Blood Plasm Research Articles

The N and C-terminal deleted variant of the dengue virus NS1 protein is a potential candidate for dengue vaccine development

NS1 is an elusive dengue protein, involved in viral replication, assembly, pathogenesis, and immune evasion. Its levels in blood plasm are positively related to disease severity like thrombocytopenia, hemorrhage, and vascular leakage. Despite its pathogenic roles, NS1 is being used in various vaccine formulations due to its sequence conservancy, ability to produce protective antibodies and low risk for inducing antibody-dependent enhancement. In this study, we have used bioinformatics tools and reported literature to develop an NS1 variant (dNS1). Molecular docking studies were performed to evaluate the receptor-binding ability of the NS1 and dNS1 with TLR4. NS1 and dNS1 (153 to 312 amino acid region) genes were cloned, expressed and protein was purified followed by refolding. Docking studies showed the binding of NS1 and dNS1 with the TLR4 receptor which suggests that N and C-terminal sequences of NS1 are not critical for receptor binding. Antibodies against NS1 and dNS1 were raised in rabbits and binding affinity of anti-dNS1 anti-NS1 sera was evaluated against both NS1 and dNS1. Similar results were observed through western blotting which highlight that N and C-terminal deletion of NS1 does not compromise the immunogenic potential of dNS1 hence, supports its use in future vaccine formulations as a substitute for NS1.

#500 Monitoring relative blood volume and calf bioimpedance in hemodialysis patients

Abstract Background and Aims Measurements of relative blood volume (RBV, %) are used to monitor the relative change in blood plasm during hemodialysis (HD). Continuous measurement of calf extracellular volume (cECV) during HD allows the calculation of relative change in cECV (RCE). RCE was defined as the ratio of cECV to its initial value, cECV0 (RCE = 100 × cECV/cECV0, %). The aim of our study was to investigate the relationship between RBV, RCE and the occurrence of intradialytic hypotension (IDH). Method Fifty-five HD patients were studied. We reduced the HD target weight stepwise by 0.1 to 0.2 kg per HD session until clinically defined IDH symptoms occurred. RBV was monitored using the blood volume monitor (BVM, Fresenius Medical Care, Germany). Whole body bioimpedance spectroscopy (wBIS) provided whole body ECV (wECV) and intracellular volume (wICV) pre and post HD. Continuous calf bioimpedance (Hydra 4200, Xitron Technologies, San Diego, CA USA) provided cECV during HD. Pre HD overhydration (OH) was calculated according to whole body composition model (Chamney et al, Am J Clin Nutr 2007; 85:80). Total number of HD sessions in all patients were divided into three groups according to the pre HD OH: 1) normal OH (NOH, OH < 1 L), 2) moderate OH (MOH, 1 ≤ OH < 4 L) and 3) severe OH (SOH, OH ≥ 4 L). Furthermore, patients were stratified by the presence or absence of IDH in each hydration group. Ultrafiltration rate (UFR), pre HD body weight, body mass index (BMI) were measured. Minimal RBV (RBVMin) and minimal RCE (RCEMin) in each HD session were recorded. These measures represent maximum reduction of fluid in the intravascular and interstitial spaces, respectively. Reducing cECV can be considered as refilling fluid from interstitial transferring to intravascular space driving by ultrafiltration. The difference between RCEMin and RBVMin was defined as vascular refilling capacity. Linear mixed-effect logistic regression (LMELR) analysis was applied to explore relationships between IDH with RBVMin, RCEMin, wECV and BMI. Results We studied 621 HD sessions in 55 patients. MOH was present in 57%, SOH in 22% and NOH in 21% of total number of HD sessions respectively. There were significant differences in age, UFR, RBVMin, RCEMin, pre HD wECV, BMI between three groups, while the IDH rate did not differ significantly (Table 1). RBVMin was inversely correlated with UFR (Fig. 1) in all measurements. Pre HD wECV was the only factor associated with IDH in overall measurement. Pre HD wECV was significantly lower in IDH than in non-IDH patients in the NOH group (Fig. 2). In the MOH group, UFR and RCEMin-RBVMin were higher in IDH than in non-IDH patients (Fig. 3a and Fig. 3b). In the SOH group, Pre HD OH is the major factor associated with IDH (Fig. 4). Conclusion This study found that the major factors leading to IDH are 1) lower hydration status; 2) relative higher UFR; and 3) the limited refilling capacity. RBVMin correlated with UFR, but it did not predict IDH. Combined analysis of RBV and calf bioimpedance may provide a means to quantitate the fluid transport between intravascular and interstitial compartments and could add to an early IDH warning system.

