Met) is the first limiting amino acid in corn and soybean meal-based diets (containing L-Met) in broiler chickens, which are often supplemented with synthetic DL-Met or DL-Hydroxy Met (OH-Met). Our objective was to quantitatively assess the efficacy of synthetic Met sources and determine differences in growth rate of broilers fed at or below requirements in response to Met intake. A systematic literature search resulted in building a database containing 480 treatment means from 39 articles published between 1985 and 2019 globally. The database was divided into starter, grower, and finisher subsets based on the age of the broilers. For each subset, linear-plateau and quadratic-plateau models were fitted to determine Met or sulfur amino acid  requirements using average daily gain as a response variable. For each phase, 4 new subsets were obtained by only retaining records with digestible Met or SAA intake at or below requirement by linear-plateau or quadratic-plateau models. Then, a linear model (without plateau) was fitted for all new subsets for each rearing phase using supplemental digestible synthetic Met or SAA intake  as independent variables. The basal diet was made of only raw materials without supplementation of any synthetic Met source. Finally, the models were extended to evaluate source of synthetic Met effects on the slope parameter. At all stages of model fitting, the inclusion of a random study effect was evaluated for each parameter. All models were fitted within a Bayesian framework, for which minimally informative priors were used. The best models, that is, the most accurate inclusion of random effects, were selected based on at least 10-point difference in leave-one-out cross-validation information criterion. Model selection criteria did not consistently favor either of the linear- and quadratic-plateau models to determine Met or SAA requirements across broiler growth phases. Extending models with covariates  did not improve any model fit. Body weight gain response of broiler chickens to the 2 sources was not different when fed at or below Met requirements for any of the growth phases.Methionine ( Met) is the first limiting amino acid in corn and soybean meal-based broiler diets for broiler chickens. Methionine plays a key role in broilers, primarily feather growth and protein synthesis (OH-Met) also known as 2-hydroxy-4-(methylthio)butanoate (HMTBA) or methionine hydroxy analogue – free acid (MHA-FA). However, animals can only utilize L-Met for protein synthesis. The other forms of Met act as precursors of L-Met which must be converted by the animals to L-Met to be utilized , and d) articles that were published in English, Portuguese or Spanish. Additionally, studies where interactions other than the Met effects such as studies with animals under thermo-stressed conditions were excluded. Studies must have been published in peer-reviewed journals. Finally, a total of 39 studies were considered eligible for this meta-analysis. The selection process is shown as PRISMA flowchart according to The papers were selected using the following inclusion criteria: a) articles that compared DL-Met and OH-Met where studies indicated the source and levels of the Met, b) articles with detailed description of diet composition, c) articles that reported at least 2 of these variables: feed intake, weight gain, or feed conversion ratio (n = 132 from 6 studies), South Korea (n = 14 from 1 study), Thailand (n = 15 from 2 studies), Iran (n = 8 from 1 study), India (n = 6 from 1 study) and Turkey (n = 5 from 1 study); Australia (n = 5 from 1 study); Europe: Germany (n = 58 from 5 studies) the Netherlands (n = 45 from 2 studies), Poland (n = 9 from 1 study), Spain (n = 9 from 1 study), France (n = 4 from 1 study); North America: United States (n = 88 from 6 studies), Canada (n = 7 from 1 study), and Mexico (n = 8 from 1 study); and South America: Brazil (n = 55 from 8 studies). The list of articles and reports included in the database are given in the supplementary material.The database included 480 records of treatments means from 39 studies conducted from 1985 to 2019 by research entities from Asia: China , experimental design , ingredient composition, and nutritional values of the experimental diets and performance information such as ADG, ADFI, and FCR.PPFR, Some studies did not report dietary nutrient composition. Thus, the nutrient composition of all diets was recalculated using the Practical Program for Formulation of Rations PPFR,  to get aBased on the age of the birds, the broiler database was subsetted into starter , grower  and finisher (≥ 21 d of age at the start of the experiment) phases. The relationships between predetermined variables  were visually assessed using scatter plots for each growth phase. This visual exploration resulted in the identification of outliers and next in the removal of the records that belonged to 4 different studies.SAA; i.e., Met+Cysteine) intake as independent variable using a 3-stage hierarchical model  in study i , i, and Nyij=ffunction :(2)fffθi=θ+The choice of prior distributions for l phases  and 4. TLOOIC; 3 iterations with the first 10 × 103 taken as burn-in period. Chains were thinned by a factor of 25, after which convergence was assessed by visually inspecting the chain traces. Upon selecting the random-effects structure per model, this selected model was refitted by running 2 chains for 50 × 103 iterations with the first 20 × 103 taken as burn-in period.The actual inclusion of a random study effect per parameter, that is, the specification of the second stage, was evaluated by fitting the hierarchical model structure to the data with a forward selection procedure. At the start of this procedure, a more parsimonious model was considered that had only one random effect in total, which could apply to any of the three parameters n LOOIC; . Subsequk estimated using digestible Met or digestible SAA intake by linear-plateau or quadratic-plateau model for each of starter, grower and finisher subset. Using subsets of a starter, grower or finisher subset, a similar hierarchical model was then used for fitting, but at the first stage the independent variable After fitting the hierarchical model structure using digestible Met or digestible SAA intake as independent variables, the starter, grower, and finisher subsets were further subsetted based on the estimated value of the requirement parameter ar model .1:(6.1)fau model . All othrding to :(7)βn=δnrstan , grower (mean 20.7 d), and finisher broilers (mean 34.9 d), respectively .The models for predicting ADG against digestible SAA intake along with the selected random effects that were fitted are graphically presented in broilers . The quaκ) determined using either linear-plateau or quadratic-plateau models. The fitted linear model was extended to evaluate the synthetic Met effect on the slope parameter  for different phases are shown in Relative efficacy of 2 most common forms of Met (DL-Met vs. OH-Met) has been well-studied and reported performance differences under certain environment and management conditions. Individual studies demonstrated that one form is favored over another under certain conditions, which has created controversy and confusion. Thus, the main goal of this study was to compile up-to-date data from the literature and compare the efficacy of 2 sources of Met using a robust Bayesian meta-analysis approach. We first divided the dataset into starter, grower and finisher age group and then determined Met requirement using either linear or quadratic-plateau models. Most importantly, our final model included the observations below the Met requirements determined using either linear or quadratic-plateau models where previous meta-analysis included all observations within certain dose limits of Met .Recent improvement in computational statistics such as development of programming languages  enables fitting complex models in Bayesian setting . In the In this meta-analysis, both linear- and quadratic-plateau models were used to determine the requirements for Met and SAA with ADG as the response variable. Previous studies used both ADG and FCR as the response variable e.g., . The choGrowth response of birds fed different Met sources depends on the Cys levels in the diet for starter phase, but it was not the case for grower or finisher phase of broiler . This inThis meta-analysis using current literature data with Bayesian approach demonstrated that growth response (ADG) of broiler was not influenced by form of the synthetic Met precursors. In agreement with our study,


