SARS-CoV-2 infection (COVID-19) in children.

Since the outbreak in December 2019, the SARS-CoV-2 pandemic virus has been a major public health problem in all countries of the world. The virus is transmitted by inhalation of respiratory droplets from the patient or asymptomatic carrier and is highly contagious. The clinical disease in children is similar to any acute respiratory infection with predominant upper respiratory symptoms, but occasionally can progress to pneumonia with acute respiratory distress syndrome and multiorgan failure. The disease is milder in children than in adults, with low mortality, and it appears that infants and young children have a somewhat more severe clinical course. Diagnosis is made by detecting the virus from respiratory samples (mainly nasopharyngeal and oropharyngeal swabs) using polymerase chain reaction. Treatment is usually symptomatic, and in severe and critical forms, the use of one of the antiviral drugs (lopinavir-ritonavir, remdesivir, hydroxychloroquine) may be consideredCopyright © 2020 Croatian Paediatric Society. All rights reserved.

Determinants of Attitude and the Intention to Stay of Employees in Low-Cost Carriers: Using Justice Theory

The purpose of this research is to examine the influential attributes of employees' attitudes and intentions to stay in the domain of human resources management in a low-cost carrier business. Using justice theory as a theoretical underpinning, financial compensation, nonfinancial compensation, coworker relationships, and procedural fairness were derived. The explained attributes of this research were attitude and intention to stay. This study used a survey and collected data on 233 employees in low-cost carriers as survey participants. To test the hypotheses, this study employed structural equation modeling. The results showed that attitude was positively impacted by financial compensation, nonfinancial compensation, coworker relationships, and procedural fairness. The results also revealed the positive effect of coworker relationships and attitudes on the intention to stay. This study sheds light on the literature by ensuring the explanatory power of justice theory in the area of low-cost carrier business.

The impact of COVID-19 epidemic on the control of important parasitic diseases.

The spread of COVID-19 has increased the risk of contact transmission between people, and imposes challenges and opportunities for the prevention and control of important parasitic diseases. There may be risks of COVID-19 infection during the implementation of the virus control, including in field investigation and intervention, contacting with fecal samples, or with asymptomatic infected people. Therefore, corresponding prevention and control measures should be taken before and during field investigation, and in sample transportation and detection to prevent the risks effectively. In this paper, the risks and corresponding countermeasures in the above processes are discussed in detail, and the opportunities that COVID-19 brought to the disease control system are also analyzed.Copyright © 2021, National Institute of Parasitic Diseases. All rights reserved.

Sleep disturbance and anxiety symptoms among asymptomatic COVID-19 carriers in Shanghai, China: the mediating role of entrapment and defeat.

BACKGROUND: The COVID-19 pandemic increases the risk of psychological problems, especially for the infected population. Sleep disturbance and feelings of defeat and entrapment are well-documented risk factors of anxiety symptoms. Exploring the psychological mechanism of the development of anxiety symptoms is essential for effective prevention. This study aimed to examine the mediating effects of entrapment and defeat in the association between sleep disturbance and anxiety symptoms among asymptomatic COVID-19 carriers in Shanghai, China. METHODS: A cross-sectional study was conducted from March to April, 2022. Participants were 1,283 asymptomatic COVID-19 carriers enrolled from the Ruijin Jiahe Fangcang Shelter Hospital, Shanghai (59.6% male; mean age = 39.6 years). Questionnaire measures of sleep disturbance, entrapment, defeat, anxiety symptoms, and background characteristics were obtained. A mediation model was constructed to test the mediating effects of entrapment and defeat in the association between sleep disturbance and anxiety symptoms. RESULTS: The prevalence rates of sleep disturbance and anxiety symptoms were 34.3% and 18.8%. Sleep disturbance was positively associated with anxiety symptoms (OR [95%CI] = 5.013 [3.721-6.753]). The relationship between sleep disturbance and anxiety symptoms (total effect: Std. Estimate = 0.509) was partially mediated by entrapment (indirect effect: Std. Estimate = 0.129) and defeat (indirect effect: Std. Estimate = 0.126). The mediating effect of entrapment and defeat accounted for 50.3% of the association between sleep disturbance and anxiety symptoms. CONCLUSION: Sleep disturbance and anxiety symptoms were prevalent among asymptomatic COVID-19 carriers. Entrapment and defeat mediate the association between sleep disturbance and anxiety symptoms. More attention is needed to monitoring sleep conditions and feelings of defeat and entrapment to reduce the risk of anxiety.

