Low Serum Concentrations Research Articles

Predictive Model for Acute Heart Failure in Patients with Acute Myocardial Infarction and Type 2 Diabetes Mellitus Based on Energy and Adipokine Metabolism Indicators

BACKGROUND: Acute heart failure (AHF) is one of the early complications of acute myocardial infarction (AMI) in diabetic patients. Evaluation of biomarkers of energy and adipokine metabolism can help in the early identification of diabetic patients at risk of AHF. AIM: The present study is aimed to predict the development of AHF in diabetic patients with AMI based on energy and adipokine metabolism parameters. METHODS: A total of 74 diabetic patients with AMI were examined between September 1, 2018, and December 31, 2020. Serum adropin, irisin, and C1q/TNF-related protein 3 (CTRP3) levels were measured by enzyme-linked immunosorbent assay. To predict AHF development in AMI patients, generalized linear mixed model (GLMM) was applied. RESULTS: The serum concentrations of adropin, irisin, and CTRP3 have been found to be reduced in diabetic patients with AMI and AHF. The accuracy of predicting AHF Killip Class 1 was 96.7%, and the accuracy of prediction for AHF Killip Class 2 was 57.1%, that is, the model was poorly sensitive to this level of complications. The prediction accuracy for AHF Killip Class 3 was 80%, that is, the model was highly sensitive to complications of this level, and for AHF Killip Class 4 – 100% being the maximum level of the model sensitivity. CONCLUSIONS: Low serum concentrations of adropin, irisin, and CTRP3 indicate an imbalance in energy and adipokine homeostasis. The constructed model predicts the probability of AHF development with high accuracy of 91.9% in diabetic patients with AMI.

AB0350 ASSESSMENT OF REAL-LIFE PATIENT HANDLING EXPERIENCE OF AVT02 ADMINISTERED SUBCUTANEOUSLY VIA AUTOINJECTOR IN PATIENTS WITH MODERATE-TO-SEVERE ACTIVE RHEUMATOID ARTHRITIS: AN OPEN-LABEL, SINGLE-ARM CLINICAL TRIAL, THEN AN EXTENSION PHASE OF AVT02 ADMINISTERED WITH A PRE-FILLED SYRINGE (ALVOPAD-PEN)

BackgroundAVT02 is an investigational biosimilar to adalimumab. It is approved in Europe, Canada, and the UK. It is not approved by the US Food and Drug Administration (FDA).ObjectivesTo assess the real-life patient handling of an autoinjector (AI) in adult patients with moderate-to-severe active rheumatoid arthritis (RA) who self-inject AVT02 subcutaneously (s.c.).MethodsThis open-label study enrolled 107 adalimumab-naïve subjects with moderate-to-severe active RA to self-inject AVT02 with the proposed AI 40 mg s.c. in Week 1, and every other week thereafter through Week 8. In an optional extension phase through Week 56, subjects were switched from AVT02 AI to pre-filled syringe and followed for safety and efficacy. The primary endpoint was the percentage of successful self-injections as reported by both the trial site and by subjects using standardized questionnaires at Week 8. Additional endpoints included ease of use and robustness of the AI at Week 8, and efficacy in RA, assessment of serum trough levels of AVT02, and detection of antidrug antibodies (ADA) throughout the study.ResultsThe AI success rate was 100% as reported by both the trial site and by subjects. No handling events were noted through Week 8. Approximately 80% (78.1–84.9%) of subjects found the AI ‘very easy’ to use and, in general, less difficulty was reported as the study progressed. The first 110 AIs used were inspected for robustness and none showed any sign of damage or malfunction.All subjects who completed Week 8 (n = 106) took part in and completed the optional extension phase through Week 56.At Week 8, 49.1%, 5.7% and 0.9% subjects achieved ACR20, -50 and -70 responses, respectively. Improvement was also reported for the SDAI, DAS28 CRP and HAQ, with scores consistently decreasing through Week 8. In the extension phase, 70.8%, 47.2% and 13.2% subjects achieved ACR20, -50 and -70 responses at Week 56. Improvement was also reported for the SDAI, DAS28 CRP and HAQ, with scores consistently decreasing through Week 56.From Baseline through Week 8 the mean serum concentrations of investigational AVT02 increased consistently at each visit, reaching a peak at Week 24. There was no significant difference in serum concentrations of AVT02 based on injection subsite (abdomen or thigh).ADAs were detected in 65.1% of subjects through Week 8, reducing to 49.1% by Week 56. Of ADA-positive subjects, most were also positive for neutralizing antibodies through Week 8 (62.3%) increasing through Week 56 (90.4%). Subjects who were ADA-positive had lower serum concentrations of study drug compared with the overall population at Week 8 and Week 56 as expected.There were no clinically relevant safety or tolerability issues. Overall, treatment-emergent adverse events reported in at least 5% of subjects at the PT level were anaemia and influenza (6 subjects [5.6%)] reported 6 events each), and no local ISRs were reported.ConclusionThe fully successful self-injection with the AI in this study supported the use of the device while administering AVT02. Furthermore, subjects typically found the AI easy to use, with ease of use increasing as time progressed.The ACR20, -50, and -70, SDAI, DAS28 CRP and HAQ results supported the use of AVT02 to treat adult patients with moderate-to-severe RA, with the treatment effect persisting beyond Week 8 and through Week 56.The safety and immunogenicity profiles were as expected for adalimumab.ClinicalTrials.gov Identifier: NCT04224194Disclosure of InterestsNemanja Damjanov: None declared, Heimo Stroissnig Employee of: Alvotech, Matjaz Steiger Employee of: Alvotech, Joanna Sobierska Employee of: Alvotech, Eric Guenzi Employee of: Alvotech, Hendrik Otto Employee of: Alvotech, Abid Sattar Employee of: Alvotech, Halimu N. Haliduola Employee of: Alvotech, Elin Edwald Employee of: Alvotech, Fausto Berti Employee of: Alvotech