Background: Antibiotics are commonly prescribed for outpatient management of cystitis. Previous evidence suggests that certain factors likely beyond the infection seem to influence the choice of antimicrobial treatment. However, studies on the specific antibiotic treatments for cystitis are lacking. This study aimed to explore the antibiotic treatments for cystitis using nationwide primary healthcare data and investigate if factors beyond the infection could be associated with fluoroquinolone treatment. Methods: This nationwide follow-up cohort study consisted of 352,507 women with cystitis. The primary aim was to investigate what specific classes of antibiotics were redeemed by patients within five days from the cystitis diagnosis. Each patient could only be included once. Logistic regression models were also used to examine the relationship between fluoroquinolone (FQ) treatment, parity, and sociodemographic factors. Results: In total, 192,065 antibiotic prescriptions were redeemed. Pivmecillinam (58.4%) followed by nitrofurantoin (22.2%), trimethoprim (12.0%), fluoroquinolone (5.6%), and cephalosporins (1.5%) were the most redeemed antibiotics. Sociodemographic factors were weakly associated with fluoroquinolone treatment; young age was inversely associated with fluoroquinolone treatment. Parity and cervical cancer history were not associated with fluoroquinolone treatment. The proportion of fluoroquinolone and trimethoprim treatments decreased over time, while pivmecillinam and nitrofurantoin increased. Conclusions: The treatment trends of antibiotics redeemed within five days from a cystitis diagnosis were similar to the national surveillance program of these antibiotics (not diagnosis linked). Fluoroquinolones were weakly associated with sociodemographic factors, which likely is only of historical relevance. Uncomplicated lower urinary tract infection (UTI), also known as uncomplicated cystitis, is one of the most common bacterial infections in women. About one in ten women have at least one infection each year and therefore cystitis is an important contributor to antibiotics exposure in otherwise healthy women in Sweden and worldwide ,2,3,4,5.However, over the last decades, safe and effective treatment options have decreased. This is partly due to an increase in antibiotic resistance, making certain antibiotics, such as fluoroquinolones, less suitable as empirical treatment options for cystitis. For example, fluoroquinolone resistance has passed 20% in several countries worldwide. Furthermore, certain UTI antibiotics have also been found to be more associated with collateral damage and severe adverse effects. For example, suspected cardiotoxicity and neuropsychiatric conditions have been related to fluoroquinolones ,18,19,20All this considered, the treatment guidelines for cystitis (including in Sweden) have turned away from certain antibiotics—mainly fluoroquinolones, reserved for more serious infections—towards more narrow-spectrum antibiotics, such as nitrofurantoin and pivmecillinam, with lower resistance rates and more suitable safety profiles for use in the treatment of cystitis ,19,21. TThe gap in previous research is most likely due to the lack of nationwide population-based data from primary healthcare, where UTIs are mostly managed. We recently identified that about 54.5% of women diagnosed with uncomplicated cystitis in Sweden redeemed an antibiotic for their prescription. Furthermore, the study found that certain factors (likely beyond the infection) were associated with antibiotic treatment for cystitis [This study presents diagnosis-linked antibiotic treatment trends for uncomplicated cystitis on a nationwide level. As expected, pivmecillinam and nitrofurantoin were the two most used antibiotics. The annual proportion of these antibiotics increased over time, as the proportion of trimethoprim and fluoroquinolones decreased. Year of infection (continuously increasing) was also inversely associated with fluoroquinolone treatment rates. These trends were in accordance with the national guidelines  and the Less than six percent of all the antibiotic treatments linked to uncomplicated cystitis were fluoroquinolones (mainly ciprofloxacin). Factors likely beyond the infection—such as sociodemographic factors and history of cervical cancer and parity—did not seem to be associated with fluoroquinolone treatment for uncomplicated cystitis to a high extent. Women from low family income backgrounds seemed to have received more fluoroquinolone treatments than women of higher socioeconomic status. For country of origin, only MENA women had lower odds of fluoroquinolone compared to Swedish-born women , while women from Asia (excluding Middle East) and Oceania, Western countries, and Latin America and the Caribbean had weak but significantly higher odds of fluoroquinolone treatment compared to Swedish-born women. The mechanisms behind these sociodemographic differences in fluoroquinolone treatment of uncomplicated cystitis, which were similar to the odds for antibiotic treatment in general , could, Limitations of this study were that we did not have access to data on the symptoms or clinical presentation in the patients, nor did we have data on adherence rates. Therefore, no firm conclusion can be made on whether the differences in fluoroquinolone treatment could have been due to unequal healthcare (differences in prescription rates) or differences in adherence (differences in pick-up rates). Further studies could examine possible mechanisms behind the found associations of factors likely beyond the infection and antibiotic treatments . The maiAntibiotics are important drugs for patients worldwide  and shouThis was a population-based cohort register study and follow-up to a recent nationwide study of ours . The timThis research was conducted at the Center for Primary Health Care Research at Lund University and Region Skåne, Malmö, Sweden. The STROBE statement for cohort studies was considered when conducting this study and in the writing process .The study population consisted of 352,507 women  between The outcome variable (redeemed antibiotic prescription) was defined as the first UTI antibiotic redeemed (measured as prescription from a physician dispensed at a pharmacy) within five days after a diagnosis of uncomplicated cystitis. The outcome is generally referred to as “antibiotic treatment” in the manuscript. The following antibiotic groups were assessed based on their ATC code: penicillins with extended spectrum (J01CA), for example, pivmecillinam; beta-lactam/b-lactam inhibitor combinations (J01CR); cephalosporins ; trimethoprim derivatives (J01EA), i.e., trimethoprim; sulphonamides/trimethoprim combinations (J01EE), i.e., trimethoprim/sulphamethoxazole; fluoroquinolones (J01MA), for example ciprofloxacin; and nitrofuran derivatives, i.e., nitrofurantoin (J01XE). Seven-level ATC codes were available during the first eight years (2006–2013) of the study. Oral drugs were exclusively assessed. Mono-sulphonamide antibiotics and fosfomycin were not considered, as these antibiotics were not widely available in Sweden at the time of the study. We did not consider either antibiotics that were not recommended ,21 to beThe predictors investigated were measured at baseline . Age groups were defined as being between 15–24, 25–34, 35–44, or 45–50 years of age. Country of origin was categorized as originating from any of the following countries/regions: Sweden; Eastern European countries; Western countries; Middle East/North Africa (MENA); Africa (excluding North Africa); Asia (excluding Middle East) and Oceania; or Latin America/the Caribbean. Countries with geographical proximity and/or cultural and economic similarities were categorized together. Both first- and second-generation immigrants were included in country groups other than Sweden. The categories for this study were based on the definition used in a previous study of ours [Socialstyrelsen), were used to identify the outcome, parity, cervical cancer, as well as comorbidities or other complicating factors not aligned with uncomplicated cystitis: [This cohort study population was identified through population-based primary healthcare data 1997–2018) accessed from 21 out of 22 administrative regions in Sweden during the study period. The coverage of these data varied by time and region based on when the patient records were digitalized. The database included around 72% of the population in Sweden in 2015 and around 90% of the population at the end of the study. The following nationwide registers, managed by the National Board of Health and Welfare  was used for all statistical analyses.Descriptive statistics on the annual treatment trends (redeemed antibiotic prescription five days after a diagnosis of uncomplicated cystitis) were calculated for each antibiotic based on the five-level-ATC-code antibiotic group for the whole study period . During The present study was a non-intervention nationwide register study of pseudonymized secondary data obtained from Swedish authorities and was approved by the Ethical Review Board in Lund. All methods were performed in accordance with the relevant guidelines and regulations.The funding sources of this study were all non-commercial and had no role in the study design; the collection, analysis, and interpretation of data; the writing of the report; or the decision to submit the paper for publication.The treatment trends of antibiotics (for urinary tract infections) redeemed within five days from a cystitis diagnosis were similar to the national surveillance program of these antibiotics (not diagnosis linked). The decreasing treatment rate of fluroquinolones over time was also in accordance with the national surveillance program. This indicates that the national surveillance program is likely representative and valid in identifying antibiotics treatments for urinary tract infections. Fluoroquinolone treatment was weakly associated with some sociodemographic factors and parity, which likely are beyond the infection but only of historical relevance, as these antibiotics seem to be used less for this infection in Sweden, in accordance with national guidelines. These findings represent important new information, particularly for healthcare planners involved with the national antibiotic surveillance of this very common bacterial infection in women.


