Changes In Plasma Proteins Research Articles

Proteomics based identification of differential plasma proteins and changes in white matter integrity as markers in early detection of mild cognitive impaired subjects at high risk of Alzheimer’s disease

Mild cognitive impairment (MCI) is an intermediate stage of cognitive decline and dementia. The absence of specific diagnostic test for identification of MCI and AD. The current study aims to find proteomics based change in plasma proteins and diffusion tensor imaging (DTI) based white matter changes in MCI for early detection of prodromal Alzheimer’s disease. Fifty cases of mild cognitive impairment and age matched control between (55–75 yrs) were screened on basis of Mini Mental State Examination (MMSE). Two dimensional gel electrophoresis and DTI imaging was performed in MCI and age matched control. The MMSE score of MCI were in the range of (28 ± 2–22.6 ± 1) as compared with healthy control (28 ± 2), DTI metrics apparent diffusion coefficient (ADC) and fractional Anisotropy (FA) has shown significant changes in fornix, corpus callosum, hippocampus, right temporal and right frontal lobe, left frontal lobe, forcep major of MCI subjects as compared with controls. The protein expression of keratin type-2 was up regulated and albumin was down regulated in MCI subjects as compared with control. The data from present study signifies that expression of Keratin type-2 and albumin along with white matter changes provides early signatures for identification of MCI at high risk of Alzheimer’s disease

Pulmonary Circulation Transvascular Fluid Fluxes Do Not Change during General Anesthesia in Dogs.

General anesthesia (GA) can cause abnormal lung fluid redistribution. Pulmonary circulation transvascular fluid fluxes (JVA) are attributed to changes in hydrostatic forces and erythrocyte volume (EV) regulation. Despite the very low hydraulic conductance of pulmonary microvasculature it is possible that GA may affect hydrostatic forces through changes in pulmonary vascular resistance (PVR), and EV through alteration of erythrocyte transmembrane ion fluxes (ionJVA). Furosemide (Fur) was also used because of its potential to affect pulmonary hydrostatic forces and ionJVA. A hypothesis was tested that JVA, with or without furosemide treatment, will not change with time during GA. Twenty dogs that underwent castration/ovariectomy were randomly assigned to Fur (n = 10) (4 mg/kg IV) or placebo treated group (Con, n = 10). Baseline arterial (BL) and mixed venous blood were sampled during GA just before treatment with Fur or placebo and then at 15, 30 and 45 min post-treatment. Cardiac output (Q) and pulmonary artery pressure (PAP) were measured. JVA and ionJVA were calculated from changes in plasma protein, hemoglobin, hematocrit, plasma and whole blood ions, and Q. Variables were analyzed using random intercept mixed model (P < 0.05). Data are expressed as means ± SE. Furosemide caused a significant volume depletion as evident from changes in plasma protein and hematocrit (P < 0.001). However; Q, PAP, and JVA were not affected by time or Fur, whereas erythrocyte fluid flux was affected by Fur (P = 0.03). Furosemide also affected erythrocyte transmembrane K+ and Cl−, and transvascular Cl− metabolism (P ≤ 0.05). No other erythrocyte transmembrane or transvascular ion fluxes were affected by time of GA or Fur. Our hypothesis was verified as JVA was not affected by GA or ion metabolism changes due to Fur treatment. Furosemide and 45 min of GA did not cause significant hydrostatic changes based on Q and PAP. Inhibition of Na+/K+/2Cl− cotransport caused by Fur treatment, which can alter EV regulation and JVA, was offset by the Jacobs Stewart cycle. The results of this study indicate that the Jacobs Stewart cycle/erythrocyte Cl− metabolism can also act as a safety factor for the stability of lung fluid redistribution preserving optimal diffusion distance across the blood gas barrier.

