Efficacy of home healthcare on the management of tracheostomy patients.

OBJECTIVES: To evaluated the home healthcare efficacy in managing tracheostomy patients at King Abdulaziz Medical City under the Ministry of National Guard Health Affairs. Home healthcare is care provided to patients in the convenience of their homes to ensure high-quality care based on healthcare providers' supervision. METHODS: This retrospective cohort study utilizing a non-probability consecutive sampling technique, including all available tracheal patients with no exclusion criteria, was carried out in Riyadh, Saudi Arabia, between January 2019 and June 2022. The collected data included patient demographic variables and respiratory settings (ventilation type, daily ventilation need, tracheostomy duration, and ventilator settings). The outcomes included mortality rate and therapeutic outcomes of tracheal management. RESULTS: Of the 183 patients in the study, the most common type of respiratory-related infection was pneumonia (53%). Unlike respiratory-related causes, The mortality rate of patients admitted to the intensive care unit that was unrelated to respiratory causes was statistically significant (57%) (p=0.003). The mortality rate of patients who used aerosol tracheal collars (34%) was markedly higher than mechanically ventilated patients (57%) (p=0.004). The mortality rate following discharge from HHC was 40%, and was higher among patients aged >70 years (47%) (p=0.04). CONCLUSION: Pneumonia was associated with the majority of ventilator-related infections and resulted in hospital readmissions. Ensuring proper practices and caregiver education is crucial to decrease the incidence of ventilator-related infections.

In silico-designed antimicrobial peptide targeting MRSA and E. coli with antibacterial and antibiofilm actions.

Antibiotic resistance is a paramount global health issue, with numerous bacterial strains continually fortifying their resistance against diverse antibiotics. This surge in resistance levels primarily stems from the overuse and misuse of antibiotics in human, animal, and environmental contexts. In this study, we advocate for exploring alternative molecules exhibiting antibacterial properties to counteract the escalating antibiotic resistance. We identified a synthetic antimicrobial peptide (AMP) by using computational search in AMP public databases and further engineering through molecular docking and dynamics. Microbiological evaluation, cytotoxicity, genotoycity, and hemolysis experiments were then performed. The designed AMP underwent rigorous testing for antibacterial and antibiofilm activities against Methicillin-Resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), representing gram-positive and gram-negative bacteria, respectively. Subsequently, the safety profile of the AMP was assessed in vitro using human fibroblast cells and a human blood sample. The selected AMP demonstrated robust antibacterial and antibiofilm efficacy against MRSA and E. coli, with an added assurance of non-cytotoxicity and non-genotoxicity towards human fibroblasts. Also, the AMP did not demonstrate any hemolytic activity. Our findings emphasize the considerable promise of the AMP as a viable alternative antibacterial agent, showcasing its potential to combat antibiotic resistance effectively.

In Vitro and In Vivo Validation of GATA-3 Suppression for Induction of Adipogenesis and Improving Insulin Sensitivity.

Impaired adipogenesis is associated with the development of insulin resistance and an increased risk of type 2 diabetes (T2D). GATA Binding Protein 3 (GATA3) is implicated in impaired adipogenesis and the onset of insulin resistance. Therefore, we hypothesize that inhibition of GATA3 could promote adipogenesis, restore healthy fat distribution, and enhance insulin signaling. Primary human preadipocytes were treated with GATA3 inhibitor (DNAzyme hgd40). Cell proliferation, adipogenic capacity, gene expression, and insulin signaling were measured following well-established protocols. BALB/c mice were treated with DNAzyme hgd40 over a period of 2 weeks. Liposomes loaded with DNAzyme hgd40, pioglitazone (positive), or vehicle (negative) controls were administered subcutaneously every 2 days at the right thigh. At the end of the study, adipose tissues were collected and weighed from the site of injection, the opposite side, and the omental depot. Antioxidant enzyme (superoxide dismutase and catalase) activities were assessed in animals' sera, and gene expression was measured using well-established protocols. In vitro GATA3 inhibition induced the adipogenesis of primary human preadipocytes and enhanced insulin signaling through the reduced expression of p70S6K. In vivo GATA3 inhibition promoted adipogenesis at the site of injection and reduced MCP-1 expression. GATA3 inhibition also reduced omental tissue size and PPARγ expression. These findings suggest that modulating GATA3 expression offers a potential therapeutic benefit by correcting impaired adipogenesis, promoting healthy fat distribution, improving insulin sensitivity, and potentially lowering the risk of T2D.