Efficient Purification of Polyhistidine-Tagged Recombinant Proteins Using Functionalized Corundum Particles.

Immobilized metal affinity chromatography (IMAC) is a popular and valuable method for the affinity purification of polyhistidine-tagged recombinant proteins. However, it often shows practical limitations, which might require cumbersome optimizations, additional polishing, and enrichment steps. Here, we present functionalized corundum particles for the efficient, economical, and fast purification of recombinant proteins in a column-free format. The corundum surface is first derivatized with the amino silane APTES, then EDTA dianhydride, and subsequently loaded with nickel ions. The Kaiser test, well known in solid-phase peptide synthesis, was used to monitor amino silanization and the reaction with EDTA dianhydride. In addition, ICP-MS was performed to quantify the metal-binding capacity. His-tagged protein A/G (PAG), mixed with bovine serum albumin (BSA), was used as a test system. The PAG binding capacity was around 3 mg protein per gram of corundum or 2.4 mg per 1 mL of corundum suspension. Cytoplasm obtained from different E. coli strains was examined as examples of a complex matrix. The imidazole concentration was varied in the loading and washing buffers. As expected, higher imidazole concentrations during loading are usually beneficial when higher purities are desired. Even when higher sample volumes, such as one liter, were used, recombinant protein down to a concentration of 1 µg/mL could be isolated selectively. Comparing the corundum material with standard Ni-NTA agarose beads indicated higher purities of proteins isolated using corundum. His6-MBP-mSA2, a fusion protein consisting of monomeric streptavidin and maltose-binding protein in the cytoplasm of E. coli, was purified successfully. To show that this method is also suitable for mammalian cell culture supernatants, purification of the SARS-CoV-2-S-RBD-His8 expressed in human Expi293F cells was performed. The material cost of the nickel-loaded corundum material (without regeneration) is estimated to be less than 30 cents for 1 g of functionalized support or 10 cents per milligram of isolated protein. Another advantage of the novel system is the corundum particles' extremely high physical and chemical stability. The new material should be applicable in small laboratories and large-scale industrial applications. In summary, we could show that this new material is an efficient, robust, and cost-effective purification platform for the purification of His-tagged proteins, even in challenging, complex matrices and large sample volumes of low product concentration.

BEMNIFOSBUVIR HAS LOW POTENTIAL TO INHIBIT P-gp, BCRP, AND OATP1B1 MEDIATED TRANSPORT

Background: Bemnifosbuvir (BEM, AT-527) is a guanosine nucleotide prodrug candidate for the treatment of COVID-19 and chronic HCV. BEM was identified in vitro as an inhibitor of drug transporters P-glycoprotein, breast cancer resistant protein (BCRP) and organic anion transporting polypeptide 1B1 (OATP1B1). Ph 1 studies in healthy participants were conducted to assess the clinical implications of these results using digoxin (DIG) and rosuvastatin (ROSU) as P-gp and BCRP/ OATP1B1 index drugs, respectively. Method(s): Both studies employed a similar design with 2 groups of 14 healthy participants: Day 1/period 1, all participants received a single dose of DIG 0.25mg or ROSU 10mg alone. In period 2, participants received DIG 0.25mg or ROSU 10mg with BEM 1100mg, simultaneously (n=14) or staggered by 2h (n=14). Serial plasma samples were collected and quantitated for DIG or ROSU concentrations. Result(s): A single dose of BEM 1100mg simultaneously administered slightly increased the Cmax of DIG (78%), yet had no effect on its AUC, consistent with the transient nature of BEM plasma PK. When dosed staggered, BEM did not affect the PK of DIG. A single dose (simultaneous or staggered) of BEM 1100mg slightly increased the plasma exposure of ROSU (20%-40%). There was no effect on vital signs, ECG, and no SAEs or drug discontinuations. Conclusion(s): A single high dose of BEM 1100mg only slightly increased the plasma exposure of the P-gp and BCRP/OATP1B1 index drugs DIG and ROSU. BEM has low potential to exhibit clinical meaningful inhibition of these transporters. No dose adjustment will be needed for drugs that are sensitive substrates of P-gp or BCRP/OAT1B1 when co-administered with BEM, staggered dosing may lessen any DDI risk.