Fabrication of a Free-Standing MWCNT Electrode by Electric Field Force for an Ultra-Sensitive MicroRNA-21 Nano-Genosensor.

Abnormal expression of microRNA-21 (miR-21) is considered to be closely associated with the pathogenesis of colorectal cancer. However, great challenges do exist for the development of ultra-sensitive biosensors to detect the abnormal expression of miR-21 due to the low concentration in serum (fm level) at the early stage of colorectal cancer. Therefore, electric field force is used to rotate and rearrange random multi-walled carbon nanotubes (MWCNTs) at the microscale to improve the active sites of the electrode in this study. The free-standing MWCNTs are densely and high-orderly embedded into the bare electrode along the direction of the electric field. Compared to the bare electrode, the peak-current response of the free-standing MWCNT electrode improves by 150times in cyclic voltammetric measurement. A nano-genosensor based on the free-standing MWCNT electrode is developed for measuring miR-21. The nano-genosensor for miR-21 shows an ultra-high sensitivity of 48.24 µA µm-1 , a wide linear range from 0.01 × 10-15 to 100 × 10-12 m, and a low detection limit of 1.2 × 10-18 m. The present nano-genosensor shows superior performance for miR-21 in human serum samples and demonstrates a potential application for the diagnosis of early stage colorectal cancer.

MO808: Indoxyl-Sulphate and Cresyl Sulphate are Respectively Linked to Calcium-Phosphate Metabolism Variables and to Cardiovascular Morbidity in Hemodialysed Patients

Abstract BACKGROUND AND AIMS Indoxyl sulphate (IS) and Cresyl sulphate (CS) are uremic toxins generated by the intestinal amino acid catabolism. Blood levels of these toxins increase in patients with CKD and are linked to cardiovascular events. Therefore, we studied the relationship between serum levels of free IS and CS and divalent ion metabolism variables and cardiovascular morbidity (stroke, heart failure, angor and myocardial infarction) in 139 haemodialysis patients (age 68 + 13 years, weight 65 + 13 kg, dialysis vintage 69 + 71 months). METHOD We divided patients according to tertiles of free serum IS and CS. RESULTS Patients in the highest tertile of serum free IS showed shorter dialysis vintage and higher body weight gain between dialysis sessions than patients in the other two tertiles. Patients in the highest tertile of IS showed lower body weight and serum concentrations of alkaline phosphatase, 1,25(OH)2D and PTH compared with the lowest tertile. Serum PTH was also measured during 2 years after the baseline; it was higher in patients in the lowest than those in the highest tertile of serum free IS. No relationships of serum free CS concentrations with phosphate and calcium metabolism variables were observed. Kaplan–Meier survival analysis shows an increased cardiovascular morbidity in patients in the CS highest tertile (blue line in the figure) compared with those in the lowest and middle tertiles taken together (red line; P = 0.01). This association was not found considering IS tertiles. CONCLUSION Our findings suggest that serum free IS may be associated with PTH and 1,25(OH)2D secretion, whereas free PCS may predict cardiovascular risk in haemodialysis patients. A point for future investigation could be the relationship between IS and adynamic bone disease.

COVID-19 Lockdown in Israel: The Environmental Effect on Ultrafine Particle Content in the Airway.

