The strategy of temperature downshift has been widely used in the biopharmaceutical industry to improve antibody production and cell-specific production rate (q p) with Chinese hamster ovary cells (CHO). However, the mechanism of temperature-induced metabolic rearrangement, especially important intracellular metabolic events, remains poorly understood. In this work, in order to explore the mechanisms of temperature-induced cell metabolism, we systematically assessed the differences in cell growth, antibody expression, and antibody quality between high-producing (HP) and low-producing (LP) CHO cell lines under both constant temperature (37°C) and temperature downshift (37°C→33°C) settings during fed-batch culture. Although the results showed that low-temperature culture during the late phase of exponential cell growth significantly reduced the maximum viable cell density (p<0.05) and induced cell cycle arrest in the G0/G1 phase, this temperature downshift leaded to a higher cellular viability, and increased antibody titer by 48% and 28% in HP and LP CHO cell cultures, respectively (p<0.001), and favored antibody quality reflected in reduced charge heterogeneity and molecular size heterogeneity. Combined extra- and intra-cellular metabolomics analyses revealed that temperature downshift significantly downregulated intracellular glycolytic and lipid metabolic pathways while upregulated TCA cycle, and particularly featured upregulated glutathione metabolic pathways. Interestingly, all these metabolic pathways were closely associated with the maintenance of intracellular redox state and oxidative stress-alleviating strategies. To experimentally address this, we developed two high-performance fluorescent biosensors, denoted SoNar and iNap1, for real-time monitoring of intracellular NAD +/NADH ratio and NADPH amount, respectively. Consistent with such metabolic rearrangements, the results showed that temperature downshift decreased the intracellular NAD +/NADH ratio, which might be ascribed to the re-consumption of lactate, and increased the intracellular NADPH amount (P<0.01) to scavenge intracellular reactive oxygen species (ROS) induced by the increased metabolic requirements for high-level expression of antibody. Collectively, this study provides a metabolic map of cellular metabolic rearrangement induced by temperature downshift and demonstrates the feasibility of real-time fluorescent biosensors for biological processes, thus potentially providing a new strategy for dynamic optimization of antibody production processes.

Background: Although Doppler evaluation using a multiplanar method is recommended to assess the severity of aortic stenosis (AS) with transthoracic echocardiography, evidence on the diagnostic significance of a non-apical method is limited. This study aimed to compare the use of the apical window (AW) with use of the right parasternal window (RW) method to evaluate AS severity and to examine the diagnostic significance of performing the RW method in addition to the AW method during the evaluation. Methods: This retrospective observational study included 287 consecutive patients (mean age: 79 ± 10 years; women, 56%) with severe AS (aortic valve area [AVA] ≤1.0cm 2). The severity of AS according to the AW method and that according to the RW for all subjects were compared, and the significance of performing the RW method in addition to the AW method was examined. Furthermore, we compared the concordance group, in which the AW and RW methods indicated matching in severity, and the discordant group, in which the AW and RW methods did not indicate matching severity. Results: Peak velocity (PV), mean pressure gradient (PG), and AVA were not significantly different between the AW and RW methods. Performing the RW method in addition to the AW method significantly decreased the number of low PG AS cases (mean PG <40 mmHg) from 71.1% to 65.0% and it increased the number of very severe AS cases (PV ≥5m/s) from 8.7% to 14.5%. Although, there was no significant difference in the Doppler angle (DA) observed using the AW method for the discordant group and the concordant group, the DA observed using the RW method was significantly smaller in the discordant group (8.8±8.2, 16.3±12.3 °, p<0.01). In the receiver-operating characteristic analysis, with the RW method, a DA of 8° was the cutoff value for discrepancies between the two groups. Conclusions: By performing the RW method in addition to the AW method to determine AS severity, different severity is observed in approximately 10% of cases. These results suggest that AS severity may be underestimated by using the AW method alone.

Pediatric Pulmonology strives to uphold the strongest ethical standards in publishing and to serve our diverse, international community of authors and readers with respect. We will be vigilant to prevent recurrence of this and other out-of-date, harmful terminology.

