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OBJECTIVES: Tobacco use is one of the greatest public health problems in the world. Prevalence of alcohol and tobacco use among lactating women ranged between 35.9-83.0% and 1.4-6.1%, respectively, in various parts of the world. Both alcohol and nicotine pass quickly into breast milk. The objective of this study is to estimate the prevalence of alcohol and tobacco use in lactating women and its association with having underweight, stunted, and wasted nursing children in India. The study also draws comparisons to identify the common associated socio-economic factors for alcohol and tobacco use among lactating women and having underweight, stunted, and/or wasted children. STUDY DESIGN: A cross-sectional analytical study involving secondary data from the National Family Health Survey-5. METHODS: A cross-sectional analytical study involving secondary data from the National Family Health Survey-5 conducted nationally in community settings in two phases during 2019-2021. Alcohol and tobacco use among lactating women was self-reported, and the nutritional status of children was assess anthropometrically during the survey and categorised into under-weight, stunted, and wasted as per standardised growth charts. RESULTS: prevalence of tobacco use among currently lactating mothers in India was 3.24%. Prevalence of alcohol use among currently lactating mothers was 0.57%. Nursing children of mothers consuming alcohol had significantly higher odds of being wasted [OR = 1.44; (95% CI = 1.07-1.92)]. Richer wealth index {compared to poor, poorer [OR = 0.74; (95% CI = 0.59-0.91)], rich [OR = 0.64; (95% CI = 0.50-0.84)], richer [OR = 0.46; (95% CI = 0.33-0.65)], richest [OR = 0.19; (95% CI = 0.11-0.33)]}, higher education status {compared to illiterate, secondary education [OR = 0.79; (95% CI = 0.63-0.97)], higher education [OR = 0.38; (95% CI = 0.24-0.62)]}, and non-tribal ethnicity [OR = 0.40; (95% CI = 0.33-0.50)] were found to be significantly associated with lower odds of alcohol and tobacco use among lactating mothers. Age, religion, residence, and occupation were also found to be significantly associated. CONCLUSIONS: Lactating women with lower education status or belonging to lower income groups needs to be prioritised for further qualitative assessment of alcohol and tobacco use during lactation. Our study reflects the national and state-level prevalence of alcohol and tobacco use among lactating women, which often masks the local and community-level intricacies. There is a need to further explore local and community-level factors affecting alcohol use during lactation and its association with child nutrition.

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Prostate cancer is the most prevalent cancer among men in the United States and is a leading cause of cancer-related death. Prostate specific membrane antigen (PSMA) has been established as a biomarker for prostate cancer diagnosis and treatment. This study aimed to develop a novel theranostic agent, PSMA-1-MMAE-Pc413, which integrates a PSMA-targeting ligand, the photosensitizer Pc413, and the microtubular inhibitor monomethyl auristatin E (MMAE) for synergistic therapeutic efficacy. In vitro uptake studies revealed that PSMA-1-MMAE-Pc413 demonstrated selective and specific uptake in PSMA-positive PC3pip cells but not in PSMA-negative PC3flu cells, with the uptake in PC3pip cells being approximately three times higher. In vitro cytotoxicity assays showed that, when exposed to light, PSMA-1-MMAE-Pc413 had a synergistic effect, leading to significantly greater cytotoxicity in PSMA-positive cells (IC50 = 2.2 nM) compared to PSMA-1-Pc413 with light irradiation (IC50 = 164.9 nM) or PSMA-1-MMAE-Pc413 without light irradiation (IC50 = 12.6 nM). In vivo imaging studies further demonstrated the selective uptake of PSMA-1-MMAE-Pc413 in PC3pip tumors. In in vivo studies, PSMA-1-MMAE-Pc413 dramatically improves the therapeutic outcome for prostate cancer by providing a synergistic effect that surpasses the efficacy of each treatment modality alone in PC3pip tumors. These findings suggest that PSMA-1-MMAE-Pc413 has strong potential for clinical application in improving prostate cancer treatment.

