Protective efficacy of pirfenidone in rats with pulmonary fibrosis induced by bleomycin.

BACKGROUND: Bleomycin causes increased production of reactive oxygen species, leads to pulmonary toxicity, fibroblast activation, and fibrosis. OBJECTIVES: This study aimed to evaluate the protective effect of pirfenidone on bleomycin-induced lung toxicity in rats. METHODS: Twenty-eight adult rats were randomly divided into 3 groups; Bleomycin (B group, n=10), Bleomycin and Pirfenidone (B-PND group, n=13), and the control group (n=5). The bleomycin regimen was administered for 9 weeks. Pirfenidone was administered at 100 mg/kg daily. Total antioxidant level (TAS), total oxidant level (TOS), tumor necrosis factor (TNF-α), transforming growth factor (TGF-ß1), matrix metalloproteinase-2 (MMP-2), plasminogen activator inhibitor (PAI) levels were studied. Histopathologically, sections were stained with Hematoxylin-eosin and Masson-trichrome for grading-scoring according to the Ashcroft score. RESULTS: Stage 3 fibrosis was observed in 50% of the B group rats, stage 3 and higher fibrosis was never detected in the B-PND group and the difference was statistically significant (p=0.003). When evaluating tissue inflammation, the inflammation was higher in the B-PND group than in the other groups (p<0.001). Pleuritis was detected in all rats in group B, while was not observed in B-PND and control group (p<0.001). The TAS level was found to be significantly higher in group B than in group B-PND (p=0.034), while no difference was found between TOS, TNF-α, MMP-2, PAI, TGF-ß1. CONCLUSIONS: Pirfenidone had a statistically significant protective effect in bleomycin-induced lung fibrosis and pleuritis in rats.  Despite the presence of inflammation in the tissue, no significant changes were observed in inflammation markers in the peripheral blood. Novel serum biomarkers are needed to indicate the presence of inflammation and fibrosis in the lung.

Plasminogen activator inhibitor-1 levels as an indicator of severity and mortality for COVID-19.

OBJECTIVE: Coronavirus disease-19 (COVID-19) is a multisystemic disease that can cause severe illness and mortality by exacerbating symptoms such as thrombosis, fibrinolysis, and inflammation. Plasminogen activator inhibitor-1 (PAI-1) plays an important role in regulating fibrinolysis and may cause thrombotic events to develop. The goal of this study is to examine the relationship between PAI-1 levels and disease severity and mortality in relation to COVID-19. METHODS: A total of 71 hospitalized patients were diagnosed with COVID-19 using real time-polymerase chain reaction tests. Each patient underwent chest computerized tomography (CT). Data from an additional 20 volunteers without COVID-19 were included in this single-center study. Each patient's PAI-1 data were collected at admission, and the CT severity score (CT-SS) was then calculated for each patient. RESULTS: The patients were categorized into the control group (n=20), the survivor group (n=47), and the non-survivor group (n=24). In the non-survivor group, the mean age was 75.3±13.8, which is higher than in the survivor group (61.7±16.9) and in the control group (59.5±11.2), (p=0.001). When the PAI-1 levels were compared between each group, the non-survivor group showed the highest levels, followed by the survivor group and then the control group (p<0.001). Logistic regression analysis revealed that age, PAI-1, and disease severity independently predicted COVID-19 mortality rates. In this study, it was observed that PAI-1 levels with >10.2 ng/mL had 83% sensitivity and an 83% specificity rate when used to predict mortality after COVID-19. Then, patients were divided into severe (n=33) and non-severe (n=38) groups according to disease severity levels. The PAI-1 levels found were higher in the severe group (p<0.001) than in the non-severe group. In the regression analysis that followed, high sensitive troponin I and PAI-1 were found to indicate disease severity levels. The CT-SS was estimated as significantly higher in the non-survivor group compared to the survivor group (p<0.001). When comparing CT-SS between the severe group and the non-severe group, this was significantly higher in the severe group (p<0.001). In addition, a strong statistically significant positive correlation was found between CT-SS and PAI-1 levels (r: 0.838, p<0.001). CONCLUSION: Anticipating poor clinical outcomes in relation to COVID-19 is crucial. This study showed that PAI-1 levels could independently predict disease severity and mortality rates for patients with COVID-19.

