Therapeutic Dose Group Research Articles

The potential role of upregulated PARP-1/RIPK1 expressions in amikacin-induced oxidative damage and nephrotoxicity in Wistar rats.

This study aimed to investigate the gene expression levels associated with nephrotoxic action of amikacin, as well as the post-treatment effect of diuretics on its nephrotoxic effects. Sixty male rats were divided equally into six groups, including the control group receiving saline intra-peritoneally (ip), and the five treated groups including therapeutic and double therapeutic dose groups, injected ip (15 and 30mg/kg b.wt./day) respectively for seven days, and another two rat groups treated as therapeutic and double therapeutic dose groups then administered the diuretic orally for seven days and the last group received amikacin ip at a rate of 15mg/kg/day for seven days, then given free access to water without diuretics for another seven days and was kept as a self-recovery group. Amikacin caused kidney injury, which was exacerbated by the double therapeutic dose, as evidenced by abnormal serum renal injury biomarkers, elevated renal MDA levels, inhibition of renal catalase and SOD enzyme activities, with renal degenerative and necrotic changes. Moreover, comet assays also revealed renal DNA damage. Interestingly, amikacin administration markedly elevated expression levels of the PARP-1, RIP1, TNF-α, IL-1β, and iNOS genes as compared to the control group. However, compared to the self-recovery group, post-amikacin diuretic treatment modulates amikacin-induced altered findings and alleviates amikacin nephrotoxic effects more efficiently. Our findings suggested the potential role of PARP-1 and RIPK1 expressions that influence the expression of proinflammatory cytokines such as IL-1β and TNF-α by exaggerating oxidative stress which may contribute to the pathogenesis of amikacin-induced nephrotoxicity.

The Effectiveness of the Intermediate and Therapeutic Doses of Enoxaparin in COVID-19 Patients: A Comparative Study of Factor Xa Inhibition

Background: Management of anticoagulant therapy in COVID-19 patients is critical. Low-molecular-weight heparin (LMWH) thromboprophylaxis is already recommended, and anti-Factor Xa (anti-FXa) monitoring has been used to titrate LMWH doses. Methods: Through a cross-sectional study, we evaluated anti-FXa activity in patients admitted to the ICU, receiving intermediate dose (30, 40, 50 mg, subcutaneously [SC], twice daily) or therapeutic dose (1 mg/kg, SC, Q12h) of enoxaparin to find whether the patients in these two groups achieved anti-FXa levels in the accepted thromboprophylaxis range. Results: The occurrence of deep vein thrombosis was 26% in the therapeutic-dose group and 17% in the intermediate-dose group. D-dimer values were nearly 3.5-fold higher in those who received a therapeutic dose of anticoagulants than in those who received intermediate-dose thromboprophylaxis. Patients in the therapeutic-dose group had significantly higher IL-6 levels (p ≤ 0.001). More than one-third of the patients in the therapeutic-dose group (n = 8; 42.18%) and approximately half of the patients in the intermediate-dose group (n = 12; 52.2%) achieved the target range level of anti-FXa. Patients who received therapeutic doses were more likely to have anti-FXa levels above the expected range (47.4 vs 13% in the intermediate-dose group; p < 0.05). Conclusion: Therapeutic dose of enoxaparin in critically ill COVID-19-infected patients did not reduce the incidence of thromboembolic events and, on the other hand, may predispose these patients to increased risk of bleeding by increasing anti-FXa activity above the desired level. Administration of intermediate-dose thromboprophylaxis is suggested to achieve anti-FXa levels in the accepted thromboprophylaxis range.

Anticoagulation strategies and risk of bleeding events in critically ill COVID-19 patients

ObjectiveTo evaluate the rate of thrombosis, bleeding and mortality comparing anticoagulant doses in critically ill COVID-19 patients. DesignRetrospective observational and analytical cohort study. SettingCOVID-19 patients admitted to the intensive care unit of a tertiary hospital between March and April 2020. Patients201 critically ill COVID-19 patients were included. Patients were categorized into three groups according to the highest anticoagulant dose received during hospitalization: prophylactic, intermediate and therapeutic. InterventionsThe incidence of venous thromboembolism (VTE), bleeding and mortality was compared between groups. We performed two logistic multivariable regressions to test the association between VTE and bleeding and the anticoagulant regimen. Main variables of interestVTE, bleeding and mortality. Results78 patients received prophylactic, 94 intermediate and 29 therapeutic doses. No differences in VTE and mortality were found, while bleeding events were more frequent in the therapeutic (31%) and intermediate (15%) dose group than in the prophylactic group (5%) (p<0.001 and p<0.05 respectively). The anticoagulant dose was the strongest determinant for bleeding (odds ratio 2.4, 95% confidence interval 1.26–4.58, p=0.008) but had no impact on VTE. ConclusionsIntermediate and therapeutic doses appear to have a higher risk of bleeding without a decrease of VTE events and mortality in critically ill COVID-19 patients.

