, and reduces circulating tumor cell counts to nearly zero. Mechanistically, Angptl7 promoted vascular permeability and supported vascular remodeling in the peri-necrotic zone. Taken together, these findings show that breast tumors actively produce factors controlling central necrosis formation and metastatic dissemination from the tumor core. Tumor dissemination events are localized spatially to dilated peri-necrotic vessels in the tumor interior, and tumor dissemination is dependent functionally on the expression of a factor, Angptl7, produced by peri-necrotic tumor cells. Our findings in breast cancer models, in conjunction with recent clinical observations, provide strong evidence for tumor dissemination from the tumor interior. Citation Format: Ami Yamamoto, Yin Huang, Brad A. Krajina, Margaux McBirney, Andrea E. Doak, Carolyn L. Wang, Michael C. Haffner, Kevin J. Cheung. Breast cancer-derived angiopoietin-like 7 regulates necrotic core formation and metastasis from the tumor interior [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr B049.

Fluid biomarkers and risk of neurodegenerative disease in retired athletes with multiple concussions: results from the International Concussion and Head Injury Research Foundation Brain health in Retired athletes Study of Ageing and Impact-Related Neurodegenerative Disease (ICHIRF-BRAIN study)

ObjectivesTo investigate the association and utility of blood plasma markers of neurodegeneration in a population of retired athletes self-reporting multiple concussions throughout a sporting career. It is hypothesised that this...

Human umbilical cord mesenchymal stem cell-derived treatment of severe pulmonary arterial hypertension

Here we report application of human umbilical cord mesenchymal stem cell (HUCMSC)-derived therapy for pulmonary arterial hypertension (PAH). A 3-year-old female presented with heritable PAH associated with hereditary hemorrhagic telangiectasia and was treated for 6 months with serial intravascular infusions of conditioned media (CM) from allogenic HUCMSCs. The treatment markedly improved clinical and hemodynamic parameters and decreased blood plasma markers of vascular fibrosis, injury and inflammation. A comparative analysis of single-cell RNA sequencing data collected from three HUCMSCs and two human umbilical vein endothelial cell (HUVEC) controls identified eight common cell clusters, all of which indicated regenerative potential specific for HUCMSCs. The properties of HUCMSCs were validated by untargeted label-free quantitation of the cell and CM proteome, suggesting increased activity of regeneration, autophagy and anti-inflammation pathways and mitochondrial function. Prostaglandin analysis demonstrated increased HUCMSC secretion of prostaglandin E2, known for its regenerative capacity. Additional prospective clinical studies are warranted to confirm and further explore the benefits of HUCMSC-derived therapy for PAH.

Single Extracellular Vesicle Analysis Using Flow Cytometry for Neurological Disorder Biomarkers

Extracellular vesicles (EVs) are membrane vesicles released from cells to the extracellular space, involved in cell-to-cell communication by the horizontal transfer of biomolecules such as proteins and RNA. Because EVs can cross the blood-brain barrier (BBB), circulating through the bloodstream and reflecting the cell of origin in terms of disease prognosis and severity, the contents of plasma EVs provide non-invasive biomarkers for neurological disorders. However, neuronal EV markers in blood plasma remain unclear. EVs are very heterogeneous in size and contents, thus bulk analyses of heterogeneous plasma EVs using Western blot and ELISA have limited utility. In this study, using flow cytometry to analyze individual neuronal EVs, we show that our plasma EVs isolated by size exclusion chromatography are mainly CD63-positive exosomes of endosomal origin. As a neuronal EV marker, neural cell adhesion molecule (NCAM) is highly enriched in EVs released from induced pluripotent stem cells (iPSCs)-derived cortical neurons and brain organoids. We identified the subpopulations of plasma EVs that contain NCAM using flow cytometry-based individual EV analysis. Our results suggest that plasma NCAM-positive neuronal EVs can be used to discover biomarkers for neurological disorders.