STAT3 mediated regulation of glucose metabolism in leukemia cells.

Cancer cells rewire metabolic pathways as they demand more ATP and building blocks for proliferation. Glucose is the most consumed nutrient by cancer cells and metabolized to lactate even in the presence of oxygen. This phenomenon is called 'aerobic glycolysis'. Also, glucose level is found lower in tumor environment. Leukemia is characterized by abnormal proliferation of hematopoietic cells. STAT3 a transcription factor and an oncogene is upregulated in many tumor types. Despite its well-defined functions, STAT3 has also been proposed as a metabolic regulator. In this study, we aimed to determine the role STAT3 activation in glucose limitation, in leukemia cell lines. K562, NB-4 and HL-60 cells were found sensitive to glucose limitation. In low glucose conditions, total and nuclear STAT3 protein was decreased in all cells. In mitochondria, S727 phosphorylated STAT3 (mitochondrial form) was determined slightly increased in K562 and NB-4 cells. On the other side, ectopically STAT3 expressing cells had increased glucose consumption and less proliferated in low glucose medium. This data suggests that aerobic glycolysis might be upregulated upon STAT3 expression in leukemia cells, in glucose limitation. Furthermore, in this study, it was found that GLUT3 expressing cells did not reduce STAT3 expression in low glucose medium. GLUT3 was previously determined as a molecular marker for cell sensitivity to glucose limitation, therefore, it could be hypothesized as GLUT3 expressing cells might not need to alter STAT3 expression in low glucose level. Overall, our data suggest that leukemia cells rewire glucose metabolism via STAT3 expression in glucose limitation. Elucidating pathways that cause differential phosphorylation of STAT3 and its interaction with other energy regulating pathways in cellular response to glucose limitation might be beneficial to design new drug targets such as STAT3 inhibitors for leukemia treatment.

The effects of serum leptin levels on thrombocyte aggregation in peritoneal dialysis patients.

OBJECTIVE: Serum leptin levels of chronic kidney disease patients have been detected higher than normal population. The aim of this study was to investigate the effects of serum leptin levels on thrombocyte aggregation in peritoneal dialysis patients. METHODS: Fourty three peritoneal dialysis patients were included in the study. Thrombocyte aggregation was calculated from the whole blood subsequently the effects of different concentrations of human recombinant leptin on thrombocyte aggregations were investigated. Four test cells were used for this process. While leptin was not added into the first test cell, increasing amounts of leptin was added into the second, third and fourth test cells to attain the concentrations of 25, 50 and 100 ng/ml respectively. RESULTS: Thrombocyte aggregation was inhibited by recombinant leptin in peritoneal dialysis patients. Thrombocyte aggregation mean values were found statistically significantly higher in first test cell when compared to leptin groups in peritoneal dialysis patients. For leptin groups we could not find any statistically significant differences for thrombocyte aggregation mean values between any of the groups. CONCLUSION: Further studies with larger number of peritoneal dialysis patients are required to prove the action of leptin on thrombocyte aggregation.

Urinary nerve growth factor and a variable solifenacin dosage in patients with an overactive bladder.

INTRODUCTION AND HYPOTHESIS: We evaluated changes in urinary nerve growth factor (NGF) and NGF/creatinine (NGF/Cr) levels after increasing the dosage of solifenacin in overactive bladder patients. METHODS: The study groups included 59 overactive bladder (OAB) patients and 20 healthy subjects as controls. We measured NGF at baseline for the patients and controls, and used the Overactive Bladder Awareness Tool (OAB-V8) to evaluate urinary symptoms. All patients received a treatment of solifenacin 5 mg for 6 weeks. The responders to treatment served as group 1 and nonresponders received solifenacin 10 mg for an additional 6 weeks. Responders and nonresponders to the 10-mg treatment were defined as groups 2 and 3 respectively. NGF was measured after each treatment using the ELISA method and normalized by the urinary creatinine levels (NGF/Cr). RESULTS: There were 21, 22 and 16 patients in groups 1, 2, and 3 respectively. At baseline, the NGF and NGF/Cr levels were higher in groups 1, 2, and 3 compared with the controls. After the solifenacin 5 mg treatment, the NGF and NGF/Cr levels of group 1 individuals decreased to those of the control level. After increasing the dosage of solifenacin to 10 mg in group 2, the NGF and NGF/Cr levels decreased to normal levels. In group 3 (patients who did not responded to any treatment), these levels remained unchanged. CONCLUSIONS: Our results suggest that urinary NGF could be a potential biomarker for monitoring the treatment of symptoms in OAB patients who are treated with solifenacin.