Proteomics in plasma of ovariectomized rats and those exposed to estradiol valerate

The menopausal period, an inevitable physiological process for women, is frequently associated with physiological and psychological dysfunction attributable to substantial fluctuation and gradual decrease in female hormones induced by ovarian failure, leading to corresponding symptoms and diseases that impact multiple systems in the body to varying degrees. As prior studies have focused primarily on menopausal syndrome-related pathophysiological changes and hormone replacement therapy effects, here we approached menopausal disease incidence risk and pathogenesis through systemic plasma proteomics analysis. Female Sprague-Dawley rats were randomly divided into sham, ovariectomized, and estrogen treatment after ovariectomy groups (n=9 per group). Tandem Mass Tag quantitative proteomics analysis of their plasma identified over 900 proteins by MS. Between group fold change of >1.2 and p<0.05 (Student’s t-test) identified 121 (including 36 up-regulated and 85 down-regulated), 117 (69 up-regulated and 48 down-regulated), and 109 (41 up-regulated and 68 down-regulated) differentially expressed proteins between groups, respectively. Of these, 5 (GHR, LIFR, apoA IV, RTN, and Lin28b) were verified by parallel reaction monitoring to be reliable. Further application of optimized screening criteria and performance of a series of bioinformatics analyses allowed the selection of 35 optimal differentially expressed proteins. Gene ontology annotation results suggested that the differentially expressed proteins are mainly annotated as protein binding, cell, and single organism process in terms of molecular function, cell composition, and biological process, respectively. KEGG pathway analysis indicated that the PI3-Akt pathway has the highest aggregation degree of differentially expressed proteins. Protein-protein interaction analysis noted GLUT4 as an important node protein. This research is the first to comprehensively analyze plasma protein changes, together with estrogen efficacy, in ovariectomized rats. The findings facilitate our understanding of the molecular mechanism of systemic menopausal changes and provide valuable clues for developing diagnostic biomarkers for menopausal dysfunctions and selecting clinical therapeutic strategies.

Plasma protein, haematologic and blood chemistry changes in African grey parrots (Psittacus erithacus) experimentally infected with bornavirus

ABSTRACTBornaviruses are considered to be the causative agent of proventricular dilatation disease (PDD) in psittacine birds. In order to detect haematological and blood chemistry changes during the development of PDD and a possible correlation with clinical signs and the virological status, six African grey parrots (Psittacus erithacus) were experimentally infected with parrot bornavirus 4 (PaBV-4) by subcutaneous route. All six parrots developed clinical signs of varying extent and successful infection was confirmed in all the birds by seroconversion or detection of RNA of the PaBV-4 infection strain. Based on population-based and intra-individual reference ranges established during 12 months prior to experimental infection, only minor haematological changes were detected in individual birds after infection. Changes in blood chemistry were restricted to aspartate aminotransferase, creatine kinase, total protein, glucose and uric acid. Plasma protein electrophoresis revealed marked changes starting 10 weeks post infection characterized by an increase in the γ-globulin fraction and a gradual decrease to normal values during weeks 22–34. Indications of an acute-phase reaction at the initial stages of infection were not detected. While three birds suffered from clinical signs of PDD, which included weight loss and neurological disorders and died before development of haematological and plasma protein changes, recovery of clinical disease was paralleled in the remaining birds by an increase in γ-globulins and bornavirus-specific antibody titres.

High-fat diet–induced plasma protein and liver changes in obese rats can be attenuated by melatonin supplementation

Obesity triggers changes in protein expression in various organs that might participate in the pathogenesis of obesity. Melatonin has been reported to prevent or attenuate such pathological protein changes in several chronic diseases. However, such melatonin effects on plasma proteins have not yet been studied in an obesity model. Using a proteomic approach, we investigated the effect of melatonin on plasma protein profiles after rats were fed a high-fat diet (HFD) to induce obesity. We hypothesized that melatonin would attenuate abnormal protein expression in obese rats. After 10weeks of the HFD, animals displayed increased body weight and fat accumulation as well as increased glucose levels, indicating an obesity-induced prediabetes mellitus–like state. Two-dimensional gel electrophoresis and liquid chromatography–mass spectrometry/mass spectrometry revealed 12 proteins whose expression was altered in response to the HFD and the melatonin treatment. The altered proteins are related to the development of liver pathology, such as cirrhosis (α1-antiproteinase), thrombosis (fibrinogen, plasminogen), and inflammation (mannose-binding protein A, complement C4, complement factor B), contributing to liver steatosis or hepatic cell death. Melatonin treatment most probably reduced the severity of the HFD-induced obesity by reducing the amplitude of HFD-induced plasma protein changes. In conclusion, we identified several potential biomarkers associated with the progression of obesity and its complications, such as liver damage. Furthermore, our findings reveal melatonin's beneficial effect of attenuating plasma protein changes and liver pathogenesis in obese rats.

