Estimation of Cytokines Involved in Acute-Phase Wound Infection with Reference to Residence Time of Patients in Hospitals

Background: Cytokines have a major role in mediating immunity as well as inflammation. The main proinflammatory cytokines are activated after injury and implicated in healing interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). High levels of IL-6 are recorded at initial inflammatory response and start decreasing down to eight day of wounding while TNF-α level remained static and IL-1α levels showed a different pattern of change following injury and consequence of infection. Methodology: This study was conducted in Al-Kindy and Al-Wasity hospitals in Baghdad on 200 patients suffering from wounds. One hundred patients were with acute wounds infection and the other 100 patients wounded but without infection and considered as control. Interleukin-1α (IL-1α), interleukin-TNF-α (TNF-α) and interleukin-6 (IL-60) were determined utilizing ELISA kit sandwich methods (Elabscience, USA). Results: The present study revealed that the values of IL-1α, and TNF-α at 48 hours of hospitalization were 23.547 and 27.177 pg/ml among patients with infected wounds respectively, and 7.05 and 28.127 pg/ml among patients without wound infections respectively. While IL-6 showed a highest level at 96 hours of residence in hospital and the value was 183.43 pg/ml for patients with infected wounds, and the value of the same interleukin was 88.696 pg/ml at 72 hours of residence of patients without wound infections. Conclusions: Interleukin-1α elevated after 24 hr of infection and then decreased. Proinflammatory cytokines (IL-6) was detectable within 24 hr of infection. The highest concentration of IL-6 was seen with mixed bacteria and followed by gram negative bacteria and this probably due to lipopolysaccharide secretion caused an increase of IL-6 in blood circulation. Irregular changes were seen in TNF-α values with durations of patients stay in hospitals.

Prevalence of Different Types of Micro-Organisms and Levels of Complement C5a in Patients with Acute-Phase Wound Infections

Background: An acute wound infection might be caused by external damage to the skin including abrasions, lacerations, bites, burns, accidents, war injuries and surgical incisions. When a wound fails to heal within a week, it should be considered a chronic type. Complement system potent inflammatory cascade in wound infection, is important and altered wound healing. Complement activation leads to the generation of many potent effectors including anaphylatoxin C5a. C5a has induced synthesis of TNF-α and IL-1β in macrophage and monocyte which are all together the goal of the present paper. Methodology: This study was conducted in Al-Kindy and Al-Wasity hospitals in Baghdad on 200 patients suffering from wounds. One hundred patients were with acute wounds infection and the other 100 patients considered as control wounds i.e. without infection. The procedure method was followed according to manufacturer’s instructions (Elabscience, USA) utilizing C5a ELISA kit for conducting the test. Blood samples were taken at 24, 48, 72, 96 and 120 hours of hospitalization of the patients. Results: It was found that the highest concentration of C5a was found at 120 hours after patients hospitalization who were with wound infection, and the mean value of C5a was 4898 pg/ml. 4661 pg/ml of C5a was recorded among patients with acute-phase infection compared to 4387 pg/ml concentration of the same complement among control group without wound infection at 96 hours post residence in hospital. Conclusions: Gram-positive bacteria were more prevalent causing wound infections. Dermacoccus nishinomiyaensis, Kocuria rosea and Kocuria kristinae were isolated for the first time in this locality. A complement 5a (C5a) revealed a very high concentration in acute-phase of wound infections. It was found that C5a was serially elevated with time of hospitalization of wounded and infected patients. C5a was highly elevated with wound infection by Gram-negative bacteria compared to infections by Gram-negatives.