In this study, wetlands in the Ili River Delta (IRD) from 1975 to 2020 were extracted based on Landsat images using the object-oriented method; changes in the wetland area, wetland landscape pattern, NDVI and NPP were analyzed; and the contributions of natural and human factors to wetland evolution were quantified. The results indicated that (1) From 1975 to 2020, the wetland area of the IRD showed an increasing trend, and changes in the wetland area were mainly found in the middle part of the delta near the Saryesik Peninsula. (2) The wetland landscape pattern in the IRD changed markedly from 1975 to 2020. The dominant patches of the wetland in the middle of the delta continued to expand; the patch aggregation index (AI) increased, and the patch fragmentation index (LFI) decreased. (3) From 2000 to 2020, the average annual normalized difference vegetation index (NDVI) and net primary productivity (NPP) in the IRD increased, which is consistent with the change in wetland expansion. (4) Inflow to the delta from the Ili River and the water level of Balkhash Lake are significantly correlated with the wetland area, which are the dominant factors driving wetland evolution; and water evaporation from the Kapchagay Reservoir and irrigation water diversion on the left bank of the reservoir obviously intensified the process of lake water level decline and wetland degradation during 1970 to 1985. These results can provide scientific background for making informed ecological protection decisions in the Ili River Delta under the impacts of climate change and human activities.

Objectives: Barium studies are routinely requested to investigate a range of conditions, however, its efficacy for several conditions is unclear. Alongside findings from our audit in a district general hospital, we aim to review current literature on the use of barium swallow as an investigation for pharyngeal pouch, globus pharyngeus, tertiary peristalsis and oesophageal malignancy in adults. Methods: A systematic literature search was conducted on three databases for papers published in English in the last ten years. Following screening, six studies were included for pharyngeal pouch, five studies were included for globus pharyngeus, seven studies were included for tertiary peristalsis, and thirteen studies were included for oesophageal malignancy. Results: Barium swallow is a useful and important investigation in diagnosing pharyngeal pouch and can be useful to investigate oesophageal dysmotility in tertiary peristalsis. Globus pharyngeus patients do not clinically benefit from undergoing barium swallow. Oesophagogastroduodenoscopy (OGD) should remain as first line in diagnosing oesophageal malignancies. Conclusion: Barium swallow should be used to investigate pharyngeal pouch and tertiary peristalsis, but not used as the main investigation for globus pharyngeus or oesophageal malignancy. There is a need to develop further guidelines for investigations requested for dysphagia.

Plants harbor a wide range of leaf-feeding insects. Insect survival and fitness are influenced by both energy-rich molecules and phytochemicals in the host foliage. Yet, how leaf chemical diversity and insect microbiota - key factors in ecological and physiological processes – shape insect nutrition and impact insect performance is still poorly understood. Here we forced Tortrix viridana larvae, an oak-specialized herbivore, to feed on two Quercus robur susceptible and resistant metabolic phenotypes (metabotypes) and examined leaf, salivary, and fecal metabolomes associated with larval performance, mortality, and fecal microbiota. We show that host chemical diversity affects larval development and that the distinct signatures of oak metabotypes are maintained in the insect digestive system. Larvae were highly efficient in nutrient assimilation and able to minimize plant chemical defenses, thanks in part to the adaptation of the gut microbiota to the different food qualities.