Выявление омик-маркеров негативных эффектов, ассоциированных с аэрогенным комбинированным воздействием соединений алюминия и фтора

At present, it is relevant to study simultaneous combined impacts exerted by chemicals on developing adverse health effects. It is also becoming vital to search for molecular indicators of adverse effects with the altered expression level. This alteration makes it possible to determine peculiarities of molecular and cellular pathogenesis mechanisms regarding a number of non-communicable diseases under exposure to a mixture of chemicals. Our research goal was to comparatively analyze and identify identical omic-markers of adverse effects under experimental and actual combined aerogenic exposure to aluminum and fluoride compounds. We substantiated molecular markers of prenosological changes by sequential implementation of an algorithm which included identifying altered proteins and peptides in blood plasma which were identical both under experimental and actual exposure; detecting and quantifying cause-effect relations between identical proteins and peptides and concentrations of aluminum and fluoride ion in urine. The research results indicate that long-term combined aerogenic exposure to aluminum and fluoride compounds in low average daily doses (0.0005 mg/(kgday) and 0.002 mg/(kg*day) accordingly) causes elevated concentrations of aluminum (by 2.8 times higher) and fluoride-ion (by 1.8 times higher) in exposed children’s urine. This fact is verified by experimental research with its focus on combined exposure to the examined chemicals. We were able to substantiate identical omic-markers, J-chain of immunoglobulin and Kelch-like protein 4 (KLHL4 gene), under simultaneous exposure to aluminum and fluoride compounds both under experimental and actual combined aerogenic exposure. We proved a cause-effect relation between levels of identical proteins and concentrations of aluminum and fluoride ion in urine under simultaneous exposure to the mixture of the examined chemicals. Identified protein markers in blood plasma give an opportunity to predict future adverse effects including developing immunoglobulins A and M deficiency with subsequent humoral immunity failure when J-chain of immunoglobulin is expressed; occurring sclerotic and inflammatory changes in vascular walls when Kelch-like protein 4 is expressed. These predicted adverse effects can be estimated as resulting from simple summated (additive) toxic impacts exerted by aluminum and fluoride under simultaneous combined aerogenic exposure to both chemicals.

On detecting omic-markers of negative effects associated with combined aerogenic exposure to aluminum and fluoride compounds

At present, it is relevant to study simultaneous combined impacts exerted by chemicals on developing adverse health effects. It is also becoming vital to search for molecular indicators of adverse effects with the altered expression level. This alteration makes it possible to determine peculiarities of molecular and cellular pathogenesis mechanisms regarding a number of non-communicable diseases under exposure to a mixture of chemicals. Our research goal was to comparatively analyze and identify identical omic-markers of adverse effects under experimental and actual combined aerogenic exposure to aluminum and fluoride compounds. We substantiated molecular markers of prenosological changes by sequential implementation of an algorithm which included identifying altered proteins and peptides in blood plasma which were identical both under experimental and actual exposure; detecting and quantifying cause-effect relations between identical proteins and peptides and concentrations of aluminum and fluoride ion in urine. The research results indicate that long-term combined aerogenic exposure to aluminum and fluoride compounds in low average daily doses (0.0005 mg/(kgday) and 0.002 mg/(kg*day) accordingly) causes elevated concentrations of aluminum (by 2.8 times higher) and fluoride-ion (by 1.8 times higher) in exposed children’s urine. This fact is verified by experimental research with its focus on combined exposure to the examined chemicals. We were able to substantiate identical omic-markers, J-chain of immunoglobulin and Kelch-like protein 4 (KLHL4 gene), under simultaneous exposure to aluminum and fluoride compounds both under experimental and actual combined aerogenic exposure. We proved a cause-effect relation between levels of identical proteins and concentrations of aluminum and fluoride ion in urine under simultaneous exposure to the mixture of the examined chemicals. Identified protein markers in blood plasma give an opportunity to predict future adverse effects including developing immunoglobulins A and M deficiency with subsequent humoral immunity failure when J-chain of immunoglobulin is expressed; occurring sclerotic and inflammatory changes in vascular walls when Kelch-like protein 4 is expressed. These predicted adverse effects can be estimated as resulting from simple summated (additive) toxic impacts exerted by aluminum and fluoride under simultaneous combined aerogenic exposure to both chemicals.