Understanding Airline Passengers during Covid-19 Outbreak to Improve Service Quality: Topic Modeling Approach to Complaints with Latent Dirichlet Allocation Algorithm.

The COVID-19 pandemic has deeply affected the airline industry, as it has many sectors, and has created tremendous financial pressure on companies. Flight bans, new regulations, and restrictions increase consumer complaints and are emerging as a big problem for airline companies. Understanding the main reasons triggering complaints and eliminating service failures in the airline industry will be a vital strategic priority for businesses, while reviewing the dimensions of service quality during the COVID-19 pandemic provides an excellent opportunity for academic literature. In this study, 10,594 complaints against two major airlines that offer full-service and low-cost options were analyzed with the Latent Dirichlet Allocation algorithm to categorize them by essential topics. Results provide valuable information for both. Furthermore, this study fills the gap in the existing literature by proposing a decision support system to identify significant service failures through passenger complaints in the airline industry utilizing e-complaints during an unusual situation such as the COVID-19 pandemic.

COVID-19 screening in asymptomatic patients scheduled for surgery. What is the role of rt-PCR in elective procedures during the pandemic?

Background: COVID-19 has changed medical practice nowadays. One of the biggest concerns has been establishing when invasive procedures such as surgery, GI endoscopy or bone marrow transplant are safe;and if it is necessary to consider screening for asymptomatic patients. Method(s): We identified asymptomatic patients that were scheduled for invasive procedures from May 2020 to April 2021 at Clinica de Marly. Patients were asked to fill a questionnaire about GI and upper respiratory symptoms and contact with possible/confirmed cases of COVID- 19 in the last 15 days. Patients taken to emergency procedures, who had symptoms or contact with probable/confirmed cases in the last 15 days were excluded. rt-PCR was performed to screen COVID-19. Result(s): A total of 1837 patients were included. 104 rt-PCRs tested positive for SARS-CoV-2, leading to a 5.66% of identified asymptomatic patients. Patients were followed-up on the 30th day after the procedure. 1733 negative patients responded to our follow-up, in which only 1 death and 2 complications were detected. 102 positive patients were followed-up and no complications or deaths were reported. Conclusion(s): We found the presence of 5.66% of asymptomatic patients with positive rt-PCR for COVID-19. Safe screening will decide if these invasive interventions can be postponed, or, if the benefit outweighs the risks.Copyright © 2022 Asociacion Colombiana de Infectologia. All rights reserved.

Coating of favipiravir (FVP) on silver nanoparticles: First principle study

Silver nanoparticles, thanks to their antiviral and antibacterial properties, have great potential in a variety of applications, such as drug-delivery carriers. The coating properties of silver nanoparticles (size range of 1.6 nm) with a well-known drug, Favipirair, were investigated in this study using quantum mechanical and classical atomistic molecular dynamics simulation in order to use as the drug delivery to treat COVID-19 disease. The drug molecule's optimized structure, frequencies, charge distribution, and electrostatic potential maps were simulated using density functional theory (DFT) at the B3LYP/6–311++g(d,p) level of theory. The coating of AgNP with each of these drugs was then studied using molecular dynamics simulation. The interaction affinity obtained from MD results agrees with the DFT results on drug adsorption on the Ag(1 1 1) slab. © 2023

A cold chain logistics traceability system framework based on the identification and resolution system for Industrial Internet

With the global outbreak of COVID-19, hundreds of pneumonias caused by cold chain products occurred worldwide, which seriously threatened the safety of people's lives and properties. To effectively prevent product quality problems caused by cold chain logistics, it is urgent to establish a cold chain logistics traceability system with interoperability of heterogeneous systems, to record, share and track the temperature, location, time, and other specific information. The traditional cold chain logistics traceability systems have many problems, such as broken cold chains, untrustworthy data, and data tampering and sharing, which hinder the coordination and interaction efficiency of cold chain logistics traceability data. This paper creatively proposes a cold chain logistics traceability system framework based on the identification and resolution system for the Industrial Internet. It establishes a general cold chain logistics traceability identification data model. The system framework and data model can effectively solve the difficulties of multi-code identification and multi-source heterogeneous system interaction, to improve the efficiency of cold chain logistics traceability, and ensure the quality of cold chain logistics products. © 2022 ACM.