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Background: Pityriasis Rosea (PR) is a common, yet enigmatic, dermatological condition characterized by a distinctive clinical presentation. Despite its prevalence, the aetiology and pathogenesis of PR remain elusive. Aims: To study the epidemiological and clinical aspects of patients with PR. To study dermoscopic findings and carry out histopathological correlation. Methods: A cross-sectional study of 50 patients was conducted. A detailed clinical history was taken and an examination was done followed by a dermoscopy. Quantitative data like age and duration of disease are presented with the help of standard deviation. Qualitative risk factors, like gender, age groups, symptomatology, site of lesion, findings or cutaneous examination, dermoscopy findings, and histopathology findings, are presented with the help of frequency and percentages. Results: PR shows male preponderance and mean age of occurrence being 30.8 ± 15.7 years. Forty per cent of patients had an atypical clinical presentation. The most frequently seen dermoscopy findings were diffuse red background (58%), peripheral collarette scale (62%), and peripheral dotted vessels (50%). On histopathology, the most common findings were spongiosis (44%), parakeratosis (38%), irregular acanthosis (34%), perivascular lymphocytic infiltrate (56%), and red blood cell extravasation (36%). Limitations: Sample size was less due to COVID. As this was a corss-sectional study follow up of patients could not be done. Conclusion: While the diagnosis of PR is clinical, it is difficult in atypical cases where dermoscopy comes to the aid. It also helps identify the age of lesions, thus helping decide the treatment strategy for patients. Biopsy remains the gold standard in ruling out other differentials of PR.

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The recent SARS-CoV-2 pandemic and associated COVID19 disease illustrates the important role of viral defence mechanisms in ensuring survival and recovery of the host or patient. Viruses absolutely depend on the host's protein synthesis machinery to replicate, meaning that impeding translation is a powerful way to counteract viruses. One major approach used by cells to obstruct protein synthesis is to phosphorylate the alpha subunit of eukaryotic translation initiation factor 2 (eIF2α). Mammals possess four different eIF2α-kinases: PKR, HRI, PEK/PERK, and GCN2. While PKR is currently considered the principal eIF2α-kinase involved in viral defence, the other eIF2α-kinases have also been found to play significant roles. Unsurprisingly, viruses have developed mechanisms to counteract the actions of eIF2α-kinases, or even to exploit them to their benefit. While some of these virulence factors are specific to one eIF2α-kinase, such as GCN2, others target all eIF2α-kinases. This review critically evaluates the current knowledge of viral mechanisms targeting the eIF2α-kinase GCN2. A detailed and in-depth understanding of the molecular mechanisms by which viruses evade host defence mechanisms will help to inform the development of powerful anti-viral measures.

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The full potential of ionizable lipid nanoparticles (LNPs) as an in vivo nucleic acid delivery platform has not yet been realized given that LNPs primarily accumulate in the liver following systemic administration, limiting their success to liver-centric conditions. The engineering of LNPs with antibody targeting moieties can enable extrahepatic tropism by facilitating site-specific LNP tethering and driving preferential LNP uptake into receptor-expressing cell types via receptor-mediated endocytosis. Obstetric conditions stemming from placental dysfunction, such as preeclampsia, are characterized by overexpression of cellular receptors, including the epidermal growth factor receptor (EGFR), making targeted LNP platforms an exciting potential treatment strategy for placental dysfunction during pregnancy. Herein, an EGFR antibody-conjugated LNP (aEGFR-LNP) platform was developed by engineering LNPs with increasing densities of antibody functionalization. aEGFR-LNPs were screened in vitro in immortalized placental trophoblasts and in vivo in non-pregnant and pregnant mice and compared to non-targeted formulations for extrahepatic, antibody-targeted mRNA LNP delivery to the placenta. Our top performing LNP with an intermediate density of antibody functionalization (1:5 aEGFR-LNP) mediated a âˆ¼twofold increase in mRNA delivery in murine placentas and a âˆ¼twofold increase in LNP uptake in EGFR-expressing trophoblasts compared to non-targeted counterparts. These results demonstrate the potential of antibody-conjugated LNPs for achieving extrahepatic tropism, and the ability of aEGFR-LNPs in promoting mRNA delivery to EGFR-expressing cell types in the placenta.