EFFECT OF THREE ANTIBIOTICS ON BYPERIMENTAL INFECTION OF BROILERS WITH MYCOPLASMA GALLISEPTICUM

EFFECT OF THREE ANTIBIOTICS ON BYPERIMENTAL INFECTION OF BROILERS WITH MYCOPLASMA GALLISEPTICUM

Exercised-enriched blood plasma rescues hippocampal impairments and cognitive deficits in an Alzheimer's disease model

Alzheimer's disease (AD) is a progressive neurodegenerative disorder, and moderate exercise holds promise in ameliorating the ongoing neurodegeneration and cognitive decline. Here, we investigated whether exercise-enriched blood plasm could yield a beneficial therapeutic effect on AD pathologies and cognitive decline in transgenic AD (P301S) mice. In this investigation, a cohort of 2-month-old C57BL/6 mice were granted continuous access to either a running wheel or a fixed wheel for 6 weeks. After that, their plasmas were extracted and subsequently injected intravenously into 4.5-month-old P301S mice biweekly over a 6-week period. A comprehensive methodology was then employed, integrating behavioral tests, pathology assessments, and biochemical analyses to unveil the potential anti-dementia implications of exercise-enriched blood plasma in P301S mice. Upon systemic administration, the findings revealed a noteworthy attenuation of hippocampus-dependent behavioral impairments in P301S mice. Conversely, blood plasma from sedentary counterparts exhibited no discernible impact. These effects were intricately associated with the mitigation of neuroinflammation, the augmentation of hippocampal adult neurogenesis, and a reduction of synaptic impairments following the administration of exercise-enriched blood plasma. These findings advance the proposition that administering exercise-enriched blood plasma may serve as an effective prophylactic measure against AD, opening avenues for further exploration and potential therapeutic interventions.

Aloe Vera Polyphenols Against Fish Stress Through Blood Glucose

This study aims to analyze the blood glucose of koi fish exposed to polyphenol fraction of A. vera. It firstly found LC50 at the treatment of 150 mg.kg-1 that caused 50% mortality, so that the doses of A. vera fraction injected into the fish muscle as follows: Control (-) no treatment, Control (+) was given tannin/synthetic polyphenolic compound as much as 3 mg.kg-1 of body weight, treatment A= 75 mg.kg-1 of body weight, B= 100mg.kg-1 of body weight, C= 125 mg.kg-1 of body weight, respectively. After 72 h of injection, the blood plasm of Koi fish was taken to observe the stress effect on the blood glucose. Results showed that for blood chemistry, glucose with increased polyphenol fraction application due to higher stress level, with increased polyphenol dose. The clinical symptom after injected with polyphenol fraction of A. vera resulted in abnormal swimming, red spots, paleness, and hemorhage.

Effect of Polyphenol Fraction Exposure of Aloe vera on the Change in Cortisol of Koi Fish (Cyprinus carpio) as Stress Response

This study aims to analyze the stress response of koi fish exposed to polyphenol fraction of A. vera, metabolic effect of cortisol. It firstly found LC50 at the treatment of 150 mg.kg-1 that caused 50% mortality, so that the doses of A. vera fraction injected into the fish muscle as follows: Control (-) no treatment, Control (+) was given tannin/synthetic polyphenolic compound as much as 3 mg.kg-1 of body weight, treatment A=75 mg.kg-1 of body weight, B= 100 mg.kg-1of body weight, C- 125 mg.kg-1 of body weight, respectively. After 72 h of injection, the blood plasm of Koi fish was taken to observe the stress effect on the cortisol. Results showed that for cortisol rose with increased polyphenol fraction application due to higher stress level.