Knowledge, attitude, and practices regarding second waves of COVID-19: A cross-sectional study among rural population in India

Coronavirus disease (COVID-19) is an infectious disease caused by a newly discovered coronavirus. Most people infected with the COVID-19 virus will experience mild to moderate respiratory illness and recover without requiring special treatment. Older people, and those with underlying medical problems like cardiovascular disease, diabetes, chronic respiratory disease, and cancer are more likely to develop serious illness. The best way to prevent and slow down transmission is to be well informed about the COVID-19 virus, the disease it causes and how it spreads. Protect yourself and others from infection by washing your hands or using an alcohol based rub frequently and not touching your face. The aim of the present study was assess the Knowledge, attitude, and practices regarding of COVID-19: among rural population.Cross-sectional questionnaire-based study was conducted on rural people A total of 200 participants hailing from rural areas were enrolled. Result(s): Most common source of information for the participants was through television (54.8%) and radio (49.2%). Only 77% claimed that they were using face mask and 72% claimed to be following social distancing. A large (60%) proportion of the participants did not know that COVID-19 could spread from asymptomatic patients. The mean (percentage) scores of knowledge, attitude, and practice were 21.26 (82%), 9.37 (92%), and 10.32 (86%), respectively. Education of participants was a key determinant for use of social distancing and face mask as a preventive tool. With increasing age, the practice of social distancing and the use of face mask were decreasing in our study. Conclusion(s): Television/Radio channels form an important source of information and need to be used more effectively to educate the people about the disease and create awareness about effective preventive measures. People need to be educated about the role of asymptomatic carriers in spreading the disease.Copyright © 2020 Ubiquity Press. All rights reserved.

Teaching Analytical Instrumentation Through Remote Access – A Gas Chromatography Perspective

An approach to online laboratory exercises for analytical chemistry students with demonstrated practical exercises through the use of remote-controlled gas chromatography (GC) instrumentation is discussed. The approach allows for a practical-based learning activity to be carried out by students who are unable to attend in-person laboratory exercises and was conducted during the COVID-19 pandemic. Learning activities focused on the operation of GC instrumentation were completed prior to a research-based analysis activity being conducted by students. At the end of this experiment, the students are expected to understand, independently operate, and learn how to achieve better separation through the manipulation of GC settings, such as split/splitless injections, carrier gas flow rate, and oven temperature, and apply principles of GC to a practical application. Additional flexibility from this approach could also be beneficial during postpandemic and/or in the circumstance where students cannot physically attend the class.

"If I lose this, I have nothing”: ‘carrier' women during the Spanish-Moroccan border closure

The closure of the Spanish Southern border between Morocco and the Spanish city of Melilla induced by Covid-19 disrupt the work and life course of thousands of women crossing merchandise on the border. This research analyzes the survival strategies of these women after the border closure and the transformations of this unequal region. Using a Grounded Theory approach, participant observation and interviews to carrier women and informants were conducted in Melilla in 2021. Migration, family support and other precarious feminized jobs were found to be the most common survival alternatives. The conclusions point out that the border closure and the changes affecting the region of Nador may be the end of this income source for these women. This analysis contributes with new data on the global feminization of survival, although further research on these strategies is needed due to the constant changes in this border. © 2023, Universidad Autonoma de Baja California. All rights reserved.

Bayesian Formulation of Time-Dependent Carrier-Borne Epidemic Model with a Single Carrier

In this paper, the time dependent carrier-borne epidemic model defined by Weiss in 1965 has been adopted into a Bayesian framework for the estimation of its parameters. A complete methodological structure has been proposed for estimating the relative infection rate and probability of survival of k out of m susceptibles after time t from the start of the epidemic. The methodology has been proposed assuming a single carrier to simplify the study of the behavioral validity of the fitted Bayesian model with respect to time and relative infection rate. Further, the proposed model has been implemented on two real data sets the typhoid epidemic data from Zermatt in Switzerland and the Covid-19 epidemic data from Kerala in India. Results show that the proposed methodology produces reliable predictions which are consistent with those of the maximum likelihood estimates and with expected epidemiological patterns. © 2023 Deo et al

A comparative DFT study on Al- and Si- doped single-wall carbon nanotubes (SWCNTs) for Ribavirin drug sensing and detection