СРАВНИТЕЛЬНОЕ ИЗУЧЕНИЕ СПЕКТРОВ ФЛУОРЕСЦЕНЦИИ БЕЛКОВ ПЛАЗМЫ И ЭРИТРОЦИТОВ КРОВИ ПРИ СТАРЕНИИ И УПОТРЕБЛЕНИИ НАРКОТИЧЕСКИХ ВЕЩЕСТВ

Aging is accompanied by intensification of oxidative degradation of proteins, including lipoproteins, proteolytic system dysfunction and accumulation of oxidized proteins. Similar changes are found in drug addiction. In this regard, we carried out a comparative study of conformational changes of plasma proteins and red blood cells of people of different age groups and people who use drugs. The study was carried out with the use of fluorescence analysis to measure the spectra of fluorescence of amino acid residues in proteins (summary fluorescence and tryptophan). In the analysis of fluorescence spectra it is preferable to use the second derivative of the fluorescence spectra as it is the most informative. The application of this research method was chosen because it has high sensitivity. It provides information on the status of living systems without damaging them; it requires large quantities of biological material; it makes it possible to differentiate between the various stages of the disease. Changes in the general phase and tryptophan fluorescence plasma proteins in older people and drug addicts indicate increase in the availability of tryptophan residues to the aqueous environment as a result of conformational changes of macromolecules and the disruption of the lipid layer due to the intensification of free radical processes.

The maternal plasma proteome changes as a function of gestational age in normal pregnancy: a longitudinal study

Pregnancy is accompanied by dramatic physiological changes in maternal plasma proteins. Characterization of the maternal plasma proteome in normal pregnancy is an essential step for understanding changes to predict pregnancy outcome. The objective of this study was to describe maternal plasma proteins that change in abundance with advancing gestational age and determine biological processes that are perturbed in normal pregnancy. A longitudinal study included 43 normal pregnancies that had a term delivery of an infant who was appropriate for gestational age without maternal or neonatal complications. For each pregnancy, 3to6 maternal plasma samples (median, 5) were profiled to measure the abundance of 1125 proteins using multiplex assays. Linear mixed-effects models with polynomial splines were used to model protein abundance as a function of gestational age, and the significance of the association was inferred via likelihood ratio tests. Proteins considered to be significantly changed were defined as having the following: (1) >1.5-fold change between 8 and 40 weeks of gestation; and (2) a false discovery rate-adjusted value of P < .1. Gene ontology enrichment analysis was used to identify biological processes overrepresented among the proteins that changed with advancing gestation. The following results were found: (1) Ten percent (112 of 1125) of the profiled proteins changed in abundance as a function of gestational age; (2) of the 1125 proteins analyzed, glypican-3, sialic acid-binding immunoglobulin-type lectin-6, placental growth factor, C-C motif-28, carbonic anhydrase 6, prolactin, interleukin-1 receptor 4, dual-specificity mitogen-activated protein kinase 4, and pregnancy-associatedplasma protein-A had more than a 5-fold change in abundance across gestation (these 9 proteins are known to be involved in a wide range of both physiological and pathological processes, such as growth regulation, embryogenesis, angiogenesis immunoregulation, inflammation etc); and (3) biological processes associated with protein changes in normal pregnancy included defense response, defense response to bacteria, proteolysis, and leukocyte migration (false discovery rate, 10%). The plasma proteome of normal pregnancy demonstrates dramatic changes in both the magnitude of changes and the fraction of the proteins involved. Such information is important to understand the physiology of pregnancy and the development of biomarkers to differentiate normal vs abnormal pregnancy and determine the response to interventions.

Potential mechanism for increased risk of premature vascular disease in long-term survivors of primary brain tumours with established hormone deficits