Inhaled ultrafine particle (UFP) content in exhaled breath condensate (EBC) was observed as an airway inflammatory marker and an indicator of exposure to particulate matter (PM). The exceptional decline in air pollution during the COVID-19 lockdown was an opportunity to evaluate the effect of environmental changes on UFP airway content. We collected EBC samples from 30 healthy subjects during the first lockdown due to COVID-19 in Israel (March–April 2020) and compared them to EBC samples retrieved during April–June 2016 from 25 other healthy subjects (controls) living in the same northern Israeli district. All participants underwent EBC collection and blood sampling. Ambient air pollutant levels were collected from the Israeli Ministry of Environmental Protection’s online database. Data were acquired from the monitoring station closest to each subject’s home address, and means were calculated for a duration of 1 month preceding EBC collection. UFP contents were measured in the EBC and blood samples by means of the NanoSight LM20 system. There was a dramatic reduction in NO, NO2, SO2, and O3 levels during lockdown compared to a similar period in 2016 (by 61%, 26%, 50%, and 45%, respectively). The specific NO2 levels were 8.3 ppb for the lockdown group and 11.2 ppb for the controls (p = 0.01). The lockdown group had higher UFP concentrations in EBC and lower UFP concentrations in serum compared to controls (0.58 × 108/mL and 4.3 × 108/mL vs. 0.43 × 108/mL and 6.7 × 108/mL, p = 0.05 and p = 0.03, respectively). In this observational study, reduced levels of air pollution during the COVID-19 lockdown were reflected in increased levels of UFP airway contents. The suggested mechanism is that low airway inflammation levels during lockdown resulted in a decreased UFP translocation to serum. Further studies are needed to confirm this hypothesis.

Metabolic Bone Disease of Prematurity.

Metabolic Bone Disease of Prematurity.

Associations between Genetically Proxied Inhibition of Lipid-Lowering Drug Targets and Serum Micronutrients among Individuals of European Descent: A Mendelian Randomization Study

BackgroundLimited and inconclusive data exist concerning the associations between lipid-lowering drugs and serum micronutrient concentrations. MethodsWe conducted Mendelian randomization (MR) analyses to explore the associations between lipid-lowering drug targets and serum micronutrients. Single-nucleotide polymorphisms in genes encoding molecular targets of LDL cholesterol-lowering therapies were selected as instrumental variables for 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR; target of statins), proprotein convertase subtilisin/kexin type 9 (PCSK9; target of PCSK9 inhibitors), and Niemann-Pick C1-Like 1 (NPC1L1; target of ezetimibe). Exposure data were extracted from a published genome-wide association study (GWAS) of lipids in 188,577 European individuals, with outcome data obtained from the Integrative Epidemiology Unit (IEU) GWAS database (https://gwas.mrcieu.ac.uk). Overall, age and sex information were not calculable from the summary-level GWAS data. MR analyses were performed using the inverse-variance weighted method and MR sensitivity analysis methods. ResultsWe found genetically proxied inhibition of HMGCR to lower iron (effect, –0.16; 95% CI: –0.27, –0.06; P value = 0.003), zinc (effect, –0.83; 95% CI: –1.36, –0.31; P value = 0.002), magnesium (effect, –0.17; 95% CI: –0.27, –0.06; P value = 0.003), potassium (effect, –0.17; 95% CI: –0.27, –0.06; P value = 0.002), genetically proxied inhibition of NPC1L1 to increase calcium (effect, 0.28; 95% CI: 0.10, 0.46; P value = 0.003), retinol (effect, 0.25; 95% CI: 0.07, 0.44; P value = 0.01), and genetically proxied inhibition of PCSK9 to increase vitamin D (effect, 0.10; 95% CI: 0.07, 0.12; P value = 1.8 × 10–19). These associations were robust in MR sensitivity analyses. However, the associations between genetically proxied inhibition of HMGCR and NPC1L1 and the micronutrients were not consistent in multiple comparisons. ConclusionsOur results provide evidence that statin use may lower serum concentrations of iron, zinc, magnesium, and potassium, PCSK9 inhibitors may increase serum vitamin D, and ezetimibe may increase serum calcium and retinol concentrations.

Colorimetric sensing strategy for detection of cysteine, phenol cysteine, and phenol based on synergistic doping of multiple heteroatomsinto sponge-like Fe/NPC nanozymes.

Nanozymes have both the high catalytic activity of natural enzymes and the stability and economy of mimetic enzymes. Research on nanozymes is rapidly emerging, and the continuous development of highly catalytic active nanozymes is of far-reaching significance. This work reports heteroatomic nitrogen (N) and phosphorus (P) double-doped mesoporous carbon structures and metallic Fe coordination generated sponge-like nanozymes (Fe/NPCs) with good peroxidase activity. On this basis, we constructed a highly sensitive colorimetric sensor with cysteine and phenol as simulated analytes using Fe/NPCs nanozymes, and the response limits reached 53.6nM and 5.4nM, respectively. Besides, the method has high accuracy in the detection of cysteine and phenol at low concentrations in serum and tap water, which lays a foundation for application in the fields of environmental protection and biosensors.