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PURPOSE: Prostate specific membrane antigen (PSMA) has been studied in human breast cancer (BCa) biopsies, however, lack of data on PSMA expression in mouse models impedes development of PSMA-targeted therapies, particularly in improving breast conserving surgery (BCS) margins. This study aimed to validate and characterize the expression of PSMA in murine BCa models, demonstrating that PSMA can be utilized to improve therapies and imaging techniques. METHODS: Murine triple negative breast cancer 4T1 cells, and human cell lines, MDA-MB-231, MDA-MB-468, implanted into the mammary fat pads of BALB/c mice, were imaged by our PSMA targeted theranostic agent, PSMA-1-Pc413, and tumor to background ratios (TBR) were calculated to validate selective uptake. Immunohistochemistry was used to correlate PSMA expression in relation to CD31, an endothelial cell biomarker highlighting neovasculature. PSMA expression was also quantified by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). RESULTS: Accumulation of PSMA-1-Pc413 was observed in 4T1 primary tumors and associated metastases. Average TBR of 4T1 tumors were calculated to be greater than 1.5-ratio at which tumor tissues can be distinguished from normal structures-at peak accumulation with the signal intensity in 4T1 tumors comparable to that in high PSMA expressing PC3-pip tumors. Extraction of 4T1 tumors and lung metastases followed by RT-PCR analysis and PSMA-CD31 co-staining shows that PSMA is consistently localized on tumor neovasculature with no expression in tumor cells and surrounding normal tissues. CONCLUSION: The selective uptake of PSMA-1-Pc413 in these cancer tissues as well as the characterization and validation of PSMA expression on neovasculature in this syngeneic 4T1 model emphasizes their potential for advancements in targeted therapies and imaging techniques for BCa. PSMA holds great promise as an oncogenic target for BCa and its associated metastases.

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BACKGROUND: Modern technologies have led to the development of new tools, practices, and digital techniques. However, their use in public health to provide adequate oral health facilities to the community is limited. One of the facilities that can help provide better oral health with minimal cost is teledentistry. The application of this approach will reduce inequalities in accessing oral healthcare. Knowledge of the use of teledentistry is of the utmost importance to its practice. Hence, the objective of this cross-sectional study is to assess the knowledge of and attitude regarding teledentistry among dental professionals in the Sangli district of Maharashtra. MATERIALS AND METHODS: A 24-unit structured online validated questionnaire with six questions regarding participants' sociodemographic information and 18 questions related to their knowledge and attitude toward teledentistry and informed consent forms were circulated via email among 100 dentists, and the responses obtained were analyzed. RESULTS: Out of 100 responses, 61 showed basic knowledge and a typical attitude toward teledentistry. Urban practitioners were more familiar with teledentistry than rural ones. CONCLUSION: This survey concludes that the branch of teledentistry still needs to be studied and publicized at a greater level to accelerate its widespread implementation in dentistry and especially to increase the outreach and time efficiency of dentistry.

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Behcet's disease is a multisystemic vasculitis. It can affect the pulmonary artery in 2% to 5% cases. We discuss a case of a young male diagnosed with Behcet's disease on immunosuppressive therapy who presented with bilateral pulmonary artery aneurysms which were closed with covered stent and other devices.