Molecular nanoparticles including polyoxometalates, proteins, fullerenes and polyhedral oligosiloxane (POSS) are nanosized objects with atomic precision, among which POSS derivatives are the smallest nanosilicas. Incorporation of molecular nanoparticles into chiral aggregates either by chiral matrices or self-assembly allows for the transfer of supramolecular chirality, yet the construction of intrinsic chirality with atomic precision in discrete molecules remains a great challenge. In this work, we present a molecular folding strategy to construct giant POSS molecules with inherent chirality. Ferrocenyl diamino acids are conjugated by two or four POSS segments. Hydrogen bonding-driven folding of diamino acid arms into parallel β-sheets facilitates the chirality transfer from amino acids to ferrocene and POSS respectively, disregarding the flexible alkyl spacers. Single crystal X-ray structures, density functional theory (DFT) calculations, circular dichroism and vibrational circular dichroism spectroscopy clearly verify the preferential formation of one enantiomer containing chiral molecular nanosilicas. The chiral orientation and chiroptical properties of POSS show pronounced dependence on the substituents of α-amino acids, affording an alternative way to control the folding behavior and POSS chirality in addition to the absolute configuration of amino acids. Through the kinetic nanoprecipitation protocol, one-dimensional aggregation enables chirality transfer from the molecular scale to the micrometer scale, self-assembling into helices in accordance with the packing propensity of POSS in a crystal phase. This work, by illustrating the construction of chiral molecular nanosilicas, paves a new way to obtain discrete chiral molecular nanoparticles for potential chiroptical applications. A molecular folding strategy is developed to construct ferrocenyl diamino acid conjugated polyhedral oligosiloxane molecules. Hydrogen bonding-driven folding facilitates the chirality transfer from the molecular scale to the micrometer scale. However, there is still plenty of room and there are also several challenges in achieving precise control over molecular chirality and supramolecular chirality in helical systems from molecular to nanoscale levels. For instance, how do we push the helical packing of giant molecular nanoobjects with atomic precision?5–9Natural organisms realize complicated functions and structures within chiral and helical environments, where noncovalent force-driven folding and hierarchical self-assembly play vital roles as a result of a long evolution process. The previous decade witnessed rapid development of helical and chiral macromolecular and supramolecular self-assembled systems across different levels, dimensions and sizes.60) and polyhedral oligosiloxanes (POSS), are widely used molecular nanoparticles, which are shape and volume-persistent objects with size over 1 nm.10 The pronounced atomic precision and specific symmetry endow these nanoobjects with superiority compared to the self-assembled particles or synthetic metal particles with polydispersity. Considering the diversified size, dimension, surface functionalization and other chemicophysical properties, molecular nanoparticles have been used as building blocks in versatile self-assembly systems.11–14 Integration between chirality and molecular nanoparticles has sparked to enable fascinating helical systems with specific topology and functions.15,16 In this regard, tubulins self-assembled into helical microtubes;17 POMs and noble metal clusters could coassemble into helical structures within chiral templates;14 C60 encapsulated in helical polymers could achieve helical orientation.8 Among them, POSS are the smallest silica nanoparticles with diameter about 1 nm or higher depending on the grafted functional groups. POSS have been intensively incorporated into ordered architectures with versatile phases such as lamellar, bcc and a special A15 phase via delicate designs.18–20 Due to the high crystallinity and apolar nature, POSS have seldom been applied to construct helical systems, and a precise control over the helical orientation of POSS has not been well accomplished to date. Foldamers are defined as synthetic polymers or oligomers adopting a compact conformation.21 Typical examples include peptoids, oligonucleotides, oligoamides, aza-aromatic oligomers and tertiary aromatic units, which undergo folding into specific conformation via hydrogen bonds or aromatic interactions.22 Through intrinsic folding, helical structures with hierarchical levels of chirality are formed, providing a potential way to control the emergence and properties of helical POSS structures.Molecular nanoparticles refer to atomically precise molecules with giant size over the nanometer scale. Metal clusters, such as polyoxometallates (POMs), proteins, fullerenes  to the 1,1′-ferrocenyl diamino acids. In the solid X-ray structures, intramolecular folding and the large size of POSS enabled the exclusive helical packing of POSS domains  via an amide condensation reaction. Nuclear magnetic resonance (NMR) and mass spectra indicated the successful synthesis , the down-left/up-right affords chiral packing  columns, where the POSS segments 1/2 and 3/4 from individual Val-POSS separately constitute a helical turn with a helical pitch of 17.6 Å , segments 1/2 and 3/4 from individual Phe-POSS constitute a staircase-like motif, which can be regarded as the tilt or 21 columnar chirality, affording the P-handed helix. The handedness of 1D packing is independent of the intramolecular folding behavior. Viewed along the a axis, a hexagonal packing of the helices can be found.The impact of amino acids on the folding and self-assembly behaviors was explored by the analysis of the solid-state structures of Phe-POSS and Met-POSS respectively . The priM- and P-helices respectively. In the packing mode, viewed along the b axis (top view), P- and M-columns pack adjacently into a supramolecular racemic phase. The major driving factors in the formation of the racemic mixture for POSS helices are associated with the existence of sulfur elements. Close contacts were found in both conformations between sulfur and methyl protons. The S⋯H interaction with distance around 3 Å hinders the close packing between POSS segments, resulting in a large lateral length up to 27.5 Å. It is believed that the homochirality originates from the transferred chirality from amino acids assisted by the