Biological Responses of Nile tilapia Oreochromis niloticus as Influenced by Dietary Florfenicol

Antibiotics are used in the treatment of bacterial diseases in commercial aquaculture. In this study, we the biological responses of Oreochromis niloticus juveniles upon dietary florfenicol (FFC) administration at 15 mg (1×) and 45 mg kg biomass−1 day−1 (3×) for 10 days in terms of feed intake, survival, biomass, hematological, erythro-morphological, serum biochemical, and histopathological aberrations as compared with controls. FFC caused a dose-dependent reduction in feed intake, survival, and biomass, with marked variations in hematology, hematological indices, and erythrocytic cellular and nuclear abnormalities, suggesting its apparent cytotoxic and nucleotoxic effects. The serum biomarkers increased significantly in a dose-dependent manner, except for calcium and chloride, which decreased significantly. The therapeutic dose (1×) group exhibited marked histopathological aberrations, such as renal tubular epithelial degeneration and a widened lumen in the kidney, as well as glycogen-type vacuolation and cytoplasmic degeneration in the liver during the dosing period. The extent of kidney and liver tissue damage was more prominent in the 3× group. The 1× serum biomarker levels became normal, with the exception of alkaline phosphatase, within 3 weeks of suspension of dosing. The recovery of the measured parameters and histopathological and erythro-morphological changes suggested that the therapeutic dietary biological responses induced by FFC are reversible and safe for O. niloticus.

Efficacy and Safety of Tinzaparin in Prophylactic, Intermediate and Therapeutic Doses in Non-Critically Ill Patients Hospitalized with COVID-19: The PROTHROMCOVID Randomized Controlled Trial.

Hospitalized patients with COVID-19 are at increased risk of thrombosis, acute respiratory distress syndrome and death. The optimal dosage of thromboprophylaxis is unknown. The aim was to evaluate the efficacy and safety of tinzaparin in prophylactic, intermediate, and therapeutic doses in non-critical patients admitted for COVID-19 pneumonia. PROTHROMCOVID is a randomized, unblinded, controlled, multicenter trial enrolling non-critical, hospitalized adult patients with COVID-19 pneumonia. Patients were randomized to prophylactic (4500 IU), intermediate (100 IU/kg), or therapeutic (175 IU/kg) groups. All tinzaparin doses were administered once daily during hospitalization, followed by 7 days of prophylactic tinzaparin at discharge. The primary efficacy outcome was a composite endpoint of symptomatic systemic thrombotic events, need for invasive or non-invasive mechanical ventilation, or death within 30 days. The main safety outcome was major bleeding at 30 days. Of the 311 subjects randomized, 300 were included in the prespecified interim analysis (mean [SD] age, 56.7 [14.6] years; males, 182 [60.7%]). The composite endpoint at 30 days from randomization occurred in 58 patients (19.3%) of the total population; 19 (17.1 %) in the prophylactic group, 20 (22.1%) in the intermediate group, and 19 (18.5%) in the therapeutic dose group (p = 0.72). No major bleeding event was reported; non-major bleeding was observed in 3.7% of patients, with no intergroup differences. Due to these results and the futility analysis, the trial was stopped. In non-critically ill COVID-19 patients, intermediate or full-dose tinzaparin compared to standard prophylactic doses did not appear to affect the risk of thrombotic event, non-invasive ventilation, or mechanical ventilation or death. Trial Registration ClinicalTrials.gov Identifier (NCT04730856). Edura-CT registration number: 2020-004279-42.

Impact of thiopurine dose in anti-tumor necrosis factor combination therapy on outcomes in inflammatory bowel disease.

Combination therapy with thiopurines and anti-tumor necrosis factor (TNF) is superior to monotherapy in Crohn's disease (CD) and ulcerative colitis (UC). The optimal dose of thiopurines in combination therapy remains unclear. We investigated the impact of thiopurine dose in combination therapy on outcomes in inflammatory bowel disease (IBD). This was a single-center, retrospective study of patients with IBD treated with thiopurine and anti-TNF combination therapy between 1/2012 and 11/2020. A therapeutic dose of thiopurines was defined as ≥1 mg/kg for 6-mercaptopurine and ≥2 mg/kg for azathioprine. The primary outcome was anti-drug antibody (ADA) formation in patients on a therapeutic thiopurine dose vs. a lower thiopurine dose group. Secondary outcomes included steroid-free clinical remission, endoscopic healing (absence of ulcers/erosions in CD and Mayo endoscopic score ≤1 for UC), and normal serum C-reactive protein (CRP) in patients who were on combination therapy. A total of 108 patients were included (median age 31.5 years; 58.3% male). A therapeutic dose of thiopurine was used in 19%. In the therapeutic thiopurine dose group, 23.8% developed ADA vs. 29.9% (P=0.58) in the lower dose group. No significant differences were noted between the therapeutic and lower dose thiopurine groups in terms of steroid-free clinical remission (57.1% vs. 60.9%, P=0.75), endoscopic healing (55% vs. 60%, P=0.69), and normal CRP (52.4% vs. 52.9%, P=0.27). In our cohort of patients with IBD on anti-TNF combination therapy, thiopurine dose was not associated with significant differences in anti-TNF immunogenicity and clinical outcomes.

