Bridging upstream and downstream for improved adenovirus 5 bioprocess.

Adenoviruses are well-known viral vectors that have been previously used in gene therapy and as a vaccine-delivery vehicle for humans and animals. During the COVID-19 pandemic, it gained renewed attention, but at the same time, it raised concerns due to side effects observed with some of the resulting vaccines administered to patients. It has been indicated that these side effects might be attributed to impurities present in the final product. Therefore, constant enhancement of the vaccine purity and further improvement of impurity detection methods are needed. In this work, we showcase an example of industry-relevant adenovirus bioprocess optimization. Our data show the effect of upstream parameters on the bioburden introduced to the downstream process. We provide an example of process optimization using a combination of the PATfix analytical method, ddPCR, infectivity, total DNA, and total protein analyses to optimize cell density, multiplicity of infection, and length of production. Additionally, we provide data illustrating the robustness of the convective interaction media quaternary amine monolithic chromatography step. This anion exchange strategy was shown to remove over 99% of protein and DNA impurities, including those unable to be addressed by tangential flow filtration, while maintaining high adenovirus recoveries.

Yoga for Migraine Prevention: An Ancient Practice with Evidence for Current Use.

PURPOSE OF REVIEW: The objective of this study is to review the recent literature on yoga for migraine prevention either as adjuvant or standalone therapy. Yoga is one of the most widely used complementary and integrative medicine (CIM) therapies; clinicians should be familiar with yoga practice so that they can best advise interested patients. It is also important to assess study design and types of yoga offered. Using PubMed and Litmaps, research published from 2018 to 2023 addressing yoga and migraine was assessed. RECENT FINDINGS: Two systematic reviews and six studies have recently been published on yoga as adjunctive migraine preventive treatment. There is class III evidence and a grade B recommendation for yoga as an adjunct migraine preventive treatment. Yoga has been shown to reduce headache frequency, disability, and likely also pain intensity and self-efficacy. Two studies (one in children and one in adults) suggested that yoga as standalone migraine preventive treatment reduces pain intensity, disability, and perceived stress. More research is needed on the long-term efficacy (including change in monthly migraine days specifically in addition to headache frequency) and adherence to yoga practice for the prevention of migraine. In addition, to our knowledge, there is no study evaluating yoga practice in the prodromal or headache phase of migraine as acute treatment.

Biological activities, identification, method development, and validation for analysis of polyphenolic compounds in Nymphaea rubra flowers and leaves by UHPLC-Q-cIM-TOF-MS and UHPLC-TQ-MS.

INTRODUCTION: Nymphaea rubra belongs to the Nymphaea family and is regarded as a vegetable used in traditional medicine to cure several ailments. These species are rich in phenolic acid, flavonoids, and hydrolysable tannin. OBJECTIVE: This study aimed to assess the biological activities of Nymphaea rubra flowers (NRF) and leaves (NRL) by identifying and quantifying their polyphenolic compounds using ultra-performance liquid chromatography coupled to quadrupole cyclic ion mobility time-of-flight mass spectrometry (UHPLC-Q-cIM-TOF-MS) and triple quadrupole mass spectrometry (UHPLC-TQ-MS). METHODOLOGY: NRF and NRL powder was extracted with methanol and fractionated using hexane, ethylacetate, and water. Antioxidant and α-glucosidase, and tyrosinase enzyme inhibitory activities were evaluated. The polyphenolic components of NRF and NRL were identified and quantified using UHPLC-Q-cIM-TOF-MS and UHPLC-TQ-MS. The method was validated using linearity, precision, accuracy, limit of detection (LOD), and lower limit of quantification (LLOQ). RESULTS: Bioactive substances and antioxidants were highest in the ethylacetate fraction of flowers and leaves. Principal component analysis showed how solvent and plant components affect N. rubra's bioactivity and bioactive compound extraction. A total of 67 compounds were identified, and among them 21 significant polyphenols were quantified. Each calibration curve had R2 > 0.998. The LOD and LLOQ varied from 0.007 to 0.09 µg/mL and from 0.01 to 0.1 µg/mL, respectively. NRF contained a significant amount of gallic acid (10.1 mg/g), while NRL contained abundant pentagalloylglucose (2.8 mg/g). CONCLUSION: The developed method is simple, rapid, and selective for the identification and quantification of bioactive molecules. These findings provide a scientific basis for N. rubra's well-documented biological effects.