duplex hydrogen bonds. Meanwhile, the bulky size of POSS moieties also contributes to the fixed chiral orientation. On account of the steric hindrance, the free rotation is forbidden even in the presence of alkyl spacers between the chiral centers and POSS segments. Thus, the specific chiral conformation is a result of the synergistic effect of hydrogen bonds, chiral centers and steric hindrance. The bulky space and relatively weak forces between POSS segments allow for swinging between the two geometries with opposite chiral orientations.Met-POSS adopted the identical β-sheet structure and Herrick's conformation, which enabled the formation of helical POSS folding as well . HoweverCompared to the above cases, Glu-POSS and Asp-POSS are special species conjugated by the four POSS segments. The conjugation of a large number of POSS with low polarity appended considerable steric hindrance with less polarity. Consequently, the crystallization of Glu-POSS and Asp-POSS is rather difficult. Density functional theory (DFT) optimization at the B3LYP/6-31g level of theory was applied to probe the geometry of Glu-POSS . Herrickl- and d-Enantiomers afforded positive and negative maximum Cotton effects at 483 nm respectively, indicating the crucial effect of the absolute chirality of amino acids on the ferrocene . The folding by hydrogen bonds as well as the steric hindrance of POSS segments allows for a structure-persistent twisted ferrocene conformation with axial chirality, contributing to large g-factors.The above studies on X-ray structures represent chirality in the solid phase. Then we studied the chiroptical responses of various species in the solution phase. In the solution phase, all the nine building units exhibit well-resolved Cotton effects corresponding to the absorbance of the ferrocene unit Fig. S21, whose ferrocene . The typund 0.01 , which a1H NMR spectroscopy .As the intramolecular hydrogen bonds are vulnerable to temperature, temperature-variable CD spectroscopy was carried out . Heatingtroscopy , raising4, chloroform, dichloromethane (DCM), tetrahydrofuran (THF), dioxane, toluene, and methyl cyclohexane (MCH). Thus, polar solvents such as acetone, acetonitrile, methanol, dimethyl formamide, dimethyl sulfoxide, and water could hardly dissolve the samples. To exclude the solvent effect in chiroptical activities, the CD spectra of l-Val-POSS with 1 mM concentration in various solvents  exhibit the same Cotton effect and similar intensity without any bathochromic or hyperchromic shift, which suggest that these solvents have a negligible effect on the chiroptical activities. The slightly changed CD intensity might be caused by the different absorption coefficients. In addition, temperature-variable CD spectra in CHCl3 were also obtained to verify the results with the above CD and NMR studies. As shown in Fig. S25,3 are in good agreement with those obtained using dioxane, further indicating the negligible solvent effect.Due to the structure-persistent property of the POSS-containing building units, the chiroptical spectra show negligible dependence on the solvent environments. Owing to the apolar nature of POSS, only few solvents with low polarity are considered good solvents, including CCl Fig. S24 were comvia intramolecular folding. To verify the chiral packing in the solution phase, VCD was applied  scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>O stretching are displayed in Fig. S30.−1 corresponds to the free N–H group. The peak at around 3284 cm−1 is assigned as the hydrogen bonded N–H group.31 These observations suggest that partial N–H of amides is involved in the hydrogen bond formation, which are in good agreement with the single crystal structure profiles and the assumptions for intramolecular hydrogen bonded folding. Furthermore, the CO stretching bands of each building unit appear from 1634 to 1628 cm−1. Such locations with low wavenumbers support the formation of the N–H⋯OC hydrogen bond. In the solution state (CHCl3), the N–H⋯OC hydrogen bond was also found  at the specific region. In solution, the presence of an active VCD signal corresponds to the intrinsic chiral POSS segments. It is in good agreement with the single crystal profile. Although there are propyl spacers between POSS and amino acids with point chirality, chirality was transferred to POSS benefiting from the rigid Herrick conformation and β-sheet structure. Heating Val-POSS from room temperature to 80 °C slightly changed the VCD intensity, which however could hardly change the VCD sign and the major absorbance , contributed by the apolar nature of POSS, whereby the chirality evolution to a macroscopic scale is achieved. Through nanoprecipitation or solvent exchange method, in a tetrahydrofuran/acetonitrile  mixture, the building units (1 mM) underwent spontaneous aggregation into a colloidal dispersion as well as precipitate phases. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were employed to inspect the morphologies. Val-POSS self-assembled into large-scale 1D fibrous structures . The aggregates were collected and deposited for the thin-film X-ray diffraction (XRD) study. Compared to the simulated powder pattern, the experimental XRD pattern shows similar peak locations with varied peak intensities  plane with the missing (001) plane. The lamellar structure found in POSS-containing building units is attributed to the selected distribution of apolar POSS tails and polar ferrocenyl diamino acid heads, and within the individual lamellas also highly crystalline arrangement was found due to the existence of multiple planes. Met-POSS exclusively showed the entire lamellar peaks from (001) to (005) planes with a perfect distance ratio of 1 : 1/2 : 1/3 : 1/4 : 1/5, with a lattice parameter of 2.88 nm. However, other lamellar cases generated a rather loose stacking with a first order d-spacing value up to 5 nm, in good accordance with the previous POSS assemblies. PGly-POSS, Asp-POSS and Glu-POSS afforded hump peaks in contrast to other entities, probably contributed by the 1D aggregates without 3D molecular aggregation. These peaks can also be assigned to the lamellar stacking with retained planes and different lattice parameters. Powder XRD patterns illustrated the general formation of lamellar structures in ferrocene-diamino acid POSS building units, whereby chirality could be transmitted to hierarchical levels.In powder XRD patterns, besides Val-POSS, Phe-POSS and Met-POSS show pronounced deviation of the