Histological effect of rifampicin drug in the kidney and liver of tissue of albino mice Mus Musclus

The aim of this study is to investigate the histopathological changes induced by the anti-tuberculosis durg (rifampicin) in the tissues of the kidneys and liver of white male mice of Mus Musculus. 25 mice were distributed into five groups that included a control group, a therapeutic dose group of 1.25 mg, and a toxic dose group of 2.5 mg. The semi-lethal LD dose group was 150 mg, and the lethal LD dose group was 240 mg.&#x0D; The results also showed behavioral changes for the treated animals, the intensity of which increased with the increase in concentrations, and the total of these changes was represented by eating less food and drinking water, lack of movement, diarrhea, rapid breathing, and red-orange urine, and these symptoms were more severe in the fatal group and ended with the animal's death.&#x0D; In the kidneys, the tissue lesions were represented by the occurrence of necrosis in the cells of the urinary tubules in some areas, the infiltration inflammatory cells clearly, the degeneration of some cells of the urinary tubules, and the swelling of a part of the cells, which led to a narrowing in the lumen of the urinary tubules. The nuclei are necrotic and disintegrated. The glomerulus was some areas enlarged and some areas atrophic. Also, fibrosis of the kidney tissue was observed, which increases with increasing dose.&#x0D; In the liver, it was represented by the occurrence of necrosis, hemorrhage and congestion, infiltration inflammatory cell thickening of the plasma membrane, widening of the sinusoids some areas and narrowing them at other areas the onset of fibrosis, kuffper cells.&#x0D; Conclusion: The administration of rifampicin led to pathological changes in the kidneys, represented by necrosis, glomerulosclerosis and fibrosis, as well as pathological changes in the liver represented by rupture of the vessel wall, nuclei enlargement and hemorrhage.

Histological effect of isoniazid drug in the kidney and liver tissues of albino mice Mus Musuclus

The aim of this study is to investigate the histopathological changes induced by the anti-tuberculosis drug (isoniazid) in the kidney and liver tissues of white male mice Mus Musculus. 25 mice were distributed into five groups that included a control group, a therapeutic dose group of 0.625 mg, a group of 1.25 mg toxic group and a dose group The semi-lethal (LD50) is 80 mg, and the lethal dose LD range is 150 mg.&#x0D; Oral administration of isoniazid led to pathological changes. In the kidneys, the tissue lesions were represented by the occurrence of necrosis in the cells of the urinary tubules in some areas, and the infiltration of inflammatory cells clearly, and the desquamation of some cells of the urinary tubules, and the swelling of some cells, which led to a narrowing in the lumen of the urinary tubules. The nuclei are necrotic and disintegrated. The glomerulus was some areas enlarged and some areas atrophic.&#x0D; As of the liver, it was represented by the occurrence of necrosis, hemorrhage and congestion, infiltration of inflammatory cell, thickening of the plasma membrane, widening of the sinusoids some araes and narrowing them at other times.&#x0D; Conclusion&#x0D; The administration of isoniazid led to pathological changes in the kidneys, including necrosis, infiltration of inflammatory cell, glomerulosclerosis, nucleoli, as well as changes in the liver, including hemorrhage, rupture of the vein wall, and hemolysis of blood cells

Efficacy and Safety of Therapeutic-Dose Heparin vs Standard Prophylactic or Intermediate-Dose Heparins for Thromboprophylaxis in High-risk Hospitalized Patients With COVID-19

Hospitalized patients with COVID-19 are at risk for venous and arterial thromboembolism and death. Optimal thromboprophylaxis dosing in high-risk patients is unknown. To evaluate the effects of therapeutic-dose low-molecular-weight heparin (LMWH) vs institutional standard prophylactic or intermediate-dose heparins for thromboprophylaxis in high-risk hospitalized patients with COVID-19. The HEP-COVID multicenter randomized clinical trial recruited hospitalized adult patients with COVID-19 with D-dimer levels more than 4 times the upper limit of normal or sepsis-induced coagulopathy score of 4 or greater from May 8, 2020, through May 14, 2021, at 12 academic centers in the US. Patients were randomized to institutional standard prophylactic or intermediate-dose LMWH or unfractionated heparin vs therapeutic-dose enoxaparin, 1 mg/kg subcutaneous, twice daily if creatinine clearance was 30 mL/min/1.73 m2 or greater (0.5 mg/kg twice daily if creatinine clearance was 15-29 mL/min/1.73 m2) throughout hospitalization. Patients were stratified at the time of randomization based on intensive care unit (ICU) or non-ICU status. The primary efficacy outcome was venous thromboembolism (VTE), arterial thromboembolism (ATE), or death from any cause, and the principal safety outcome was major bleeding at 30 ± 2 days. Data were collected and adjudicated locally by blinded investigators via imaging, laboratory, and health record data. Of 257 patients randomized, 253 were included in the analysis (mean [SD] age, 66.7 [14.0] years; men, 136 [53.8%]; women, 117 [46.2%]); 249 patients (98.4%) met inclusion criteria based on D-dimer elevation and 83 patients (32.8%) were stratified as ICU-level care. There were 124 patients (49%) in the standard-dose vs 129 patients (51%) in the therapeutic-dose group. The primary efficacy outcome was met in 52 of 124 patients (41.9%) (28.2% VTE, 3.2% ATE, 25.0% death) with standard-dose heparins vs 37 of 129 patients (28.7%) (11.7% VTE, 3.2% ATE, 19.4% death) with therapeutic-dose LMWH (relative risk [RR], 0.68; 95% CI, 0.49-0.96; P = .03), including a reduction in thromboembolism (29.0% vs 10.9%; RR, 0.37; 95% CI, 0.21-0.66; P < .001). The incidence of major bleeding was 1.6% with standard-dose vs 4.7% with therapeutic-dose heparins (RR, 2.88; 95% CI, 0.59-14.02; P = .17). The primary efficacy outcome was reduced in non-ICU patients (36.1% vs 16.7%; RR, 0.46; 95% CI, 0.27-0.81; P = .004) but not ICU patients (55.3% vs 51.1%; RR, 0.92; 95% CI, 0.62-1.39; P = .71). In this randomized clinical trial, therapeutic-dose LMWH reduced major thromboembolism and death compared with institutional standard heparin thromboprophylaxis among inpatients with COVID-19 with very elevated D-dimer levels. The treatment effect was not seen in ICU patients. ClinicalTrials.gov Identifier: NCT04401293.