Ion mobility mass spectrometry in the omics era: Challenges and opportunities for metabolomics and lipidomics.

Researchers worldwide are taking advantage of novel, commercially available, technologies, such as ion mobility mass spectrometry (IM-MS), for metabolomics and lipidomics applications in a variety of fields including life, biomedical, and food sciences. IM-MS provides three main technical advantages over traditional LC-MS workflows. Firstly, in addition to mass, IM-MS allows collision cross-section values to be measured for metabolites and lipids, a physicochemical identifier related to the chemical shape of an analyte that increases the confidence of identification. Second, IM-MS increases peak capacity and the signal-to-noise, improving fingerprinting as well as quantification, and better defining the spatial localization of metabolites and lipids in biological and food samples. Third, IM-MS can be coupled with various fragmentation modes, adding new tools to improve structural characterization and molecular annotation. Here, we review the state-of-the-art in IM-MS technologies and approaches utilized to support metabolomics and lipidomics applications and we assess the challenges and opportunities in this growing field.

Prevalence of carbapenemases in Enterobacterales from urine specimens in an university hospital in Istanbul, Turkey.

We aimed to investigate the prevalence of carbapenemases in Enterobacterales strains isolated from urine specimens between July 2019 and July 2020.CIM and modified CIM tests were applied as well as detection of blaOXA-48, blaNDM, blaVIM, blaKPC and blaIMP genes was performed by multiplex PCR.One hundred fifty of 3,242 Enterobacterales strains were found to be carbapenem resistant and 46 were included in the study. Forty five (98%) of the 46 strains included in the study were Klebsiella spp. and one (2%) of them was Escherichia coli. Susceptibility to ceftazidime-avibactam, amikacin and gentamicin was 97%, 11% and 9%, respectively. Forty three (94%) isolates were found positive at 2 and 4 h with CIM test. Forty four (97%) strains were found positive at 4 h and 43 (94%) strains were found positive at 2 h with modified CIM test.While blaOXA-48, blaNDM and blaOXA-48 with blaNDM association were found in Klebsiella spp. isolates in 55%, 27% and 11%, respectively, blaVIM, blaKPC, blaIMP were not found. Only blaOXA-48 and blaNDM-1 were detected in the E. coli strain.None of the investigated genes were detected in three Klebsiella strains but with whole genome analysis the combination of blaOXA-534, blaCMY-99 and blaKPC-3 was found in the first strain, blaOXA-370 in the second strain and no resistance gene was found in the third strain.Ceftazidime-avibactam was found to be active against 97% of strains, and the most common resistance genes were blaOXA-48 and blaNDM-1. Previously undetected resistance genes have been identified in our country.

Analyzing Citramalic Acid Enantiomers in Apples and Commercial Fruit Juice by Liquid Chromatography-Tandem Mass Spectrometry with Pre-Column Derivatization.

Optically active citramalic acid (CMA) is naturally present as an acidic taste component in fruits, such as apples. The absolute configuration of CMA in such fruits was investigated by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) following pre-column derivatization with a chiral reagent, benzyl 5-(2-aminoethyl)-3-methyl-4-oxoimidazolidine-1-carboxylate. The developed LC-MS/MS method successfully separated the enantiomers of CMA using an octadecylsilica column with a resolution and separation factor of 2.19 and 1.09, respectively. Consequently, the R-form of CMA was detected in the peel and fruit of three kinds of apple at concentrations in the 1.24-37.8 and 0.138-1.033 mg/wet 100 g ranges, respectively. In addition, R- CMA was present in commercial apple juice, whereas no quantity was detected in commercial blueberry, perilla, or Japanese apricot juice.