BackgroundNumber of medicines and medicines appropriateness are often used as outcome measures to evaluate the effectiveness of deprescribing interventions. The aim of this study was to evaluate changes in prescribing, potentially inappropriate prescriptions (PIP) and prescribing of low-value medicines during the SPPiRE trial.MethodsWe retrospectively analysed trial prescription data from 51 general practices with 404 participants aged ≥65 years and prescribed ≥15 repeat medicines. Repeat medications at baseline and follow-up (~1 year later) were assigned Anatomical Therapeutic Classification (ATC) codes. Outcomes were the most commonly prescribed and potentially inappropriately prescribed drug groups, the most frequently discontinued or initiated drug groups and the number of changes per person between baseline and follow-up.Results There were 7,051 medicines prescribed to 404 participants at baseline. There was a median of 17 medicines (IQR 15-19) at baseline and 16 (IQR 14-19) at follow-up. PIP represented 17.1% of prescriptions at baseline and 15.7% (n=6,777) at follow-up. There were reductions in the prescription of most drug groups with the largest reduction in antiplatelet prescriptions. Considering medication discontinuations, initiations and switches, there was a median of five medication changes per person (range 0-30, IQR 3-9) by follow-up. There were 95 low-value prescriptions at baseline reducing to 78 at follow-up.ConclusionThe number of medication changes per person was not reflected by summarising medication count at two time points, highlighting the complexity of prescribing for patients with polypharmacy. Frequent medication changes has potentially important implications for patients in terms of adherence and medication safety.Key words: Multimorbidity, polypharmacy, cluster randomised controlled trial, deprescribing, potentially inappropriate prescribingThe SPPiRE trial was registered prospectively on the ISRCTN registry (ISRCTN12752680).

Toxic Epidermal Necrolysis (TEN) is potentially life threatening cutaneous disorder usually induced by drugs. Non steroidal anti-inflammatory drugs (NSAIDS) are one of the common groups implicated in drug induced TEN. Aceclofenac is a relatively new NSAID exhibiting preferential cyclooxygenase-2 inhibition. It is widely used in management of musculoskeletal disorders. This report describes TEN induced by aceclofenac in a 58 year old female who self medicated herself with higher dose of aceclofenac for knee joint pain.

Aim: Antegrade Selective Cerebral Perfusion (ASCP) with lower body circulatory arrest (LBCA) used in aortic arch surgery can lead to postoperative ischemic organ dysfunctions if it lasts long enough. We aimed to evaluate methods that can provide early detection of spinal cord ischemia during aortic arch surgery. Methods: Thirty consecutive patients were prospectively enrolled and Near infrared spectrometry (NIRS) data obtained from the 5 th and 10 th thoracic vertebral region, S100β protein, lactate blood levels during various operative phases and postoperative neurological outcomes were evaluated. Results: A total of 30 patients underwent elective hemi arch (73.29%) or total arch (23.31%) replacement and with a mean ASCP period of 25.1 ± 19.0 (limits 10-90) minutes. In-hospital mortality was 6.66% (two patients). Paraparesis developed in one patient (3,33%). Thoracic T5 and T10 NIRS values were lowest during the ASCP period (p<0.001) with a good correlation between them (r=0.853, p<0.001). However, a significant difference between the T5 and T10 levels was observed during the same period (55.40 vs 51.07 respectively, p=0.001). A moderately negative correlation between the lactate levels in descending aorta and NIRS values at the T10 level was found during ASCP (r =-0.514, p = 0.004). Conclusion: Thoracic 5 th and 10 th level NIRS monitoring for spinal cord oxygenation were significantly lower during ASCP period compared to the other periods of aortic arch surgery with T10 values being lower than T5 values during the same period indicating a more significant flow disturbance at this level. Measuring lactate levels with thoracic NIRS monitoring seems promising for future studies with larger volumes and longer ASCP periods.

Hippophaerhamnoides is widely known for its important ecological, economic, and social benefits. It is known as the pioneer plant of soil and water conservation, with homology in food and medicine. Here we used occurrence data and environmental (climate and soil) variables to simulate and predict the habitat distribution for H. r. sinensis and H. r. trkestanica in China, both at the current time and in the 2050s (2041-2060). Our aim was to analyze the dominant factors effecting its distribution using MaxEnt and the spatial analysis of geographic information system. The results indicated that H. r. sinensis is mainly distributed in Shaanxi, Shanxi, Sichuan, Qinghai, Gansu, Ningxia, Tibet, and Inner Mongolia, and is mainly affected by bio13 (precipitation of the wettest month), bio11 (mean temperature of the coldest quarte) and bio3 (Isothermality). The suitable habitat of H. r. trkestanica is mainly distributed in Xinjiang, and Tibet, and is mainly affected by bio13 (precipitation of the wettest month), bio2 (mean diurnal range) and bio15 (precipitation seasonality). Although, the two subspecies tend to expand and migrate toward lower latitude under future climate scenarios, there are some differences. H. r. sinensis will migrate westward, while H. r. trkestanica will migrate eastward as a whole. They have a high stability of suitable habitat and are not at risk of extinction in the future. The study’s findings help to clarify the resource reserve of Hippophaerhamnoides L. in China, which will help to guide the protection of wild resources and to popularize artificial planting in suitable areas, and provides scientific basis for the protection of ecological environment.

