The production of proteins in sufficient amounts is key for their study or use as biotherapeutic agents. Escherichia coli is the host of choice for recombinant protein production given its fast ...growth, easy manipulation, and cost‐effectiveness. As such, its protein production capabilities are continuously being improved. Also, the associated tools (such as plasmids and cultivation conditions) are subject of ongoing research to optimize product yield. In this work, we review the latest advances in recombinant protein production in E. coli.
Atherosclerosis and its clinical manifestation as ischaemic heart disease remains a considerable health burden. Given that many factors contribute to ischaemic heart disease, a multifactorial ...approach to prevention is recommended, starting with lifestyle advice, smoking cessation, and control of known cardiovascular risk factors, such as blood pressure and lipids. Within the lipid profile, the principal target is lowering LDL cholesterol, firstly with lifestyle interventions and subsequently with pharmacological therapy. Statins are the recommended first-line pharmacological treatment. Some individuals might require further lowering of LDL cholesterol or be unable to tolerate statins. Additional therapies targeting different pathways in cholesterol metabolism are now available, ranging from small molecules taken orally, to injectable therapies. Examples include ezetimibe, which targets Niemann-Pick C1-like protein, and monoclonal antibodies that target PCSK9. Phase 3 trials have also been completed for bempedoic acid (targeting ATP-citrate lyase) and inclisiran (an interference RNA-based therapeutic targeting hepatic PCSK9 synthesis). In addition to LDL cholesterol, mendelian randomisation studies support a causal role for lipoprotein(a) and triglycerides in ischaemic heart disease. In this Series paper, we appraise currently available and emerging therapies for lowering LDL cholesterol, lipoprotein(a), and triglycerides for prevention of ischaemic heart disease.
Obesity-related glomerulopathy (ORG) is characterized by glomerulomegaly accompanied in many patients by lesions of focal and segmental glomerulosclerosis. Slowly increasing subnephrotic proteinuria ...is the commonest presentation of ORG. Occasionally, massive proteinuria (>5-10 g/day) is detected, but the typical findings of nephrotic syndrome are characteristically absent even in patients with massive proteinuria. Superimposed obesity can fuel the progression of other renal diseases, and a reduced number of functioning nephrons (of congenital or acquired causes) synergizes with obesity to induce end-stage renal disease. Weight loss, either induced by diet or bariatric surgery, and renin-angiotensin blockers are effective treatments to slow progression, but a significant proportion of cases will develop end-stage renal disease.
Starch edible films (EFs) have been widely studied due to their potential in food preservation; however, their application is limited because of their poor mechanical and barrier properties. Because ...of that, the aim of this work was to use the extrusion technology (ExT) as a pretreatment of casting technique to change the starch structure in order to obtain EFs with improved physicochemical properties. To this, corn starch and a mixture of plasticizers (sorbitol and glycerol, in different ratios) were processed in a twin screw extruder to generate the starch modification and subsequently casting technique was used for EFs formation. The best conditions of the ExT and plasticizers concentration were obtained using response surface methodology. All the response variables evaluated, were affected significatively by the Plasticizers Ratio (Sorbitol:Glycerol) (PR (S:G)) and Extrusion Temperature (ET), while the Screw Speed (SS) did not show significant effect on any of these variables. The optimization study showed that the appropriate conditions to obtain EFs with the best mechanical and barrier properties were ET = 89 °C, SS = 66 rpm and PR (S:G) = 79.7:20.3. Once the best conditions were obtained, the optimal treatment was characterized according to its microstructural properties (X‐ray diffraction, Scanning Electron Microscopy and Atomic Force Microscopy) to determine the damage caused in the starch during ExT and casting technique. In conclusion, with the combination of ExT and casting technique were obtained EFs with greater breaking strength and deformation, as well as lower water vapor permeability than those reported in the literature.