Soluble Programmed Death 1 (PD-1) Is Decreased in Patients With Immune Thrombocytopenia (ITP): Potential Involvement of PD-1 Pathway in ITP Immunopathogenesis.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by dysregulation of T cells. Programmed death (PD) 1 and programmed death 1 ligand 1 (PD-L1) are cosignaling molecules, and the major role of the PD-1 pathway is the inhibition of self-reactive T cells and to protect against autoimmune diseases. We measured levels of serum soluble PD 1 (sPD-1) and serum soluble PD-L1 (sPD-L1) in 67 patients with ITP (24 newly diagnosed ITP [ndITP], 43 chronic ITP [cITP]) and 21 healthy controls (HCs). We determined decreased serum sPD-1 levels both in patients with ndITP and in patients with cITP when compared to HC. Moreover, there was a positive correlation between sPD-1 levels and platelet counts. The sPD-L1 levels were decreased in patients with ndITP when compared to patients with cITP. This is the first study investigating PD-1 signaling pathway in ITP. Decreased sPD-1 levels may have a role in ITP pathogenesis as without the inhibitory regulation of PD-1, sustained activation of T cells may cause inflammatory responses which is the case in ITP.

Diagnostic value of S100B protein in the differential diagnosis of acute vertigo in the emergency department.

OBJECTIVES: Vertigo is a common presenting complaint resulting from central or peripheral etiologies. Because central causes may be life-threatening, ascertaining the nature of the vertigo is crucial in the emergency department (ED). With a broad range of potential etiologies, distinguishing central causes from benign peripheral causes is a diagnostic challenge. Cranial magnetic resonance imaging (MRI) is the recommended neuroimaging method when clinical findings are ambiguous. However, MRI scanning for every patient with an uncertain diagnosis may not be efficient or possible. Therefore, to improve ED resource utilization for patients with vertigo, there is a need to identify the subset most likely to have MRI abnormalities. It has previously been shown that S100B protein provides a useful serum marker of stroke, subarachnoid hemorrhage, and traumatic brain injury. This study evaluated whether S100B levels could predict central causes of vertigo as identified by cranial MRI in the ED. METHODS: This prospective, observational study was conducted with adult patients with acute-onset vertigo (within 6 hours) in the ED of a teaching hospital in Kocaeli, Turkey. Patients with nausea or dizziness complaints without previously known vertigo or cranial pathology, and who agreed to participate in the study, were included. Patients with trauma or with neurologic findings that developed concurrent with their symptoms were excluded. Serum levels of S100B were measured with an electrochemiluminescence immunoassay kit. All subjects underwent cranial MRI. The predictors of positive MRI results were evaluated using logistic regression analysis. Sensitivity and specificity of S100B​ levels for identifying subjects with central causes of vertigo on MRI were calculated with receiver operating characteristic (ROC) curve. RESULTS: Of the 82 subjects included in the study, 48 (58.5%) were female, and the mean (±SD) age was 51 (±16) years. Thirty-one (37.8%) subjects had positive MRI results. Median (with interquartile range [IQR]) serum S100B levels were significantly different between MRI-negative and MRI-positive groups (median = 27.00 pg/mL, IQR = 10.00 to 44.60 vs. median = 60.94 pg/mL, IQR = 38.25 to 77.95, respectively; p = 0.04). In logistic regression analysis, subjective "he or she is spinning" (p = 0.030, odds ratio [OR] = 1.63, 95% confidence interval [CI] = 1.38 to 2.49), systolic blood pressure (sBP; p = 0.045, OR = 1.044, 95% CI = 1.021 to 1.080), and serum S100B level (p = 0.042, OR = 1.22, 95% CI = 1.018 to 1.445) were found to be independent predictors of MRI abnormalities. In the ROC analysis, S100B > 30 pg/mL predicted the clinical outcome with 83.9% sensitivity (95% CI = 66.3% to 94.5%) and 51.0% specificity (95% CI = 36.6% to 65.2%). The area under the ROC curve was 0.774 (95% CI = 0.666 to 0.881). CONCLUSIONS: To the best of our knowledge this is the first study assessing the utility of serum S100B levels for diagnosis of acute-onset vertigo. Serum S100B levels are associated with the presence of central causes of vertigo on cranial MRI. However, serum S100B levels are not sufficiently sensitive to exclude candidates from cranial MRI.