Anticoagulation strategies and risk of bleeding events in critically ill COVID-19 patients.

ObjectiveTo evaluate the rate of thrombosis, bleeding and mortality comparing anticoagulant doses in critically ill COVID-19 patients.DesignRetrospective observational and analytical cohort study.SettingCOVID-19 patients admitted to the intensive care unit of a tertiary hospital between March and April 2020.Patients201 critically ill COVID-19 patients were included. Patients were categorized into three groups according to the highest anticoagulant dose received during hospitalization: prophylactic, intermediate and therapeutic.InterventionsThe incidence of venous thromboembolism (VTE), bleeding and mortality was compared between groups. We performed two logistic multivariable regressions to test the association between VTE and bleeding and the anticoagulant regimen.Main variables of interestVTE, bleeding and mortality.Results78 patients received prophylactic, 94 intermediate and 29 therapeutic doses. No differences in VTE and mortality were found, while bleeding events were more frequent in the therapeutic (31%) and intermediate (15%) dose group than in the prophylactic group (5%) (p < 0.001 and p < 0.05 respectively). The anticoagulant dose was the strongest determinant for bleeding (odds ratio 2.4, 95% confidence interval 1.26–4.58, p = 0.008) but had no impact on VTE.ConclusionsIntermediate and therapeutic doses appear to have a higher risk of bleeding without a decrease of VTE events and mortality in critically ill COVID-19 patients.

Therapeutic Anticoagulation May Be Associated with Reduced 14-day Mortality in Mechanically Ventilated Patients with COVID-19.

Purpose: To assess the impact of therapeutic dose versus prophylactic dose anticoagulation regimens on outcomes in mechanically ventilated patients with COVID-19. Methods: We performed a retrospective cohort analysis of consecutive mechanically ventilated adult patients with COVID-19 admitted to the intensive care unit (ICU) and initiated on anticoagulation from February 1 st to May 31 st , 2020. The primary endpoint was 14-day mortality. Secondary endpoints included 30-day mortality, hospital length of stay (LOS), duration of mechanical ventilation, major bleeding, and new thromboembolic event. Results: Of the 121 mechanically ventilated patients with COVID-19, 33 in the therapeutic-dose group and 34 patients in the prophylactic-dose group were included in the final analysis. The therapeutic-dose group had a decreased 14-day mortality compared to the prophylaxis dose group (9.1% vs 41.2%, p=0.004). In addition, 30-day mortality was also lower in the therapeutic anticoagulation group (24.2% vs. 52.9%, p=0.024). A longer hospital LOS (45.7 vs 26 days, p=0.003) and duration of mechanical ventilation (33.9 vs 13.3 days, p<0.001) were observed in patients on therapeutic anticoagulation in comparison to the prophylaxis dosing group. A higher rate of major bleeding was observed in patients who received therapeutic anticoagulation. Conclusion: In this analysis of mechanically ventilated COVID-19 patients in the ICU, therapeutic dose anticoagulation was associated with a significantly lower 14-day mortality, but increased bleeding.

The Role of D-Dimer for Optimal Thromboprophylaxis Strategy in Patients with COVID-19