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PURPOSE: To examine the current falls prevention knowledge, beliefs and practices of physiotherapists providing clinical care to people with breast cancer. METHODS: A cross-sectional online survey of currently registered, practising Australian physiotherapists was conducted. The survey was developed and reported using the Checklist for Reporting Results of Internet E-Surveys (CHERRIES) checklist, with data analysed descriptively or using bivariate tests. Free-text responses to open-ended questions were classified into key themes for analysis. RESULTS: Forty-two physiotherapists completed the survey, of which 55% (23/42) believed that people with breast cancer had a higher risk of falls compared to the general population. Whilst most respondents received prior training in assessing and managing falls risk factors (30/42; 71%), they reported only moderate confidence in assessing and delivering falls prevention care to people with breast cancer (median 6; IQR 4). Only half of respondents (20/38; 53%) routinely asked about falls history although 61% assessed standing balance (23/38) either through an overall functional assessment (16/38; 42%) or using specific balance measures (7/38; 18%). CONCLUSIONS: Further resources and training for physiotherapists may be required to optimise their skills and confidence, and to embed best-practice falls prevention strategies into the physiotherapy care of people with breast cancer.IMPLICATIONS FOR REHABILITATIONThere is an opportunity to better address falls in routine breast cancer care.Falls screening and prevention activities should be included in the care pathways for breast cancer.More resources are required for physiotherapists to optimise their skills and confidence to facilitate the uptake of best-practice falls prevention strategies.

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BACKGROUND: Coordination between arm movements and postural adjustments is crucial for reaching-while-stepping tasks involving both anticipatory postural adjustments (APAs) and compensatory movements to effectively propel the whole-body forward so that the hand can reach the target. Stroke impairs the ability to coordinate the action of multiple body segments but the underlying mechanisms are unclear. Objective. To determine the effects of stroke on reaching performance and APAs during whole-body reaching. METHODS: We tested arm reaching in standing (stand-reach) and reaching-while-stepping (step-reach; 15 trials/condition) in individuals with chronic stroke (n = 18) and age-matched healthy subjects (n = 13). Whole-body kinematics and kinetic data were collected during the tasks. The primary outcome measure for step-reach was "gain" (g), defined as the extent to which the hip displacement contributing to hand motion was neutralized by appropriate changes in upper limb movements (g = 1 indicates complete compensation) and APAs measured as spatio-temporal profiles of the center-of-pressure shifts preceding stepping. RESULTS: Individuals with stroke had lower gains and altered APAs compared to healthy controls. In addition, step onset was delayed, and the timing of endpoint, trunk, and foot movement offset was prolonged during step-reach compared to healthy controls. Those with milder sensorimotor impairment and better balance function had higher gains. Altered APAs were also related to reduced balance function. CONCLUSIONS: Altered APAs and prolonged movement offset in stroke may lead to a greater reliance on compensatory arm movements. Altered APAs in individuals with stroke may be associated with a reduced shift of referent body configuration during the movement.

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Recombinant adeno-associated viruses (rAAVs) are valuable viral vectors for in vivo gene transfer, also having significant ex vivo therapeutic potential. Continued efforts have focused on various gene therapy applications, capsid engineering, and scalable manufacturing processes. Adherent cells are commonly used for virus production in most basic science laboratories because of their efficiency and cost. Although suspension cells are easier to handle and scale up compared to adherent cells, their use in virus production is hampered by poor transfection efficiency. In this protocol, we developed a simple scalable AAV production protocol using serum-free-media-adapted HEK293T suspension cells and VirusGEN transfection reagent. The established protocol allows AAV production from transfection to quality analysis of purified AAV within two weeks. Typical vector yields for the described suspension system followed by iodixanol purification range from a total of 1 × 1013 to 1.5 × 1013 vg (vector genome) using 90 mL of cell suspension vs. 1 × 1013 to 2 × 1013 vg using a regular adherent cell protocol (10 × 15 cm dishes). Key features • Adeno-associated virus (AAV) production using serum-free-media-adapted HEK293T suspension cells. • Efficient transfection with VirusGEN. • High AAV yield from small-volume cell culture. Graphical overview.

