Opportunities and challenges to implementing mRNA-based vaccines and medicines: lessons from COVID-19.

The messenger RNA (mRNA) platform emerged at the forefront of vaccine development during the COVID-19 pandemic, with two mRNA COVID-19 vaccines being among the first authorized globally. These vaccines were developed rapidly. Informed by decades of laboratory research, and proved to be safe and efficacious tools for mitigating the global impact of the COVID-19 pandemic. The mRNA platform holds promise for a broader medical application beyond COVID-19. Herein, we provide an overview of this platform and describe lessons learned from the COVID-19 pandemic to help formulate strategies toward enhancing uptake of future mRNA-based interventions. We identify several strategies as vital for acceptance of an expanding array of mRNA-based vaccines and therapeutics, including education, accurate and transparent information sharing, targeted engagement campaigns, continued investment in vaccine safety surveillance, inclusion of diverse participant pools in clinical trials, and addressing deep-rooted inequalities in access to healthcare. We present findings from the Global Listening Project (GLP) initiative, which draws on quantitative and qualitative approaches to capture perceptions and experiences during the COVID-19 pandemic to help design concrete action plans for improving societal preparedness for future emergencies. The GLP survey (>70,000 respondents in 70 countries) revealed tremendous disparities across countries and sociodemographic groups regarding willingness to accept novel mRNA vaccines and medicines. The comfort in innovations in mRNA medicines was generally low (35%) and was marginally lower among women (33%). The GLP survey and lessons learnt from the COVID-19 pandemic provide actionable insights into designing effective strategies to enhance uptake of future mRNA-based medicines.

Angiographic characteristics of culprit lesions in infarct related artery and correlation of TIMI score with SYNTAX score to predict extent and severity of coronary artery disease in patients undergoing primary percutaneous coronary interventions.

Objective: The current study was designed to explore the relationship of TIMI and SYNTAX risk score to predict the CAD extent and severity in STEMI patients. Methods: For this cross-sectional study, 304 STEMI patients undergoing PPCI were enrolled at Department of Interventional Cardiology NICVD Karachi from September 2021 to January 2022. and the TIMI risk score was determined at enrolment. Based on these scorings, the patients were grouped as low, intermediate, and high risk, i.e., a score of ≤ 3, 4 to 7, and ≥ 8, respectively. The SYNTAX scores were utilized to assess the extent of CAD. Results: Statistically significant difference was found in symptoms to balloon time (p=0.001), history of diabetes (p=0.006), angina (p=0.011), obesity (p=0.048), STEMI type (p=0.003), Killip classes (p=0.000), Infarct-Related Artery (p=0.006), number of diseased vessels (p<0.01), LMS > 50% (p=0.000), PCI type (p<0.01), collateral circulation (p<0.01), In-hospital mortality (p<0.01), LV support (p<0.01), and post-procedural TIMI flow (p=0.013), among the three TIMI risk groups. Significant correlation was found among TIMI risk score and SYNTAX score. Conclusion: It is observed that the TIMI risk scores are highly correlated with the SYNTAX Score in predicting the CAD severity in STEMI patients.

Tuning the Structural and Magnetic Properties in Mixed Cation MnxCo2-xP2S6.

The metal thiophosphates (MTP), M2P2S6, are a versatile class of van der Waals materials, which are notable for the possibility of tuning their magnetic properties with the incorporation of different transition-metal cations. Further, they also offer opportunities to probe the independent and synergistic role of the magnetically active cation sublattice when coupled to P2Q6 polyhedra. Herein, we report the structural, magnetic, and electronic properties of the series of MTPs, MnxCo2-xP2S6 (x = 0.25, 0.5, 1, 1.5, 1.75) synthesized by the P2S5 flux method. Structural and elemental analysis indicates a homogeneous stoichiometry in the MnxCo2-xP2S6 compounds. We observe that a correlation is apparent between the intensities of specific Raman modes and Raman shifts with respect to the alloying ratio between Mn and Co. Magnetic susceptibility measurements indicate that the alloyed systems adopt an ordered antiferromagnetic (AFM) configuration with a dependence of the Néel temperature on the alloying ratio. A possible magnetic frustration behavior was observed for the composition MnCoP2S6 due to magnetic moment compensation as the alloying ratio between Mn and Co approaches parity. Interestingly, mixed oxidation states of the metal cation species are also observed in MnxCo2-xP2S6 along with a linear dependence of the work function on the alloying ratio of Mn and Co.