Background:Novel coronavirus infection (SARS CoV-2 or COVID-19) is associated with a high risk of thrombotic complications, including macro- and micro-thrombi in major organs, leading to increased morbidity and mortality. Anticoagulant use, mainly heparin, which has both anticoagulant and anti-inflammatory properties, has been suggested as potentially beneficial. However, the optimal dose of anticoagulant for patients with COVID-19 is unknown. Establishing the optimal thromboprophylaxis strategy and determining the role of biomarkers for patient risk stratification may help to improve outcomes in COVID-19.Methods:This single-center retrospective cohort study is part of an ongoing Quality Improvement project on the use of an anti-factor Xa-driven heparin protocol, which includes a low-dose intravenous (IV) unfractionated heparin (UFH) option, being conducted at our medical center. Data on the type, dose, and indication for anticoagulation as well as outcomes including thrombosis, bleeding and survival was collected for inpatients diagnosed with COVID-19 between mid-March and June 15, 2020. To address COVID-coagulopathy we developed a d-dimer-based anticoagulation protocol for patients with COVID-19 (Figure 1). We recorded anticoagulant use as either standard prophylactic, escalated prophylactic (low-dose intravenous unfractionated heparin titrated to achieve an anti-factor Xa level of 0.1-0.3 anti-Xa units or enoxaparin 0.5mg/kg subcutaneously every 12 hours) or standard therapeutic dose used during the hospitalization. The primary endpoints assessed were ISTH-defined major and clinically relevant non-major bleeding (CRNMB) events and survival. Secondary endpoints included incidence of breakthrough thrombosis and duration of hospitalization.Results:A total of 263 patients with COVID-19 were reviewed. Of these, 68.44% of patients received prophylactic, 12.55% escalated prophylactic and 19.01% therapeutic dosage. Of total, 129 (49%) were receiving ICU level of care. No major bleeding events were observed. The incidence of CRNMB was 4.56% in the whole cohort, which did not differ significantly between the escalated prophylactic and therapeutic groups (12% and 12.12%, respectively). Patients treated with standard prophylaxis had less CRNMB (1.11%), but this was not statistically significant in a multivariate analysis that included other confounding factors such as age, sex, ethnicity, BMI, comorbidity, HASBLED bleeding risk, and sepsis induced coagulopathy score (SICS).The mortality rate was 12.6% in the whole cohort (7.22%, 21.21% and 26% in prophylactic, escalated prophylactic, and therapeutic dosage, respectively). Factors significantly associated with increased mortality included age and ICU level of care (HR 1.10, 95%CI [1.05, 1.15] and HR 20.42, 95%CI [2.84, 146.72], respectively). The use of therapeutic dose heparin and high-flow nasal cannula demonstrate a survival benefit in multivariate analysis (HR 0.13, 95%CI [0.04,0.44] and HR 0.23, 95%CI [0.07, 0.72], respectively; Figure 2).Breakthrough thrombosis occurred in 7 (2.66%) patients; 1 (0.56%), 1 (3.03%) and 5 (10%)) in prophylactic, escalated prophylactic and therapeutic dosage, respectively but very few diagnostic tests were performed during this time period. Duration of hospitalization was significantly longer in the therapeutic dose group when compared to escalated prophylaxis and standard prophylactic groups.Conclusion:In this cohort of inpatients with COVID-19, there were no major bleeding events related to any dose of heparin or LMWH prophylaxis. By multivariate analysis, implementation of a d-dimer-titrated anticoagulation strategy was not associated with increased CRNMB. Therapeutic dose heparin based on a d-dimer-driven anticoagulation protocol was associated with a survival benefit in COVID19-infected patients. Limitations of this study include the retrospective observational nature and a lack of a uniform diagnostic protocol for patients with suspected VTE. Although no significant difference in bleeding events were observed in our study subgroups, randomized clinical trials are necessary to determine optimal thromboprophylaxis strategy in the COVID-19 population.DisclosuresNo relevant conflicts of interest to declare.

Toxopathological and Cytogenetic Effects of Commercial Sweetener Aspartame after Chronic Oral Administration in Rat Pups

Objective: This study was done to evaluate the potential toxic effect of aspartame (APS) artificial sweetener after chronic oral administration in rat pups as a model for children that consumption high quantities of different types of sweets and juices from the peddler, regularly and daily. Methods: Twenty-four rat pups were divided equally into three groups. Therapeutic dose group (T1) and double dose group(T2) received 0.08, 0.16 g/kg. body weight(BW), respectively, while control group (C) received distilled water orally for 90 (d)days. Results: The result of present study revealed that there was an increased consumption of food, water and arising nervous signs(aggressive) in T1 and T2 groups comparing with control. The cytogenetic study includes the mitotic index and blast index showing a substantial decline relative to the control one in both treated groups. While the blast index showed a significant reduction(P<0.05) in T2 relative to T1 and the control group. No chromosomal aberration observed in all groups exposed to aspartame. Different histopathological lesions were recorded in the liver of T1 group represented by inflammatory cell (neutrophil & mononuclear cell) aggregation around the blood vessels. While noted in a double dose group (0.16g/ kg.bw), a granulomatous lesion with mild change in fat is shown. Conclusion: Used daily for lengthy periods ASP has cytogenetic and pathological risks.

Pharmacokinetics, pharmacodynamics and safety of a novel extrafine BDP/FF/GB combination delivered via metered-dose inhaler in healthy Chinese subjects