Comparison of impact of various application methods for the blood plasm, enriched by the thrombocytes, on healing of the burn and scalped wounds in experiment

Objective. Comparison, in conditions of experiment between impact of the blood plasm, enriched by thrombocytes, on the terms of the burn and scalped wounds healing and morphological characteristics of the cicatrices formatted in its injectional or noninjectional introduction, using the method of electrophoresis.
 Materials and methods. The investigations were conducted on 30 white rats, which were divided into three groups with 10 animals in every group, median body mass of the animals have constituted (275.5 ± 15.1) gr. After achievement of medicinal sleep (intraperitoneal introduction of 1.0 ml of 0.5% solution of sodium thiopental in combination with 0.2 ml of propofol 1% solution the scalped skin incisions with the 1.5 × 0.5 cm dimensions were performed to the animals. Besides that, one of the incisions was processed with diathermy up to the scab crust formation. Both wounds were left open, without sutures. To the control group animals after the wounding conduction no procedures were performed, the wounds have been healed in primary pattern. To the Group I animals during two consequent weeks the blood plasm, enriched by thrombocytes, was introduced, using intracutaneous injections twice a week in quantity of 10 ml, puncturing the injured zone around. To the investigation Group II animals the electrophoresis in accordance to own procedure through napkin, soaked in the blood plasm, enriched by thrombocytes, was performed with the same rate. The blood plasm, enriched by thrombocytes, was prepared from the whole venous blood of human Group A(0), using centrifugation for achievement of targeted quantity of thrombocytes 850 000 in 1 ml. Morphological features of cicatrices formatted and their width were studied, using the methods of light microscopy and morphometry on the 21th day of experiment.
 Results. Application of the blood plasm, enriched by thrombocytes, was accompanied by statistically significant reduction of the cicatrices width, which were formatted on the 21-th day in locations of the scalped wounds simulation (p < 0.05 and p < 0.01 accordingly, for injectional and electrophoretic methods of application in comparison with the control group) and the burn (p < 0.01 and p < 0.005, accordingly, for the injectional and electrophoretic methods of application in comparison to the control group) wound. At the same time the cicatrices width in electrophoretic application of the blood plasm, enriched by thrombocytes, was statistically significantly lesser (p < 0.05), than in its injectional introduction, not depending on the wound type. Besides that, the cicatrices connective tissue while application of the blood plasm, enriched by thrombocytes, in a moment of the animals evacuation from the experiment have looked like more mature and differed by enhanced organization of the collagen fascicles localization.
 Conclusion. The method of noninjectional introduction of the blood plasm, enriched by thrombocytes, which was proposed, owes certain advantages over its standard injectional application, which constitute the enhanced degree of maturation of connective tissue, more organized localization of the collagen fascicles, lesser width of the cicatrices evolved and the procedure painlessness. Noninjectional introduction of the blood plasm, enriched by thrombocytes, using method of electrophoresis is free from postinjectional complications, typical for standard methods of its introduction.

Highly sensitive detection of anti-cancer drug based on bimetallic reduced graphene oxide nanocomposite

Herein, we describe a high-performance electrochemical sensor for the detection of regorafenib (REG) using bimetallic Pd–Ru nanoparticles anchored on pomegranate peel extract (PPE) derived reduced graphene oxide (Pd–Ru/rGO). PPE was employed to neutralize the extremely acidic graphene then cast-off along with the metal precursor for the duration of the chemical reduction to accomplish well dispersed Pd–Ru nanoparticles. Bimetallic Pd–Ru/rGO nanocomposites were synthesized using a facile chemical reduction method. Under optimal conditions, based on the differential pulse voltammetric studies it has been confirmed that the fabricated sensors has good electrocatalytic activity toward the detection of REG, spanning over the linear dynamic range of 0.5–300 nM. Moreover, the sensor exhibited a low limit of detection of 1.6 nM and a limit of quantification of 4.8 nM. The electrochemical sensor unveiled admirable selectivity and sensitivity, reproducibility, and repeatability. The fabricated sensor was suitable for real sample analysis (pharmaceutical tablet, human blood plasm, wastewater) with satisfactory recovery. The strategy presented herein can be employed in the development of electrochemical sensors for other target analytes.