In this work, we have presented a comparative study on Ribavirin (RBV) drug sensing and detection on the pristine and functionalized single-wall carbon nanotubes (f-SWCNTs) by Density Functional Theory (DFT) method. The pristine and metal-doped zigzag (4,0) and (6,0) SWCNTs were first considered for the RBV adsorption. All the probable positions of RBV adsorption were investigated to find which one is energetically favourable. The topology analysis of the Quantum theory of atoms in a molecule (QTAIM) with non-covalent interactions (NCI-RDG), Frontier molecular orbitals (FMO), Density of states (DOS), and non-linear optical (NLO) analysis were carried out to understand the molecular structure, electrical, electronic and optical properties of complexes. The charge analysis indicates that charge transfer is from the adsorbed RBV to the pristine and metal-doped (4,0) and (6,0) SWCNTs. The highest values of adsorption energies for Al-, Si-doped and pristine (4,0) SWCNTs were determined as −34.688, −87.999 and −10.382 kcal/mol, respectively, whereas corresponding values for metal-doped and pristine (6,0) SWCNTs are about −43.592, −20.661 and −12.414 kcal/mol, respectively. The results suggest that those bare and metal-doped (4,0) SWCNTs and (6,0) Si-SWCNTs can serve as promising sensors in practical applications to detect, recognize and carrier RBV drug for its medicinal drug delivery applications. Based on the NLO properties of (6,0) Si-SWCNTs and pristine (6,0) SWCNT (with an acceptable recovery time of 279s and first hyper polarizability value of β = 229.25 × 10−30 cm5 esu−1), those nanotubes may be possible candidates to be used as the optoelectronic sensor for RBV drug. The appropriate short length of nanotubes was obtained. © Elsevier Ltd

Epidemiological study and health management of an imported incident of COVID-19 asymptomatic cases in Haikou, China: a retrospective case series

Background: To control the imported risks brought by all international arrivals, China Customs has implemented strict closed-loop health management policy called "three checks, three screenings and one transfer". This study provides epidemiological evidence for prevention and control measures on imported cases of asymptomatic infections and describes the current COVID-19 prevention and control system on imported risks in China. Methods: We retrospectively analyzed an imported incident of three asymptomatic carriers. Serum SARS-CoV-2 IgM and IgG antibodies were detected by chemiluminescence and gold immnnochromatography(GICA). Results: Three cases were reported positive for SARS-CoV-2 nucleic acid on their arrival, total antibodies and IgG, but negative for IgM. The Ct values of cases A, B and C were 34/36/36, 32/33/32 and 25/31/29, respectively. There were 10726434 pair-end reads sequenced for case C, and approximate 80% reads were aligned to the hCoV-19/Wuhan/IVDC-HB-01/2019 genome (EPI_ISL_402119). The viruses of case A and C were homologous and came from the SARS-CoV-2 variant. Conclusion: Serum antibody IgM and IgG tests are recommended for international travelers from epidemic areas. The "three checks, three screenings and one transfer" policy implemented at custom's entry points was effective in COVID-19 prevention and control.

Non-Pharmaceutical Interventions are Measures to Control Coronavirus Disease-2019 (COVID-19) Transmission in India

The coronavirus disease 19 (COVID-19) is a pathogenic and transmittable viral disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and this disease emerged in December 2019 in Wuhan, China. It was later spread across the world, including the United States, Great Britain, Brazil, Russia, Germany, Spain, Italy, Iran, Pakistan, India and other countries. The whole genome of SARS-CoV-2 shared some similarity with the SARS virus, hence bat was a suspected source of the spread of this virus in China. However, human to human transfer was reported. In India, more than 300 COVID-19 cases were reported on March 21, 2020, and these numbers were increased as 892551 on July 13, 2020. These numbers of COVID-19 cases were detected from a total of 11806256 samples according to the report given by the Government of India. On July 13, Maharashtra state of India recorded a total of 254427 cases and 10289 deaths were registered. Tamilnadu was another state that screened more number of COVID-19 cases, followed by Maharashtra. On July 6, 114978 COVID-19 cases were reported in Tamilnadu and 1571 deaths were recorded. In Tamilnadu alone, over 3000 COVID-19 cases were reported daily from July first week to the second week and total cases were 142798 on July 13, 2020. Among these cases, 78573 COVID-19 cases were detected in Chennai city alone accounted for about 62% of total COVID-19 cases. It was earlier believed that patients over aged 60 and less than 10 were highly vulnerable to this disease. However, this disease affected all age groups. To prevent COVID-19, no approved antiviral vaccine or drug has yet been discovered. COVID-19 is transmitted through droplets during coughing and sneezing by symptomatic and asymptomatic carriers. Non-pharmaceutical interventions such as washing hands and social distancing are effective measures to control COVID-19 transmission in India. The present study was designed to analyze the control measures of COVID-19 infection and ground reality to control the COVID-19 outbreak.Copyright © 2021 Bentham Science Publishers.