BackgroundLong-term survivors of childhood-onset brain tumours have increased mortality rates, even after excluding deaths related to recurrence or regression of the original tumour. In particular, there is an increased risk of premature vascular disease by mechanisms that remain unclear. We tested our hypothesis that a thrombotic fibrin network profile is one mechanism for the increased vascular risk in this population. MethodsWe undertook a cross-sectional study in 33 patients (19 men, mean age 31·5 years [SD 14·2]) with previous history of primary brain tumours of either childhood or adulthood onset and 33 age-matched and sex-matched healthy controls. We performed clot structure analysis using a validated turbidimetric assay and also assessed plasma concentrations of thrombotic and inflammatory vascular markers including fibrinogen, C-reactive protein (CRP), and complement C3. The mean time from brain tumour diagnosis to the time of the study was 8·9 years (SD 6·2). FindingsAll patients had cranial radiotherapy, whereas 25 (76%) and 13 (39%) had additional surgery and chemotherapy, respectively. 31 patients (94%) had growth hormone deficiency, five (15%) had deficiences in luteinising hormone and follicle-stimulating hormone, four (12%) in adrenocorticotropic hormone, and two (6%) in thyroid-stimulating hormone. Patients had raised clot maximum absorbance (a measure of clot density) compared with controls (mean 0·412 arbritary units [SD 0·10] vs 0·277 [0·09], p<0·0001), increased clot lysis time (an indicator of fibrinolysis potential) (3695 s [731] vs 2720 [636], p<0·0001), and larger lysis area (a complex measure of clot formation time, clot density, and lysis potential) (1949 arbitrary units [1390] vs 906 [704], p<0·0001). Despite differences in fibrin network characteristics, fibrinogen concentrations were similar in patients and controls. Plasma CRP was higher in patients than in controls (3·08 mg/L [4·25] vs 0·78 [1·08], p=0·006) with similar findings for C3 (0·75 mg/mL [0·13] vs 0·67 [0·13], p=0·027). InterpretationWe demonstrate, for the first time to our knowledge, that survivors of brain tumours with hypopituitarism display a thrombotic fibrin network profile, providing one mechanism for the increased vascular risk in this population. These changes are not related to altered fibrinogen concentrations suggesting that other plasma proteins or qualitative changes in the proteins might contribute to the observed differences. Further longitudinal studies are required to clarify whether optimisation of the hormonal profile results in amelioration of the thrombotic environment in this population. FundingPfizer (investigator-initiated research grant).

Pharmacokinetics and pharmacodynamics of cisatracurium in patients undergoing surgery with two hemodilution methods

ObjectiveTo investigate the pharmacokinetics and pharmacodynamics of cisatracurium in patients undergoing surgery under acute normovolemic hemodilution (ANH) and acute hypervolemic hemodilution (AHH). MethodsNinety patients with orthopedic surgery were divided into ANH, AHH and control groups, which received hemodilution by hydroxyethyl starch 130/0.4 transfusion, Voluven transfusion and regular transfusion and infusion during surgery, respectively. Each group was divided into 3 sub-groups, administrated with cisatracurium at dosage of 0.1, 0.2 and 0.3mg/kg respectively. The changes in plasma protein, pH and electrolytes from the hemodilution beginning to surgery finish were monitored. Before and after cisatracurium administration, the phamocodynamic indicators of muscle relaxant were observed, and the plasma drug concentration was measured. ResultsAfter hemodilution or regular transfusion, all three groups experienced a distinct drop in total plasma protein, albumin and pH. Compared with control group, the plasma concentrations of both K+ and Ca2+ in ANH and AHH groups significantly dropped (P<0.05), and those in each group after administration of cisatracurium also dropped, compared with before (P<0.05). After administration with cisatracurium, the onset time of muscle relaxation in AHH group was extended notably, compared with AHH and control groups (P<0.05), with no distinct difference of residual pharmacodynamics parameters (P>0.05). In the same hemodilution or regular infusion, with increase of drug dosage, the onset time of muscle relaxation was shortened, and the period of no response to train-of-four stimulation, muscle relaxation blockade duration and action time of muscle relaxation blockade in body were extended (P<0.05). ConclusionWhen using cisatracurium under AHH, the dosage should be increased appropriately, while it need not be adjusted under ANH.