In spite of the discovery of various medicines to heal post-surgical malignant wounds caused by the recurrence of breast cancer cells after chemotherapy, still it’s a serious problem to heal malignant wounds completely for surgeons and this has to be cured in order to avoid further risk of wound issues. This study is one such attempt taken to give a solution to heal surgical breast wounds for the future analysts. For this purpose, we have synthesized a novel bio-compatible fluorescent quantum dot using curcumin-loaded Zinc oxide bionanocomposite (Cur-ZnOBC). The ZnOBC is composed of neem essential oil-Aloe-vera gel complex which acted as a nanoconjugate for the formation of Cur-ZnOBC.Then it is converted into curcumin-based quantum dots (Cur-ZnOQDs) using microwave-assisted method in order to improve the drug delivery purpose. The resulting Cur-ZnOQDs was treated in the malignant wounds of breast cancer cells post-surgery for wound healing activity. The synthesized Cur-ZnOQDs was characterized to confirm its size and morphology. Thus, prepared Cur-ZnOQDs showed low cytotoxicity (IC50 0.17 µg/mL) to human breast cancer cells. Cell death was noticed at a very low concentration of 0. 6mg/mL itself. It was also found that wound closure area was increased from 22.3% at 24 h to 44.2% at 48 h which was 20 times greater cell migration when compared to control (untreated cells) and at a very low concentration of Cur-ZnOQDs (0.1mg/mL). Hence, Cur-ZnOQDs act as bio-enhancers assisting in the improvement of outcomes associated with breast cancer and, ultimately, improving the survival rate.

To report a case of Vogt-Koyanagi-Harada disease (VKH) in a 27-year-old male two weeks proceeding COVID-19 infection onset. Severe complications of VKH can be avoided by early diagnosis and adequate treatment with corticosteroids and immunosuppressants. It is possible that COVID-19 was a potential immunological trigger of VKH in our patient.

Computer software was used for implant planning and guided surgery. A provisional prosthesis was loaded on the day of surgery; the definitive prosthesis with a milled titanium framework was loaded one year later. There were no clinical or radiographic changes around the dental implants at two-years post- surgery.

The effectiveness of patent foramen ovale closure for migraine has been previously discussed. By contrast, very few studies have reported the association between migraine and atrial septal defect closure. Here we report a case in which atrial septal defect closure effectively relieved migraine headaches.

A document by Steffen Mickenautsch. Click on the document to view its contents.

This Supporting Information includes: a comparison of the REAL (Robot Ear Accomplished by Laser) with a typical vibration measuring system (Laser Doppler Vibrometers, LDV), frequency response of various materials on REAL and real-time analysis of REAL audio neural network model. Xiaoping Hong  Email:    hongxp@sustech.edu.cn

Pichia pastoris as an efficient host for the production of recombinant proteins is mostly cultivated in fed-batch mode in which the cell’s environment is continuously changing. Therefore, to fine-tune bioreactor performance in respect to the associated metabolic changes of the microorganism, it is crucial to understand the influence of feeding strategy parameters on the intracellular reaction network. In this study, dynamic flux balance analysis (DFBA) integrated with transcriptomics data was used to simulate the recombinant P.pastoris (Muts) growth during induction phase for 3 fed-batch µ-stat strategies. The induction phase was divided into equal time intervals and the correlated reactions with protein yield were identified in the 3 fed-batch strategies using the Pearson correlation coefficient. Subsequently, Principal Component Analysis was applied to cluster induction phase time intervals and identify the role of correlated reactions on metabolic differentiation of time intervals. It was found that increasing fluxes through the methanol dissimilation pathway increased protein yield. By adding a methanol assimilation pathway inhibitor (HgCl2) to the shake flask medium containing 10% (v/v) glycerol, the protein titer increased by 60%. Using the DFBA revealed that the higher the dimensionless flux of methanol, the higher amounts of protein yield. Finally, a novel feeding strategy was developed so that the dimensionless methanol flux increased compared to the performed cultivations. Protein titer increased by 16% compared to the optimally performed cultivation, while production yield increased by 85%.