Expression of lipopolysaccharide binding protein and its receptor CD14 in experimental alcoholic liver disease

AIM: To evaluate the relationship between the expression of lipopolysaccharides (LPS) binding protein (LBP) and CD14 mRNA and the severity of liver injury in alcohol-fed rats. METHODS: Twenty Wistar rats were divided into two groups:ethanol-fed group (group E) and control group (group C). Group E was fed with ethanol(5-12 g x kg(-1) x d(-1)) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 weeks and 8 weeks. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed under light and electronic microscopy. Expressions of LBP and CD14 mRNA in liver tissues were determined by RT-PCR analysis. RESULTS: Plasma endotoxin levels were increased more significantly in group E(129+/-21) ng x L(-1) and (187+/-35) ng x L(-1) at 4 and 8 wk than in control rats(48+/-9) ng x L(-1) and (53+/-11) ng x L(-1), respectively (P【0.05). Mean values of plasma ALT levels were (1867+/-250) nkat x L(-1) and (2450+/-367) nkat x L(-1) in Group E. The values were increased more dramatically in ethanol-fed rats than in Group C after 4 and 8 weeks. In liver section from ethanol-fed rats, there were marked pathological changes (steatosis, cell infiltration and necrosis). In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels compared with the control group (P【0.05). CONCLUSION: Ethanol administration led to a significant increase in endotoxin levels in serum and LBP and CD14 mRNA expressions in liver tissues. The increase of LBP and CD14 mRNA expression might wake the liver more sensitive to endotoxin and liver injury.

Prospects of diagnostic and prognostic biomarkers of pyometra in canine

Pyometra is one of the most common uterine pathologies of intact bitch at middle to advanced age. In the early stages, the disease shows subtle changes, making diagnosis a challenge. In contrast, at later stages, it manifests as potentially life-threatening systemic inflammatory response syndrome. Ultrasonographic examination of the uterus aids in the diagnosis, although it has limitation in ascertaining the clinical severity of pyometra. Moreover, differentiation of cystic endometrial hyperplasia from pyometra could not be discerned with greater accuracy. Therefore, false negative diagnosis of pyometra patients leads to development of systemic inflammatory response, which delays administration of therapies and results in deaths during early course of treatment. Further, indiscriminate use of broad-spectrum antimicrobials at higher dose in false positive cases considerably contributes to the rising pool of drug resistant pathogens, thereby increasing the risk of case fatality due to sepsis in a long-term. Monitoring the circulating pro-inflammatory cytokines, acute phase proteins, endotoxin, growth factors and inflammatory mediators is the current trend in pyometra research, especially for developing diagnostic and prognostic biomarkers. The present review deals with the prospects of developing diagnostic and prognostic biomarkers in the canine pyometra.

Microvascular pathological features and changes in related injury factors in a rat acute blood stasis model

OBJECTIVE: To examine the microvascular pathological characteristics and changes in related injury factors in a rat model of acute blood stasis.METHODS: A total of 75 Sprague-Dawley rats were divided randomly and equally into a control group and four experimental groups assessed at different times after the induction of stasis(0, 1, 3 or 6 h after stasis)(n = 15). The acute blood stasis model was established through rat tail-vein injection of high-molecular-weight dextran. After Electrocardiograph(ECG) detection at predetermined times(0,1, 3 and 6 h after induction of stasis), the rats were sacrificed and blood and cardiac samples were harvested for analysis. Hematoxylin-eosin(HE) staining and transmission electron microscopy were used for histopathological detection; an enzyme linked immunosorbent assay(ELISA) was used to detect thromboxane B2(TXB2) and 6-keto-prostaglandin F1α(6-Keto-PGF1α) concentrations; a real-time polymerase chain reaction(PCR) reaction system was used to detect intercellular adhesion molecule1(ICAM-1) and vascular cell adhesion molecule1(VCAM-1) m RNA expression; western blotting was used to detect vascular endothelial cadherin(VE-cadherin) protein expression.RESULTS: The ST segment in the ECG showed gradual elevation after induction of stasis and continued elevation at a high level at 3 and 6 h. The HE staining showed changes in myocardial cell necrosis and tissue dissociation after the induction of stasis, along with inflammatory infiltration. Results of transmission electron microscopy showed immediate changes in blood stasis and lumen occlusion in the microvasculature, along with endothelial cell swelling. After the induction of stasis, TXB2 concentrations gradually increased while 6-Keto-PGF1αconcentrations were immediately significantly reduced. The TXB2/6-Keto-PGF1αratio was maintained at a high level. ICAM-1 m RNA expression showed an unstable elevation while VCAM-1 m RNA expression was significantly reduced after the induction of stasis. Compared with the control group, VE-cadherin protein expression increased at 0 and 3 h after the induction of stasis, while no change occurred at 1 and 6 h.CONCLUSION: The pathological manifestations of acute blood stasis are microvascular blood retention, lumen stenosis and even occlusion. The condition is also called "blood coagulation and weep" in Traditional Chinese Medicine. The blood stasis model resulted in the injury and necrosis of endothelial cells and cardiomyocytes, along with the presence of an imbalance of vasomotor factor levels, platelet activation, and increases in the expression of adhesion molecules and endothelial barrier dysfunction,which corresponds to "blood failed to nourish" in Traditional Chinese Medicine.