High Rate of SARS-CoV-2 Transmission Due to Choir Practice in France at the Beginning of the COVID-19 Pandemic.

OBJECTIVE: To document the case of a high transmission rate of SARS-CoV-2 generating a cluster linked to an indoor choir rehearsal hold at the beginning of the COVID-19 pandemic in France. METHOD: Case study. Data were obtained via a questionnaire. RESULTS: Twenty-seven participants, including 25 singers, 1 conductor and 1 accompanist attended a choir practice on March 12, 2020. The practice was indoor and took place in a non ventilated space of 45 m2. No choir member reported having been symptomatic for COVID-19 between March 2 and March 12.The mean age of the participants was 66.9 (range 35-86) years. The secondary attack rate was 70%: 19/27 participants were diagnosed with COVID-19 from 1 to 12 days after the rehearsal with a median of 5.1 days. Thirty-six percent of the cases needed a hospitalization (7/19), and 21% (4/19) were admitted to an ICU. The index cases were asymptomatic and possibly multiple. CONCLUSION: In the absence of valid barrier measures to prevent COVID-19 transmission, indoor choir practice should be suspended during the SARSCoV-2 surging phases. Transmission of the virus among gatherings from asymptomatic cases is a crucial issue and a main challenge to COVID-19 control.

How big of an impact do asymptomatic people have on the dynamics of an epidemic?

Asymptomatic carriers serve as a potential source of transmission of epidemic diseases. Exposed people who develop symptoms only get tested and remain isolated in their homes or sometimes in hospitals when needed. In contrast, the asymptomatic individuals go untested and spread the disease silently as they roam freely throughout their entire infectious lifetime. The work intends to explore the role of asymptomatic carriers in the transmission of epidemic diseases and investigate suitable optimal control strategies. We propose a SEIAQR compartmental model subdividing the total population into six different compartments. To illustrate the model's implication, we estimate the number of asymptomatic individuals using COVID-19 data during June 9–July 18, 2021 from Bangladesh. We then analyze the model to explore whether the epidemic subsides if the asymptomatic individuals are tested randomly and isolated. Finally, to gain a better understanding of the potential of this unidentified transmission route, we propose an optimal control model considering two different control strategies: personal protective measures and isolation of asymptomatic carriers through random testing. Our results show that simultaneous implementation of both control strategies can reduce the epidemic early. Most importantly, sustained effort in identifying and isolation of asymptotic individuals allows relaxation in personal protective measures. © 2023

Assembly of Immunogenic Protein Particles toward Advanced Synthetic Vaccines.

Immunogenic carrier proteins such as the non-toxic diphtheria toxin variant, cross-reacting material 197 (CRM197), are widely used in subunit vaccine formulations to boost immunogenicity of chemically conjugated antigens. Conjugate vaccines are inherently expensive due to laborious manufacturing steps. Here, this work develops a particulate vaccine platform based on using engineered Escherichia coli to assemble CRM197-antigen fusion proteins into discrete submicron-sized particles. This approach enables precise loading of diverse antigens and epitopes enhancing their immunogenicity. A cost-effective, high-yield, and scalable biomanufacturing process is developed. Purified particulate CRM197-antigen vaccines are ambient-temperature stable. CRM197 particles incorporating pathogen-specific antigens or epitopes from SARS-CoV-2, Streptococcus pyogenes (group A), and Mycobacterium tuberculosis induced cell-mediated and humoral immune responses mediating protective immunity in respective animal models of infection. The CRM197 particle vaccine platform is versatile, enabling co-delivery of selected antigens/epitopes together with immunogenic CRM197 as discrete stable particles avoiding laborious manufacture of soluble CRM197 and antigen followed by chemical conjugation.