Practical Application
Starch is becoming an environmentally friendly alternative to produce films due to its low cost, high biodegradability, and thermoplastic properties. However, native starches have many disadvantages, limiting their broader applications and industrial use. For this reason, the use of ExT as a pretreatment combined with the conventional method (casting technique) to elaborate EFs, could be an alternative to change the structure of starch in order to generate films with improved mechanical and barrier properties.
This study aimed to describe updated information on the basic reproductive biology of Sphoeroides annulatus females and potential use of the gonadosomatic index IG as input data to assess length at ...50% maturity (L50) and identify the breeding season. From April 2014 to December 2015, 767 females were analysed along the north‐west coast of the Mexican Pacific Ocean. Both immature and mature females were represented in the total length (LT), which ranged from 81 to 440 mm, and the smallest mature female was 92 mm (new record for the species). Gonadal maturity phases were assigned through histological analysis. A IG threshold value Icut was established using the lower whisker value for maturity phase spawning capable (2.06%). The monthly variation of IG supported the annual spawning seasonality, and the presence of reproductively active mature females IG≥Icut coincided with the observed periods with histological data and the peaks for both seasons 2014 and 2015. Also the presence of reproductively inactive IG<Icut mature females confirmed the nonreproductive season observed from January to April 2015. Estimates of L50 were based on the cumulative proportion of reproductively active mature females: both datasets showed a ‘bumpy trajectory’. Considering the functional form of data and models, a second objective was to verify if each dataset presented a simple or double sigmoid trajectory and select the most suitable model through the Akaike information criterion. For both datasets, the best fitted model was the double sigmoidal MDS, while the single sigmoidal MSS had low performance and did not have statistical support. The MDS provided a L50 value ca. 254 mm LT for both data sets. A trade‐off between the model's correct fit and a reliable L50 value was found, but this fact is not always considered when calculating the maturity, resulting in biased parameters. Correct data fitting may seem trivial, but the consequences for the conservation of immature individuals could be dramatic, affecting the population recruitment. In conclusion, the histologically calibrated Icut was suitable for classifying females of S. annulatus into reproductively inactive or active types, with an agreement percentage of 99%. The Icut = 2.06% can be useful for subsequent studies for this species.
OBJECTIVE: Dapagliflozin, a novel inhibitor of renal sodium-glucose cotransporter 2, allows an insulin-independent approach to improve type 2 diabetes hyperglycemia. In this multiple-dose study we ...evaluated the safety and efficacy of dapagliflozin in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: Type 2 diabetic patients were randomly assigned to one of five dapagliflozin doses, metformin XR, or placebo for 12 weeks. The primary objective was to compare mean change from baseline in A1C. Other objectives included comparison of changes in fasting plasma glucose (FPG), weight, adverse events, and laboratory measurements. RESULTS: After 12 weeks, dapagliflozin induced moderate glucosuria (52-85 g urinary glucose/day) and demonstrated significant glycemic improvements versus placebo (ΔA1C -0.55 to -0.90% and ΔFPG -16 to -31 mg/dl). Weight loss change versus placebo was -1.3 to -2.0 kg. There was no change in renal function. Serum uric acid decreased, serum magnesium increased, serum phosphate increased at higher doses, and dose-related 24-h urine volume and hematocrit increased, all of small magnitude. Treatment-emergent adverse events were similar across all groups. CONCLUSIONS: Dapagliflozin improved hyperglycemia and facilitates weight loss in type 2 diabetic patients by inducing controlled glucosuria with urinary loss of ~200-300 kcal/day. Dapagliflozin treatment demonstrated no persistent, clinically significant osmolarity, volume, or renal status changes.