A systematic literature review and network meta-analysis feasibility study to assess the comparative efficacy and comparative effectiveness of pneumococcal conjugate vaccines.

Background: No head-to-head studies are currently available comparing pneumococcal non-typeable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV) with 13-valent pneumococcal conjugate vaccine (PCV-13). This study explored the feasibility of using network meta-analysis (NMA) to conduct an indirect comparison of the relative efficacy or effectiveness of the two vaccines.Methods: A systematic literature search was conducted for published randomized controlled trials (RCTs) and non-RCT studies reporting data on vaccine efficacy or effectiveness against invasive pneumococcal disease in children aged <5 years receiving 7-valent pneumococcal conjugate vaccine (PCV-7), PHiD-CV or PCV-13. Study quality was evaluated using published scales. NMA feasibility was assessed by considering whether a connected network could be constructed by examining published studies for differences in study or patient characteristics that could act as potential treatment effect modifiers or confounding variables.Results: A total of 26 publications were included; 2 RCTs (4 publications), 7 indirect cohort studies, and 14 case-control studies (15 publications). Study quality was generally good. The RCTs could not be connected in a network as there was no common comparator. The studies differed considerably in design, dose number, administration schedules, and subgroups analyzed. Reporting of exposure status and subject characteristics was inconsistent.Conclusion: NMA to compare the relative efficacy or effectiveness of PHiD-CV and PCV-13 is not feasible on the current evidence base, due to the absence of a connected network across the two RCTs and major heterogeneity between studies. NMA may be possible in future if sufficient RCTs become available to construct a connected network.

Quercetin protects against stress-induced anxiety- and depression-like behavior and improves memory in male mice.

The present study evaluates the protective role of Quercetin (Quer), against immobilization stress- induced anxiety, depression and cognition alteration in mice using behavioral and biochemical parameters. 24 adult Albino mice were distributed into 2 groups vehicle (n=12; 1 ml/kg) and Quer injected (n=12; 20 mg/kg/ml). The animals received their respective treatment for 14 days. On day 15, after the drug administration, animals were sub-divided into 4 groups (n=6); (i) unstressed + vehicle; (ii) stressed + vehicle; (iii) unstressed + Quer; (iv) stressed + Quer. On day 16, 24 h after the immobilization stress behavioral activities (light-dark activity, elevated plus maze, Morris water maze, and forced swim test) monitored and then animals were decapitated 1 h after the drug administration. Brain samples were collected for biochemical (antioxidant enzymes, AChE, ACh, 5-HT and its metabolite) analysis. The present study indicates the Quer reversed the stress-induced anxiety and depression, in addition, memory performance was more enhanced in stressed group. Following the treatment of Quer, stress-induced elevation of lipid peroxidation and suppression of antioxidant enzymes were also reversed. Administration of Quer decreased AChE in unstressed, while levels of acetylcholine were increased in vehicle and Quer treated stressed animals. The metabolism of 5-HT was increased in Quer treated stressed than unstressed animals. In conclusion, the present finding showed that Quer could prevent the impairment of antioxidant enzymes and also regulate the serotonergic and cholinergic neurotransmission and produce antianxiety, antidepressant effect and enhance memory following 2 h immobilization stress in mice.

Diallel cross analysis of plesiomorphic traits in Triticum aestivum L. genotypes.