Involvement of kisspeptin in androgen-induced hypothalamic endoplasmic reticulum stress and its rescuing effect in PCOS rats

Endoplasmic reticulum (ER) stress, with adaptive unfolded protein response (UPR), is a key link between obesity, insulin resistance and type 2 diabetes, all of which are often present in the most common endocrine-metabolic disorder in women of reproductive age, polycystic ovary syndrome (PCOS), which is characterized with hyperandrogenism. However, the link between excess androgen and endoplasmic reticulum (ER) stress/insulin resistance in patients with polycystic ovary syndrome (PCOS) is unknown. An unexpected role of kisspeptin was reported in the regulation of UPR pathways and its involvement in the androgen-induced ER stress in hypothalamic neuronal cells. To evaluate the relationship of kisspeptin and ER stress, we detected kisspeptin and other factors in blood plasm of PCOS patients, rat models and hypothalamic neuronal cells. We detected higher testosterone and lower kisspeptin levels in the plasma of PCOS than that in non-PCOS women. We established a PCOS rat model by dihydrotestosterone (DHT) chronic exposure, and observed significantly downregulated kisspeptin expression and activated UPR pathways in PCOS rat hypothalamus compared to that in controls. Inhibition or knockdown of kisspeptin completely mimicked the enhancing effect of DHT on UPR pathways in a hypothalamic neuronal cell line, GT1–7. Kp10, the most potent peptide of kisspeptin, effectively reversed or suppressed the activated UPR pathways induced by DHT or thapsigargin, an ER stress activator, in GT1–7 cells, as well as in the hypothalamus in PCOS rats. Similarly, kisspeptin attenuated thapsigargin-induced Ca2+ response and the DHT- induced insulin resistance in GT1–7 cells. Collectively, the present study has revealed an unexpected protective role of kisspeptin against ER stress and insulin resistance in the hypothalamus and has provided a new treatment strategy targeting hypothalamic ER stress and insulin resistance with kisspeptin as a potential therapeutic agent.

Species-specific identification of porcine blood plasma in heat-treated chicken meatballs.

This study was conducted to detect the presence of chicken and porcine DNA in meatballs using mitochondria DNA (mtDNA) of cytochrome b (cyt b) and nuclear DNA (nDNA) short interspersed nuclear element (SINE) species-specific primers, respectively. While, the mtDNA primers targeted transfer RNA-ATP8 (tRNA-ATP8) gene was used for 1 and 5% (w/w) chicken meatball spiked with commercial porcine blood plasm. Chicken meatballs spiked with 1% and 5% (v/w) fresh and commercial porcine blood plasma, respectively were prepared and heat-treated using five (n = 5) cooking methods: boiling, pan-frying, roasting, microwaving and autoclaving. Two pairs of mtDNA and nDNA primers used, produced 129 and 161 bp amplicons, respectively. Whereas, tRNA-ATP8 primers produced 212 bp of amplicon. Electrophoresis analysis showed positive results for porcine DNA at 1% and 5% (w/w or v/v) for all of the different cooking techniques, either for fresh or commercial blood plasma using SINE primers but not for tRNA-ATP8 primers. The present study has highlighted the useful of species-specific primers of SINE primers in PCR analysis for detecting porcine DNA blood plasma in heat-treated chicken meatballs.

Lymphodynamics and adrenergic innervation of the kidney and renal lymph nodes in toxic hepatitis