Diagnostic levels of sepsis biomarkers in children with oncological diseases.

The data on the diagnostic levels of sepsis markers in blood plasma in 117 patients with oncological diseases at the age from 1 to 18 years are presented. The patients were divided into 4 groups depending on the outcome of the clinical course of the inflammatory process or infectious complications: group 1 - patients with no complications of anticancer treatment (n = 13/11.1%), group 2 - the presence of a systemic inflammatory response in patients (n = 64/54.7%), group 3 - patients with sepsis (n = 27/23.1%), group 4 - patients with septic shock (n = 13/11.1%). The threshold level of presepsin between groups 1 and 2 was 202 pg/ml, 371 pg/ml between groups 1 and 3, 604 pg/ml between groups 2 and 3 and 1500 pg/ml between groups 3 and 4. For procalcitonin, the threshold level between groups 1and 2 was 0.23 ng/ml, 0.48 ng/ml between groups 1 and 3, 0.51 ng/ml between groups 2 and 3 and 3.9 ng/ml between groups 3 and 4. The threshold value of C-reactive protein in patients with solid tumors was 12.6 g/l between groups 1 and 2. In patients with oncohematological diseases, the threshold level of C-reactive protein was 43.4 g / L between groups 2 and 3, 77.1 g / L between groups 2 and 4. According to the ROC analysis, presepsin was superior to procalcitonin and C-reactive protein in the diagnosis of septic complications.

Застосування есенціальних фосфоліпідів для лікування жирової хвороби печінки

Передумови. З урахуванням схожості гістопатологічного патерну неалкогольної жирової хвороби печінки та алкогольної хвороби печінки було запропоновано обидва захворювання об’єднати в одну нозологічну форму під загальною назвою «жирова хвороба печінки». Методологія. При виконанні даного огляду був проведений повторний аналіз 25 досліджень, включених в огляд K.J. Gundermann зі співавт. (2016). У цих роботах вивчали ефективність і безпечність есенціальних фосфоліпідів, що були отримані з соєвих бобів і містили не менше 72 % фосфатидилхоліну. Результати. При аналізі 20 досліджень із неалкогольної жирової хвороби печінки було з’ясовано, що есенціальні фосфоліпіди зменшували вираженість симптомів, у тому числі загального нездужання, стомлюваності, нудоти, частоту болю в правому підребер’ї, здуття живота, закрепів, індекс ожиріння та товщину шкірної складки. Було відзначено позитивний вплив есенціальних фосфоліпідів на лабораторні показники плазми крові (функціональні тести печінки — аланінамінотрансфераза, аспартатамінотрансфераза, гамма-глутамілтранспептидаза, лужна фосфатаза; показники ліпідного обміну — загальний холестерин, тригліцериди, холестерин ліпопротеїнів низької щільності). Есенціальні фосфоліпіди покращували дані ультразвукового дослідження та комп’ютерної томографії печінки в 60–75 % пацієнтів. Одночасно спостерігалася ревертація гістопатологічних змін у печінці: зменшувалася вираженість стеатозу, балонування, інтраацинарного та портального запалення, що призводило до зниження стадії стеатогепатиту за класифікацією Brunt. Загальна ефективність лікування есенціальними фосфоліпідами становила 80,5 %, тоді як у групах плацебо або відсутності призначення препарату — 27,9 % (p < 0,05). При вивченні 5 досліджень з алкогольної хвороби печінки були зафіксовані подібні результати, однак для їх верифікації необхідно виконати більшу кількість досліджень. Дані всіх названих вище досліджень свідчать про те, що лікування есенціальними фосфоліпідами було безпечним. Висновки. У 25 клінічних дослідженнях були підтверджені ефективність і безпека есенціальних фосфоліпідів при лікуванні жирової хвороби печінки.