Dysbiosis and intestinal inflammation caused by Salmonella Typhimurium in mice can be alleviated by preadministration of a lytic phage

Many studies have shown the efficacy of phage therapy in reducing intestinal pathogens. However, phage-based probiotic treatment is poorly studied in view of effects on the gut microbiota and intestinal inflammation. In this study, a lytic or a temperate phage (each at 4 ×108 PFU per day) or a streptomycin solution (40 mg per day) were administered to mice via drinking water for 31 days. Subsequently, mice were challenged with Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium). S. Typhimurium does not serve as the host bacterium and is not lysed by both phages. For intestinal inflammation evaluation, mice were given one dose of streptomycin for 24 h before the S. Typhimurium challenge. High-throughput sequencing analysis revealed that the phylum Firmicutes became the most abundant in mice pretreated with phages. The alpha diversity of gut bacteria was higher in phage treated than in streptomycin treated mice. Moreover, pretreatment with the lytic and the temperate phage before the S. Typhimurium challenge increased two beneficial genera, Lactobacillus and Bifidobacterium. According to the pathological analysis of ileum, cecum, and serum, temperate or lytic gut phage pretreatment of mice markedly reduced intestinal inflammation, concomitant with lower serum concentration of lipopolysaccharides (LPS) and diamine oxidase (DAO). The oral pretreatments of mice (ST, Lyt, Lys, SM) generally caused an increased expression of IL-1β, TNF-α, IFN-γ, IL-4, and IL-10 compared to the matching control. However, in mice pretreated with the lytic phage, the mRNA expression for the pro-inflammatory cytokine TNF-α was not significantly higher than that of the control group. No significant differences were detected for peripheral blood B lymphocytes, CD3 +T cells, and the CD4 + /CD8 + ratio in mice pretreated with the lytic and lysogenic phage. This study demonstrated that even lytic phages not targeting the pathogenic serovar Salmonella Typhimurium alleviated intestinal dysbiosis and inflammation in challenged mice.

Pro-inflammatory immune profile mediated by TNF and IFN-γ and regulated by IL-10 is associated to IgG anti-SARS-CoV-2 in asymptomatic blood donors.

The SARS-CoV-2 virus has infected and killed millions of people, but little is known about the risk factors that lead to the development of severe, mild or asymptomatic conditions after infection. The individual immune response and the balance of cytokines and chemokines have been shown to be important for the prognosis of patients. Additionally, it is essential to understand how the production of specific antibodies with viral neutralizing capacity is established. In this context, this study aimed to identify positive individuals for IgG anti-SARS-CoV-2 in a large population of blood donors (n = 7837) to establish their immune response profile and to evaluate its viral neutralization capacity. The prevalence found for IgG anti-SARS-CoV-2 was 5.6% (n = 441), with male blood donors (61.9%) being more prevalent among the positive ones. The results showed that positive individuals for IgG anti-SARS-CoV-2 have high serum concentrations of chemokines, TNF, IFN-γ and IL-10. The analyses showed that the positivity index for IgG anti-SARS-CoV-2 is associated with the neutralizing capacity of the antibodies, which, in turn, is significantly related to lower serum concentrations of CCL5 and CXCL10. The results allow us to hypothesize that the development and maintenance of IgG anti-SARS-CoV-2 antibodies in infected individuals occurs in a pro-inflammatory microenvironment well regulated by IL-10 with great capacity for recruiting cells from the innate and adaptive immune systems.

516 Phosphorus Requirements in Patients with Severe Thermal Injuries Requiring High-Volume Hemofiltration

IntroductionPatients with thermal injuries have increased metabolic demands, requiring increased phosphate supplementation. Evidence is scant depicting incidence of hypophosphatemia and repletion requirements in patients with thermal injuries treated with high-volume hemofiltration (HVHF) and a high-flux membrane. The objective of this study was to determine the incidence of hypophosphatemia and characterize repletion requirements in this population.MethodsThis study was a case-control, retrospective chart review. Patients were included if sustained at least 20% total body surface area (TBSA) thermal injuries and required continuous HVHF (prescribed doses ≥ 35 mL/kg/hr). A randomly selected cohort (matched to age, TBSA, and inhalation injury) without acute kidney injury (AKI) was used to compare phosphorus requirements over an initial 14 day period. An a priori sample size was calculated (n = 26) to detect a minimum difference of 0.3 mmol/kg/day. Repeated measures ANOVA was used to compare requirements and concentrations. Demographics, diet, and variables affecting phosphorus concentrations were compared utilizing Fisher's exact, Student's t-test, or Mann-Whitney test depending on type and distribution.ResultsOne thousand sixty-six patients were screened. Most were excluded from the HVHF group for TBSA < 20% (58%) or not a burn injury (29%). Sixteen patients were included in each group. The average age was 60.2 ± 15.1 vs 53.3 ± 16.4 (p = 0.22) with median TBSA (p = 0.73) of 30% (23.4, 56.3) vs 29% (26.4, 33.9). All patients in the study group were started on HVHF for AKI, utilizing a 1.6m2 polyethersulfone membrane (mean delivered prefilter dose of 54.7 ± 1.5 ml/kg/hr), and had statistically higher potassium and phosphorous laboratory values at baseline. Parenteral phosphorus replacements were 2 fold higher in the HVHF group (p = 0.02), but not statistically different after accounting for estimated enteral intake. Despite providing 0.75 mmol/kg/day of phosphorous supplementation (vs 0.66 mmol/kg/day in control, p = 0.45), the HVHF group experienced more days with hypophosphatemia (49.6 ± 12.4 % vs 29.3 ± 16.3 %, p = 0.012). By 72h, every HVHF patient experienced at least one episode of hypophosphatemia. Patients on longer durations of therapy had increasing risk of hypophosphatemia. There was a significant difference in days requiring mechanical ventilation (p < 0.001)ConclusionsThis study demonstrates thermally injured patients receiving HVHF for AKI are at increased risk for hypophosphatemia and require higher phosphate supplementation to maintain lower average serum concentrations, as compared to the controls with similar burns but without acute kidney injury.