Soluble Triggering Receptor Expressed on Myeloid Cells-1 and Inflammatory Markers in Colorectal Cancer Surgery： A Prospective Cohort Study

Background： Major abdominal surgery, including colorectal cancer （CRC） surgery, leads to systemic inflammatory response syndrome that can be detected and monitored with inflammatory markers testing. The aims of the study were to evaluate the usefulness of soluble triggering receptor expressed on myeloid cells-l （sTREM-1 ）, interleukin-6 （IL-6）, procalcitonin （PCT）, and C-reactive protein （CRP） in following the inflammatory response in CRC surgery and postoperative period, as well as to determine if duration of the surgery and the time that the colon has been opened during the surgery （open colon time [OCT]） refect a larger surgical stress through inflammatory markers rise. Methods： The study included 20 patients who underwent CRC surgery and 19 healthy volunteers from June 2011 to September 2012. We determined inflammatory markers 1 day before surgery （T0）, 24 h （T1）, 48 h （T2）, and 7 days after the surgery （T3）. All statistical analyses were calculated using MedCalc Statistical Software version 14.8.1 （MedCalc Software bvba, Ostend, Belgium）. Results： Concentrations ofCRP, PCT, and I L-6 in all measurement times were statistically different and sTREM- 1 did not yield statistical significance. A weak positive correlation was/bund between l L-6 in T 1 and T2 with the duration of the surgery （T 1 ： r= 0.4060, P 〈 0.0001 ; T2：r =0.3430, P〈0.0001）andOCT（T1：r= 0.3640, P〈0.0001,T2：r=0.3430, P〈0.0001）.AweakpositivecorrelationbetweenCRP in T2 and OCT （r = 0.4210, P 〈 0.0001 ） was also found. The interconnectivity of tested parameters showed a weak positive correlation between CRP and IL-6 in T1 （r= 0.3680; P 〈 0.0001 ）, moderate positive correlation in T2 （r = 0.6770; P 〈 0.0001）, and a strong positive correlation in T3 （r = 0.8651; P 〈 0.0001）. Conclusions： CRP, IL-6, and PCT were shown to be reliable for postoperative monitoring. Simultaneous determination of CRP and IL-6 might not be useful as they follow similar kinetics, sTREM- 1 might not be useful in CRC postoperative monitoring.

Impact of dietary supplementation of b-hydroxybutyric acid on performance,nutrient digestibility,organ development and serum stress indicators in early-weaned goat kids