We conducted a 5 x 5 complete diallel cross experiment in bread wheat (Triticum aestivum) with the genotypes 6309, Chkwal-50, Dhrabi, Bhkhar-02, and FS-08. Our objective was to evaluate the type of gene action and the general and specific combining abilities required for various morphological traits in wheat. The results of analysis of variance revealed highly significant differences among genotypes for all the investigated traits. The results of joint regression analysis showed that the data for all the investigated traits fitted a simple additive dominance model. Graphical representation of variance and covariance suggested that most of the investigated traits were controlled by overdominance gene action. However, the peduncle length and plant height were controlled by additive gene action. Variety 6309 carried the highest number of dominant genes for the number of spikelets per spike, number of tillers per plant, plant height, number of fertile tillers per plant, and grain yield per plant. Chakwal-50 carried the highest number of recessive genes for grain yield per plant, number of tillers per plant, number of grains per spike, number of fertile tillers per plant, and plant height. Chakwal-50 and 6309 were the best general combiners for number of spikelets per spike, number of grains per spike, grain yield per plant, 1000-grain weight, number of fertile tillers per plant, and number of tillers per plant. On other hand, 6309 performed well in specific crosses with Chakwal-50, FS-08, and Bhakhar-02 for spike length and number of tillers per plant.

High-mobility and air-stable single-layer WS2 field-effect transistors sandwiched between chemical vapor deposition-grown hexagonal BN films.

An emerging electronic material as one of transition metal dichalcogenides (TMDCs), tungsten disulfide (WS2) can be exfoliated as an atomically thin layer and can compensate for the drawback of graphene originating from a gapless band structure. A direct bandgap, which is obtainable in single-layer WS2, is an attractive characteristic for developing optoelectronic devices, as well as field-effect transistors. However, its relatively low mobility and electrical characteristics susceptible to environments remain obstacles for the use of device materials. Here, we demonstrate remarkable improvement in the electrical characteristics of single-layer WS2 field-effect transistor (SL-WS2 FET) using chemical vapor deposition (CVD)-grown hexagonal BN (h-BN). SL-WS2 FET sandwiched between CVD-grown h-BN films shows unprecedented high mobility of 214 cm(2)/Vs at room temperature. The mobility of a SL-WS2 FET has been found to be 486 cm(2)/Vs at 5 K. The ON/OFF ratio of output current is ~10(7) at room temperature. Apart from an ideal substrate for WS2 FET, CVD-grown h-BN film also provides a protection layer against unwanted influence by gas environments. The h-BN/SL-WS2/h-BN sandwich structure offers a way to develop high-quality durable single-layer TMDCs electronic devices.

Design and preclinical characterization of a novel vaccine adjuvant formulation consisting of a synthetic TLR4 agonist in a thermoreversible squalene emulsion.

We describe the development, analytical characterization, stability and preclinical efficacy of AF04, a combination adjuvant comprising the synthetic toll-like receptor 4 (TLR4) agonist, E6020, formulated in AF03, a thermoreversible squalene emulsion. By using AF04 with the recombinant major outer membrane protein of Chlamydia trachomatis (Ct-MOMP) and with the recombinant surface glycoprotein gB from human cytomegalovirus (CMV-gB) as model antigens, we show that AF03 and E6020 can synergize to augment specific antibody and Th-1 cellular immune responses in mice. In terms of formulation, we observe that the method used to incorporate E6020 into AF03 affects its partition between the oil and water phases of the emulsion which in turn has a significant impact on the tolerability (IV pyrogenicity test in rabbits) of this novel adjuvant combination.

Photocurrent response of MoS2 field-effect transistor by deep ultraviolet light in atmospheric and N2 gas environments.