Long-term opioid treatment and endocrine measures in patients with cancer-related pain: asystematic review.

Opioid analgesics are the main stay for cancer pain management; however, long-term opioid treatment (L-TOT) may suppress the endocrine system. This systemic review aimed at investigating effects of L-TOT on the endocrine system in patients with cancer-related pain. A search on MEDLINE, EMBASE and Web of Science databases was performed. Inclusion criteria were clinical studies investigating endocrine measures in adult patients with cancer-related pain in L-TOT (≥4weeks). Outcomes and quality of evidence were assessed. A total of 252 abstracts were identified; out of which 247 were excluded and five cross-sectional studies were included and analyzed. L-TOT was associated with lower serum concentration levels of total- and free testosterone in males, follicular stimulating hormone in females, and luteinizing hormone in both sexes. Moreover, higher morphine equivalent daily doses (MEDDs) were correlated with higher levels of cortisol and lower levels of LH in both sexes, and lower levels of total- and free testosterone in males. Sexual dysfunction was associated with low sex hormone levels. Level of evidence was low/very low. The studies identified demonstrated that patients with cancer-related pain in L-TOT may have gonadal hypofunction causing sexual dysfunction, which may be correlated with opioid dose level. In addition, high serum concentrations of cortisol were positively correlated with high opioid dose levels. However, the evidence was weak and further research is necessary. PROSPERO, ID-number: CRD42020213059.

Cardiometabolic risk factors and renin-angiotensin system polymorphisms in young individuals with various metabolic phenotypes

Background. The increased prevalence of obesity and associated cardiometabolic diseases attract attention worldwide. Renin-angiotensin system can link obesity and cardiovascular and metabolic diseases.Objective. To access a comprehensive assessment of cardiometabolic risk factors and gene polymorphisms of the renin-angiotensin system in metabolic phenotypes among young individuals.Design and methods. The sample consisted of 251 individuals, who were divided into four groups: group 1 — metabolically healthy individuals with normal body mass index (BMI) (n = 62); group 2 — metabolically unhealthy individuals with normal BMI (n = 57); group 3 — metabolically healthy overweight/obese individuals (n = 16); group 4— metabolically unhealthy overweight/ obese individuals (n = 116). All participants answered a questionnaire designed for this study. Anthropometric, clinical and biochemical parameters were assessed. The following polymorphisms were evaluated:, A1166C polymorphism of the angiotensin II type 1 receptor gene (rs5186), M235T polymorphism of the angiotensinogen gene (rs699), T174M polymorphism of the angiotensinogen gene (rs4762), I/D polymorphism of the angiotensinconverting enzyme gene (rs4340).Results. In young individuals with metabolically unhealthy overweight/ obesity, a higher frequency of coexistent abdominal obesity and hypertension was found in combination with a higher frequency of the allele T of AGT 235M/T. The greater differences in carbohydrate and lipid metabolism in combination with a higher serum levels of leptin and low serum concentrations of adiponectin were also found in young individuals with metabolically unhealthy overweight/obesity.