simulated patterns from single crystal files, indicating that the solution-assembly adopted different ways of aggregation. It is interesting that the patterns of Ala-POSS, Leu-POSS, Pro-POSS and Phe-POSS possess 4–5 peaks ranging from 6 to 11 2theta degree, which could be assigned to the crystalline lamellar phase.In conclusion, we illustrated the design and fabrication of molecular chiral nanosilicas, chiroptical properties and self-assemblies. POSS segments were conjugated to the ferrocenyl diamino acids. The resultant Herrick conformation formation and folded parallel β-sheet structure enabled the asymmetric orientation of POSS segments disregarding the flexible spacers. The chirality of POSS was determined from the absolute chirality of amino acids with a pronounced substituent effect. The intrinsic chiral POSS molecules exhibited active VCD signals, which were persistent at high temperatures. Single crystal structures indicated the formation of helical motifs, which underwent aggregation into 1D helices with homo-chirality. This work reported the unprecedented formation of intrinsic helical silica nanoparticles with atomic precision in solid and solution states, creating new possibilities in the precise control of chirality and supramolecular chirality using molecular nanoparticles.l-valine methyl ester hydrochloride (1500 mg) were placed in a 250 mL flask. Then, hydroxybenzotriazole , 4-dimethylaminopyridine , 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide  and triethylamine  were added. The mixture was stirred in DCM (60 mL) at room temperature for 8 hours. When the reaction was complete, the mixture was poured into acidic water, extracted with DCM three times, and then reversely extracted twice, and finally purified using a silica gel column to obtain the pure product. Other compounds were synthesized using the same procedure.The synthesis route is shown in Scheme S1.The synthesis route is shown in Scheme S2.2 (2.0 g) with HOBT (200 mg), DMAP (200 mg), EDC (1200 mg), and TEA (1 mL) were stirred in DCM (60 mL) at room temperature for 8 h. After the reaction was complete, the DCM solution was poured into acidic water, extracted with DCM and reversely extracted twice. The crude product was purified using a silica gel column and finally an orange-yellow powder was obtained by rotary evaporation. Other compounds were synthesized using the same procedure.The synthesis route is shown in Scheme S3.Ala-POSS was prepared as an orange-yellow solid in total 31% yield . 1H NMR  δ 8.75 , 6.63 , 4.87 , 4.80–4.57 , 4.49 , 4.27 , 3.50 , 3.13 , 1.85 , 1.65 , 1.46 , 0.95 , 0.60 . 13C NMR  δ 174.91, 71.84, 71.04, 70.52, 70.05, 49.50, 42.28, 25.70, 25.68, 23.90, 23.85, 23.10, 22.50, 22.47, 17.41, 9.56. HRMS (TOF) m/z [M + H]+, calcd for C80H159FeN4O28Si16+, 2127.6793; found, 2127.5887.Val-POSS was prepared as an orange-yellow solid in 32% yield . 1H NMR  δ 8.36 , 5.98 , 4.84 , 4.78–4.72 , 4.47 , 4.29 , 4.06–3.97 , 3.49 , 3.19 , 2.31–2.23 , 1.85 , 1.66 , 1.07 , 0.95 , 0.60 . 13C NMR  δ 173.56, 71.92, 71.37, 69.90, 42.47, 28.94, 25.73, 25.72, 23.92, 23.87, 23.03, 22.49, 22.46, 20.24, 19.88, 9.73. HRMS (TOF) m/z [M + H]+, calcd for C84H167FeN4O28Si16+, 2183.7419; found, 2183.7245.d-Val-POSS was prepared as an orange-yellow solid in 33% yield . 1H NMR  δ 8.41 , 6.23 , 4.88–4.79 , 4.77 , 4.47 , 4.29 , 4.05 , 3.50 , 3.18 , 2.39–2.19 , 1.85 , 1.67 , 1.07 , 0.95 , 0.60 . 13C NMR  δ 173.57, 170.78, 76.17, 71.68, 71.16, 70.00, 69.83, 61.49, 42.36, 28.89, 25.70, 25.69, 23.90, 23.85, 23.01, 22.50, 22.47, 20.15, 19.81, 9.69. HRMS (TOF) m/z [M + H]+, calcd for C84H167FeN4O28Si16+, 2183.7419; found, 2183.7177.Leu-POSS was prepared as an orange-yellow solid in 31% yield . 1H NMR  δ 8.73 , 6.22 , 4.88 , 4.53 , 4.28 , 3.45 , 3.16 , 1.91–1.80 , 1.65 , 0.95 , 0.60 . 13C NMR  δ 71.92, 71.13, 70.34, 70.15, 52.82, 42.36, 40.17, 25.74, 25.71, 24.87, 23.92, 23.87, 23.33, 23.10, 22.49, 22.48, 21.37, 9.65. HRMS (TOF) m/z [M + H]+, calcd for C86H171FeN4O28Si16+, 2211.7732; found, 2211.7567.Pro-POSS was prepared as an orange-yellow solid in 30% yield . 1H NMR  δ 9.09 , 4.86 , 4.49 , 4.28 , 3.73 , 3.64 , 3.22 , 1.83 , 0.93 , 0.69–0.52 . 13C NMR  δ 171.00, 167.47, 78.14, 73.44, 70.17, 69.26, 67.26, 62.40, 46.22, 41.71, 31.15, 24.67, 22.85, 22.82, 21.88, 21.45, 21.41, 20.50, 8.68. HRMS (TOF) m/z [M + H]+, calcd for C84H163FeN4O28Si16+, 2179.7106; found, 2179.7111.Met-POSS was prepared as an orange-yellow solid in 31% yield . 1H NMR  δ 8.71 , 6.44–6.20 , 4.89 , 4.79 , 4.69 , 4.52 , 4.30 , 3.40 , 3.23 , 2.63 , 2.18 , 2.09 , 1.96 , 1.90–1.81 , 1.66 , 0.95 , 0.60 . 13C NMR  δ 172.81, 169.97, 74.41, 71.01, 70.36, 69.20, 52.20, 41.44, 29.99, 29.72, 24.71, 24.69, 24.66, 22.88, 22.83, 22.82, 21.91, 21.44, 14.47, 8.62. HRMS (TOF) m/z [M + H]+, calcd for C84H167FeN4O28S2Si16+, 2247.6860; found, 2247.6077.Phe-POSS was prepared as a light yellow solid in 30% yield . 1H NMR  δ 8.81 , 7.37–7.18 , 5.93 , 4.79 , 4.49 , 4.29 , 3.25 , 1.91–1.77 , 1.46 , 1.02–0.87 , 0.59 . 13C NMR  δ 172.37, 169.67, 136.60, 128.28, 127.67, 125.86, 74.75, 70.88, 70.26, 69.12, 55.62, 41.47, 36.47, 24.69, 24.66, 22.86, 22.83, 21.70, 21.48, 21.43, 8.52. HRMS (TOF) m/z [M + H]+, calcd for C92H167FeN4O28Si16+, 2279.7419; found, 2279.7478.PGly-POSS was prepared as an orange-yellow solid in 32% yield . 1H NMR  δ 8.68 , 7.40 , 5.97 , 5.74 , 4.87 , 4.51 , 4.33 , 3.20 , 3.02 , 1.90–1.79 , 1.53 , 0.98–0.89 , 0.59 . 13C NMR  δ 170.13, 168.98, 136.73, 127.83, 127.45, 127.18, 75.16, 70.96, 70.63, 70.46, 68.91, 55.79, 41.51, 24.68, 24.66, 22.85, 22.82, 21.75, 21.43, 21.39, 8.41. HRMS (TOF) m/z [M + H]+, calcd for C90H163FeN4O28Si16+, 2251.7106; found, 2251.7398.Asp-POSS was prepared as a light yellow solid in 25% yield . 1H NMR  δ 8.08 , 6.68 , 4.85 , 4.52 , 4.32 , 3.39–3.08 , 2.83 , 2.62 , 1.85 , 1.56 , 1.06–0.81 , 0.59 . 13C NMR  δ 173.93, 170.62, 70.80, 51.77, 43.43, 43.16, 26.48, 26.46, 24.66, 24.63, 24.61, 23.62, 23.47, 23.26, 23.21, 23.16, 10.42. HRMS (TOF) m/z [M + H]+, calcd for C144H297FeN6O54Si32+, 3926.2639; found, 3926.2296.Glu-POSS was prepared as a yellow solid in 26% yield . 1H NMR  δ 8.63–8.42 , 6.97–6.69 , 5.84 , 4.83 , 4.52 , 4.30 , 3.37–3.10 , 2.39 , 2.17 , 1.99 , 1.84 , 1.69–1.50 , 1.03–0.85 , 0.60 . 13C NMR  δ 174.38, 173.20, 171.55, 78.12, 71.97, 33.43, 53.69, 43.28, 33.43, 26.50, 26.47, 24.66, 24.61, 23.64, 23.27, 23.24, 23.21, 10.45, 10.28. HRMS (TOF) m/z [M + H]+, calcd for C146H301FeN6O54Si32+, 3954.2952; found, 3954.2988.The data that support the findings of this work are included in the ESI.†P. Xing and Y. Zhao conceived the research idea. Z. Zong carried out the experiments and collected the data. A. Hao participated in the analysis and discussions. All authors contributed to the final manuscript.There are no conflicts to declare.SC-013-D2SC00793B-s001SC-013-D2SC00793B-s002