Plasma viscosity during normal pregnancy

The changes in plasma viscosity and plasma proteins were serially measured in 14 nulli- and 10 multiparae, during their normal pregnancies. As compared to the non-pregnant value plasma viscosity decreased in the first trimester, but increased during the second and third trimester until term. At 37 weeks, the plasma viscosity value exceeded the non-pregnant value, the difference being statistically significant. The increase in plasma viscosity during the second and third trimester of pregnancy was determined by the rise in fibrinogen concentration, which overruled the plasma viscosity lowering influence of the falling total serum protein concentration

Mechanisms of toxicity and resilience of an in vitro 3D human dopaminergic model to Rotenone

The expression of plasma proteins changes dramatically as a result of cytokine induction, particularly interleukin-6, and their levels are used as clinical markers of inflammation. miRNAs are important regulators of gene expression and play significant roles in many inflammatory diseases and processes. The interactions between miRNAs and the genes that they regulate during the acute phase response have not been investigated. We examined the effects of IL-6 stimulation on the transcriptome and miRNome of human and mouse primary hepatocytes and the HepG2 cell line. Using an integrated analysis, we identified differentially expressed miRNAs whose seed sequences are significantly enriched in the 3′ untranslated regions of differentially expressed genes, many of which are involved in inflammation-related pathways. Our finding that certain miRNAs may de-repress critical acute phase proteins within acute timeframes has important biological and clinical implications.

Changes in the plasma proteome at asymptomatic and symptomatic stages of autosomal dominant Alzheimer's disease.

The autosomal dominant form of Alzheimer’s disease (ADAD) is far less prevalent than late onset Alzheimer’s disease (LOAD), but enables well-informed prospective studies, since symptom onset is near certain and age of onset is predictable. Our aim was to discover plasma proteins associated with early AD pathology by investigating plasma protein changes at the asymptomatic and symptomatic stages of ADAD. Eighty-one proteins were compared across asymptomatic mutation carriers (aMC, n = 15), symptomatic mutation carriers (sMC, n = 8) and related noncarriers (NC, n = 12). Proteins were also tested for associations with cognitive measures, brain amyloid deposition and glucose metabolism. Fewer changes were observed at the asymptomatic than symptomatic stage with seven and 16 proteins altered significantly in aMC and sMC, respectively. This included complement components C3, C5, C6, apolipoproteins A-I, A-IV, C-I and M, histidine-rich glycoprotein, heparin cofactor II and attractin, which are involved in inflammation, lipid metabolism and vascular health. Proteins involved in lipid metabolism differed only at the symptomatic stage, whereas changes in inflammation and vascular health were evident at asymptomatic and symptomatic stages. Due to increasing evidence supporting the usefulness of ADAD as a model for LOAD, these proteins warrant further investigation into their potential association with early stages of LOAD.

Long-term heat stress induces the inflammatory response in dairy cows revealed by plasma proteome analysis

In this work we employed a comparative proteomic approach to evaluate seasonal heat stress and investigate proteomic alterations in plasma of dairy cows. Twelve lactating Holstein dairy cows were used and the treatments were: heat stress (n = 6) in hot summer (at the beginning of the moderate heat stress) and no heat stress (n = 6) in spring natural ambient environment, respectively. Subsequently, heat stress treatment lasted 23 days (at the end of the moderate heat stress) to investigate the alterations of plasma proteins, which might be employed as long-term moderate heat stress response in dairy cows. Changes in plasma proteins were analyzed by two-dimensional electrophoresis (2-DE) combined with mass spectrometry. Analysis of the properties of the identified proteins revealed that the alterations of plasma proteins were related to inflammation in long-term moderate heat stress. Furthermore, the increase in plasma tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) directly demonstrated that long-term moderate heat stress caused an inflammatory response in dairy cows.

The role of vitamin E or clay in growing Japanese quail fed diets polluted by cadmium at various levels