[ICD-11: New revision and impact of this classification in psychiatry]. / CIM-11 (OMS) : nouvelle révision et impact de la classification onusienne en psychiatrie.

OBJECTIVES: After more than 20 years of work, the World Health Organization's efforts have culminated in the adoption of the 11th revision of the International Classification of Diseases (ICD-11). The process has been guided by the principles of global applicability, scientific validity, and clinical utility. The update of the chapter on mental, behavioral, and neurodevelopmental disorders (MBND) within the ICD-11 has generated widespread interest worldwide. This has raised various questions about the status of mental disorder nosology, changes to diagnostic guidelines, and the potential implications for clinical practice. METHODS: We conducted a narrative analysis of the literature in four different languages to track the various stages of the ICD-11 revision and to highlight the major changes. We searched databases such as PubMed, EMBASE, MEDLINE, and Google Scholar, and consulted the official websites of the WHO, APA, and UNICEF. A total of 79 articles from 40 different editorials and websites were analyzed and included in this study. RESULTS: The new chapters on mental disorders in the ICD-11 include 21 groups, as opposed to the 11 in the ICD-10. The changes aim to align the diagnoses with those of the DSM-5 and introduce a new chapter structure, new diagnostic categories, modifications to diagnostic criteria, and advancements in dimensionality. For the first time in the history of the ICD, sleep and wakefulness disorders, as well as disorders related to sexual health, have been addressed in separate chapters of the international classification. Four new diagnoses have been added: complex post-traumatic stress disorder (PTSD), gaming disorder, prolonged grief disorder, and compulsive sexual behavior which replaces "excessive sexual activity" in the ICD-10. Moreover, the ICD-11 revision has brought about a fundamental change in the clinical conceptualization of addictive behaviors, introducing a distinction between substance use disorders and addictive behaviors. The criteria for many existing conditions have been revised, particularly those related to bipolar disorders, eating disorders, and gender identity disorders. CONCLUSIONS: The revision process for mental, behavioral, and neurodevelopmental disorders in the ICD-11 has witnessed unprecedented participation in the history of mental disorder classification. These changes could have a significant impact on clinical practice in psychiatry. However, it is crucial to examine the advantages and limitations of this new classification compared to previous versions.

A Preliminary Contribution towards a Risk-Based Model for Flood Management Planning Using BIM: A Case Study of Lisbon.

Preparing a city for the impact of global warming is becoming of major importance. Adopting climate-proof policies and strategies in response to climate change has become a fundamental element for city planning. To this end, this research considers a multidisciplinary approach, at the local scale, able to connect urban planning and architecture, as a vital base for considering a coastal cities' ability to control the consequences of climate change, specifically floods. So far, there is a scarcity of research connecting sea ground and land surveys, and this study could become a foundational reference for coastline settlement management using BIM. We found in BIM (Building Information Modeling) a possible tool for managing coastal risk, since it can combine crowdsourced data for geometric and information modeling of the city. The proposed BIM model includes a topography used for 3D thematic maps, a riverbed model, and a waterway model. This model aims to facilitate coordination across separate actors and interests since the urban area model is always updatable and improvable. Focusing on a case study of Lisbon, we developed risk-based 3D maps of the area close to the shoreline of the Tagus River.

Migraine-relevant sex-dependent activation of mouse meningeal afferents by TRPM3 agonists.