Fluorescent immunosensor based on fluorescence resonance energy transfer between CdSe/ZnS quantum dots and Au nanorods for PRRSV detection

An ultrasensitive and selective sandwich-type fluorescent immunosensor based on fluorescence resonance energy transfer (FRET) between CdSe/ZnS quantum dots (QDs) and Au nanorods (AuNRs) was developed for the rapid detection of porcine reproductive and respiratory syndrome virus (PRRSV). Modified CdSe/ZnS QDs and AuNRs were bioconjugated with primary antibodies M and GP5, respectively. When the target antigen was present, a well-known sandwich-type form CdSe/ZnS QDs-M antibody-antigen-GP5 antibody-AuNRs was produced, and thus reducing the distance between the QDs and NRs. As a result, FRET occurred and the fluorescence intensity of CdSe/ZnS QDs decreased, and this decrease was used to determine the antigen concentration. Ultrasensitive detection of PRRSV at low concentrations in swine serum was achieved with a limit of detection of 0.55 TCID50/mL and a linear detection range of 101–3.5 ​× ​104 TCID50/mL. Therefore, the fluorescent immunosensor is selective and highly sensitive; specifically, it can detect PRRSV at low concentrations in swine serum over a large concentration range. This proposed detection method can facilitate the development of high-performance fluorescent immunosensors for the detection of PRRSV and other antigens.

Overactivation of hepatic mechanistic target of rapamycin kinase complex 1 (mTORC1) is associated with low transcriptional activity of transcription factor EB and lysosomal dysfunction in dairy cows with clinical ketosis

Ketosis occurs most frequently in the peripartal period and is associated with liver injury and steatosis. Lysosomes serve as the terminal degradative station and contribute to liver homeostasis through their role in the digestion of dysfunctional organelles and lipid droplets. Transcription factor EB (TFEB) has been identified as a master regulator of lysosomal function. Thus, the objective of the present study was to investigate the status of lysosomal function and TFEB transcriptional activity and potential changes in abundance of upstream effectors of TFEB identified in nonruminants, including mechanistic target of rapamycin kinase complex 1 (mTORC1), protein kinase B (Akt), glycogen synthase kinase β (GSK3β), and extracellular signal-regulated kinase1/2 (ERK1/2), and to explore which factor induces the above changes. Liver and blood samples were collected from healthy cows (n = 10) and ketotic cows (n = 10) that had a similar number of lactations (median = 3, range = 2-4) and days in milk (median = 6 d, range = 3-9 d). Calf hepatocytes were isolated from Holstein calves and treated with 10 ng/mL growth hormone (GH), 3.0 mM β-hydroxybutyrate (BHB), 1.5 ng/mL interleukin-18 (IL-18), 0.15 ng/mL tumor necrosis factor-α (TNF-α), or 1.2 mM free fatty acid (FFA) for 12 h. Serum levels of FFA and activities of alanine aminotransferase and aspartate aminotransferase were greater in ketotic cows, whereas glucose was lower. Additionally, ketotic dairy cows exhibited higher serum concentrations of GH, IL-18, and TNF-α, and lower serum concentration of insulin. The lower protein abundance of lysosome-associated membrane protein 1 (LAMP1) and mRNA abundance of LAMP1 indicated that hepatic lysosomal mass was lower in ketotic cows. Furthermore, lower protein abundance of cathepsin D (CTSD) and mRNA abundance of CTSD and V0 domain of the vacuolar ATPase along with lower activity of β-N-acetylglucosaminidase indicated impairment in hepatic lysosomal function due to ketosis. The lower nuclear abundance, total protein, and mRNA abundance of TFEB and peroxisome proliferator-activated receptor γ coactivator 1 α along with greater phosphorylated (p)-TFEB in the liver of ketotic cows indicated an impairment of hepatic TFEB transcriptional activity. The protein abundances of phosphorylated mTOR (p-mTOR) and its downstream effectors ribosomal protein S6 kinase B (RPS6KB) and eukaryotic factor 4E-binding protein 1 (EIF4EBP1) were greater, whereas p-Akt, p-GSK3β, and p-ERK1/2 were lower in the liver of ketotic cows. Importantly, elevated phosphorylation of mTOR, RPS6KB, and EIF4EBP1 was observed in calf hepatocytes treated with GH, BHB, IL-18, TNF-α, and FFA. Moreover, BHB, TNF-α, and FFA, not GH and IL-18, reduced TFEB transcriptional activity and impaired lysosomal function in calf hepatocytes. Taken together, these data suggest that BHB, TNF-α, and FFA overactivate the hepatic mTORC1 signaling pathway during ketosis and further impaired TFEB transcriptional activity and lysosomal function, which may contribute to liver injury and steatosis.