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BACKGROUND: Avelumab, a programmed death ligand-1 inhibitor, has shown success in providing durable responses for difficult-to-treat Merkel cell carcinomas (MCCs). OBJECTIVE: Evaluate the efficacy and safety of avelumab in the treatment of advanced MCC. METHODS: Studies reporting the use of avelumab as a monotherapy or in combination with other agents in the treatment of stage III or IV (advanced) MCC were included. The primary outcomes were overall response rate, overall survival (OS), and treatment-related adverse events. RESULTS: A total of 48 studies were included, involving 1,565 patients with advanced MCC. Most patients were male (1,051, 67.3%) with stage IV MCC (517, 97.0%). The overall response rate was 46.1% (partial response-25.4% and complete response-20.7%) after a mean follow-up period of 9.5 months. Kaplan-Meier survival curves for the pooled stage III and IV group demonstrated OS rates of 58% at 1 year, 47% at 2 years, and 28% at 5 years after completion of treatment with avelumab (median OS: 23.1 months). The most common treatment-related adverse events consisted of constitutional (44%), gastrointestinal (19%), and dermatologic (12%) symptoms. CONCLUSION: Avelumab monotherapy and combination therapy have shown success in the overall response rate and survival for patients with advanced MCC.

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BACKGROUND: The beneficial role of dual anti-platelet therapy (DAPT) in coronary artery disease is well established. However, there is limited data describing the effects of DAPT in patients with atherosclerotic peripheral artery disease (PAD). The aim of this meta-analysis is to compare clinical outcomes associated with DAPT versus single anti-platelet therapy (SAPT) in patients with symptomatic PAD. METHODS: We performed a literature search for studies assessing the risk of adverse cardiovascular and limb events in cohorts receiving either DAPT or SAPT. The primary endpoint was all cause mortality. The secondary endpoints included graft failure, amputation, total bleeding, severe bleeding and fatal bleeding. The search included the following databases: Ovid MEDLINE, EMBASE, Web of Science, and Google Scholar. The search was not restricted to time or publication status. RESULTS: A total of 11 studies with 54,331 participants (24,449 on SAPT and 29,882 on DAPT) were included. Patients with PAD treated with SAPT had higher all-cause mortality compared to patients treated with DAPT (OR 1.37, 95 % CI 1.09-1.74; p < 0.01). There was no difference in risk of graft failure or amputation between patients treated with SAPT or DAPT (OR 0.9, 95 % CI 0.77-1.06; p = 0.19; OR 1.11, 95 % CI 0.88-1.41; p = 0.37). Patients treated with SAPT had lower total bleeds compared to patients treated with DAPT (OR 0.53, 95 % CI 0.36-0.77; p < 0.01). However, For SAPT plus AC vs SAPT, a total of 8 studies with 17,100 participants (3447 with SAPT plus AC and 8619 with only SAPT) were included. Patients on SAPT plus AC did not have a statistically significant difference in risk for all-cause mortality, (OR 0.91, 95 % CI 0.67-1.24; p = 0.56). SAPT plus AC had significantly lower risk of MI (OR 0.82, 95 % CI 0.69-0.97; p = 0.02), amputation (OR 0.72, 95 % CI 0.53-0.97; p = 0.03), and graft failure (OR 0.66, 95 % CI 0.48-0.93; p = 0.02). There was no significant different in risk of fatal bleeding be-tween the two groups (OR 1.60, 95 % CI 0.76-3.35; p = 0.22). CONCLUSIONS: In patients with symptomatic PAD, a strategy of DAPT may confer a mortality benefit when compared to SAPT without significantly increasing the risk of serious bleeding events.