BACKGROUND: Migraine is a common brain disorder that predominantly affects women. Migraine pain seems mediated by the activation of mechanosensitive channels in meningeal afferents. Given the role of transient receptor potential melastatin 3 (TRPM3) channels in mechanical activation, as well as hormonal regulation, these channels may play a role in the sex difference in migraine. Therefore, we investigated whether nociceptive firing induced by TRPM3 channel agonists in meningeal afferents was different between male and female mice. In addition, we assessed the relative contribution of mechanosensitive TRPM3 channels and that of mechanosensitive Piezo1 channels and transient receptor potential vanilloid 1 (TRPV1) channels to nociceptive firing relevant to migraine in both sexes. METHODS: Ten- to 13-week-old male and female wildtype (WT) C57BL/6 J mice were used. Nociceptive spikes were recorded directly from nerve terminals in the meninges in the hemiskull preparations. RESULTS: Selective agonists of TRPM3 channels profoundly activated peripheral trigeminal nerve fibres in mouse meninges. A sex difference was observed for nociceptive firing induced by either PregS or CIM0216, both agonists of TRPM3 channels, with the induced firing being particularly prominent for female mice. Application of Yoda1, an agonist of Piezo1 channels, or capsaicin activating TRPV1 channels, although also leading to increased nociceptive firing of meningeal fibres, did not reveal a sex difference. Cluster analyses of spike activities indicated a massive and long-lasting activation of TRPM3 channels with preferential induction of large-amplitude spikes in female mice. Additional spectral analysis revealed ​a dominant contribution of spiking activity in the α- and ß-ranges following TRPM3 agonists in female mice. CONCLUSIONS: Together, we revealed a specific mechanosensitive profile of nociceptive firing in females and suggest TRPM3 channels as a potential novel candidate for the generation of migraine pain, with particular relevance to females.

Clinical and neurophysiological characterization of muscular weakness in severe COVID-19.

OBJECTIVE: To report clinical and electroneuromyographic (ENMG) characteristics of patients affected by severe COVID-19 infection, evaluated for muscular weakness. MATERIALS AND METHODS: ENMGs performed for evaluation of diffuse weakness in patients who could not be discharged from semi-intensive care COVID unit because of difficulties in ventilation weaning were reviewed. Patients with severe COVID-19 infection who had undergone endotracheal intubation and able to co-operate were considered. ENMG protocol was focused on neurophysiological items that excluded or confirmed critical illness polyneuropathy (CIP), myopathy (CIM), or polyneuromyopathy (CIPM). Standardized clinical evaluation was performed using Medical Research Council (MRC) sum score. RESULTS: Eight patients were included in the study. All presented known risk factors for intensive care unit-acquired weakness (ICU-AW), and none of them had history of underlying neuromuscular disorders. ENMG findings were normal in two patients, while only two patients had an altered MRC sum score (< 48). Neuromuscular involvement was diagnosed in 6/8 patients (75%): 2 had CIP, 1 had possible CIM, 1 had CIPM, while 1 patient, with clinically evident weakness but equivocal ENMG findings, was classified as ICU-AW. Finally, 1 patient was diagnosed with acute demyelinating neuropathy. Patients with neuromuscular involvement were those with longer intubation duration and higher levels of IL-6 at admission. CONCLUSION: Neuromuscular complications are frequent in severe COVID-19 and cannot be excluded by MRC sum scores above 48. Standardized ENMG is helpful in guiding diagnosis when clinical evaluation is not reliable or possible. Elevated IL-6 at admission may be a predictor biomarker of ICU-AW in COVID-19.

Genetic architecture of Al3+ toxicity tolerance in rice F2:3 populations determined through QTL mapping.