•Polymeric nanoparticles (PNp) and radionuclide-based theranostic systems (nano-radiopharmaceuticals) merge the advantages of radiopharmaceuticals in theranostic applications.•The use of derivatives ...such as DOTA as bifunctional chelating agent decreases the specific activity of the radiolabelled-PNp.•The formation of the complexes Lu(HAsc)(OH)2(H2O)4, Lu(HAsc)(OH)2(PLGA)2 and Lu(HAsc)(OH)2(γ-PGA)2 with a probable coordination number of eight and a coordination sphere of eight oxygen atoms is proposed.•The preparation of PNp, the Lu(HAsc)(OH)2(γ-PGA)2 is very attractive for preparing theranostic nanoradiopharmaceuticals with 177Lu.
Radiolabelled nanoparticles (nanoradiopharmaceuticals) merge the advantages of radiopharmaceuticals and nanoparticles. Among them, polymeric nanoparticles (PNp) can be designed and prepared with architecture, surface charge and functionalization according to applications. Lu-177 is a radionuclide widely used for the preparation of nanoradiopharmaceuticals due to its theranostic properties. PNp are loaded with a therapeutic agent while 177Lu is usually indirectly bonded to PNp-surface, mostly through bifunctional chelating agents such as DOTA and its derivatives. However, the use of these derivatives typically does not yield a high specific activity of the radiolabelled-PNp. A strategy to increase the specific activity of these nanosystems is implementing direct radiolabelling methodologies which bind 177Lu into the PNp surface. The aim of this study was to design and propose, aided by computational chemistry tools, a labelling strategy for PNp systems. Molecules selected for complexing Lu were bidentate and tridentate ligands. Such ligands together should close the 177Lu coordination sphere to form Lu-complexes with near Lu-DOTA stability constant. The combinations of ligands (glycine, diethylenetriamine, lactic acid, glycolic acid, ascorbic acid, poly(lactic-co-glycolic acid) (PLGA) and poly(L-γ-glutamic acid) (γ-PGA) have the desired characteristics and also are frequently used in the preparation of PNp, another feature sought in the ligand selection. The potential formation of Lu-complex using the aforementioned compounds were simulated (with non-radioactive atom) under standard conditions and in aqueous media at different temperatures. Four of the simulated complexes were prepared in acid media (also with a non-radioactive atom) and their potentiometric and spectrophotometric responses were followed during titration with NaOH. Experimental results showed the formation of 3 different complexes with probable coordination number 8, one of which shows possibilities as a potential candidate to assemble 177Lu-PNp.
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Hematuria is a cardinal symptom in IgA nephropathy, but its influence on the risk of disease progression has been scarcely investigated. We followed a cohort of 112 patients with IgA nephropathy for ...a mean±SEM period of 14±10.2 years, during which clinical and analytic risk factors (including urine sediment examination) were regularly recorded. According to the magnitude of time-averaged hematuria, we classified patients as those with persistent hematuria and those with negative or minimal hematuria. We also classified patients according to the magnitude of time-averaged proteinuria (>0.75 or ≤0.75 g/d). The proportion of patients reaching ESRD or a 50% reduction of renal function was significantly greater among patients with persistent hematuria than patients with minimal or negative hematuria (30.4% and 37.0% versus 10.6% and 15.2%, respectively;
=0.01). Multivariable analysis revealed time-averaged hematuria, time-averaged proteinuria, renal function at baseline, and the presence of tubulointerstitial fibrosis on renal biopsy as independent predictors of ESRD. After hematuria disappearance, which occurred in 46% of the patients, the rate of renal function decline changed from -6.45±14.66 to -0.18±2.56 ml/min per 1.73 m
per year (
=0.001). Patients with time-averaged proteinuria >0.75 g/d had significantly poorer renal survival than those with time-averaged proteinuria ≤0.75 g/d. However, on further classification by time-averaged hematuria, only those patients with time-averaged proteinuria >0.75 g/d and persistent hematuria had significantly worse renal survival than those in the other three groups. In conclusion, remission of hematuria may have a significant favorable effect on IgA nephropathy outcomes.