Introduction . The lymphatic system maintains a steady state of organism and plays crucial role in a volume regulation of fluid surrounding the organs. The researching goal is to study the fluid redistribution process and to reveal adrenoceptive renal innervation of its lymphatic nodes in toxic hepatitis. Materials and methods . We studied the rat kidneys and their renal lymphatic nodes. The lymph flow of renal nodes and its adrenergic innervation were evaluated. Results . Rats with toxic hepatitis showed the decreasig fluorescence of adrenergic nerves and absence of the single neurons in kidney tissue and pelvis. Nerves in the renal parenchyma adjacent to the lymphatic nodes had a weak fluorescence. The average decreasing of lymph flow, volume of blood plasm and diuresis was 35, 21 and 60 %, correspondently. The disturbance of fluid redistribution, whole protein and electrolytes were observed. Conclusions . Toxic hepatitis leads to the destruction of adrenergic innervation in the tissues and blood vessels of the kidneys and renal lymph nodes, the disappearance of small terminals of nerves and part of varicose thickenings, the depots of catecholamines. This causes the diffusion of catecholamines into the surrounding tissues, suppressing the efferent signaling of nervous processes. The rats with hepatitis have had an impaired water-salt homeostasis resulted in a decreasing volume of plasma, circulating blood and lymph flow. The lymphatic system in hepatitis promotes the uptake of potassium and calcium ions released from the bloodstream, preventing them from being excreted in the urine.

Effect of antibiotics, hormones and anthelmintic on high molecular weight protein fractions in the common carp

We assumed that effect of sulfanilamide, chlortetracycline, nandrolone, and albendazole on the carp fishes depends on substance concentration in the water and associated with changes in the fractional composition of plasma protein. We found that sulfanilamide in concentration of 0.015 mg/dm3 in water practically had no effect, while at concentrations of 0.15 and 0.30 mg/dm3 it changed the protein spectrum of carp blood plasma, reduced the level of proteins with molecular weights of 260 and 140 kDa. Chlortetracycline in the concentration of 1.10 mg/dm3 did not affect the protein content of macromolecular fractions in the blood plasma of fish, and at the doses of 3.15 and 6.30 mg/dm3 it reduced the content of proteins with molecular weights of 450, 340, 260 and 140 kDa. Nandrolone in the concentration of 1.10 mg/dm3 increased the content of individual proteins in carp plasma compared to the control, and in the concentration of 0.10 and 0.50 mg/dm3 it did not affect the fractional composition of proteins with high molecular weight. Albendazole in the concentrations of 0.2, 0.5, and 1.00 mg/dm2 reduced the protein content of macromolecular fractions in carp plasma. Such xenobiotics, like sulfanilamide, chlortetracycline, and albendazole in different concentrations did not affect the total protein content of fish plasma, while the nandrolone increased it. The number of respiratory movements, behavior, body surface, and pathomorphological parameters of the main internal organs in the fish from the experimental groups did not differ from the control. Our results indicated the important role of proteins of high molecular weight fractions in carp blood plasm as these proteins determine the fish adaptation to the water xenobiotics. Keywords: carp, blood plasma, proteins, sulfanilamide, chlortetracycline, nandrolone, albendazole.

C-reactive protein and procalcitonin - laboratory markers of degrees of severity and complications of an acute necrotic pancreatitis

Objective. To conduct comparative estimation of efficacy of C-reactive protein and procalcitonin as laboratory markers for stratification of the patients severity state suffering an acute necrotic pancreatitis.
 Materials and methods. Prospective cohort investigation, including 151 patients with an acute necrotic pancreatitis, was conducted. Clinical, laboratory and bacteriological investigations were accomplished. The levels of C-reactive protein and procalcitonin were determined in the blood plasm.
 Results. The necrotic accumulations infectioning was diagnosed in 89 (58.9%) patients: local purulent complications - in 27, sepsis - in 33,septic shock - in 29. In 62 patients with sterile pancreonecrosis a C-reactive protein concentration have raised from (5.6 ± 0.89) to (206 ± 29) mg/l (p˂0.001). Development of purulent-septic complications was accompanied by significant and trustworthy (p<0.01) elevation of procalcitonin concentration: in the patients with sepsis - up to (5.05 ± 0.92) ng/ml, in the patients with septic shock - up to (7.25 ± 2.15) ng/ml.
 Conclusion. Simultaneous measurement of levels of C-reactive protein and procalcitonin in the blood plasm in patients, suffering acute necrotic pancreatitis, gives permission to determine the inflammatory process character and stratify the disease severity in its early terms.