Despite decades of stewardship efforts to combat antimicrobial resistance and quantify changes in use, the quality of antibiotic use in British Columbia (BC) remains unknown. As the overuse and misuse of antibiotics drives antibiotic resistance, it is imperative to expand surveillance efforts to examine the quality of antibiotic prescriptions. In late 2019, Canadian expected rates of antibiotic prescribing were developed for common infections. These rates were utilized to quantify the gap between the observed rates of prescribing and Canadian expected rates for antibiotic use for the province of BC. The prescribing data were extracted and matched to physician billing systems using anonymized patient identifiers from 1 January 2000 to 31 December 2018. Outpatient prescribing was further subdivided into community and emergency department settings and stratified by the following age groups: <2 years, 2–18 years, and ≥19 years. The proportions of physician visits that received antibiotic prescription were compared against the Canadian expected rates to quantify the unnecessary use for 18 common indications. Respiratory tract infections (RTI), including acute bronchitis, acute sinusitis, and acute pharyngitis, reported significant levels of overprescribing. Across all ages and health care settings, prescribing for RTI indications occurred at rates 2–8 times higher than the expected rates recommended by a group of expert Canadian physicians. Understanding the magnitude of unnecessary prescribing is a first step in delineating the provincial prescribing quality. The quantification of antibiotic overuse offers concrete targets for provincial stewardship efforts to reduce unnecessary prescribing by an average of 30% across both outpatient and emergency care settings. The misuse of antibiotics is a global crisis. Over 30% of the antibiotic prescriptions in the United States were deemed inappropriate, and one third of the antibiotic prescriptions for upper respiratory tract infections (URTI) in Europe had no justification for their use ,2. ThereAntibiotic use is ubiquitous globally, as well as within Canada, and, while effective for specific indications, the overuse and misuse of antibiotics has been strongly associated with the increasing rates of antimicrobial resistance (AMR) ,7. AntibOver the 19-year study period, we had a total of 3,490,585 unique patients, with an average of 447,107 unique patients per year prescribed an antibiotic for one of the 18 included indications across both community and emergency care .The data for 78,495,265 physician encounters were extracted between 1 January 2000 and 31 December 2018. Outpatient care was responsible for 77,754,597 of the cases over the 19-year study period, with 7,708,016 unique patients. Emergency care accounted for an additional 740,668 physician encounters between 2012 to 2018, attributed to 289,954 unique patients. Across both healthcare settings, 57% of all antibiotic dispensation records linked to an indication within the study scope. Antibiotics were more often prescribed for females in urban settings, particularly in the Fraser health authority, and a negative association was observed between increased antibiotic use and income quintile.The most common diagnoses in outpatient care were the common cold, UTI, asthma, and purulent SSTI, making up 53% of the cases . In contThe potential unnecessary antibiotic prescribing is shown in By age groups in outpatient care: patients <2 years of age were the most overprescribed for acute pharyngitis (59%), while bronchitis led overprescribing for those aged 2–18 years, with 49.8% unnecessary. For adult patients (≥19 years), dental infections (56%) received the highest rate of unnecessary antibiotic prescription. The trends by age in emergency care varied from the community prescribing patterns. The patients aged <2 years were overprescribed for AOM at the highest rate (45%), while dental infections (65%) were overprescribed the most in those aged 2–18 years. The highest rate of unnecessary antibiotic use in adults was 76%, for dental conditions. The magnitudes of unnecessary antibiotic use varied across healthcare settings with similar directional trends across indications. Elevated, unnecessary antibiotic use was most pronounced in respiratory tract indications (RTI) overall, including: acute bronchitis (53%), acute sinusitis (46%), and acute pharyngitis (46%), wherein all three age categories were prescribed in excess of at least 30%. Of these RTI conditions, children aged less than 2 years received the most unnecessary prescriptions for acute pharyngitis, while adult patients led bronchitis and sinusitis. Moreover, as respiratory tract infections accounted for over 60% of all the physician visits, and 55% of all the associated antibiotic use, the scale of unnecessary use for these indications translates to high levels of inappropriate prescription in both the community and emergency healthcare settings. Across all the indications and both healthcare settings, prescribing for adults was 29% unnecessary, with 28% unnecessary for children aged 2–18 years and 24% unnecessary for children aged less than 2 years.Our study corroborates findings from several studies across jurisdictions operating under similar universal healthcare systems, such as the United Kingdom, and other provinces within Canada ,16. WhenAlthough dental conditions account for a minority of the total prescribing (1%), the elevated proportion of unnecessary prescribing in adults is concerning. BC emergency care had a 75% rate of overprescribing for dental conditions, far more than the 32% seen in ON outpatient care or the 50% in BC outpatient care. However, as dental conditions are a tier 2c indication, with a 4% expected rate of prescription across all ages, unnecessary antibiotic use is significant for this diagnosis regardless of the healthcare setting. While most medical care is publicly funded in BC and essentially universal in Canada, dental services remain a privately insured sector of healthcare. As such, dentists in BC do not bill MSP directly, nor were they explicitly identified within NACRS (emergency) analyses. The