Aluminum (Al3+) toxicity is one of the factors limiting crop production in acidic soils. Identifying quantitative trait loci (QTLs)/genes for tolerance to Al3+ toxicity at seed germination can aid the development of new tolerant cultivars. The segregating population derived from Pak Basmati (Indica) × Pokkali (Indica) was used for mapping QTLs linked with tolerance to Al3+ toxicity ranging from 0 to 20 mM at pH 4 ± 0.2 at germination. The favorable alleles for all new QTLs were analyzed based on germination traits, i.e., final germination percentage (FG%), germination energy (GE), germination speed (GS), germination index (GI), mean germination time (MGT), germination value (GV), germination velocity (GVe), peak value of germination (GPV), and germination capacity (GC), and growth traits, such as root length (RL), shoot length (SL), total dry biomass (TDB) and germination vigor index (GVI). The phenotypic evolution showed transgressive variations. For genome-wide mapping, 90 polymorphic SSRs with 4 gene-specific markers and Win QTL Cart were used for QTL analysis. In all, 35 QTLs for germination and 11 QTLs for seedling growth were detected in distinct chromosomal regions by composite interval mapping (CIM), and multiple interval mapping (MIM) confirmed the pleiotropy at region RM128 on chromosome 1. Based on our genetic mapping studies, the genes/QTLs underlying tolerance to Al3+ toxicity could differ for both the germination and seedling stages in segregated populations. The QTLs identified in this study could be a source of new alleles for improving tolerance to Al3+ toxicity in rice.

Covert Information Mapped Spatial and Directional Modulation toward Secure Wireless Transmission.

Recently, the concept of spatial and direction modulation (SDM) has been developed to reap the advantages of both spatial modulation (SM) and directional modulation (DM). On the one hand, DM ensures the transmission security at the expected direction. On the other hand, the structure of SM-aided distributed receivers can enhance the security even if the eavesdropper is located in the same direction as the legitimate receiver. However, the above advantages are achieved based on the assumption that the eavesdropper is not equipped with distributed receivers. On the other hand, the information security can no longer be guaranteed when the eavesdropper is also equipped with distributed receivers. To alleviate this problem, we considered a joint design of SDM and covert information mapping (CIM) in order to conceive of a more robust structure of CIM-SDM. Furthermore, both the detection performances at the eavesdropper and the legitimate user were quantified through theoretical derivation. In general, both the analysis and simulation results supported that the proposed CIM-SDM structure provides more robust secure performance compared to the original SDM, even if the extreme condition of distributed receivers at the eavesdropper is considered, at the cost of moderate performance loss at the legitimate user.

Post-concussion syndrome: Still forgotten? From the disappearance of the DSM-5 to a multidisciplinary consultation.

OBJECTIVES: Following the presence of both post-traumatic stress disorder (PTSD) and post-concussion syndrome (PCS) in the nosography since the publication of DSM-IV, large-scale studies investigated the links between these two entities: exposure to a mild traumatic brain injury was correlated with the presence of PTSD and vice versa, and the strongest factor associated with PCS was the presence of PTSD. But PCS entity was recently suppressed from the 5th edition of the American diagnostic and statistical classification of neuropsychiatric disorders (DSM-5, 2013). In the 11th edition of the CIM, PCS is also likely to be omitted. This elimination raises more questions if we take into consideration the emancipation of PTSD, which now includes the full category of "disorders related to trauma and stressors" to which PCS could have legitimately been added. METHODS: We discuss current scientific literature and clinical practices with a socio-anthropological point of view. RESULTS: Post-concussion and post-traumatic clinical entities often show similar anamnestic temporalities, with an initial acute phase where memory (amnesia following TBI; dissociative post-traumatic amnesia) and consciousness (initial loss of consciousness secondary to TBI; peri-traumatic psychic dissociation) impairments predominate, followed by a pauci-symptomatic latency phase. Finally, a symptomatic phase occurs in which similar symptoms for both entities are observed (sleep disorders, anxiety and depression, irritability, fatigue, attention disorder, tendency to avoidance). If similar therapies (pharmacological and psychological) are effective in treating the clinical consequences of head and mental trauma, this suggests that they have common etiopathogenic origins. CONCLUSIONS: Yet, post-concussion syndrome remains a clinical-biological reality. If a diffusion tensor imaging MRI in the acute phase is likely to provide predictive elements for subsequent cognitive dysfunctions, it would appear useful to consider combining biomarkers, and linguistics markers, with the creation of a clinical-radio-bio- neuropsychological score in order to differentiate benign outcomes from neuro- and/or psycho-traumatic disorders.