Correlation of Cystatin-C and radionuclidic measurement method of glomerular filtration rate in patients with lung cancer receiving cisplatin treatment.

OBJECTIVE: Cisplatin is a chemotherapeutic agent which affects renal functions adversely. The best indicator of renal functions is glomerular filtration rate (GFR) measurement. Cystatin-C appears to be a good alternative to existing methods of measuring GFR. However, it is controversial whether Cystatin-C demonstrates GFR correctly for patients receiving chemotherapy. This study aimed to investigate the correlation between GFR values calculated by Cystatin-C based formulas, radionuclidic method (multiple blood sampling) and blood Cystatin-C values in patients with lung cancer, receiving cisplatin treatment in both pre-treatment and post-treatment periods. MATERIALS AND METHODS: Thirty-six patients with lung cancer who were going to receive cisplatin treatment were included in this study. However, the evaluation was performed with 20 patients since 16 of them could not complete the treatment. Blood Cystatin-C values, GFR values calculated via Cystatin-C based formulas, and radionuclidic method were investigated before and after the cisplatin treatment. RESULTS: After treatment significant decreases were detected in GFR values, obtained via radionuclidic measuring method. However, there was no significant difference in Cystatin-C values between pre-treatment and post-treatment periods. Also GFR values obtained by Cystatin-C based formulas were not significantly different in pre-treatment and post-treatment periods. There were meaningful correlations between radionuclidic method and Cystatin-C values and Cystatin-C based formulas before treatment. However, all correlations disappeared after the treatment. CONCLUSION: GFR values, calculated by Cystatin-C may not be reliable in following renal functions in patients receiving chemotherapy. When reliable monitoring of the renal functions is necessary radionuclidic method may be preferred in these patients.

Aqueous humor and serum erythropoietin levels in patients with pseudoexfoliation syndrome and pseudoexfoliative glaucoma.

A prospective, non-randomized, comparative study was carried out to investigate aqueous humor and serum erythropoietin (EPO) levels in patients with pseudoexfoliation syndrome (PXS), pseudoexfoliative glaucoma (PXG), and, as controls, senile cataract without glaucoma. Aqueous humor was aspirated at the beginning of cataract or combined glaucoma-cataract surgery from 24 patients with PXS, 24 patients with PXG, and 24 patients with cataract only. Venous blood samples were obtained from all patients at the same time. Levels of EPO were measured in aqueous humor and venous blood using a sandwich enzyme-linked immunosorbent assay (ELISA) kit. There was no statistically significant difference between the mean aqueous humor EPO concentrations in eyes with PXS (10.70 ± 7.18 mU/ml), PXG (10.18 ± 7.20 mU/ml) and controls (9.75 ± 5.04 mU/ml) (P = 0.732). There was no statistically significant difference between the mean serum EPO concentrations in eyes with PXS (6.32 ± 2.86 mU/ml), PXG (7.06 ± 4.35 mU/ml) and controls (7.30 ± 4.15 mU/ml) (P = 0.672). Contrary to predictions based on the relation with ischemic processes, no difference was revealed between levels of EPO in aqueous humor and serum in patients with PXS and PXG.

Protective effect of a potent antioxidant, pomegranate juice, in the kidney of rats with nephrolithiasis induced by ethylene glycol.