Social media addiction and emotions during the disaster recovery period-The moderating role of post-COVID timing.

BACKGROUND: Social media addiction, a recently emerged term in medical science, has attracted the attention of researchers because of its significant physical and psychological effects on its users. The issue has attracted more attention during the COVID era because negative emotions (e.g., anxiety and fear) generated from the COVID pandemic may have increased social media addiction. Therefore, the present study investigates the role of negative emotions and social media addiction (SMA) on health problems during and after the COVID lockdown. METHODS: A survey was conducted with 2926 participants aged between 25 and 45 years from all eight divisions of Bangladesh. The data collection period was between 2nd September- 13th October, 2020. Partial Least Square Structural Equation Modelling (PLS-SEM) was conducted for data analysis by controlling the respondents' working time, leisure time, gender, education, and age. RESULTS: Our study showed that social media addiction and time spent on social media impact health. Interestingly, while anxiety about COVID increased social media addition, fear about COIVD reduced social media addition. Among all considered factors, long working hours contributed most to people's health issues, and its impact on social media addiction and hours was much higher than negative emotions. Furthermore, females were less addicted to social media and faced less health challenges than males. CONCLUSION: The impacts of negative emotions generated by the COVID disaster on social media addiction and health issues should be reconsidered. Government and employers control people's working time, and stress should be a priority to solve people's social media addiction-related issues.

Human rights violations and associated factors of the Hijras in Bangladesh-A cross-sectional study.

BACKGROUND: Hijras in Bangladesh face considerable discrimination, stigma, and violence despite the 2013 legislation that recognized Hijras as a third gender. There is a dearth of published literature describing the extent of human rights violations among this population and their associated factors. METHODS: A questionnaire was administered to 346 study participants aged 15 years and older, living in five urban cities of Bangladesh who self-identified as Hijra, in 2019. The six human rights violation indicators (Economic, Employment, Health, Education, Social and Civic and Political Right) assessed were categorized as binary. Associations between sociodemographic characteristics and the six human rights violations were tested using univariate and multivariate logistic regression. RESULTS: Human right violations including economic, educational, political, employment, health and social/civil right violations were reported in 73.3%, 59.3%, 58.5%, 46.4%, 42.7%, and 34.4% of the participants, respectively. Economic rights violations were associated with bisexuality (Adjusted odds ratios [AOR] 3.60, 95%CI: 1.57, 8.26) and not living with family (AOR 2.71, 95%CI: 1.21, 6.09), while Hijras who earned more than 10,000 Bangladesh Taka experienced higher odds of educational (AOR 2.77, 95%CI: 1.06, 7.19) and political rights violations (AOR 4.30, 95%CI: 1.06, 7.44). Living in Dhaka city was associated with a reduced odds for economic and political rights violation while experiencing violations of one human right could lead to violation of another in the Hijra community. CONCLUSION: Human rights violations were common in Bangladesh Hijras, particularly the Bisexual Hijras. Media and educational awareness campaigns are needed to address the underlying roots of a violation. Programs focused on the families, young people and high-income earners of this community are needed in Bangladesh.

Think like a Virus: Toward Improving Nanovaccine Development against SARS-CoV-2.

There is no doubt that infectious diseases present global impact on the economy, society, health, mental state, and even political aspects, causing a long-lasting dent, and the situation will surely worsen if and when the viral spread becomes out of control, as seen during the still ongoing coronavirus disease 2019 (COVID-19) pandemic. Despite the considerable achievements made in viral prevention and treatment, there are still significant challenges that can be overcome through careful understanding of the viral mechanism of action to establish common ground for innovating new preventative and treatment strategies. Viruses can be regarded as devil nanomachines, and one innovative approach to face and stop the spread of viral infections is the development of nanoparticles that can act similar to them as drug/vaccine carriers. Moreover, we can use the properties that different viruses have in designing nanoparticles that reassemble the virus conformational structures but that do not present the detrimental threats to human health that native viruses possess. This review discusses the current preventative strategies (i.e., vaccination) used in facing viral infections and the associated limitations, highlighting the importance of innovating new approaches to face viral infectious diseases and discussing the current nanoapplications in vaccine development and the challenges that still face the nanovaccine field.

Tocilizumab effectiveness in mechanically ventilated COVID-19 patients (T-MVC-19 Study): a multicenter real-world evidence.