KADAR SENYAWA TOKSIK DALAM DARAH IKAN NILA (Oreochromis niloticus) SAAT MUSIM UP-WELLING DI DANAU MANINJAU, SUMATRA BARAT

Maninjau lake is the largest center for the freshwater floating-net-cage fisheries system in West Sumatra. Every year, during the up-welling season, a huge number of fish suffered to death in the lake including nile tilapia (Oreochromis niloticus), a common species cultivated in the floating-net cages. This current study aimed to determine the factors contributing to the death of fish by measuring the concentration of toxic chemical substances in the blood plasm of nile tilapia as well as in the subtrate collected from the bottom of the lake. The samples were collected in the sites with higher number of floating-net cages as well as higher number of mass death of fish (Tanjung Sani, Pakan Raba’a dan Maninjau) during the up-welling season. The result revealed that the levels of phosphate, nitrate, nitrit, and amonium were higher in the blood plasm of the fish. Moreover, the levels of those chemical substances were also higher in the substrate of the lake. The higher concentration of the toxic chemical substances might contribute to the mass death of fish living in the floating-net cages during the up-welling season.

Detection of the State of Thrombotic Readiness in Rats under One-Time Suprathreshold Physical Activity of Various Durations by Means of Thromboelastography

Suprathreshold physical activity causing distress in the organism can lead to the damage of various organs and systems including the hemostatic system. A modern integrated method of the hemostatic system assessment is thromboelastography. The purpose of the present study was to evaluate the state of the hemostatic system under one-time suprathreshold physical activity of various durations by means of thromboelastography. Experimental groups of rats were exposed to 4-hour and 8-hour physical activities in the form of forced running on a moving platform with the speed of 6-8 m/min. Immediately after the one-time physical activity, blood samples taken from rats were examined using the thrombelastograph in the Natem mode for 35 minutes. The 4-hour physical activity caused a reduction in coagulation time (CT) and an increase in the alpha angle and the maximum clot firmness (MCF). After the 8-hour activity, the thrombelastograph registered a reduction in coagulation time (CT), an increase in the alpha angle, a decrease in the clot formation time (CFT), a decrease in the maximum clot firmness (MCF), and a reduction in the maximum clot lysis (ML). The 4-hour physical activity resulted in partial activation of the hemostatic system without changing the fibrinolytic activity of blood plasm. The changes revealed in thromboelastography parameters indicate a high risk of the development of thrombotic readiness. The 8-hour physical activity causes a shift of the hemostatic system parameters in rats towards the increased clot formation: hypercoagulation, fibrinogen and platelet consumption, inhibition of fibrinolysis. The combination of changes in thromboelastogram parameters is indicative of the development of thrombotic readiness

Escherichia coli BL21(DE3) expression system using TorA signal peptide for Recombinant Human Albumin (rHA) secretion

Human serum albumin (HSA) is the most abundant protein in blood plasm. This protein consisted of 585 amino acids with a molecular weight of 66 kDa and 17 disulfide bonds. HSA obtained from conventional technique allow viral or prion contamination. For that reason, recombinant DNA technology becomes a promising alternative. Because of its well-known genetic, simplicity, and capacity to accommodate many foreign protein, Escherichia coli remains the most widely used in the production of recombinant proteins. But, overproduction of protein may lead to the formation of inclusion bodies and proteolytic degradation. These problems can be overcome by using protease-deficient strain and protein secretion into periplasmic space. The objective of this research is to secrete recombinant HA on E. coli BL21(DE3) using TorA signal peptide and proved using SDS-PAGE. This research method begins with the preparation of competent cell and transformation of E. coli BL21(DE3), expression of recombinant HA in E. coli BL21(DE3), and characterization of expression result by using SDS-PAGE. The result of this study was rHSA can be secreted into extracellular medium using TorA signal peptide with a molecular weight of ± 66.5 kDa.