A WiFi scanner in conjunction with disposable multiplex paper assay for the quantitation of disease markers in blood plasma.

Herein we report a quantitative, multiplex assay for disease markers in plasma based on an integrated setup of a portable scanner and a disposable paper-based analytical device (PAD). The quantitative analysis relies on the digital colorimetric reading of the three-layer PAD with 30 assay sites for performing respective chromogenic reactions for plasma uric acid, glucose, and triglyceride, which are considered as important risk factors for cardiovascular diseases. A portable scanner with WiFi transmission capability was used to produce high-quality color images of the PADs and wirelessly transfer them to a smartphone or other mobile devices for data processing. The concentrations of biomarkers in both standard solutions and plasma samples can be directly obtained using acustom-designed smartphone app that is also capable of constructing calibration curves. The detection limits of uric acid, glucose, and triglyceride were determined to be 0.50mg/dL, 0.84mmol/L, and 14mg/dL, respectively, which are below the normal limits and adequate for clinical validation. Owing to the distinct advantages-simple, portable, and cost-effective-this mobile assay protocol can be used for point-of-care (POC) settings or resource-limited situations, and potentiallyfor the diagnosis and prevention of infectious diseases.

Association of Immunosuppression and Viral Load With Subcortical Brain Volume in an International Sample of People Living With HIV

Despite more widely accessible combination antiretroviral therapy (cART), HIV-1 infection remains a global public health challenge. Even in treated patients with chronic HIV infection, neurocognitive impairment often persists, affecting quality of life. Identifying the neuroanatomical pathways associated with infection in vivo may delineate the neuropathologic processes underlying these deficits. However, published neuroimaging findings from relatively small, heterogeneous cohorts are inconsistent, limiting the generalizability of the conclusions drawn to date. To examine structural brain associations with the most commonly collected clinical assessments of HIV burden (CD4+ T-cell count and viral load), which are generalizable across demographically and clinically diverse HIV-infected individuals worldwide. This cross-sectional study established the HIV Working Group within the Enhancing Neuro Imaging Genetics Through Meta Analysis (ENIGMA) consortium to pool and harmonize data from existing HIV neuroimaging studies. In total, data from 1295 HIV-positive adults were contributed from 13 studies across Africa, Asia, Australia, Europe, and North America. Regional and whole brain segmentations were extracted from data sets as contributing studies joined the consortium on a rolling basis from November 1, 2014, to December 31, 2019. Volume estimates for 8 subcortical brain regions were extracted from T1-weighted magnetic resonance images to identify associations with blood plasma markers of current immunosuppression (CD4+ T-cell counts) or detectable plasma viral load (dVL) in HIV-positive participants. Post hoc sensitivity analyses stratified data by cART status. After quality assurance, data from 1203 HIV-positive individuals (mean [SD] age, 45.7 [11.5] years; 880 [73.2%] male; 897 [74.6%] taking cART) remained. Lower current CD4+ cell counts were associated with smaller hippocampal (mean [SE] β = 16.66 [4.72] mm3 per 100 cells/mm3; P < .001) and thalamic (mean [SE] β = 32.24 [8.96] mm3 per 100 cells/mm3; P < .001) volumes and larger ventricles (mean [SE] β = -391.50 [122.58] mm3 per 100 cells/mm3; P = .001); in participants not taking cART, however, lower current CD4+ cell counts were associated with smaller putamen volumes (mean [SE] β = 57.34 [18.78] mm3 per 100 cells/mm3; P = .003). A dVL was associated with smaller hippocampal volumes (d = -0.17; P = .005); in participants taking cART, dVL was also associated with smaller amygdala volumes (d = -0.23; P = .004). In a large-scale international population of HIV-positive individuals, volumes of structures in the limbic system were consistently associated with current plasma markers. Our findings extend beyond the classically implicated regions of the basal ganglia and may represent a generalizable brain signature of HIV infection in the cART era.