BACKGROUND: This study aimed to evaluate the effectiveness of tocilizumab in mechanically ventilated patients with coronavirus disease 2019 (COVID-19). RESEARCH DESIGN AND METHODS: This retrospective multicenter study included adults (≥18 years) diagnosed with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by real-time polymerase chain reaction (RT-PCR) from nasopharyngeal swab, and requiring invasive mechanical ventilation during admission. Survival analyses with inverse propensity score treatment weighting (IPTW) and propensity score matching (PSM) were conducted. To account for immortal bias, we used Cox proportional modeling with time-dependent covariance. Competing risk analysis was performed for the extubation endpoint. RESULTS: A total of 556 (tocilizumab = 193, control = 363) patients were included. Males constituted the majority of the participants (69.2% in tocilizumab arm,74.1% in control arm). Tocilizumab was not associated with a reduction in mortality with hazard ratio [(HR) = 0.82,95% confidence interval (95%CI): 0.62-1.10] in the Inverse propensity score weighting (IPTW) analysis and (HR = 0.86,95% CI: 0.64-1.16) in the PSM analysis. However, tocilizumab was associated with an increased rate of extubation (33.6%) compared to the control arm (11.9%); subdistributional hazards (SHR) = 3.1, 95% CI: 1.86-5.16). CONCLUSIONS: Although tocilizumab was not found to be effective in reducing mortality, extubation rate while on mechanical ventilation was higher among tocilizumab treated group.

Recent Developments in Nanomaterials-Based Drug Delivery and Upgrading Treatment of Cardiovascular Diseases.

Cardiovascular diseases (CVDs) are the leading causes of morbidity and mortality worldwide. However, despite the recent developments in the management of CVDs, the early and long outcomes vary considerably in patients, especially with the current challenges facing the detection and treatment of CVDs. This disparity is due to a lack of advanced diagnostic tools and targeted therapies, requiring innovative and alternative methods. Nanotechnology offers the opportunity to use nanomaterials in improving health and controlling diseases. Notably, nanotechnologies have recognized potential applicability in managing chronic diseases in the past few years, especially cancer and CVDs. Of particular interest is the use of nanoparticles as drug carriers to increase the pharmaco-efficacy and safety of conventional therapies. Different strategies have been proposed to use nanoparticles as drug carriers in CVDs; however, controversies regarding the selection of nanomaterials and nanoformulation are slowing their clinical translation. Therefore, this review focuses on nanotechnology for drug delivery and the application of nanomedicine in CVDs.

Covid-19 detection in chest X-ray through random forest classifier using a hybridization of deep CNN and DWT optimized features.

Chest X-ray image contains sufficient information that finds wide-spread applications in diverse disease diagnosis and decision making to assist the medical experts. This paper has proposed an intelligent approach to detect Covid-19 from the chest X-ray image using the hybridization of deep convolutional neural network (CNN) and discrete wavelet transform (DWT) features. At first, the X-ray image is enhanced and segmented through preprocessing tasks, and then deep CNN and DWT features are extracted. The optimum features are extracted from these hybridized features through minimum redundancy and maximum relevance (mRMR) along with recursive feature elimination (RFE). Finally, the random forest-based bagging approach is used for doing the detection task. An extensive experiment is performed, and the results confirm that our approach gives satisfactory performance compare to the existing methods with an overall accuracy of more than 98.5%.

Internet Use Impact on Physical Health during COVID-19 Lockdown in Bangladesh: A Web-Based Cross-Sectional Study.

Previous studies on internet use frequency were focused on mental health impact, with little known about the impact on physical health during the COVID-19 lockdown. This study examined the impact of internet use frequency on self-reported physical health during the COVID-19 lockdown in Bangladesh. A web-based cross-sectional study on 3242 individuals was conducted from 2 August-1 October 2020. The survey covered demographics, internet use frequency and self-reported physical health questions. Linear regression analyses were used to examine the impact of internet use frequency on physical health. 72.5%, 69.9%, 65.1% and 55.3% respondents reported headache, back pain, numbness of the fingers and neck pain, respectively. The analyses showed increased physical health impact among regular (coefficient ß = 0.52, 95% confidence interval [CI]: 0.18-0.85, p = 0.003), frequent (ß = 1.21, 95% CI: 0.88-1.54, p < 0.001) and intense (ß = 2.24, 95% CI: 1.91-2.57, p < 0.001) internet users. Other important predictors were gender, income, occupation, regions, and working status. Frequent and extensive uses of the internet were strong predictors of physical health problems, and our findings suggest the need for increased awareness about the physical health problems that can be triggered by excessive internet usage.

Activation and Contraction of Human "Vascular" Smooth Muscle Cells Grown From Circulating Blood Progenitors.