A mathematical examination of the SIR model under mutation is presented in this paper, by integrating an incubation time lag and a general nonlinear incidence rate. When the virus mutates, the recovered population loss its immunity. A time lag gives for a grace period before people become vulnerable once more. At the rate ‘c,’ they become vulnerable, which is the recovery rate, depending upon their status at (t-τau). The three-state variable are S (Susceptible population), I (Infected population) and R (Recovered population). A non-zero equilibrium point has been found. Stability and Directional analysis are performed about this non zero equilibrium. Hopf-Bifurcation occurred when the delay parameter τ goes beyond a critical point value. Sensitivity analysis is performed by using direct method and Directional analysis is performed by using K. R. Schneider, ‘Hassard, B. D. [22]. Numerical simulation is done to support analytical results using MATLAB.

In this study, the regulatory effect of exogenous melatonin on the expression of genes related to the AsA-GSH cycle and nitrogen metabolism in seedlings under NaCl stress was determined; the homology between enzyme genes and similar genes in other species was analyzed by applying different concentrations of exogenous melatonin to rice seedlings under NaCl stress; and the morphology and physiological indicators of seedlings were determined. Exogenous melatonin was found to upregulate the expression of OsGR3, OsGPX1, OsNR2, OsGS1, and OsGOGAT1 in seedlings under NaCl stress, which enhanced the enzyme activities of the AsA-GSH cycle and nitrogen metabolism, ultimately reducing the content of reactive oxygen species (ROS) and improving the photosynthetic rate and nitrogen efficiency. A high homology was found when rice OsGR3, OsGPX1, OsNR2, OsGS1, and OsGOGAT1 were compared with similar genes in Arabidopsis thaliana, Hordeum vulgare, Zea mays, and Sorghum bicolor. Further, the location of the gene on the chromosome, the similar gene structure, GO annotation, and KEGG metabolic pathways were explained by the functions and metabolic processes of OsGR3, OsGPX1, OsNR2, OsGS1, and OsGOGAT1. Overall, our findings indicate that exogenous melatonin participates in the regulation of enzyme gene expression during the AsA-GSH cycle and nitrogen metabolism, ultimately affecting the activities of related enzymes and alleviating the oxidative damage induced by ROS in seedlings.

Niclosamide is a FDA-approved anthelminthic drug for the treatment of parasitic infection. However, over the past few years, the increasing evidence has shown that Niclosamide could treat diseases beyond parasitic diseases, which include metabolic diseases, immune system diseases, bacterial and viral infections, other types of diseases and cancer. Therefore, we systematically review the pharmaceutical activities and therapeutic prospects of Niclosamide in human disease and cancer, and summarize the related molecular mechanisms and signaling pathways, which provides theoretical support for the new therapeutic value of old drug.

Introduction: topical rapamycin has been established as an effective and safe therapy for facial angiofibromas in tuberous sclerosis. Different formulations have been tested for this skin disease, most using an ointment as a vehicle. Purpose: to improve the classical formulation of topical rapamycin and to determine the validity period of the proposed options based on chemical, physical and microbiological stability studies. Methods: four different 0.4% rapamycin formulations were prepared (ointment, emulsion, gel and liposomes). The stability studies for each formulation were: chemical (extraction with lipophilic solvents and high-performance liquid chromatography assay), physical (pH, uniformity, extensibility, absence of crystals, absence of phase separation and only for liposomal formulation was determined particle size, zeta potential and encapsulation efficiency) and microbiological (culture samples in blood-agar media) during 56 days. Results: only liposomes were chemically, physically and microbiologically stable after 8 weeks. Ointment, emulsion and gel formulations lost their chemical and physical stability before 56 days. Conclusions: this study describes a new four formulations to improve the previously treatment for facial angiofibromas in tuberous sclerosis. It also provides favorable stability data only for liposomes. However, more dermokinetic and clinical studies are needed to confirm that liposomes are most appropriate to ensure effectiveness, safety and high patient satisfaction.