This study was conducted to verify whether vitamin (Vit) E or natural clay as feed additives has the potential to modulate the deleterious effects resulting from exposure to cadmium (Cd) in growing Japanese quail. 648 Japanese quail chicks (1 week old) were used to evaluate the effects of dietary Cd (0, 40, 80 and 120 mg/kg diet) and two levels of Vit E (0, 250 mg/kg diet) or two levels of natural clay (0 and 100 mg/kg diet) to study the influences of Cd, Vit E, clay or their different combinations on growth performance, carcass traits, some blood biochemical components and Cd residues in muscles and liver. Live BW and weight gain of quails were linearly decreased with increasing dietary Cd levels. Moreover, feed conversion was significantly worsened with increasing Cd level. Mortality percentage was linearly increased as dietary Cd level increased up to 120 mg/kg diet. Carcass percentage was linearly decreased as dietary Cd level increased. While, giblets percentage were linearly and quadratically differed as dietary Cd level increased. Cd caused significant changes in total plasma protein, albumin, globulin, A/G ratio, creatinine, urea-N and uric acid concentrations as well as ALT, AST and ALP activities. Increasing dietary Cd level was associated with its increase in the muscles and liver. Dietary supplementation with 250 mg of Vit E/kg diet or 100 mg clay/kg improved live BW, BW gain and feed conversion when compared with the un-supplemented diet. Quails fed diet contained 250 mg Vit E/kg and those fed 100 mg clay/kg had the highest percentages of carcass and dressing than those fed the un-supplemented diet. Blood plasma biochemical components studied were better when birds received 250 mg of Vit E/kg diet and those received 100 mg clay/kg. Cd residues in the muscles and liver were significantly less in the birds had 250 mg of Vit E/kg or those received 100 mg clay/kg diet than those un-supplemented with Vit E. Growth performance traits and blood plasma biochemical components studied were significantly affected linearly by the interactions among Cd and each of Vit E and clay levels. In conclusion, the present results indicate that the deleterious effects induced by Cd plays a role in decreasing the performance of Japanese quail and that dietary supplementation with natural clay or Vit E may be useful in partly alleviating the adverse effects of Cd.

Longitudinal Protein Changes in Blood Plasma Associated with the Rate of Cognitive Decline in Alzheimer's Disease.

Biomarkers of Alzheimer's disease (AD) progression are needed to support the development of urgently needed disease modifying drugs. We employed a SOMAscan assay for quantifying 1,001 proteins in blood samples from 90 AD subjects, 37 stable mild cognitive impaired (MCI) subjects, 39 MCI subjects converting to AD within a year and 69 controls at baseline and one year follow up. We used linear mixed effects models to identify proteins changing significantly over one year with the rate of cognitive decline, which was quantified as the reduction in Mini Mental State Examination (MMSE) scores. Additionally, we investigated proteins changing differently across disease groups and during the conversion from MCI to AD. We found that levels of proteins belonging to the complement cascade increase significantly in fast declining AD patients. Longitudinal changes in the complement cascade might be a surrogate biomarker for disease progression. We also found that members of the cytokine-cytokine receptor interaction pathway change during AD when compared to healthy aging subjects.

Physiological changes in the peri-partum period and colostral IgG transfer in prolific D'man sheep: effects of parity and litter size.

The aim of this work was to assess maternal and neonatal changes in plasma proteins, glucose and cortisol and to quantify the colostral immunoglobulin G (IgG) transfer in the peri-partum period in D'man sheep, a prolific breed, taking into account the parity of the ewe. The concentrations of proteins and glucose were high in the ewes on day 7 and at lambing before decreasing. Likewise, cortisol plasma concentration was maximal during the 6h following lambing and dropped at 12h. Protein and glucose concentrations were low in lambs at 1h of birth after which they increased. By contrast, cortisol level was the highest during the first 12h of birth and then decreased. The colostral IgG level was high at lambing and dropped by over 87% from 1 to 48h post-partum. In the newborn, the plasma IgG concentration was lowest at birth and increased rapidly during the first 24h of birth. Parity influenced maternal physiology with multiparous ewes having the lowest concentrations of proteins, glucose, IgG and cortisol, but the highest colostrum IgG level. Accordingly, lambs born from primiparous ewes had lower protein, glucose and plasma IgG concentrations than lambs born from multiparous ewes. The main outcome of this study was that lambs born from primiparous ewes are characterized by the lowest physiological indices and this may influence their survival chance.

Determination of changes in bovine plasma and milk proteins during naturally occurring Escherichia coli mastitis by comparative proteomic analysis

The aim of this study was to investigate changes in plasma and milk proteins in response to Escherichia coli infection in cows. Plasma and milk were collected from healthy cows, cows suffering from mild E. coli mastitis, and cows suffering from severe E. coli mastitis. Protein composition was examined by two-dimensional gel electrophoresis coupled with mass spectrometry. Plasma haptoglobin and α-1 acid glycoprotein demonstrated greater expression in mastitic cows compared with controls, but there were no difference between mildly and severely mastitic cows. Milk from mildly mastitic cows showed increased albumin and casein variants. Severely mastitic cows showed lower casein levels and increased anti-microbial and acute phase proteins. Milk α-1 acid glycoprotein and cathelicidins were associated with severe mastitis. A greater number of β-casein fragments that corresponded to β-casein isoforms were found in milk from mildly mastitic cows. These results suggest that caseins levels decreased and the concentrations of anti-microbial and acute phase proteins increased corresponding to the degree of E. coli mastitis. Nevertheless, further studies are needed to determine whether cathelicidin could serve as a diagnostic marker for mastitis.