Involvement of Kisspeptin in Androgen-Induced Hypothalamic Endoplasmic Reticulum Stress and Its Rescuing Effect in PCOS Rats

Background: Endoplasmic reticulum (ER) stress, with adaptive unfolded protein response (UPR), is a key link between obesity, insulin resistance and type 2 diabetes, all of which are often present in the most common endocrine-metabolic disorder in women of reproductive age, polycystic ovary syndrome (PCOS), which is characterized with hyperandrogenism. However, the link between excess androgen and Endoplasmic reticulum (ER) stress/insulin resistance in patients with polycystic ovary syndrome (PCOS) is unknown. Methods: An unexpected role of kisspeptin was reported in the regulation of UPR pathways and its involvement in the androgen-induced ER stress in hypothalamic neuronal cells. To evaluate the relationship of kisspeptin and ER stress, We detected Kisspeptin and other factors in blood plasm of PCOS patients, rat models and hypothalamic neuronal cells. Findings: We detected higher testosterone and lower kisspeptin levels in the plasma of PCOS than that in non-PCOS women. We established a PCOS rat model by dihydrotestosterone (DHT) chronic exposure, and observed significantly down-regulated kisspeptin expression and activated UPR pathways in PCOS rat hypothalamus compared to that in controls. Inhibition or knockdown of kisspeptin completely mimicked the enhancing effect of DHT on UPR pathways in a hypothalamic neuronal cell line, GT1-7. Kp10, the most potent peptide of kisspeptin, effectively reversed or suppressed the activated UPR pathways induced by DHT or thapsigargin, an ER stress activator, in GT1-7 cells, as well as in the hypothalamus in PCOS rats. Similarly, Kisspeptin attenuated thapsigargin-induced Ca2+ response and the DHT- induced insulin resistance in GT1-7 cells. Interpretation: Our data has revealed an unexpected protective role of kisspeptin against ER stress and insulin resistance in the hypothalamus and provided a new treatment strategy targeting hypothalamic ER stress and insulin resistance with kisspeptin as a potential therapeutic agent. Funding Statement: National Basic Research Program of China, National Natural Science Foundation of China (2012CB944902, 2012CB944703, 2013CB967404, 2013CB967401, 2013CB967403, 81571403, 81471429) and State Key laboratory of Reproductive Medicine (SKLRM-K201805). Declaration of Interests: The authors have no conflicts of interest about this work. Ethics Approval Statement: The study was approved by the Ethics Committee of Tongji University and all patients gave informed consent. All procedures in this study were approved by the Animal Experimentation Ethics Committee of the Chinese University of Hong Kong, and the methods were used in accordance with the guidelines of the Chinses University of Hong Kong.

Eltrombopag Combined with G-CSF and Cyclosporine Could Effect for Severe Acquired Aplastic Anemia

BackgroudAcquired aplastic anemia (AA) is a potential life-threatening hematopoietic stem cell (HSC) disorder resulting in cytopenia. The first line therapy for AA is HSC transplantation for young patients who have suitable donors and immunosuppressive therapy (IST) with antithymocyte globulin and cyclosporine for the remaining patients. However, about 30% of patients are refractory to IST or relapse after IST. IST with antithymocyte globulin and cyclosporine result in severe complication and mortality infection. To reduce the mortality infection and increase the response of IST for AA is still problem. Eltrombopag, a thrombopoietin mimetic, demonstrated efficacy in restoring trilineage hematopoiesis, has recently emerged as an encouraging and promising agent for patients with refractory AA. To explore the effect of eltrombopag for severe acquired aplastic anemia, we treated seven severe AA patients with eltrombopag combined with cyclosporine and G-CSF. Herein we report initial results of the eltrombopag combined with cyclosporine and G-CSF for severe AA.MethodsThe diagnostic of AA patient consisted of a complete blood count, a bone marrow biopsy, bone marrow karyotype analysis and assessment of a paroxysmal nocturnal hemoglobinuria (PNH) clone. Patients with SAA aged ≥18 years old who without suitable donors received eltrombopag 75mg/d, cyclosporine 6mg/kg by oral, and G-CSF 300ug/d by subcutaneous injection from diagnosis. Red blood was infused to maintained HB more than 60g/L. Platelet were infused to maintained PLT more than 20x109/L. G-CSF was administered until neutrophil count more than 1.0x109/L. Vale concentration of cyclosporine were maintained more than 100ug/ml in blood plasm and maintained two years. Eltrombopag was taper down when platelet was more than 100x109/L. Eltrombopag was given at least three months. Antibacterial was administered when patient was high fever. Posaconazole were given for fungal infections prophylaxsis. Hematologic improvements were assessed by the National Institutes of Health (NIH) response criteria for AA.ResultsThe median age of 7 patients with SAA was 44 years old (range 19-68 yr). Full hematologic improvements were achieved in 3 patients. All patients achieved platelet and RBC infusion independence. The median time from the first eltrombopag therapy to platelet infusion independence was 35 days (range 33-46d). The median time from the first eltrombopag therapy to RBC infusion independence was 40 days (range 30-50d). Median 6 units (1200ml) (range 3-10U, 600ml -2000ml) RBC and 7 units (2.5x109/unit) platelet were infused. With median 8 months follow-up (3-12 months),3 patients are still full hematologic improvements and 4 platelet and RBC infusion independence. No severe fugual infection was observed in this group patients. ALT slightly elevate in one patient. No other severe adverse effect was observed.ConclusionsTreatment of SAA patients with G-CSF、cyclosporine combined with eltrombopag is feasible and effect. Our results deserve further research and confirmation in larger samples. DisclosuresNo relevant conflicts of interest to declare.