overprescribing reported is not related to dental practitioners but rather physicians treating dental conditions—as further indicated by the reduced number of overall physician visits for dental conditions in comparison to other indications. However, inappropriate antibiotic use in the treatment of dental conditions, in both community and emergency settings, has been well documented across BC, the United States, the United Kingdom, as well as Australia ,19,20. PThe expected Canadian rates of prescribing were engaged within this study as maximal rates throughout analyses—wherein only instances of excess prescribing were characterized as unnecessary. The decision to disregard the rates of prescribing below expected was founded in the principles of antimicrobial stewardship ,22. In tOur limitations are similar to other retrospective studies conducted using administrative data. The linkage of PharmaNet dispensing records and physician billing data may exclude refill prescriptions as the patient would not visit the clinic. Further, our rates do not account for unfilled prescriptions issued, and the levels of compliance to the dispensed medications are unknown. Throughout the study period, 57% of all the antibiotic use was linked to an indication within the study scope, with an additional 22,193,615 antibiotic dispensations unlinked and excluded from further analyses. These prescriptions can be attributed to indications beyond the study scope, hospital discharge records, prescription refills, and/or non-physician healthcare providers . Moreover, the records of indications are reliant on accurate coding by billing physicians; in the absence of lab data to confirm the bacterial etiology, our use of ICD-9/10 codes may be subject to misclassification bias. Although it is notable that Canadian primary-care physician claims data have a high positive-predictive value for the diagnosis of some common infections, including acute non-bacterial upper-respiratory infections, concerns regarding the specificity of ICD-9/10 codes remain ,25. FurtE. coli, Klebsiella spp., and other Enterobacteriae, continues to rise due to the acquisition of extended spectrum beta-lactamases (ESBL) and carbapenenases; interventions to protect the efficacy of these essential medications is paramount [Research has identified a strong association between antibiotic overprescribing and subsequent antimicrobial resistance ,29,30,31aramount ,33,34. Maramount . With tharamount ,37. Higharamount . As furtThe Ministry of Health in British Columbia houses several healthcare-related databases, which have comprehensive information on BC residents (population: 5 million) . Antibion < 5 were excluded from subsequent analyses to preserve subject anonymity.Our study included all BC residents from 1 January 2000 to 31 December 2018. Physician visit and indication data were pulled from both MSP (outpatient care) and NACRS (emergency care) indices, and then matched to antibiotic dispensations extracted from PharmaNet using anonymized patient identifiers. A prescription and diagnosis were linked using an algorithm that matched the date on which the medication was dispensed to a practitioner service date within 5 days prior. If a practitioner service date was associated with more than one diagnostic code, or multiple service dates fell within a single 5-day period of a prescription dispensing date, then a three-tiered hierarchy was applied to link only the most relevant diagnostic code to the prescription. If multiple diagnoses were listed from the same tier, the first physician code was selected for analysis. Multiple prescriptions per subject were permitted in our analyses. Antibiotic data that did not match to a physician visit record for an indication within study scope were not included in the analyses. All data outputs with A study by Wu et al., published in 2020, presented Canadian expected rates of outpatient prescribing for the incidence of various common infections . These rWe have mirrored their methodology to generate comparable rates of observed prescribing in BC. Furthermore, data were extracted for 18 of the same common clinical conditions they reviewed: acute otitis media (AOM), acute pharyngitis, acute sinusitis, asthma, bronchitis, chronic sinusitis, common cold, dental conditions, epididymo-orchitis, influenza, non-purulent soft tissue skin infections (SSTI), otitis externa, pneumonia, prostatitis, purulent SSTI, pyelonephritis, reproductive tract infections, and urinary tract infections (UTI); the exception was chronic obstructive pulmonary disease (COPD) as their expected prescribing rate is for acute exacerbations of COPD, which is not well-captured by billing codes. These indications were further organized into 5 tiers based on the proportion of expected prescribing. Tier 1 included diagnoses for which antibiotics are routinely used . Tier 2 were diagnoses for which antibiotics are frequently indicated (tier 2a: 51–99% expected prescribing rate), sometimes indicated (tier 2b: 21–50% expected prescribing rate), or rarely indicated (tier 2c: 1–20% expected prescribing rate). Finally, tier 3 included diagnoses for which antibiotic use is never indicated (0% expected prescribing rate).Antibiotics were classified based on the Anatomical Therapeutic Chemical (ATC) classification system developed by WHO . ConsumpOverall rates of total antibiotic use were examined and then stratified by age group . Observed rates were compared against maximal references, extracted from Canadian expected rates, and the difference in prescribing rates were calculated for each indication across both outpatient and emergency department care. Unnecessary antibiotic use was identified using the magnitude of difference between expected and observed prescribing by indication, age, and healthcare setting. Prescribing below expected rates was not classified as inappropriate, and all negative magnitudes of difference were subsequently reported to be: 0% prescribing.This study reports elevated levels of unnecessary antibiotic use for RTI indications across all ages and two distinct healthcare settings. These diagnoses, commonly of viral etiology, do not often warrant the use of antibiotic treatment (tier 2c and/or 3), yet antibiotics continue to be prescribed at rates far above the expert and guideline recommendations when compared to the expected rates. The magnitudes of the reported unnecessary prescribing offer new, actionable targets for provincial stewardship efforts.