Dr Daniel GraceMBBS BSc (Hons) Dip EWM DTMH MRCGP (2017) MFTM RCPS(Glasg) FRGS Portfolio GP, Travel Health Physician and Expedition DoctorAbertawe Bro Morgannwg University Health Boarddaniel.grace@nhs.netCompeting interests: noneFinancial contributions: none

Background and Purpose: Selective peroxisome proliferator-activated receptors (PPARs) are widely used to treat metabolic complications; however, the limited effect of PPARα agonists on glucose metabolism and the adverse effects associated with selective PPARγ activators have stimulated the development of novel pan-PPAR agonists to treat metabolic disorders. Here, we synthesised a new prenylated benzopyran (BP-2) and evaluated its PPAR-activating properties, anti-inflammatory effects and impact on metabolic derangements. Experimental Approach: BP-2 was used in transactivation assays to evaluate its agonism to PPARα, PPARβ/δ and PPARγ. A parallel-plate flow chamber was employed to investigate its effects on TNFα-induced leucocyte-endothelium interactions, and flow cytometry and immunofluorescence were used to determine its effects on the expression of endothelial cell adhesion molecules (CAMs) and chemokines and p38-MAPK/NF-κB activation. PPARs/RXRα interactions were determined using a gene silencing approach. Analysis of its impact on metabolic abnormalities and inflammation was performed in ob/ob mice. Key Results: BP-2 displayed strong PPARα activity, with moderate and weak activity against PPARβ/δ and PPARγ activity, respectfully. In vitro, BP-2 reduced TNFα-induced endothelial ICAM-1, VCAM-1 and fractalkine/CX3CL1 expression, suppressed mononuclear cell arrest via PPARβ/δ-RXRα interactions and decreased p38-MAPK/NF-κB activation. In vivo, BP-2 improved the circulating levels of glucose and triglycerides in ob/ob mice, suppressed T-lymphocyte/macrophage infiltration and proinflammatory markers in the liver and white adipose tissue, but increased the expression of the M2-like macrophage marker CD206. Conclusion and Implications: BP-2 emerges as a novel pan-PPAR lead candidate to normalise glycaemia/triglyceridaemia and minimise inflammation in metabolic disorders, likely preventing the development of further cardiovascular complications.

Background: Cancer patients were considered to be at high risk for COVID-19 because they are more likely to develop serious infections. Children and adolescents with cancer are also more vulnerable to COVID-19. This study aimed to describe the impact of COVID-19 on clinical outcomes in a cohort of children and adolescents with cancer in NE-Brazil. Methods: This is a survey of SARS-CoV-2 infection in a cohort of cancer patients hospitalized between April 2020 and September 2021 at the public pediatric oncology service of Sergipe. All children and adolescents underwent detection of SARS-CoV-2 through RT-PCR. Results: One hundred fifteen patients were hospitalized but three were transferred to other services and they were excluded from this study. The final sample consisted of 112 children. Had at least one COVID-19 positive test 54.46% and 40.18% among these patients died. There was difference between death and positive test (p=0.03; odds ratio [OR], 2.32, 95% confidence interval [CI], 1.06-5.09), observing higher mortality rate among patients with coronavirus positive test, having them or not COVID-19 symptoms. There was also difference between outcome and patients’ oncological diagnosis (p=0.02; OR, 3.65; 95% CI, 1.16-11.44). Conclusion: COVID-19 infection and fatality rates among pediatric oncology patients were significantly higher than those found in previous studies. There was also a higher number of deaths in the group with a positive test and with solid tumors, with the predominant cause of death being the progression of the oncological disease.