Molybdenum disulfide (MoS2), which is one of the representative transition metal dichalcogenides, can be made as an atomically thin layer while preserving its semiconducting characteristics. We fabricated single-, bi-, and multilayer MoS2 field-effect transistor (FET) by the mechanical exfoliation method and studied the effect of deep ultraviolet (DUV) light illumination. The thickness of the MoS2 layers was determined using an optical microscope and further confirmed by Raman spectroscopy and atomic force microscopy. The MoS2 FETs with different number of layers were assessed for DUV-sensitive performances in various environments. The photocurrent response to DUV light becomes larger with increasing numbers of MoS2 layers and is significantly enhanced in N2 gas environment compared with that in atmospheric environment.

Immunoglobulin-specific responses to Chlamydia elementary bodies in individuals with and at risk for genital chlamydial infection.

Renewed interest in chlamydia vaccination has revealed the need for a greater understanding of the seroprevalence of chlamydial infection in US populations. We used a Chlamydia trachomatis elementary body (EB)-based enzyme-linked immunosorbent assay to define the characteristics of the humoral immune response and to determine seroprevalence. Two groups were analyzed: one consisting of patients with current, laboratory confirmed, genital chlamydial infection (n = 98) and one group of individuals whose chlamydia infection history was unknown (n = 367). C. trachomatis seropositivity was detected in 90% of the infected group and in 31% of the chlamydia-unknown group. IgG1 and IgG3 comprised the predominant anti-Chlamydia serum antibody responses. Serum IgA1 responses were variably positive, and individuals were rarely positive for anti-chlamydia IgG2, IgG4 or IgA2. The magnitude of the IgG1 and IgG3 responses was greatest in female and African American individuals and was sustained for at least 6 months. Antibody responses were not serovar restricted or confounded by Chlamydia pneumoniae cross-reactivity.

Splenic injury after blunt abdominal trauma: an unusual presentation.

BACKGROUND: The spleen is the most commonly injured viscus in blunt abdominal trauma. Abdominal pain with left upper quadrant tenderness or signs of peritonitis in a patient with history of trauma is the most common presentation of this condition. OBJECTIVE: To describe an unusual presentation of splenic rupture in a patient with history of motor vehicle crash and blunt abdominal trauma. CASE REPORT: A young man was brought to the Emergency Department with a history of being in a motor vehicle crash 10 h earlier. He experienced gradually worsening difficulty breathing while sitting or lying down for the previous 4 h, although he was asymptomatic in the upright position. He was transported to the hospital standing upright, supported by two men, on the open back of a vehicle normally used to transport cattle. The patient was found to have left upper quadrant abdominal tenderness on examination and free fluid in the pelvis on the focused abdominal sonography for trauma examination done while the patient was standing. A grade III splenic injury with hemoperitoneum was diagnosed on computed tomography scan and the patient was treated with splenectomy. CONCLUSION: We report an unusual presentation of a splenic injury in a young man who had symptoms only in the supine position.

Elevated elafin/trappin-2 in the female genital tract is associated with protection against HIV acquisition.

OBJECTIVES: Globally, heterosexual intercourse is the primary route of HIV-1 (HIV) transmission. It follows that mechanisms that protect against HIV infection are likely operative at the genital mucosa. In HIV-resistant Kenyan sex workers who are highly exposed to HIV infection yet remain uninfected, protection correlates with HIV-specific immune responses and genetic factors. However, these factors do not entirely explain this model of natural immunity to HIV. We hypothesized that protection may be mediated by innate immune proteins in the genital tract of HIV-resistant sex workers. DESIGN AND METHODS: The genital proteome of mucosal secretions from HIV-resistant women was examined using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Cervical lavage samples were collected from 315 HIV-resistant, HIV-uninfected and HIV-infected commercial sex workers. RESULTS: Univariate analysis identified a 6 kDa biomarker of HIV resistance in genital secretions from these women. This protein was identified by tandem mass spectrometry as elafin and was found to be overexpressed in HIV-resistant women compared with HIV-uninfected (P = 0.001) and infected (P = 0.002) women. The elevated levels of elafin/trappin-2 in HIV-resistant women were confirmed using ELISA. The prospective association of elevated cervicovaginal elafin/trappin-2 levels with protection from HIV acquisition was then confirmed in an independent cohort of high-risk female sex workers. CONCLUSION: Using a unique proteomics approach in a large scale, cross-sectional cohort study, we identified elafin/trappin-2 as a novel innate immune factor, which is highly associated with resistance. This association was confirmed within an independent, prospective cohort study. Genital tract elafin/trappin-2 levels constitute a natural correlate of HIV protection in humans.