The impact of anesthesia method on the severity of oxidative stress during coronary artery bypass grafting with cardiopulmonary bypass

The objective : to investigate the impact of anesthesia method on the severity of oxidative stress in patients after coronary artery bypass grafting with cardiopulmonary bypass. Subjects and methods . Patients were randomized (seed 6556 as of 04.01.2016, www.randomization.com) to Volatile Induction and Maintenance Anesthesia ( VIMA ) with Sevoflurane group ( n = 65) and Total Intravenous Anesthesia with Propofol and Fentanyl ( TIVA ) group ( n = 66). The changes in oxidative stress markers in blood plasma were studied: carbonylated proteins, nitrotyrosine, oxidized forms of lo w-d ensity lipoproteins – ox y- L DL . Results . At the critical stage with 24 hours after the surgery, statistically significant differences in the carbonyls blood levels were found between VIMA with Sevoflurane and TIVA with Propofol groups: 0.88 (0.79–0.96) nmo l/m g protein (nmo l/m g) in TIVA group vs 0.81 (0.75–0.91) nmo l/m g in VIMA group, p = 0.01; and ox y- L DL levels 0.96 ± 0.40 m g/m l vs 0.83 ± 0.33 m g/m l, respectively, p = 0.04. Nitrotyrosine demonstrated no diagnostic value. Conclusion. It has been suggested that sevoflurane possesses antioxidant properties that can be regarded as a positive quality of VIMA in coronary artery bypass grafting with cardiopulmonary bypass.

Altered motor, anxiety-related and attentional task performance at baseline associate with multiple gene copies of the vesicular acetylcholine transporter and related protein overexpression in ChAT::Cre+ rats

Transgenic rodents expressing Cre recombinase cell specifically are used for exploring mechanisms regulating behavior, including those mediated by cholinergic signaling. However, it was recently reported that transgenic mice overexpressing a bacterial artificial chromosome containing choline acetyltransferase (ChAT) gene, for synthesizing the neurotransmitter acetylcholine, present with multiple vesicular acetylcholine transporter (VAChT) gene copies, resulting in altered cholinergic tone and accompanying behavioral abnormalities. Since ChAT::Cre+ rats, used increasingly for understanding the biological basis of CNS disorders, utilize the mouse ChAT promotor to control Cre recombinase expression, we assessed for similar genotypical and phenotypical differences in such rats compared to wild-type siblings. The rats were assessed for mouse VAChT copy number, VAChT protein expression levels and for sustained attention, response control and anxiety. Rats were also subjected to a contextual fear conditioning paradigm using an unconditional fear-inducing stimulus (electrical foot shocks), with blood samples taken at baseline, the fear acquisition phase and retention testing, for measuring blood plasma markers of hypothalamic–pituitary–adrenal gland (HPA)-axis activity. ChAT::Cre+ rats expressed multiple mouse VAChT gene copies, resulting in significantly higher VAChT protein expression, revealed anxiolytic behavior, hyperlocomotion and deficits in tasks requiring sustained attention. The HPA-axis was intact, with unaltered circulatory levels of acute stress-induced corticosterone, leptin and glucose. Our findings, therefore, reveal that in ChAT::Cre+ rats, VAChT overexpression associates with significant alterations of certain cognitive, motor and affective functions. Although highly useful as an experimental tool, it is essential to consider the potential effects of altered cholinergic transmission on baseline behavior in ChAT::Cre rats.