Acute Phase Responses to Novel, Investigational Vaccines in Toxicology Studies: The Relationship Between C-Reactive Protein and Other Acute Phase Proteins.

The objective of this study was to determine the effects of investigational vaccine candidates on acute-phase proteins (APPs) as determined in GLP toxicology studies. Sixty-four GLP toxicity studies, which were submitted to the Food and Drug Administration from 2008 to 2012 in support of proposed clinical investigations, were reviewed and entered into a database. These studies employed the intramuscular route of injection and were conducted using New Zealand White rabbits. A retrospective review of these GLP toxicity studies was conducted to evaluate the changes in plasma levels of C-reactive protein (CRP), fibrinogen, and albumin as APPs following the administration of various investigational vaccines. The incidence and intensity of responses associated with acute-phase responses both positive and negative were observed to increase in animals when treated with vaccines containing more potent immunological components such as novel adjuvants that activate Toll-like receptors in the investigational vaccine products. Changes in plasma levels of CRP were prominent among these responses and provided a basis to propose a classification scheme of H, M, L, and N responding groups. These changes in plasma proteins reflect an activation of the acute-phase response and indicate increasing levels of systemic inflammation, which potentially may be correlated with important clinical adverse events.

Integrated analysis of mRNA and miRNA expression in response to interleukin-6 in hepatocytes

The expression of plasma proteins changes dramatically as a result of cytokine induction, particularly interleukin-6, and their levels are used as clinical markers of inflammation. miRNAs are important regulators of gene expression and play significant roles in many inflammatory diseases and processes. The interactions between miRNAs and the genes that they regulate during the acute phase response have not been investigated. We examined the effects of IL-6 stimulation on the transcriptome and miRNome of human and mouse primary hepatocytes and the HepG2 cell line. Using an integrated analysis, we identified differentially expressed miRNAs whose seed sequences are significantly enriched in the 3′ untranslated regions of differentially expressed genes, many of which are involved in inflammation-related pathways. Our finding that certain miRNAs may de-repress critical acute phase proteins within acute timeframes has important biological and clinical implications.

Evaluation of In Vivo Toxicity of Dichloromethane: Methanolic Leaf Extracts of Prosopis juliflora in Female Wistar Albino Rats

Prosopis juliflora (Mathenge) is an exotic, evergreen leguminous lant found in the dry Coastal, Rift Valley and Northern parts of Kenya. It is tolerant to extreme environmental conditions, rated among top 100 most invasive species worldwide. The species leaf extracts is used in folk cure to various ailments and have promising pharmacological properties however; information on their toxicity in animals and human is insufficient. The study assessed phytochemical composition of P. juliflora leaf extracts, effects on body weights, organ weights, hematological parameters, liver function markers and histopathology in major organs of Wistar albino rats. Acute toxicity test was carried out at 2000 mg/kg body weight followed by a 28 days sub chronic toxicity study at 100, 350 and 1000 mg/kg body weight extracts dosages. The control animals were administered with normal saline. Animals were monitored for physical and behavioral changes including death. They were fasted overnight on 28th day and sacrificed on anesthesia on 29th day. Blood was collected by cardiac puncture. Hematological and liver functions tests were done. Tissue sample of selected organs were processed for histopathology. Data from control and treated animals groups were analyzed by ANOVA and Dunnett’s test. Phytochemicals confirmed included alkaloids, flavonoids, phenols, tannins, terpenoids and saponnins but no cardiac glycosides. Median lethal dose was estimated at above 2000 mg/kg body weight. Dose related transient toxicity symptoms included wheezing, decreased activity, and pilo erection. No significant toxicity effects on hematological parameters were noticed except in mean platelets volume. Similarly, no significant adverse effects occurred in liver function tests except at 1000 mg/kg body weight dosage. No significant adverse changes in plasma proteins, body weights and absolute organ weights were observed except in kidneys and spleen. Histological examination of sample tissues showed mild effects in spleen and kidneys but no adverse pathology in other organ tissues.