Mucosal innate immunity as a determinant of HIV susceptibility.

Most human immunodeficiency virus (HIV) is acquired during sex, across a mucosal membrane. Despite many advances in our understanding of HIV pathogenesis, the initial events during mucosal transmission have been poorly characterized, and a better understanding of these events will probably be a key to the development of successful microbicide(s) and/or a preventative HIV vaccine. While a vast majority of mucosal HIV exposures do not result in productive infection, implying that innate mucosal immune defenses are highly protective, failure of these mucosal defenses resulted in over 3 million new HIV infections in 2006. We review recent findings regarding HIV mucosal immunopathogenesis, emphasizing the importance of innate immunity in natural protection from infection, and examine how natural or induced perturbations in the mucosal innate system may underpin HIV transmission. Given the great challenges to the development of HIV microbicides and vaccines, identification and enhancement of 'natural' innate immune defenses present attractive avenues for development of safe, non-toxic microbicides.

Elevated T cell counts and RANTES expression in the genital mucosa of HIV-1-resistant Kenyan commercial sex workers.

The initial site of exposure to human immunodeficiency virus (HIV)-1 during heterosexual transmission occurs in the genital tract. Although the majority of immunological studies have focused on the immune response to HIV-1 at the systemic level, our understanding of tissue-specific immunity is deficient. The goal of the present study was to characterize T cell populations found in the cervix of women shown to be resistant to infection by HIV-1. Levels of both systemic and cervical mucosal lymphocytes were compared between HIV-1-resistant, HIV-1-uninfected, and HIV-1-infected commercial sex workers (CSWs) as well as HIV-1-uninfected non-CSW control subjects at low risk for exposure. The HIV-1-resistant CSWs had increased cervical CD4+ and CD8+ T cell counts, compared with the HIV-1-uninfected CSWs; importantly, these increases were not reflected in the systemic lymphocyte compartment. There was a 2-fold increase in CD4+ T cell counts in the HIV-1-resistant CSWs, compared with both the HIV-1-infected and the HIV-1-uninfected CSWs. Expression of the HIV-1 coreceptors CCR5 and CXCR4 was also determined, and cytokine and beta chemokine levels in the genital mucosa were assessed. The HIV-1-resistant CSWs had a 10-fold increase in RANTES expression, compared with the HIV-1-uninfected CSWs. This is the first study to show elevated levels of beta chemokines and CD4+ T cells in the genital tracts of women who are exposed to HIV-1 and yet are uninfected.

Comparison of retrobulbar anesthesia and intracameral anesthesia using preservative free bupivacaine hydrochloride 0.5% in phacoemulsification with posterior chamber intraocular lens implantation.