Diversity and phylogeny of basidiomycetous yeasts from plant leaves and soil: Proposal of two new orders, three new families, eight new genera and one hundred and seven new species.

Nearly 500 basidiomycetous yeast species were accepted in the latest edition of The Yeasts: A Taxonomic Study published in 2011. However, this number presents only the tip of the iceberg of yeast species diversity in nature. Possibly more than 99 % of yeast species, as is true for many groups of fungi, are yet unknown and await discovery. Over the past two decades nearly 200 unidentified isolates were obtained during a series of environmental surveys of yeasts in phyllosphere and soils, mainly from China. Among these isolates, 107 new species were identified based on the phylogenetic analyses of nuclear ribosomal DNA (rDNA) [D1/D2 domains of the large subunit (LSU), the small subunit (SSU), and the internal transcribed spacer region including the 5.8S rDNA (ITS)] and protein-coding genes [both subunits of DNA polymerase II (RPB1 and RPB2), the translation elongation factor 1-α (TEF1) and the mitochondrial gene cytochrome b (CYTB)], and physiological comparisons. Forty-six of these belong to 16 genera in the Tremellomycetes (Agaricomycotina). The other 61 are distributed in 26 genera in the Pucciniomycotina. Here we circumscribe eight new genera, three new families and two new orders based on the multi-locus phylogenetic analyses combined with the clustering optimisation analysis and the predicted similarity thresholds for yeasts and filamentous fungal delimitation at genus and higher ranks. Additionally, as a result of these analyses, three new combinations are proposed and 66 taxa are validated.

Implementation of a gait center training to improve walking ability and vital parameters in inpatient neurological rehabilitation- a cohort study.

BACKGROUND: Many studies showed that robot-assisted gait training might improve walking of patients after stroke. The question remains whether patients with other neurological diagnoses can improve their ability to walk by training in a gait center. Aim of the present study was therefore to investigate the effects of a gait center training in inpatient neurological rehabilitation on walking ability. METHODS: We implemented a gait center training in addition to individual inpatient rehabilitation. Our primary outcome was walking ability based on the Functional Ambulation Categories (FAC). Our secondary outcomes were vital capacity and blood pressure. We predefined subgroups of patients with ischemic and hemorrhagic stroke and critical illness myopathy (CIM) and polyneuropathy (CIP). RESULTS: We included 780 patients from our inpatient rehabilitation center in our cohort study. We analyzed 329 patients with ischemic, 131 patients with hemorrhagic stroke and 74 patients with CIP/ CIM. A large number of patients were able to improve their ability to walk. At the end of rehabilitation, patients with ischemic stroke and FAC 3 = increased theirFAC scores by 5%, FAC 4 = 4% and FAC 5 = 7%. Patients with hemorrhagic stroke and FAC 3 = increased by 5%, FAC 4 = 11% and FAC 5 = 9% and patients with CIP/CIM increased by FAC 3 = 3%, FAC 4 = 22% and FAC 5 = 26%. The largest improvement in walking ability during rehabilitation had patients with a FAC = 1 at baseline who improved by a median of 1.4 FAC points (p < 0.001). After adjusting for the number of gait training sessions, the largest improvement in walking ability during rehabilitation had patients with a FAC = 0 at baseline who improved by 1.8 FAC points (p < 0.001). CONCLUSIONS: Implementation of an additional gait center training may significantly improve walking ability in neurological rehabilitation.

Binding Kinetics of Ruthenium Pyrithione Chemotherapeutic Candidates to Human Serum Proteins Studied by HPLC-ICP-MS.