PURPOSE: We aimed to study the protective effects of pomegranate juice (PJ) on ethylene glycol (EG)-induced crystal deposition in renal tubules, renal toxicity, and inducible nitric oxide synthase (iNOS) and nuclear factor-kappaB activities in rat kidneys. MATERIALS AND METHODS: Fifty-six rats were divided into four equal groups: Control, EG, EG + 50 microL PJ/d (PJ50), and EG + 100 microL PJ/d (PJ100). Rats were sacrified on days 10 and 45. Tissue sections were evaluated under light and polarized microscopy for the presence and degree of crystal deposition and toxicity in the kidneys. Crude extracts of the cortex were used to determine reduced gluthatione (GSH), nitric oxide (NO), and malondialdehyde (MDA) levels. RESULTS: In the EG group, crystal depositions were more evident and mild crystalization was observed in proximal tubules on day 10; severe crystalization and granulovacuolar epithelial cell degeneration were observed on day 45. There was limited or no crystal formation in the EG + PJ-given groups. There were completely normal renal and tubular structures in the control group. There was no significant difference between the four groups in serum levels of sodium, potassium, blood urea nitrogen, and creatinine in any sampling time. Hyperoxaluria, a marked increase in MDA and NO levels, and decrease of GSH were observed in the EG-given groups compared with the others. There were marked iNOS and p65 expressions in only the EG-given rats compared with control and PJ groups, immunohistochemically. CONCLUSION: This experiment shows the protective effect of PJ in the EG-induced crystal depositions in renal tubules.

Biochemical effects of experimental epidural hematoma on brain parenchyma of rats.

INTRODUCTION: The management of epidural hematoma is classified into surgical or conservative treatment according to clinical and radiologic parameters. In the recent years, the number of paper suggesting conservative management has been increasing. The experimental works that have been performed are based on especially the effects of epidural hematomas. Basic pathophysiologic factors on ischemia result of brain trauma are based on biochemical mediators. Nitric oxide (NO) and malondialdehyde (MDA) are the substances that play important roles in brain damage after trauma. MATERIAL AND METHOD: In this study, 36 rats are divided into three groups (n = 12/group). Epidural hematoma was achieved by 0.1 ml autolog blood in rat epidural space with balloon model. Early and late phase biochemical effects on parenchyma of epidural hematoma operated in a volume which neither alters intracranial pressure (ICP) nor creates shift effect were observed. Biochemical changes of NO and MDA levels were examined in each of three experimental groups. RESULTS: NO values increased significantly in the early group (6 hours) compared with those in the control group. Difference of NO values between the control and late groups was not significant. An increase has been found in MDA values in the control group compared with those in the early group. MDA values of the late group (30 days) were closer to that of the control group. CONCLUSION: In this study, considering biochemical results, we have found that conservative volumes which neither increase ICP nor cause brain shift do not lead to permanent changes on brain.

Rapid communication: protective effect of a nuclear factor kappaB inhibitor, pyrolidium dithiocarbamate, in the kidney of rats with nephrolithiasis induced by ethylene glycol.

PURPOSE: To study the protective effects of a selective nuclear factor kappa B (NF-kappaB) inhibitor, pyrolidium dithiocarbamate (PDTC), on ethylene glycol-induced crystal deposition in the renal tubules, renal toxicity, as well as inducible nitric oxide synthase (iNOS) and NF-kappaB activities in rat kidneys. MATERIALS AND METHODS: Rats were divided into three equal groups: control, ethylene glycol-treated (EG), and ethylene glycol + PDTC treated (EG+PDTC). Rats were sacrificed on day 7, 15, or 45, and tissue sections were evaluated under light and transmission electron microscopy for the presence and degree of crystal deposition and toxicity in the kidneys. The iNOS and NF-kappaB activity were evaluated immunohistochemically, with p65 being stained to define NF-kappaB activity. Crude extracts of the cortex were used to determine reduced glutathione (GSH), nitric oxide (NO), and malondialdehyde (MDA) concentrations. RESULTS: Crystal depositions were more evident in the proximal tubules on day 7 in the EG than in the other groups. Mild crystallization was observed on day 15, and severe crystallization and granulovacuolar epithelial-cell degeneration were observed on day 45. There was limited or no crystal formation in the EG+PDTC group and completely normal renal and tubular structures in the control group. Whereas ethylene glycol administration stimulated iNOS and NF-kappaB/p65 activity in renal tubules, PDTC inhibited it. Rats given only vehicle demonstrated no significant alterations. Hyperoxaluria, a marked increase in MDA and NO concentrations, and a decrease in GSH were observed in the EG group. CONCLUSION: This experiment has shown the role of transcription factors, NF-kappaB, and iNOS in ethylene glycol-induced crystal depositions in renal tubules.