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The optimal surveillance and management strategies for breast cancer patients receiving anthracycline therapy are limited by our incomplete understanding of the role of biomarkers heralding the onset of cardiotoxicity. The purpose of this study was to determine whether there is a temporal correlation between cardiac biomarkers and subclinical left ventricular dysfunction in breast cancer patients receiving anthracycline chemotherapy. Thirty-one females between 46 and 55 years old with breast cancer treated with anthracycline chemotherapy were prospectively enrolled. Cardiac biomarkers were correlated with echocardiography with speckle tracking at baseline, post-anthracycline therapy, and 6 months post-anthracycline chemotherapy. Subclinical cardiotoxicity was defined as ≥ 10% reduction in global longitudinal strain (GLS). There was a relative reduction in left ventricular ejection fraction (LVEF) ≥ 10% in 5/30 (17%) and 7/27 (26%) patients post-anthracycline therapy and 6 months post-anthracycline therapy, respectively. Subclinical cardiotoxicity was noted in 8/30 (27%) and 10/26 (38%) patients post-anthracycline and 6 months post-anthracycline therapy, respectively. Baseline N-terminal pro B-type natriuretic peptide (NT-proBNP) was the strongest predictor of LVEF (ρ = -0.45; p = 0.019), with post-therapy NT-proBNP values illustrating similar predictive value (ρ = -0.40; p = 0.038). Interim changes in suppression of tumorigenicity 2 (ST2) and galectin-3 correlated with a 6-month change in LVEF (ρ = -0.48; p = 0.012 and ρ = -0.45; p = 0.018, for ST2 and galectin-3, respectively). Changes in galectin-3 from baseline to mid-therapy paralleled changes in GLS. NT-proBNP, ST2, and galectin-3 correlate with reduced LVEF among breast cancer patients receiving anthracycline therapy. Additional trials focusing on a cardiac biomarker approach may provide guidance in the early diagnosis and management of anthracycline-induced cardiotoxicity.

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Organophosphate poisoning (OPP) results from the occupational, accidental, or suicidal intake of organophosphate pesticides (OPs). There is a huge limitation in the available literature on OPP cases and the role of physiotherapy in such cases. We report a case of a 33-year-old farmer, found in a state of ingested insecticide and admitted to the emergency department. The patient had two to three episodes of vomiting, associated with continuous tremors in his hands and legs. Immediately infused with atropine, the patient's general condition deteriorated, and he was intubated with an endotracheal tube. With the signs of long-term intubation, a tracheostomy was done. A respiratory therapy consult was taken for indications of intermediate syndrome and to achieve early weaning. The patient's referral was received in view of the long-term requirement of bronchial hygiene and the need for early ambulation as well as psychological support. Informed consent was taken from the family prior to the commencement of the treatment. Respiratory therapy interventions included body positioning, bronchial hygiene techniques, chest proprioceptive neuromuscular facilitation (PNF) techniques, and mobility exercises to achieve early ambulation. Physiotherapists have the appropriate training, knowledge, and skills to deliver the exercise components and help patients return to their activities of daily living. Significant levels of improvement have been seen in the general condition of the patient. The overall functioning of the patient's health was seen as improved on the scales of consciousness, early mobility in the step-down unit, and quality of life.

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Background: Immune-related adverse events (irAEs) challenge the use of immune checkpoint inhibitors (ICIs). We performed a retrospective study to evaluate response to infliximab for immune-related adverse event management, and infliximab's effect on progression-free survival (PFS) and overall survival (OS) with a focus on melanoma and genitourinary cancers. Methods: We retrospectively reviewed records of all cancer patients exposed to infliximab after immune checkpoint inhibitor (ICI) treatment from 2004 to 2021 at the MD Anderson Cancer Center. Survival was assessed utilizing the Kaplan-Meier method. Univariate and multivariate logistic regression was utilized to evaluate predictors of infliximab response, OS, and PFS. Results: We identified 185 cancer patients (93 melanoma and 37 genitourinary cancers) treated with ICI and who received infliximab to treat irAEs. Within 3 months of treatment initiation, 71% of the patients responded to infliximab, 27% had no response, and 2% had unknown response. Among different irAEs, colitis was associated with increased response to infliximab at 3 months, irrespective of the type of malignancy. We evaluated best tumor response before and after infliximab in the entire cohort and again in the melanoma and genitourinary (GU); the findings were similar in the melanoma cohort and the entire cohort, where best tumor response before and after infliximab was not significantly different. In the melanoma cohort, acute kidney injury (AKI) was associated with increased risk of death, p = 0.0109, and having response to infliximab was associated with decreased risk of death, p = 0.0383. Interestingly in GU cancer patients, myositis was associated with increased risk of death, p = 0.0041, and having a response to infliximab was marginally associated with decreased risk of death, p = 0.0992. As regards PFS, in a multivariate Cox regression model, having a history of cardiovascular disease remained significantly associated with shorter PFS in the melanoma cohort. For patients with GU cancers, response to infliximab was associated with longer PFS. Conclusions: Our study is among the largest retrospective analyses of infliximab use for irAE management. Patients with colitis were the best responders to infliximab. AKI before initiation of infliximab in the melanoma subcohort and myositis in GU subcohort are associated with higher risk of death. Our results indicate no association between infliximab and cancer progression with the exception of genitourinary cancers.