The objective of this study was to determine the effect of dietary supplementation of b-hydroxybutyric acid(BHBA)on performance,nutrient digestibility,organ development,and serum composition in earlyweaned goat kids.Sixty-four goat kids at 30 d of age were assigned to 4 treatments in a completely randomized design:1)control(basal diet);2)low(basal diet with 3 g/d per animal BHBA);3)medium(basal diet with 6 g/d per animal BHBA;and 4)high(basal diet with 9 g/d per animal BHBA).Subsequently,48(6 kids per treatment)goat kids were randomly selected and slaughtered at 60 and 90 d of age.Compared with the control group,BHBA at low and high doses increased body weight(P<0.05),average daily gain(P<0.01),and average daily starter intake(P<0.01).The BHBA improved organ development,especially at the lowest dose(P<0.01).The digestibility of dry matter and crude protein increased with age(P<0.05).However,BHBA did not affect nutrient digestibility.Compared with the control group,serum ceruloplasmin increased(P<0.05)with high BHBA level at 90 d of age.However,the serum creatinine(P<0.05)increased over time but was not affected by BHBA.The serum total antioxidant capacity and superoxide dismutase decreased with the high dose of BHBA at 90 d of age(P<0.01).In contrast,the serum glutathione peroxidase and malondialdehyde increased with the high doses of BHBA(P<0.01).Overall,low doses of BHBA were positive for growth performance,organ development,and health status against weaning stress.Whereas high doses of BHBA in the long term could negatively affect antioxidant status.

Alpha 1-antitrypsin and alpha 1-acid glycoprotein reduce the sensitivity of human dermal fibroblast to endotoxin

Objective: To test the hypothesis that acute phase reactants, such as alpha 1-antitrypsin and alpha 1-acid glycoprotein, could protect mammalian cells from further damage. Methods: Human dermal fibroblasts (5×10 4) were cultured with DMEM plus 10% FBS at 37℃ in a 5% CO 2 incubator. Different doses of LPS (lipopolysaccharide) and/or acute phase reactants were added. After 24 hours, the cultured supernatant was aspirated, the cells were washed, fixed and stained by methylene blue. The unbound stain was washed off. The stained cells were solubilized in 0.1 ml of 1% Triton X-100. The absorbance of each well was measured using an ELISA spectrophotometer. The concentration of LPS which decreased the absorbance to 70% of the control (LPS-free) cultures was defined as LD 30. Results: In order to achieve LD 30 in the presence of acute phase proteins, it was necessary to alter the LPS concentrations. The LD 30 of LPS treated with 0, 0.5, 2, 10 mg/ml antitrypsin and 0, 0.5, 2, 10 mg/ml glycoprotein was 5.4, 6.5, 7.6, 14.2 mg/ml and 5.2, 5.9, 6.9, 10.5 mg/ml, respectively. Statistically, with the treatment of more than 2 mg/ml antitrypsin or glycoprotein, LD 30 increased significantly. Conclusions: Our data show that fibroblasts are susceptible to the direct toxicity of LPS. Alpha 1-antitrypsin and alpha 1-acid glycoprotein can reduce the toxicity and/or increase the tolerance of mammalian cells to LPS.

Hypoalbuminemia: an underestimated, vital characteristic of hospitalized COVID-19 positive patients?

The COVID-19 pandemic has led to the greatest worldwide health crisis in decades.The number of infected patients with severe SARS-CoV-2(COVID-19)disease has overwhelmed the capacity of almost all health care systems around world.Hypoalbuminemia has now been reported in patients with severe disease seeking help in the emergency room because of COVID-19 infection.In the past,hypoalbuminemia was considered to be a negative prognostic marker,not only in patients with chronic liver disease,but also in patients with SARS and MERS infections.Albumin is the major serum protein synthesized by the liver.A low serum albumin level is an ominous clinical sign.Introduction of amino acids to a patient's diet is of fundamental importance to hepatic albumin synthesis in different clinical situations.This highlights the importance of nutritional support during the early phases of COVID-19-infection.Furthermore,albumin synthesis in the hepatocyte is downregulated at a pretranslational level by the direct interaction of the major acute-phase cytokines which are released into the circulation during the cytokine"storm"induced by the viral effects on the lungs.Both mechanisms contribute to severe hypoalbuminemia which,combined with massive fluid losses due to the fever,is responsible for severe hypovolemia and shock commonly observed in patients with COVID-19 in critical care settings.