The development of ruthenium-based complexes for cancer treatment requires a variety of pharmacological studies, one of them being a drug's binding kinetics to serum proteins. In this work, speciation analysis was used to study kinetics of ruthenium-based drug candidates with human serum proteins. Two ruthenium (Ru) complexes, namely [(η6-p-cymene)Ru(1-hydroxypyridine-2(1H)-thionato)Cl] (1) and [(η6-p-cymene)Ru(1-hydroxypyridine-2(1H)-thionato)pta]PF6 (2) (where pta = 1,3,5-triaza-7-phosphaadamantane), were selected. Before a kinetics study, their stability in relevant media was confirmed by nuclear magnetic resonance (NMR). Conjoint liquid chromatography (CLC) monolithic column, assembling convective interaction media (CIM) protein G and diethylamino (DEAE) disks, was used for separation of unbound Ru species from those bound to human serum transferrin (Tf), albumin (HSA) and immunoglobulins G (IgG). Eluted proteins were monitored by UV spectrometry (278 nm), while Ru species were quantified by post-column isotope dilution inductively coupled plasma mass spectrometry (ID-ICP-MS). Binding kinetics of chlorido (1) and pta complex (2) to serum proteins was followed from 5 min up to 48 h after incubation with human serum. Both Ru complexes interacted mainly with HSA. Complex (1) exhibited faster and more extensive interaction with HSA than complex (2). The equilibrium concentration for complex (1) was obtained 6 h after incubation, when about 70% of compound was bound to HSA, 5% was associated with IgG, whereas 25% remained unbound. In contrast, the rate of interaction of complex (2) with HSA was much slower and less extensive and the equilibrium concentration was obtained 24 h after incubation, when about 50% of complex (2) was bound to HSA and 50% remained unbound.

Impact of Long-Term Copper Exposure on Growth, Photosynthesis, Antioxidant Defence System and Artemisinin Biosynthesis in Soil-Grown Artemisia annua Genotypes.

The effects of copper (Cu) exposure on growth and physiological characteristics of three genotypes (CN-12, Cim-Sanjeevani and Cim-Arogya) of Artemisia annua L. were elucidated. The plants were grown under naturally illuminated greenhouse conditions and were harvested after physiological maturity (120 days after sowing). Results suggest that 10 mg kg- 1 Cu significantly enhanced the growth and physiological parameters like enzyme activities, photosynthesis. At higher concentrations, Cu inhibited the growth, biomass, photosynthetic parameters; while increased lipid peroxidation in all the genotypes. The activities of antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase were upregulated by the Cu stress. The highest applied concentration of Cu (60 mg kg- 1) proved most toxic for plants. Moreover, artemisinin content was increased upto 10 mg kg- 1 of Cu treatment, compared with control, however, the artemisinin accumulation decreased at higher doses of Cu in all the genotypes. On the basis of studied parameters, Cim-Arogya was found to be most tolerant among all for Cu toxicity.

Essential and selective role of SNX12 in transport of endocytic and retrograde cargo.

The endosomal protein-sorting machineries play vital roles in diverse physiologically important cellular processes. Much of the core membrane-sorting apparatus is conserved in evolution, such as retromer, which is involved in the recycling of a diverse set of cargoes via the retrograde trafficking route. Here, in an RNAi-based loss-of-function study, we identified that suppression of SNX12 leads to a severe blockage in CIM6PR (also known as IGF2R) transport and alters the morphology of the endocytic compartments. We demonstrate that SNX12 is involved in the early phase of CIM6PR transport, and mediates receptor recycling upstream of the other well-established SNX components of retromer. Ultra-structural analysis revealed that SNX12 resides on tubulo-vesicular structures, despite it lacking a BAR domain. Furthermore, we illustrate that SNX12 plays a key role in intraluminal vesicle formation and in the maturation of a subpopulation of early endosomes into late endosomes, thereby regulating selective endocytic transport of cargo for degradation. This study therefore provides evidence for the existence of early endosomal subpopulations that have differential roles in the sorting of the cargoes along endocytic degradative pathways.