BackgroundBDP/FF/GB pMDI is a novel triple fixed-dose combination of extra-fine inhalation aerosol beclomethasone dipropionate (BDP)/formoterol fumarate (FF)/glycopyrronium bromide (GB). Limited data on the pharmacokinetic (PK) and pharmacodynamic (PD) properties of BDP/FF/GB fixed-dose combination in healthy subjects was available. PurposesThis study aimed to evaluate the pharmacokinetics, pharmacodynamics and safety of BDP/FF/GB pMDI in healthy Chinese subjects. MethodsThis is an open-label, parallel-group, randomized, single and multiple dose study. In the single dose group, subjects received single supra-therapeutic inhaled dose of BDP/FF/GB pMDI (BDP/FF/GB 400/24/50 µg). In the multiple dose group, subjects received therapeutic inhaled dose of BDP/FF/GB pMDI (BDP/FF/GB 200/12/25 µg), twice daily, for 7 consecutive days. Plasma BDP, B17MP, formoterol and GB were determined by a validated ultra performance liquid chromatography method with tandem mass spectrometric detection (UPLC/MS–MS). Heart rate (HR), QTcF, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were evaluated as the surrogate indicators of pharmacodynamic effects. ResultsA total of 24 subjects were randomized and 22 (11 in each group) completed the study. The dose adjusted pharmacokinetic profiles of BDP, beclomethasone-17-monopropionate (B17MP, the most active metabolite of BDP), formoterol and GB were overall similar in therapeutic and supra- therapeutic dose group, showing dose proportional increase of the systemic exposure to BDP, B17MP, formoterol and GB. The pharmacodynamic variables were within the normal range and showed no significant difference between the two groups. All the treatment-emergent adverse events (TEAEs) were mild and no severe TEAE was reported. ConclusionsDose adjusted PK profiles were similar between therapeutic and supra-therapeutic dose for all compounds, nearly dose proportional systemic exposure to B17MP, formoterol and GB after BDP/FF/GB pMDI administration in healthy Chinese subjects. BDP/FF/GB pMDI was safe and well tolerated in healthy Chinese subjects. The PK profiles were comparable to previously published data from Western European healthy Caucasian subjects.

Monosodium Glutamate Plus Artemether-lumefantrine Overdose Altered Malondialdehyde, Total Protein and Albumin Concentration in Rats

This study aimed at assessing alterations in malondialdehyde, MDA, total protein and albumin concentration in the serum and liver homogenate of monosodium glutamate (MSG)-challenged rats co-treated with artemether-lumefantrine, AL. Methods involving colourimetric estimation were employed in thirty rats randomly grouped into six (n = 5) and for seven consecutive days, fed feed and water (Group A), AL therapeutic dose (Group B), AL overdose (therapeutic dose × 5) (Group C), MSG (8000 mg/kg body weight) (Group D), AL therapeutic dose plus MSG (Group E) or AL overdose plus MSG (Group F). Total protein concentration (2.64±0.09 g/dL, 2.81±0.14g/dL, respectively) in the liver homogenate of rats exposed to MSG (group D) or MSG plus AL overdose (group F) and malondialdehyde concentration in the liver homogenate of MSG plus AL overdose-fed rats (0.45±0.04 mg/ml) lowered (P&lt;0.05) as against other groups. However, serum albumin concentration in MSG (2.59±0.13 g/dl) or AL overdose plus MSG (3.24±0.12 g/dl) fed rats was higher (P&lt;0.05) compared to the control (2.02±0.04 g/dl). The Total protein: Albumin ratio lowered while the Albumin: total protein ratio increased in rats in MSG, AL overdose plus MSG or AL overdose groups compared with the control. Thus, the apparent MSG plus AL overdose-induced adverse influence on the studied parameters and samples of non-malarial infested rats could be via compromised liver-mediated protein metabolism capacity and bio-functions following possibly enhanced protein-malondialdehyde adduct formation in the rats.

Investigation of Efficacy Enhancing and Toxicity Reducing Mechanism of Combination of Aconiti Lateralis Radix Praeparata and Paeoniae Radix Alba in Adjuvant-Induced Arthritis Rats by Metabolomics.

Combination of Aconiti Lateralis Radix Praeparata (FZ) and Paeoniae Radix Alba (BS) shows a significant effect in rheumatoid arthritis (RA). This study aimed to investigate the efficacy enhancing and toxicity reducing mechanism of combination of them in adjuvant-induced arthritis (AIA) rats by metabolomics. Rats were randomly divided into seven groups, including A (healthy control), B (model control), C1 (therapy group), C2 (efficacy enhancing group), D1 (toxicity group), and D2 (toxicity reducing group), and dexamethasone group was used as positive control. The plasma biochemical indexes showed that therapeutic dose of lipid-soluble alkaloids of FZ could significantly inhibit the concentrations of IL-1β, TNF-α, and IFN-γ in AIA rats, and combination with total glucosides of peony could further reduce the concentration of IL-1β. Then, UPLC-LTQ/Orbitrap MS with untargeted metabolomics was performed to identify the possible metabolites and pathways. Through multivariate data analysis of therapeutic dose groups (A vs. B vs. C1 vs. C2) and multivariate data analysis of toxic dose groups (A vs. B vs. D1 vs. D2), 10 and 7 biomarkers were identified based on biomarker analysis, respectively. After inducing AIA model, the plasma contents of spermidine, vanillylmandelic acid, catechol, and linoleate were increased significantly, and the contents of citric acid, L-tyrosine, L-phenylalanine, leucine, L-tryptophan, and uridine 5'-monophosphate (UMP) were decreased significantly. High dose of lipid-soluble alkaloids of FZ could increase the plasma contents of L-lysine, L-arginine, and deoxycholic acid, while the plasma contents of UMP, carnitine, N-formylanthranilic acid, and adenosine were decreased significantly. The pathway analysis indicated that therapeutic dose of lipid-soluble alkaloids of FZ could regulate energy and amino acid metabolic disorders in AIA rats. However, toxic dose could cause bile acid, fat, amino acid, and energy metabolic disorders. And combination with total glucosides of peony could enhance the therapeutic effects and attenuate the toxicity induced by lipid-soluble alkaloids of FZ.