Blood outgrowth smooth muscle cells (BO-SMCs) offer the means to study vascular cells without the requirement for surgery providing opportunities for drug discovery, tissue engineering, and personalized medicine. However, little is known about these cells which meant that their therapeutic potential remains unexplored. Our objective was to investigate for the first time the ability of BO-SMCs and vessel-derived smooth muscle cells to sense the thromboxane mimetic U46619 by measuring intracellular calcium elevation and contraction. U46619 (10-6 M) increased cytosolic calcium in BO-SMCs and vascular smooth muscle cells (VSMCs) but not in fibroblasts. Increased calcium signal peaked between 10 and 20 s after U46619 in both smooth muscle cell types. Importantly, U46619 (10-9 to 10-6 M) induced concentration-dependent contractions of both BO-SMCs and VSMCs but not in fibroblasts. In summary, we show that functional responses of BO-SMCs are in line with VSMCs providing critical evidence of their application in biomedical research.

Psychological Impact of COVID-19 Pandemic in Bangladesh: Analysis of a Cross-Sectional Survey.

The COVID-19 pandemic has generated fear, panic, distress, anxiety, and depression among many people in Bangladesh. In this cross-sectional study, we examined factors associated with different levels of psychological impact as a result of COVID-19 in Bangladesh. From April 1 to 30, 2020, we used a self-administered online questionnaire to collect data from 10,609 respondents. Using the Impact of Event Scale-Revised to assess the psychological impact of the COVID-19 pandemic on respondents, we categorized the levels of impact as normal, mild, moderate, or severe. Ordinal logistic regression was used to examine the associated factors. The prevalence of mild, moderate, and severe psychological impact was 10.2%, 4.8%, and 45.5%, respectively. Multivariate analysis revealed that the odds of reporting normal vs mild, moderate, or severe psychological impact were 5.9 times higher for people living in the Chittagong Division, 1.7 times higher for women with lower education levels, 3.0 times higher among those who were divorced or separated, 1.8 times higher for those working full time, and 2.4 times higher for those living in shared apartments. The odds of reporting a psychological impact were also higher among people who did not enforce protective measures inside the home, those in self-quarantine, those who did not wear face masks, and those who did not comply with World Health Organization precautionary measures. Increased psychological health risks due to COVID-19 were significantly higher among people who experienced chills, headache, cough, breathing difficulties, dizziness, and sore throat before data collection. Our results showed that 1 in 2 respondents experienced a significant psychological impact as a result of the COVID-19 pandemic. Public health researchers should consider these factors when targeting interventions that would have a protective effect on the individual's psychological health during a pandemic or future disease outbreak.

Prevalence and Factors Associated with Mental Health Impact of COVID-19 Pandemic in Bangladesh: A Survey-Based Cross-Sectional Study.

Background: Feelings of isolation, insecurity, and instability triggered by COVID-19 could have a long-term impact on the mental health status of individuals. Objectives: The aim of this study was to examine the prevalence of mental health symptoms (anxiety, depression, and stress) in Bangladesh and the factors associated with these symptoms during the COVID-19 pandemic. Methods: From 1 to 30 April 2020, we used a validated self-administered questionnaire to conduct a cross-sectional study on 10,609 participants through an online survey platform. We assessed mental health status using the Depression, Anxiety, and Stress Scale (DASS-21). The total depression, anxiety, and stress subscale scores were divided into normal, mild, moderate, severe, and multinomial logistic regression was used to examine associated factors. Findings: The prevalence of depressive symptoms was 15%, 34%, and 15% for mild, moderate, and severe depressive symptoms, respectively. The prevalence of anxiety symptoms was 59% for severe anxiety symptoms, 14% for moderate anxiety symptoms, and 14% for mild anxiety symptoms, while the prevalence for stress levels were 16% for severe stress level, 22% for moderate stress level, and 13% for mild stress level. Multivariate analyses revealed that the most consistent factors associated with mild, moderate, and severe of the three mental health subscales (depression, anxiety, and stress) were respondents who lived in Dhaka and Rangpur division, females, those who self-quarantined in the previous seven days before the survey, and those respondents who experienced chills, breathing difficulty, dizziness, and sore throat. Conclusion: Our results showed that about 64%, 87%, and 61% of the respondents in Bangladesh reported high levels of depression, anxiety, and stress, respectively. There is a need for mental health support targeting women and those who self-quarantined or lived in Dhaka and Rangpur during the pandemic.