OBJECTIVE: To compare the retrobulbar anesthesia and intracameral anesthesia using preservative-free bupivacaine hydrochloride 0.5% in terms of effectiveness, complications and comfort to the patient during phacoemulsification with posterior chamber intraocular lens implantation. METHODS: This was a hospital based comparative study of two methods of anesthesia, conducted at LRBT Free Eye and Cancer Hospital, Lahore from January to July 2000. Study included 200 patients with uncomplicated age-related cataract, equally divided in two groups on simple random basis. Group A (100 patients) received the retrobulbar anesthesia and Group B (100 patients) received the intracameral anesthesia with bupivacaine hydrochloride 0.5% for phacoemulsification with posterior chamber intraocular lens implantation. Outcome measures like pain, visual acuity, intraocular pressure and anterior chamber reaction were compared. RESULTS: On day 1, 79% of the patients in group A and 82% patients in group B had unaided visual acuity ranging between 6/6-6/18. On day 7, this was 88% in group A and 89% group B. On day 1, 99% in group A and 98% in group B had <1+ cells in the anterior chamber while on day 7 this increased up to 100%. On day 1, 97% in group A and 98% group B had intraocular pressure less than 20 mmHg. On day 7, it increased up to 100% in both groups. 97% patients in group A and 96% patients in group B had painless surgery. Results were analysed using computer software SPSS version 10.0. Results showed no significant statistical difference between two groups in terms of pain, visual acuity, intraocular pressure, anterior chamber reaction and patient comfort. CONCLUSION: In the hands of expert surgeons and in selected patients, intracameral anesthesia with preservative-free bupivacaine hydrochloride 0.5% is a safe and effective technique of ocular anesthesia for phacoemulsification with posterior chamber intraocular lens implantation.

IL-4 primes human endothelial cells for secondary responses to histamine.

Interleukin-4 (IL-4) is a multifunctional cytokine, which is involved in numerous disease states, including atopic asthma. IL-4 not only induces direct responses in cells but can also prime for secondary responses to stimuli. Little is known about the priming effects of IL-4 on endothelial cells; therefore, we chose to examine the ability of IL-4 to prime endothelial cells for platelet-activating factor (PAF) synthesis and prostaglandin E(2) (PGE(2)) release. IL-4 alone did not enhance PAF synthesis or PGE(2) release; however, pretreatment with IL-4 primed for PAF synthesis and PGE(2) release in response to subsequent stimulation with histamine. In contrast, tumor necrosis factor alpha (TNF-alpha), oncostatin M (OSM), and IL-1beta did not prime endothelial cells for PAF synthesis in response to histamine. The priming effects of IL-4 occurred without any detectable changes in the requirement for signaling pathways upstream of PGE(2) release. IL-4 treatment increased the expression of mRNA for histamine receptor 1 (HR1) and shifted the inhibition curve for pyrilamine, a specific HR1 antagonist. In addition, the dose-response curve for histamine-induced elevations in intracellular calcium was shifted following IL-4 stimulation. Together, these data indicate that HR1 is up-regulated in IL-4-stimulated human umbilical vein endothelial cells (HUVEC) and suggest that this up-regulation may contribute to the enhanced responsiveness of IL-4-stimulated HUVEC to histamine.

Therapeutic angiogenesis for patients with limb ischemia by utilization of fibrin meshwork. Pilot randomized controlled study.

AIM: The aim of this paper is to assess the feasibility and efficacy of fibrin in inducing neovascularization as an angiogenic substance and/or carrier for vascular endothelial growth factor (VEGF) in patients with limb ischemia. METHODS: Twenty-three patients with limb ischemia and referred for below the knee amputations were randomized for treatment: group 1, 7 patients received only a saline injection; group 2, 9 received intramuscular injection of fibrin and group 3, 7 received the fibrin composition with deferoxamine and added VEGF(165). The fibrin meshwork was introduced into the popliteal area of the diseased limbs using a dual syringe system (1 contained thrombin solution [1 mg, 5000 U] and 1 contained fibrinogen [1 mg, Baxter Hyland Immuno] solution). In group 3, Deferoxamine (100 microg) and 500 microg of VEGF(165) were added to the fibrinogen solution. RESULTS: In the control group 5 patients had undergone below the knee amputation at the 3-6 month follow-up. Only 1 patient from Group 2 had below the knee amputation 5 months following Fibrin only administration. Clinical improvement was noted in all 8 patients following fibrin administration and in all 7 patients injected with the fibrin meshwork, deferoxamine and VEGF combination. CONCLUSION: IM injection of fibrin is safe and appears to be an efficient method to treat limb ischemia. These findings indicate that use of fibrin may be a novel and simple method for inducing therapeutic angiogenesis.