The Children’s Hospital Association’s Improving Pediatric Sepsis Outcomes (IPSO) collaborative is a multi-center quality improvement (QI) learning collaborative of 61 U.S. children’s hospitals that seeks to improve sepsis outcomes through collaborative learning and reliable implementation of evidence-based interventions in pediatric emergency departments, intensive care units, general care units, and hematology/oncology units. Specifically, IPSO’s goals are to decrease sepsis-attributable mortality and prevent hospital-onset sepsis among children.The following 10 abstracts represent a select group of projects undertaken by IPSO participating hospitals that were presented at one of three collaborative events in 2020 and 2021. IPSO’s Research Workgroup reviewed all submitted abstracts and selected the top 10 for inclusion in this Supplement Bundled resuscitation including timely fluid resuscitation and antibiotic administration improves mortality in children with sepsis. Two of the central challenges of implementing a hospital-wide quality improvement initiative are addressing the unique obstacles in widely varying care environments and delivering timely performance data.In April 2019, our sepsis team implemented a unit-based scorecard reporting system in an effort to provide unit directors with personalized and actionable data regarding the care of septic children in their particular unit.The scorecards contain hospital-wide as well as unit-specific performance data for four of the five key Improving Pediatric Sepsis Outcomes process measures—huddles, order set utilization, time to first IV antibiotic, and time to first fluid bolus Fig. . SpecifiBefore implementation, we met or exceeded the less than 60-minute threshold for intravenous antibiotic administration for 1 month and the less than 20-minute threshold for first fluid bolus in zero months. Following implementation, we met or exceeded the antibiotic threshold in six out of 9 months (67%) and the fluid bolus threshold in three out of 9 months (33%). The average time to first antibiotic decreased by 28% (89 to 64 minutes) after implementation. The average time to first fluid bolus decreased by 32% (38 to 26 minutes) after implementation.Providing timely, specific, and actionable information at the unit level can lead to meaningful system wide improvements in care delivery to septic children.


Continuous processing is without doubt the most effective and economically viable processing mode, because it optimally uses the installed assets. While most chemical industrial processes are designed and executed already in continuous operation, the biotechnology industry still relies on batchwise production, due to multiple unknown variables of complex biological systems.Recent reports have demonstrated, specifically in the area of cell culture processes, that continuous biomanufacturing (CBM) has significant economic advantages. Continuous processes significantly influence the ecological and economic footprint, as facilities become smaller and versatile with the application of modular design approaches. Continuous cultivation approaches have the need to tightly control process variability along prolonged processing times, calling for accurate process analytics and robust control strategies.However, establishing integrated processes for production with dedicated microbial systems is far more complicated, although of course continuous processes are standard, e.g., in the areas of biogas production, wastewater treatment and bioleaching. The complication is due to the fact that, especially in the area of the production of defined recombinant proteins, the protein often accumulates in the cell, which represents a natural physical barrier contradicting continuous operational strategies. In addition, common induction systems have a highly time dependent effect and may lead to high temporary metabolic load. Both effects lead to limitations in prolonged recombinant protein production, seen in an irreversible drop in protein productivity. Hence, the lack of basic knowledge about the regulatory mechanisms leading to population evolution during prolonged recombinant protein production prevents the industrial application of continuous biomanufacturing. But also in other microbial processes, with both engineered and non-engineered single-species microbial cell factories, continuous operation has so far rarely been realized.So what are the current challenges and scientific enablers for advancing continuous biomanufacturing with microbial cells? Several challenges needs to be answered by scientific knowledge (- How to control metabolic load for sustained recombinant product production, for example by the assessment of population stability, analyzing phenotypic, and genotypic instabilities- How to change the product location to circumvent the internal physical limit of the cell?- How to achieve prolonged time invariant processing providing stable productivity and product quality to the downstream process- How to efficiently integrate upstream and down stream processing?The goal of this electronic article collection is to give an overview on recent scientific advances addressing above points.The selected contributions in this Research Topic show significant scientific advancements to microbial CBM and can be classified in the following three aspects (Cellular mechanismsIntensified bioprocessingProduct recovery.Those contributions address well the challenges mentioned above.Risager Wright et al. addresses the importance that population heterogeneity should be taken into account for CBM design and analyzed the suitability of current analytical devices. The contribution “The Lazarus Escherichia coli Effect: Recovery of Productivity on Glycerol/Lactose Mixed Feed in Continuous Biomanufacturing” by Kittler et al. addresses a sudden phenomenon of productivity drop and suggests that the mixed feed of Glycerol and Lactose, but also Galactose as a light inducer, are responsible for population dynamics. Graf et al., with their contribution “Revisiting the Growth Modulon of Corynebacterium glutamicum Under Glucose Limited Chemostat Conditions,” analyze the control mechanisms on transcriptomic and metabolic level, with the conclusion, that glycolysis, pentose-phosphate pathway and citric acid cycle are predominately metabolically controlled under glucose-limiting chemostat conditions and that transcriptional regulation takes control over glycolysis once glucose-rich growth conditions are installed.It is important to understand the population dynamics and its mechanisms of the cell being cultivated in the bioreactor. The perspective article “Single-Cell Technologies to Understand the Mechanisms of Cellular Adaptation in Chemostats” by How to achieve prolonged sustained productivities in the bioreactor? Challenges such as population heterogeneity, loss of the catalyst need to be overcome. In cell culture processes, perfusion techniques are the method of choice currently.E. coli” by Kopp et al. found that a repetitive fed-batch for E. coli leads to a higher space-time yield compared to a single-cycle fed-batch and can potentially outperform continuous biomanufacturing by classical chemostats. Raveschot et al., with their contribution “Integrated Continuous Bioprocess Development for ACE-Inhibitory Peptide Production by Lactobacillus helveticus Strains in Membrane Bioreactor” developed an integrated continuous process for peptide production with a membrane bioreactor, which led to a 3 fold increased peptide productivity compared to batch production. In the contribution “The Impact of Glass Material on Growth and Biocatalytic Performance of Mixed-Species Biofilms in Capillary Reactors for Continuous Cyclohexanol Production,” Heuschkel et al. growth and catalytic performance of mixed-species biofilms for cyclohexanol was investigated. Four phases of the biofilm cultivation could be distinguished and the cyclohexanol production rate could be significantly improved.The research article “Repetitive Fed-Batch: A Promising Process Mode for Biomanufacturing With How to deliver continuous product to the downstream? In addition to the production mode, the product must be efficiently be separated from the cells in order to integrate smoothly to the subsequent down stream process.Pekarsky and Spadiut with their review “Intrinsically Magnetic Cells: A Review on Their Natural Occurrence and Synthetic Generation” investigate the state of the art of Natural intrinsically magnetic cells which can be used to separate the cells during CBM. The research article “Selective release of recombinant periplasmic protein from E. coli using continuous pulsed electric field treatment” by Schottroff et al. focusses on novel process methods to circumvent product accumulation in E. coli and triggering product secretion to the extracellular medium by applying pulsed electric fields. Thakur et al., with their contribution “Automation of Dead End Filtration: An Enabler for Continuous Processing of Biotherapeutics” demonstrated that Dead end filtration, which is conventionally done in batch mode, can be automated and can therefore server as a potential tool for continuous biomanufacturing.We believe, this topic issue made a significant contribution to advance microbial CBM to become reality. However, we are aware that many more aspects need to be addressed to complete the picture, such as PAT, modeling, and control aspects!CH designed and wrote the manuscript. FD, PN, and CS revived and revised the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.