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East Coast fever (ECF) is a cattle disease caused by a protozoan parasite called Theileria parva (T. parva). Theileria parva is transmitted among cattle by ticks. It is endemic in parts of central, eastern, and southern Africa and imposes an economic burden through illness and death of approximately a half of a billion U.S. dollars annually. This paper reviews existing science on the economics of ECF. We utilize a conceptual model that defines primary categories of economic costs due to ECF and use it to organize a synthesis of the literature on aggregate and micro level direct costs of the disease and the costs and benefits related to various ECF management strategies. We then identify knowledge gaps to motivate for future research.

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During healthy pregnancy, the placenta develops to allow for exchange of nutrients and oxygen between the mother and the fetus. However, placental dysregulation can lead to several pregnancy disorders, such as preeclampsia and fetal growth restriction. Recently, lipid nanoparticle (LNP)-mediated delivery of messenger RNA (mRNA) has been explored as a promising approach to treat these disorders. Here, iterative libraries of LNPs with varied excipient molar ratios are screened in vitro for enhanced mRNA delivery to placental cells with minimal cytotoxicity when compared to an LNP formulation with a standard excipient molar ratio. LNP C5, the top formulation identified by these screens, demonstrates a fourfold increase in mRNA delivery in vitro compared to the standard formulation. Intravenous administration of LNP C5 to pregnant mice achieves improved in vivo placental mRNA delivery compared to the standard formulation and mediates mRNA delivery to placental trophoblasts, endothelial cells, and immune cells. These results identify LNP C5 as a promising optimized LNP formulation for placental mRNA delivery and further validates the design of experiments strategy for LNP excipient optimization to enhance mRNA delivery to cell types and organs of interest.

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Prodigiosin pigment is a secondary metabolite produced by many bacterial species and is known for its medicinal properties. A few of these prodigiosin-producing bacteria are also reported to be entomopathogenic. It is intriguing to unravel the role of prodigiosin in insecticidal activities and its mode of action. In this study, we have shown the production and characterization of prodigiosin from the Serratia rubidaea MJ 24 isolated from the soil of the Western Ghats, India. Further, we assessed the effect of this pigment on the lepidopteran agricultural pest, Helicoverpa armigera. Prodigiosin-fed H. armigera indicated defective development of insect growth upon treatment. Due to defective early development, about 50% mortality and 40% reduction in body weight were observed in insects fed on a 500 ppm prodigiosin-containing diet. The transcriptomic analysis of these insects indicated significant dysregulation of Juvenile hormone synthesis and response related genes. In addition, dopamine related processes and their resultant melanization and sclerotization processes were also found to be affected. The changes in the expression levels of the key transcripts were further validated using real-time quantitative PCR. The metabolome data confirmed the developmental dysregulation of precursors and products of differentially regulated genes due to prodigiosin. Therefore, the corroborated data suggests that prodigiosin majorly affects H. armigera development through dysregulation of the Juvenile hormone-dopamine system and can be considered as a bioactive scaffold to design insect-pest management compounds. This study provides the first report of in-depth analysis of insecticidal system dynamics in H. armigera insects upon prodigiosin feeding via gene expression and metabolic change via omics approach.

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Here, we describe a fast and cost-effective procedure to generate a large array of mutant proteins and immediately screen for those with altered protein function. This protocol is a modification from three existing approaches: fusion PCR, Saccharomyces cerevisiae in-yeast recombination, and semi-quantitative growth assays. We also describe a mating step to reduce the occurrence of false positive findings due to ectopic mutations. The only requirement is that the protein elicits a phenotype in Saccharomyces cerevisiae.

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