Direct ß-Lactam Inactivation Method: a New Low-Cost Assay for Rapid Detection of Carbapenemase- or Extended-Spectrum-ß-Lactamase-Producing Enterobacterales Directly from Positive Blood Culture Bottles.

We validate and evaluate a new phenotypic assay, named the direct ß-lactam inactivation method (dBLIM), for the rapid and simultaneous detection of carbapenemase or extended-spectrum-cephalosporinase activity directly from Enterobacterales (EB)-positive blood cultures (BCs). It originates from the carbapenem inactivation method (CIM), an inexpensive and highly sensitive assay for carbapenemase activity detection. dBLIM cutoff values to detect extended-spectrum ß-lactamase (ESBL) and carbapenemase activities resulted in diameters of ≤12 mm for a 5-µg-cefotaxime disk and for a 10-µg-meropenem disk. dBLIM assessment was determined with both aerobic and anaerobic BC bottles spiked with 422 characterized EB strains, classifiable into the following 4 phenotypic groups: (i) ESBL/AmpC-type ß-lactamase (ACBL)/carbapenemase (CARB)-nonproducing (np-ESBL/ACBL/CARB) EB (n = 116), (ii) ESBL-producing EB (n = 111), (iii) AmpC-ß-lactamase-producing EB (n = 33), and (iv) carbapenemase-producing EB (n = 162). No false-positive results were obtained in any of the np-ESBL/ACBL/CARB EB, ESBL, and AmpC groups, demonstrating an overall assay specificity of 100%. There were no significant discrepancies in dBLIM performance between aerobic and anaerobic BCs across all groups, except with VIM-type carbapenemase-expressing EB. Interestingly, among BCs spiked with blaVIM-harboring EB, the sensitivity rates of the assay in anaerobic and aerobic bottles were 53.6% and 100%, respectively. In contrast, dBLIM performance was deemed excellent for the KPC, OXA-48, and NDM carbapenemase producers regardless of the type of bottle being tested, with a sensitivity rate ranging between 99% and 100%. Concerning the detection of the extended-spectrum cephalosporinases of the ESBL-producing and AmpC types, dBLIM sensitivities was 100% and 84 to 87%, respectively. dBLIM may be a cost-effective and highly robust phenotypic screening method for the reliable detection of carbapenemases or extended-spectrum cephalosporinases directly from BCs on the same day of bottle positivity detection.

Systematic Comparison of Four Methods for Detection of Carbapenemase-Producing Enterobacterales Directly from Blood Cultures.

Early identification of infections caused by carbapenemase-producing Enterobacterales (CPE) can help to optimize patient treatment and improve outcome. In this study, protocols for rapid detection of carbapenemase production directly from positive blood cultures were developed applying a concentration and hemolysis step before a test for carbapenemase production was performed. Four different methods (three modified colorimetric assays [ß-Carba, bcCarba NP, and NeoRapid Carb] and a variation of the carbapenem inactivation method [CIM] test with blood cultures [bcCIM]) were assessed on blood cultures spiked with 185 different molecularly characterized Enterobacterales isolates. The challenge collection included 81 carbapenemase-negative isolates and 104 CPEs (OXA-48 [n = 25], NDM [n = 20], KPC [n = 18], VIM [n = 25], GIM [n = 5], OXA-48-like [n = 9], and OXA-48-like plus NDM [n = 2]). The sensitivity/specificity was 99.0%/95.1% for bcCarba NP, 99.0%/91.4% for NeoRapid Carb, 100%/95.1% for ß-Carba and 100%/100% for bcCIM. Weakly hydrolyzing carbapenemases (e.g., OXA-48-like) were also well detected by the assays. The time to result was 20 to 45 min for ß-Carba, 2 to 3 h for bcCarba NP, 2.5 to 2 h for NeoRapid Carb, and 18 to 24 h for bcCIM. In conclusion, all assays demonstrated good detection of CPE. The protocols can be easily implemented in any clinical microbiology laboratory and could help to optimize therapy early in bloodstream infections by CPE.