Defective IGF-1 prohormone N-glycosylation and reduced IGF-1 receptor signaling activation in congenital disorders of glycosylation

The insulin-like growth factor-1 (IGF-1) signaling pathway is crucial for the regulation of growth and development. The correct processing of the IGF-1Ea prohormone (proIGF-1Ea) and the IGF-1 receptor (IGF-1R) peptide precursor requires proper N-glycosylation. Deficiencies of N-linked glycosylation lead to a clinically heterogeneous group of inherited diseases called Congenital Disorders of Glycosylation (CDG). The impact of N-glycosylation defects on IGF-1/IGF-1R signaling components is largely unknown. In this study, using dermal fibroblasts from patients with different CDG [PMM2-CDG (n = 7); ALG3-CDG (n = 2); ALG8-CDG (n = 1); GMPPB-CDG (n = 1)], we analyzed the glycosylation pattern of the proIGF-1Ea, IGF-1 secretion efficiency and IGF-1R signaling activity. ALG3-CDG, ALG8-CDG, GMPPB-CDG and some PMM2-CDG fibroblasts showed hypoglycosylation of the proIGF-1Ea and lower IGF-1 secretion when compared with control (CTR). Lower IGF-1 serum concentration was observed in ALG3-CDG, ALG8-CDG and in some patients with PMM2-CDG, supporting our in vitro data. Furthermore, reduced IGF-1R expression level was observed in ALG3-CDG, ALG8-CDG and in some PMM2-CDG fibroblasts. IGF-1-induced IGF-1R activation was lower in most PMM2-CDG fibroblasts and was associated with decreased ERK1/2 phosphorylation as compared to CTR. In general, CDG fibroblasts showed a slight upregulation of Endoplasmic Reticulum (ER) stress genes compared with CTR, uncovering mild ER stress in CDG cells. ER-stress-related gene expression negatively correlated with fibroblasts IGF-1 secretion. This study provides new evidence of a direct link between N-glycosylation defects found in CDG and the impairment of IGF-1/IGF-1R signaling components. Further studies are warranted to determine the clinical consequences of reduced systemic IGF-1 availability and local activity in patients with CDG.

Low serum concentration of zinc, selenium, calcium, potassium and high serum concentration of iron, sodium are associated with myocardial infarction

BackgroundNowadays, myocardial infarction (MI) is one of the major causes of death in developing countries including Bangladesh. Trace elements and macro-minerals play an important role in the pathogenesis of MI. This study aimed to determine the trace elements and macro-minerals in Bangladeshi patients with MI. MethodsThis case control study included 74 MI patients and 74 healthy individuals as control group. Flame atomic absorption spectroscopy and graphite furnace atomic absorption spectroscopy were used to measure serum hint factors (Zn, Fe, and Se) and macro minerals (Ca, K, and Na). Statistical evaluation was done using an independent sample t- test and Pearson's correlation test. ResultsMean serum concentrations of Zn, Fe, Se, Ca, K, and Na in patients with MI were 0.26 ± 0.01(Zn), 0.61 ± 0.05(Fe), 0.01 ± 0.0004(Se), 33.75 ± 1.39(Ca), 67.63 ± 5.25(K), and 3189.30 ± 47.63(Na)mg/L, whereas in control group those were 0.82 ± 0.04(Zn), 0.15 ± 0.01(Fe), 0.06 ± 0.004(Se), 87.39 ± 4.17(Ca), 166.90 ± 3.71(K), and 3000 ± 137.20(Na) mg/L, respectively. Serum concentrations of Zn, Se, Ca, and K became remarkably (P < 0.001) lower but the concentration of Fe and Na turned significantly (P < 0.001) and insignificantly (P = 0.215) higher in the patient group, respectively. Imbalanced homeostasis among affected persons was observed using inter- element relationship analysis. ConclusionsThis study found decreased levels of Zn, Se, Ca, and K but an increased level of serum Fe and Na in patients with MI. The findings suggest that MI sufferers have a considerably lower degree of critical hint factors in serum and as a consequence might need an increased intake of important trace elements for better clinical management.

Common variant p.D19H of the hepatobiliary sterol transporter ABCG8 increases the risk of gallstones in children

Gallstones are increasingly common in children. Genetic analyses of adult cohorts demonstrated that the sterol transporter ABCG8 p.D19H and Gilbert UGT1A1*28 variants enhance the odds of developing gallstones. The genetic background of common lithiasis in children remains unknown. Overall, 214 children with gallstone disease (1month-17years, 107 boys) were inclueded. The control cohorts comprised 214 children (age 6-17years, 115 boys) and 172 adults (age 40-92years, 70 men) without gallstones. The ABCG8 p.D19H and UGT1A1*28 polymorphisms as well as ABCB4 (c.504C>T rs1202283, c.711A>T rs2109505) and NPC1L1 variants (p.V1296V rs217434, c.-18C>A rs41279633) were genotyped using TaqMan assays. Serum concentrations of plant sterols and cholesterol precursors were measured by gas chromatography/mass spectrometry. The ABCG8 risk allele was associated with an increased risk of stones (OR=1.82, p=.03). Children carrying the p.19H allele presented with lower serum concentrations of surrogate markers of intestinal cholesterol absorption and decreased ratios of phytosterols to the cholesterol precursor desmosterol. Carriers of the common NPC1L1 rs217434 allele had an increased gallstone risk compared with stone-free adults (OR 1.90, p<.01). This variant also affected the ratio of phytosterols to cholesterol precursors (p=.03). Other tested variants were not associated with gallstone risk. The p.D19H ABCG8 and, to a lesser extent, NPC1L1 rs217434 variants increase the risk of early-onset gallstone formation. These results point to the presence of a common lithogenic pathway in children and adults.