Optimisation and investigations into the effect of a phosphorylated tocopherol mixture on growth performance, meat quality and plasma inflammatory biomarkers in broilers

This study investigated the effect of a novel phosphorylated tocopherol mixture (TPM) on the growth performance, nutrient digestibility, meat quality, and blood plasma markers of inflammation in broilers reared in normal temperatures (NT) or cyclical high temperatures (CHT). TPM has previously been shown to increase the oral bioavailability of lipid soluble nutrients. Following a preliminary dose optimisation study, three hundred and sixty Ross 308 day old broilers were housed in groups of 5 in cages (n = 12). Treatments were arranged in a factorial design encompassing 3 dietary TPM levels and ± CHT. From day of placement until d 35, broilers were assigned to 1 of 3 vitamin E-adequate, wheat and soybean meal-based diets containing TPM at 0 (Control), 10 or 20 mg/kg diet. From 21-35 d of age, birds were exposed to either normal temperatures (NT; 22 ± 1 °C; 60% RH) or cyclical high temperature (CHT; 32 ± 1 °C; 8 h; 80–90% RH and 16 h at 22 ± 1 °C; 60% RH). Implementing CHT negatively impacted feed intake (FI), body weight (BW) gain, feed conversion ratio (FCR) and final BW (d 35), apparent ileal digestibility (AID) and shear force of cooked breast muscle. Under both NT and CHT regimes, the TPM 10 group had improved BW gain (d 0-35), FCR (d 0-21) and final BW (d 35) when compared with Control diet groups containing TPM at 0. There was an interaction between diet and temperature regime on AID. The TPM 20 + CHT group had decreased AID of nitrogen (N) and gross energy (GE) when compared with the TPM 20 + NT group. However, there was no effect of the Control or TPM 10 on AID under either temperature regime. Breast muscle from CHT birds had increased temperature and shear force when cooked, and decreased pH and drip loss when compared with NT birds. The TPM 10 group tended to have lower drip loss (P = 0.057) under both temperature regimes when compared with the Control group. Implementing CHT increased the plasma concentration of select cytokines and chemokines (INFγ, IL-6, IL-10, IL-16, IL-21 and CCL5) when compared with the NT treatment. Birds offered the TPM 10 diets had numerically lower concentrations of all plasma chemokines and cytokines when compared with the Control diets. In summary, CHT depressed performance and nutrient digestibility and negatively impacted breast muscle quality. The TPM 10 treatment improved growth performance variables under both NT and CHT regimes and tended to reduce muscle drip loss.

The protective effect of curcumin against aluminum chloride-induced oxidative stress and hepatotoxicity in rats

Aluminum is recognized as a public health concern because of its potential toxic effects on human health. Therefore, the present experiment was undertaken to determine the effectiveness of curcumin (CUR) in reducing the hepatotoxicity induced by aluminum chloride (AlCl3) in animals. In this experimental study, forty male rats were allocated to five groups (N = 8), viz. no treatment (control), solvent (DMSO or distilled water), CUR (10 mg/kg B.W.), AlCl3 (10 mg/kg B.W.), and CUR+AlCl3 (each with 10 mg/kg B.W.). Treatments were performed by intra-peritoneal injections for 28 days. On the final day, animals were sacrificed, and liver function markers in blood plasma, hepatic antioxidants, and lipid peroxidation index in liver homogenate were estimated. AlCl3 treatment resulted in a significant increase in plasma AST, ALT, ALP and LDH activities with decreased total protein compared to control. AlCl3 significantly reduced superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels but increased malondialdehyde (MDA) level in the liver compared to control. AlCl3 also caused various histopathological changes in the livers of rats. Curcumin could normalize nearly all these parameters. CUR improved levels of changes in different parameters when was combined with AlCl3. It is concluded that CUR has beneficial effects being able to antagonize AlCl3 toxicity.