Persistency of high proinflammatory cytokine levels from colostrum to mature milk in preeclampsia.

OBJECTIVES: Recent evidence suggests a role of an excessive maternal inflammatory response in the pathogenesis of preeclampsia. Whether this imbalance can be transferred from mother to breast milk remains to be established. DESIGN AND METHODS: 15 preeclamptic and 15 healthy pregnant women were recruited in this study. Colostrum and milk samples were collected postpartum in the first 48 h and at 30 days, respectively. Samples were analyzed for interleukin (IL)-1beta, IL-6, IL-8, tumor necrosis factor (TNF)-alpha and soluble IL-2R (sIL-2R) levels with chemiluminescence enzyme immunometric assays. RESULTS: Colostrum cytokine levels corrected for gestational age and type of delivery were not significantly different in the two groups. Cytokine levels significantly decreased in mature milk versus colostrum in the control group (P < 0.05), but did not significantly decrease in the preeclampsia group (P > 0.05), except for TNF-alpha (P < 0.05). Mature milk IL-8 and TNF-alpha levels were higher in the preeclampsia group versus controls (P < 0.05). CONCLUSION: Results of this study show that proinflammatory cytokines in breast milk exhibit biological variation at different periods of human lactation. In preeclampsia, high cytokine levels persist at least for 30 days. These results suggest that preeclampsia may affect milk cytokine balance and offer an immunological signal for the host defense in high-risk neonates.

Decreased adrenomedullin and total nitrite levels in breast milk of preeclamptic women.

OBJECTIVE: To observe the adrenomedullin (AM) and total nitrite levels in the milk of preeclamptic and normal pregnant women. DESIGN AND METHODS: Fifteen women with preeclampsia and 15 normal pregnant women were included in the study. Total nitrite was quantitated by Griess reaction, while AM was measured by HPLC. RESULTS: The levels of AM and total nitrite in colostrum and 30th-day breast milk were decreased in preeclamptics. Total nitrite levels (micromol/l) were 56.09 +/- 11.18 vs. 82.20 +/- 12.01, P < 0.05, in colostrum of preeclamptics and controls, respectively. The level of total nitrite was 37.75 +/- 12.10 vs. 53.28 +/- 10.25, P < 0.05, in 30th-day milk of same patients. AM levels (pg/ml) were 11.18 +/- 1.11 vs. 16.59 +/- 1.24, P < 0.0001, in colostrum of preeclamptics and controls, respectively. In 30th-day milk of same patients, AM levels were 8.41 +/- 1.39 vs. 12.18 +/- 1.48, P < 0.005, respectively. CONCLUSION: This report shows for the first time that human milk has decreased levels of AM and total nitrite in preeclampsia.

Plasma lipid and lipoprotein concentrations in pregnancy induced hypertension.

OBJECTIVES: Aim of this study was to evaluate implication of pregnancy induced hypertension on maternal plasma lipid, lipoprotein, apolipoprotein concentrations and lipid peroxidation products by a comparison of normal pregnancy vs. preeclampsia. DESIGN AND METHODS: Thirty-four women with preeclampsia and 32 healthy pregnant women (controls) in the third trimester were recruited for this study. RESULTS: In the preeclamptic group plasma total triglyceride, low density lipoprotein cholesterol (LDL-C), malondialdehyde (MDA) and apolipoprotein B (apo-B) were significantly increased, while plasma high density lipoprotein cholesterol (HDL-C) was significantly decreased compared to that of control group. There was no significant difference in total cholesterol and apolipoprotein A1 (apo-A1) concentrations. CONCLUSION: Our findings suggest that preeclampsia share some metabolic characteristics with coronary artery disease such as dislipidemia and increased lipid peroxidation. However lipoprotein concentrations may be better biochemical markers of dislipidemia in the preeclamptic state than the corresponding apolipoproteins.