Ефективність вживання вуглеводного напою перед ортопедичними хірургічними втручаннями у дітей

Мета. Визначити вплив передопераційного вживання вуглеводного напою на метаболічні показники стресової відповіді на ортопедичні хірургічні втручання у дітей.
 Матеріали і методи. У дослідній групі 30 дітей віком старше 1 року вживали напій, що містив 25 мг харчового вуглеводу мальтодекстрину на 100 мл води, за дві години до індукції анестезії. У контрольній групі 30 дітям призначали стандартне передопераційне голодування. Порівнювали концентрації глюкози, інсуліну, кортизолу у плазмі крові, індекси інсулінорезистентності та чутливості до інсуліну та наявність кетонових тіл у сечі після операції.
 Результати. Перед операцією показники концентрації глюкози у плазмі крові в дослідній групі були вище, ніж у контрольній групі (р = 0,02). Наприкінці операції в дослідній групі були вище показники індексу чутливості до інсуліну (р = 0,01), нижче показники концентрації інсуліну у плазмі крові (р=0,008) та індексу інсулінорезистетності (р = 0,01). У контрольній групі індекс інсулінорезистентності перевищував норму у 30% хворих. Кетонові тіла в сечі в дослідній групі спостерігали у 20%, в контрольній - у 50% хворих.
 Висновки. Вживання вуглеводного напою за дві години до хірургічного втручання покращувало метаболічний профіль у дітей, яким виконували ортопедичні операції, та зменшувало прояви хірургічного стресу.

Short Communication: Genetic diversity of Ongole Grade Cattle of Rembang District, Central Java, Indonesia, based on blood protein polymorphism

Sutiyono, Sutopo, Ondho YS, Setiatin ET, Samsudewa D, Suryawijaya A, Lestari DA, Kurnianto E. 2018. Genetic diversity of Ongole Grade Cattle of Rembang District, Central Java, Indonesia, based on blood protein polymorphism. Biodiversitas 19: 1429-1433. The objective of this study was to identify genetic diversity in Ongole Grade cattle of Rembang District, Central Java, Indonesia, based on blood-protein polymorphism. A total of 34 blood samples were collected from unrelated Ongole Grade cattle and the blood plasm was used to identify the blood proteins. Blood protein analysis was performed using Sodium Dodecyl Sulfate-Polyacrilamide Gel Electrophoresis (SDS-PAGE). Observed bands were used to perform allele interpretation of four loci, namely Albumin (Alb), Post-albumin (Po-Alb), Ceruloplasmin (Cp) and Amylase-1 (Am-1). Results showed the presence of polymorphism in all of the four blood-protein loci, average heterozigosity value (H) being 0.506 and almost all of the blood-protein loci studied showed deviation from Hardy-Weimberg Equilibrium (HWE). In conclusion, the study indicated that population of Ongole Grade cattle in Rembang, Indonesia has high genetic diversity.