•The environmental niche of chub mackerel was analyzed via minimum volume ellipsoid.•Model calibration with climatic extremes may improve prediction of spatial patterns.•High temporal resolution ...climatic data describe well the estimation of species niche.•Chub mackerel environmental suitability areas could reduce in future scenarios.
To understand and characterize the Pacific chub mackerel (Scomber japonicus) spatiotemporal distribution in the northern Peruvian Current System, this study characterized the ecological niche using the minimum volume ellipsoid (MVE) approach considering two meteorological conditions jointly: El Niño (EN) and La Niña (LN). For this purpose, the species presence records collected by the Peruvian onboard observer program from 1997 to 2018 were used. All presence records were matched with the date (day/month/year) and location (longitude, latitude) of the corresponding oceanographic variables (sea surface temperature SST, sea surface salinity SSS, chlorophyll, and oxygen). Also, the future projected climate change effects on chub mackerel spatiotemporal distribution under Representative Concentration Pathway (RCP) 4.5 and 8.5 scenarios by mid- (2040–2050) and end‐of‐the‐century (2090–2100) were investigated. Ellipsoid evaluations demonstrated that models calibrated with SST and SSS had good performance. Projections for EN conditions predicted areas with high environmental suitability close and along the Peruvian coast. Contrastingly, during LN these areas were extended to the oceanic zone. The projected future scenarios showed that for the RCP 4.5 scenario, environmental suitability maps were similar to those found in LN but with persistent distribution in south-central Peru. For the RCP 8.5 scenario, habitats tended to be coastal. Based on climate refugia –areas where the bulk of the species may aggregate under future environmental change with the potential to re-expand once the stress abates–, the RCP 4.5 scenario did not show changes in the hotspots; conversely, the RCP 8.5 scenario predicted a reduction of around 47% of hotspots from 2040–2050 to 2090–2100 at 10° S-18° S. Finally, the consideration of extreme conditions (EN and LN) in model calibration can potentially generate models with good performance allowing to better characterize species niche when the data is restricted by political boundaries. Nevertheless, further testing must be performed to assert this idea.
Flaviviruses are representative arboviruses carried by arthropods and/or vertebrates; these viruses can pose a public health concern in many countries. By contrast, it is known that a novel virus ...group called insect-specific flaviviruses (ISFs) also infects arthropods, although no such virus has yet been isolated from vertebrates. The characteristics of ISFs, which affect replication of human-pathogenic flaviviruses within co-infected mosquito cells or mosquitoes without affecting the mosquitoes themselves, mean that we should pay attention to both ISFs and human-pathogenic flaviviruses, despite the fact that ISFs appear not to be directly hazardous to human health. To assess the risk of diseases caused by flaviviruses, and to better understand their ecology, it is necessary to know the extent to which flaviviruses are harbored by arthropods.
We developed a novel universal primer for use in a PCR-based system to detect a broad range of flaviviruses. We then evaluated its performance. The utility of the novel primer pair was evaluated in a PCR assay using artificially synthesized oligonucleotides derived from a template viral genome sequence. The utility of the primer pair was also examined by reverse transcription PCR (RT-PCR) using cDNA templates prepared from virus-infected cells or crude supernatants prepared from virus-containing mosquito homogenates.
The novel primer pair amplified the flavivirus NS5 sequence (artificially synthesized) in all samples tested (six species of flavivirus that can cause infectious diseases in humans, and flaviviruses harbored by insects). In addition, the novel primer pair detected viral genomes in cDNA templates prepared from mosquito cells infected with live flavivirus under different infectious conditions. Finally, the viral genome was detected with high sensitivity in crude supernatants prepared from pooled mosquito homogenates.
This PCR system based on a novel primer pair makes it possible to detect arthropod-borne flaviviruses worldwide (the primer pair even detected viruses belonging to different genetic subgroups). As such, an assay based on this primer pair may help to improve public health and safety, as well as increase our understanding of flavivirus ecology.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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