Antibiotic-mediated changes in the fecal microbiome of broiler chickens define the incidence of antibiotic resistance genes

BackgroundAntimicrobial agents have been widely used in animal farms to prevent and treat animal diseases and to promote growth. Antimicrobial agents may change the bacterial community and enhance the resistome in animal feces. We used metagenome-wide analysis to investigate the changes in bacterial community, variations in antibiotic resistance genes (ARGs), and their bacterial hosts in the feces of broiler chickens over a full-treatment course of chlortetracycline at low and therapeutic dose levels.ResultsThe effects of chlortetracycline on resistome were dependent on the specific ARG subtypes and not simply the overall community-level ARGs. Therapeutic dose of chlortetracycline promoted the abundance of tetracycline resistance genes (tetA and tetW) and inhibited multidrug resistance genes (mdtA, mdtC, mdtK, ompR, and TolC). The therapeutic dose of chlortetracycline led to loss of Proteobacteria mainly due to the decrease of Escherichia/Shigella (from 72 to 58%). Inhibition of Escherichia by chlortetracycline was the primary reason for the decrease of genes resistant to multiple drugs in the therapeutic dose group. The ARG host Bifidobacterium were enriched due to tetW harbored by Bifidobacterium under chlortetracycline treatment. Escherichia was always the major host for multidrug resistance genes, whereas the primary host was changed from Escherichia to Klebsiella for aminoglycoside resistance genes with the treatment of therapeutic dose of chlortetracycline.ConclusionsWe provided the first metagenomic insights into antibiotic-mediated alteration of ARG-harboring bacterial hosts at community-wide level in chicken feces. These results indicated that the changes in the structure of antibiotic-induced feces microbial communities accompany changes in the abundance of bacterial hosts carrying specific ARGs in the feces microbiota. These findings will help to optimize therapeutic schemes for the effective treatment of antibiotic resistant pathogens in poultry farms.Graphical abstractResistome variations in faecal microbiome of chickens exposed to chlortetracycline

Pregabalin administration induces alterations in neural tube development during early embryonic stages.

Antiepileptic drugs (AEDs) have been known to cause congenital malformations, including neural tube defects. However, inadequate data are available regarding the effects of recently developed AEDs, such as pregabalin. The present study was conducted to evaluate the effect of pregabalin on neural tube development using early chick embryos. Experiments were conducted on specific pathogen-free Leghorn chick embryos, which were equally divided into control and pregabalin-treated (therapeutic dose, 600 mg; supratherapeutic dose, 1200 mg) groups. The embryos were macroscopically and microscopically evaluated following pregabalin administration. Expression levels of mammalian target of rapamycin (mTOR), c-Jun N-terminal kinase (JNK), and microtubule-associated proteins 1A/1B light chain 3 (LC3) in the embryos were observed. The embryos in the therapeutic dose group appeared more curved than those in the control group. The vesicles in the supratherapeutic dose group were more distinct but smaller than those in the control and therapeutic dose groups. mTOR expression was high in the control group and low in the therapeutic and supratherapeutic dose groups. JNK expression was low in the control and therapeutic dose groups and moderate in the supratherapeutic dose group. LC3 expression was moderate in the control and therapeutic dose groups and strong in the supratherapeutic dose group. Pregabalin administration induced neural tube defects and fetal abnormalities in the chick embryos through increased autophagy due to enhanced apoptosis in the prenatal fetus.

Sub-therapeutic doses of fluvastatin and valsartan are more effective than therapeutic doses in providing beneficial cardiovascular pleiotropic effects in rats: A proof of concept study

BackgroundStatins and sartans can, in therapeutic doses, induce pleiotropic cardiovascular effects. Similar has recently been shown also for sub-therapeutic doses. We thus explored and compared the cardiovascular pleiotropic efficacy of sub-therapeutic vs. therapeutic doses. MethodsWistar rats were randomly divided into 7 groups receiving fluvastatin, valsartan and their combination in sub-therapeutic and therapeutic doses, or saline. After 6weeks, the animals were euthanised, their hearts and thoracic aortas isolated, and blood samples taken. Endothelium-dependent relaxation of the thoracic aortae and ischaemic-reperfusion injury of the isolated hearts were assessed along with the related serum parameters and genes expression. ResultsFluvastatin and valsartan alone or in combination were significantly more effective in sub-therapeutic than therapeutic doses. The sub-therapeutic combination greatly increased thoracic aorta endothelium-dependent relaxation and maximally protected the isolated hearts against ischaemia-reperfusion injury and was thus most effective. Beneficial effects were accompanied by increased levels of nitric oxide (NO) and decreased levels of asymmetric dimethylarginine (ADMA) in the serum (again prominently induced by the sub-therapeutic combination). Furthermore, nitric oxide synthase 3 (NOS3) and endothelin receptor type A (EDNRA) genes expression increased, but only in both combination groups and without significant differences between them. In the therapeutic dose groups, fluvastatin and valsartan decreased cholesterol values and systolic blood pressure. ConclusionSub-therapeutic doses of fluvastatin and valsartan are more effective in expressing cardiovascular pleiotropic effects than therapeutic doses of fluvastatin and/or valsartan. These results could be of significant clinical relevance.