Population Pharmacokinetic and Exposure–Response Model Simulations: Predicted Exposure and Efficacy for Maintenance Doses of Intravenous Golimumab Every 6 or 8 Weeks in Patients With Moderately to Severely Active Rheumatoid Arthritis

Golimumab is approved to treat moderate-to-severe active rheumatoid arthritis when given intravenously at weeks 0 and 4, then every 8 weeks (Q8W) with concomitant methotrexate. These analyses assessed whether a shorter dosing interval could ameliorate diminished efficacy experienced by a small proportion of patients toward the end of the dosing interval. Population pharmacokinetic and exposure-response modeling simulations were performed for intravenous golimumab 2 mg/kg at weeks 0 and 4, then Q8W or every 6 weeks (Q6W) through 1 year. A 2-compartment pharmacokinetic model with linear clearance developed based on GO-FURTHER (A Multicenter, Randomized, Double-blind, Placebo-controlled Trial of Golimumab, an Anti-TNFα Monoclonal Antibody, Administered Intravenously, in Patients With Active Rheumatoid Arthritis Despite Methotrexate Therapy) study data was used for pharmacokinetic simulations. A latent-variable indirect exposure-response model developed based on GO-FURTHER American College of Rheumatology (ACR) 20%/50%/70% improvement (ACR20, ACR50, and ACR70, respectively) data was used to predict clinical endpoints of ACR20/ACR50/ACR70 response rates. For Q6W and Q8W dosing, respectively, predicted median golimumab steady-state trough (Ctrough,ss) concentrations were 0.57 and 0.24 µg/mL, and Cmax at steady state values were 33.1 and 32.9µg/mL. Predicted peak median ACR20 steady-state response rates were 76.7% (Q6W) and 75.6% (Q8W). Predicted median ACR20 response rates at Ctrough,ss increased by 4.7 percentage points with Q6W (73.7%) versus Q8W (69.0%) dosing. Greater improvement in ACR20 response rates at trough time points was predicted in patients with lower golimumab trough serum concentrations. Consistent findings were observed for ACR50/ACR70 response rates. These simulations suggest that intravenous golimumab Q6W dosing increases golimumab Ctrough,ss, which may improve clinical response in the small proportion of patients with rheumatoid arthritis with waning efficacy at the end of the standard dosing interval. gov identifier: NCT00973479. Clinicaltrialsregister.eu: EudraCT 2008-006064-11.

Effects of prepartum supplementation of β-carotene in Holstein cows

Whether supplemental dietary β-carotene affects periparturient cows and vitamins A and E in cows when dietary vitamin A is adequate remains uncertain. Our objective was to assess the effect of β-carotene supplementation during the close-up dry period in a herd with adequate status of vitamins A and E but low in β-carotene. The study was conducted on a large commercial dairy farm in Indiana during early summer of 2015. Ninety-four multiparous Holstein cows were assigned to either control (CON; n = 47) or β-carotene (BC; n = 47) treatments. When locked in headgates each morning, each cow received a topdress of β-carotene (Rovimix, 8 g/d; provided 800 mg of β-carotene) or carrier from 21 d before expected calving until calving. Blood samples were collected at 21 ± 1 d (mean ± standard deviation) before expected calving (before treatments began), 7 ± 1 d before calving, immediately following parturition, and 7 ± 1 d postpartum. Blood serum was analyzed for vitamins A and E, β-carotene, cholesterol, and other metabolites and enzymes. Data were analyzed using the MIXED procedure in SAS (SAS Institute Inc.). Cows had low β-carotene concentrations (0.85 μg/mL) in blood serum before treatments began. Compared with CON cows, BC cows had higher overall mean concentrations of β-carotene (2.87 μg/mL vs. 0.73 μg/mL) and retinol (165 vs. 143 ng/mL). Cows fed BC had lower α-tocopherol in serum than cows fed CON (2.26 vs. 2.46 µg/mL). Cows fed BC had lower peak milk than cows fed CON (50.9 vs. 55.3), but total lactation milk yield did not differ significantly. No effects of BC were observed on days to conception (100 d) or times bred (2.4). Treatments did not affect incidences of ketosis, retained placenta, displaced abomasum, off feed, lameness, footrot, mastitis, or metritis. In conclusion, in pregnant cows already receiving adequate vitamin A but with low serum β-carotene concentration, supplementation of β-carotene increased concentrations of β-carotene and vitamin A in blood serum, but did not affect production, reproduction, or health.