Determination of Thiodiglycol and Its Oxide in Biomedical Samples by Gas Chromatography–Mass Spectrometry

A unified procedure is proposed for the determination of thiodiglycol (TDG) and its oxide (TDGO) as markers of exposure to sulfur mustard in biomedical samples (urine, blood plasma, hairs) by gas chromatography–electron ionization mass spectrometry. The limit of detection of the procedure is comparable with the lowest level of endogenous TDG in biosamples (1 ng mL–1). Solid-phase microextraction of the derivatized analytes from the reaction mixture decreases the matrix effect. The procedure was tested in the analysis of rat biosamples obtained after in vivo exposure to sulfur mustard. The procedure was found to offer a higher potential in the retrospective determination of the total TDG and TDGO level in rat urine (within 7 days after exposure to 2 mg kg–1 of sulfur mustard) compared to the determination of the same markers in blood plasma. Blood plasma can be used for confirmatory analysis, provided the samples are taken within 2 days after exposure. Hair was found to be the least suitable biomatrix in view of the variability of the TDG background level in these samples and the possibility of contamination with TGD from other sources.

THE EFFECT OF CURCUMIN ON LIVER FIBROSIS IN THE RAT MODEL OF MICROSURGICAL CHOLESTASIS

We aimed to determine the effects of curcumin on liver fibrosis and to clarify the role of nuclear factor-κB (NF-κB) and inducible nitric oxide synthase (iNOS) in a model of microsurgical cholestasis in the early stage of extrahepatic biliary atresia. Twelve-week-old Wistar rats were divided into four groups (n = 8): sham-operated rats (received olive oil after laparotomy); a curcumin group (received curcumin, 200 mg kg(-1) per day, after laparotomy); a biliary duct ligated group (BDL, received olive oil after operation); and a biliary duct ligated/curcumin group (BDL curc, received curcumin, 200 mg kg(-1) per day, after operation). After 3 weeks of treatment, curcumin did not modify blood plasma markers as well as the expressions of iNOS and NF-κB (p65) in the livers of the sham group. Interestingly, there was a significant increase in the extent of both liver and kidney fibrosis. On the other hand, despite a decrease in the expression of iNOS and NF-κB (p65), treatment with curcumin did not affect fibrosis enlargement due to bile duct ligation in the liver. In the BDL group, treatment with curcumin decreased the level of blood plasma markers investigated. In conclusion, treatment with curcumin was able to improve the functional properties of hepatocytes and to inhibit the upregulations of both NF-κB and iNOS in the BDL group; however, no beneficial effect was observed on the liver fibrosis developed in this model of cholestasis. Thus, in the studied model of microsurgical cholestasis, other factors different from NF-κB and iNOS are responsible for fibrotic processes in the liver.

Galanin and its N-terminal fragments reduce acute myocardial infarction in rats

Agonists and antagonists for galanin receptor subtypes GalR1-3 can be used as putative therapeutics targets for the treatment of various human diseases. However, effects of galanin and its N-terminal fragments on myocardial ischemia/reperfusion injury remain unclear. This study was designed to assess the ability of the full-length galanin (GWTLNSAGYLLGPHAIDNHRSFSDKHGLT-NH2, G1), the natural fragments WTLNSAGYLL-NH2 (G2) and WTLNSAGYLLGPHA (G3), and their modified analogs WTLNAAGYLL (G4) and WTLNSAGYLLGPβAH (G5) to limit acute myocardial infarction in rats in vivo. The peptides G2-5 were synthesized by the automatic solid phase method using Fmoc technology, purified by preparative HPLC and identified by 1H NMR spectroscopy and MALDI −TOF mass spectrometry. The peptides G1-5 were administered by i.v. bolus injection at the onset of reperfusion at doses of 0.25, 0.50, 1.0, 2.0 or 3.0 mg/kg. The optimal doses of the peptides G1-5 significantly reduced the infarction area and decreased the activity of CK-MB and LDH in blood plasma at the end of reperfusion compared with the control. Among the peptides studied, G5 showed high efficacy in reducing the infarct size and the activity of necrosis markers in blood plasma with no significant effect on hemodynamic parameters. The results suggest that a novel agonist for galanin receptors G5 may be a promising tool for the treatment of myocardial ischemia/reperfusion (I/R) injury. Further studies are warranted to explore the stability of this peptide in blood plasma and mechanisms that contribute to its cardioprotective effects.