Low-Dose Heparin in Critically Ill Patients Undergoing Extracorporeal Membrane Oxygenation - the Help-ECMO Pilot Randomised Controlled Trial

Extra corporeal membrane oxygenation (ECMO) is a rescue therapy for reversible cardiac and/or respiratory failure. Despite improvement in management of patients undergoing ECMO, mortality remains high. Due to thrombosis risk, which includes arterial and venous thrombosis as well as in the extracorporeal circuit and components, variable intensity systemic anticoagulation with unfractionated heparin is routinely used. However, bleeding is one of the most frequent complications, can be severe and is independently associated with worse outcomes. Optimal anticoagulation to prevent thrombosis whilst minimising bleeding in adults on ECMO remains unknown. Before conducting a large randomised controlled trial to determine whether lower anticoagulation intensity is safe and effective compared with therapeutic anticoagulation, we aimed to determine the feasibility of randomising ECMO patients to these anticoagulation protocols.Methods:The HELP-ECMO pilot study (ACTRN12613001324707) is a randomised, controlled, unblinded trial at two Australian intensive care units (ICUs). Inclusion criteria were ICU patients who required ECMO (venous-venous [VV] or venous-arterial [VA]). Patients who did not meet any exclusion criteria were randomised to receive either therapeutic anticoagulation with heparin (target activated partial thromboplastin time [aPTT] between 50 and 70 seconds) or low dose heparin (12000 units/24 hours aiming for aPTT <45 seconds). Paired aPTT and anti-Xa assays were taken according to protocol, and at a minimum once a day. The primary endpoints for feasibility were difference in mean heparin dose, aPTT and anti-Xa levels between both study groups. Secondary endpoints included incidence of thromboembolic events, ECMO circuit thrombosis and bleeding events. All primary outcomes were log-transformed prior to analysis and reported as geometric means (95%CI) with overall differences determined using Repeat measures ANOVA.Results:Between May 2014 and March 2016, 31 patients who underwent ECMO (9 (29%) VA and 22 (71%) VV) were enrolled; 16 were randomised to low dose and 15 to therapeutic dose heparin. The groups were similar in age (mean 41 years [SD 16.8] vs 43 [SD 17.6] p=0.75), gender (68% vs 80% male, p=0.47), type of ECMO (31% vs 27% VA, p=0.78) and Acute Physiology and Chronic Health Evaluation III illness severity score (mean 65.4 [SD 23.5] vs 61.8 [SD 30.1], p=0.72) and sepsis-related organ failure assessment score (mean 10 [SD 3.6] vs 10 [SD 3.3], p=1.0). The mean duration of ECMO support was 9.33 days (SD 5.97) in the low dose and 9.79 days (SD 4.77) in the therapeutic dose group (p=0.82).For the primary outcomes, there was a significant difference in the daily mean aPTT (48.1 [95% CI 43.5-53.3] vs 56.2 [95% CI 50.7-62.3], p=0.03), daily mean anti-Xa (0.11 [95% CI 0.07-0.18] vs 0.30 [IQR 0.19-0.46], p=0.003) and daily mean heparin dose (11784 units [95% CI 8693-15972] vs 22050 [IQR 16262-29899], p=0.004) in the low dose compared to therapeutic group.There was no difference in thrombotic complications with regard to DVT (2 [12.5%] vs 3 [20%], p=0.57), PE (1 [6.3%] vs 0 [0%], p=0.33), stroke (no events), intracardiac thrombus (1 [6.3%] vs 2 [13.3%], p=0.51), acute pump (1 [6.3%] vs 1[6.7%], p=0.96) and distal perfusion cannula thrombosis (2 [12.5%] vs 0 [0%], p=0.16) in low dose compared with therapeutic group, respectively.With regard to bleeding, there was no difference in intracranial haemorrhage (ICH) (0 [0%] vs 1 [6.7%], p=0.29), retroperitoneal bleeding (1 [6.3%] vs 1 [6.7%], p=0.96), gastrointestinal bleeding (0 [0%] vs 2 [13.3%] , p=0.13), or haemoptysis (1 [6.5%] vs 1 [6.7%], p=0.96) in low dose compared with therapeutic group, respectively.Conclusion: In this pilot trial, administration of a low dose heparin protocol was feasible, and resulted in a significant difference in mean heparin dose administered and daily aPTT and anti-Xa levels between groups. Low dose heparin was not associated with an increase in thrombotic events nor a decrease in bleeding events; however the study was not powered for these outcomes. Our findings support the feasibility of a larger phase III study to evaluate the safety and efficacy of low-dose anticoagulation compared with therapeutic heparin with regard to thrombotic and bleeding events in patients receiving ECMO. DisclosuresNo relevant conflicts of interest to declare.