Dengue in Dhaka, Bangladesh: Hospital-based cross-sectional KAP assessment at Dhaka North and Dhaka South City Corporation area.

Dengue, the most important mosquito-borne viral disease of humans is a recurring global health problem. In Bangladesh, dengue outbreaks are on the increase despite the efforts of government and it is not clear what the understanding of the general Dhaka population towards dengue fever is. Knowledge, attitude and practice (KAP) studies are essential guides in public health interventions. Hence, using KAP, this study aims to assess patient-perspectives with regards to factors associated with dengue, as well as investigate the associated factors between the two corporations in Dhaka. A Hospital-based cross-sectional study of 242 fever patients from two city-corporations in Dhaka (Dhaka North City Corporations, DNCC (n = 91, 37.6%) and Dhaka South City Corporation, DSCC (n = 151, 62.4%) was conducted using pre-tested KAP items. Wilcoxon's Rank Sum was used to determine the KAP by DNCC, DSCC and both corporations and multivariate Poisson regression analyses. The two corporations were analysed separately due to the differences in income distribution, concentration of slums, hospitals and clinics. The study found that more than half of the study population were knowledgeable about dengue (mean percentage scores was 52%), possess an appropriate and acceptable attitude towards the disease (69.2%), and about two thirds of the respondents (71.4%) engaged in practices towards its prevention. After adjusting for the potential cofounders, the factors associated with KAP about dengue fever varied between DNCC and DSCC; with duration of residency and use of mosquito nets were associated with knowledge in the north while income class and age were associated with knowledge and attitude in the south. In the pooled analysis (combining both corporations), knowledge of dengue was associated with good practice towards dengue fever among the respondents. The duration of residence in Dhaka (10+ years), not using mosquito nets and length of time spent in the hospital (7+ days) due to dengue, and decreased knowledge (Adjusted coefficient (ß) = -0.01, 95%CI: -0.02, -0.01) were associated with attitude towards dengue in DNCC. On the other hand, middle-high income class, age (40+ years) and increased knowledge were associated with practice towards dengue in DSCC (ß = 0.02, 95%CI: 0.01, 0.03). Efforts to increase knowledge about dengue fever through education by the administrations of both corporations would benefit from targeting these high-risk groups for a more sustainable outcome.

COVID-19 Detection from Chest X-ray Images Using Feature Fusion and Deep Learning.

Currently, COVID-19 is considered to be the most dangerous and deadly disease for the human body caused by the novel coronavirus. In December 2019, the coronavirus spread rapidly around the world, thought to be originated from Wuhan in China and is responsible for a large number of deaths. Earlier detection of the COVID-19 through accurate diagnosis, particularly for the cases with no obvious symptoms, may decrease the patient's death rate. Chest X-ray images are primarily used for the diagnosis of this disease. This research has proposed a machine vision approach to detect COVID-19 from the chest X-ray images. The features extracted by the histogram-oriented gradient (HOG) and convolutional neural network (CNN) from X-ray images were fused to develop the classification model through training by CNN (VGGNet). Modified anisotropic diffusion filtering (MADF) technique was employed for better edge preservation and reduced noise from the images. A watershed segmentation algorithm was used in order to mark the significant fracture region in the input X-ray images. The testing stage considered generalized data for performance evaluation of the model. Cross-validation analysis revealed that a 5-fold strategy could successfully impair the overfitting problem. This proposed feature fusion using the deep learning technique assured a satisfactory performance in terms of identifying COVID-19 compared to the immediate, relevant works with a testing accuracy of 99.49%, specificity of 95.7% and sensitivity of 93.65%. When compared to other classification techniques, such as ANN, KNN, and SVM, the CNN technique used in this study showed better classification performance. K-fold cross-validation demonstrated that the proposed feature fusion technique (98.36%) provided higher accuracy than the individual feature extraction methods, such as HOG (87.34%) or CNN (93.64%).

Studies on metal-organic framework (MOF) nanomedicine preparations of sildenafil for the future treatment of pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is an incurable disease, although symptoms are treated with a range of dilator drugs. Despite their clinical benefits, these drugs are limited by systemic side-effects. It is, therefore, increasingly recognised that using controlled drug-release nanoformulation, with future modifications for targeted drug delivery, may overcome these limitations. This study presents the first evaluation of a promising nanoformulation (highly porous iron-based metal-organic framework (MOF); nanoMIL-89) as a carrier for the PAH-drug sildenafil, which we have previously shown to be relatively non-toxic in vitro and well-tolerated in vivo. In this study, nanoMIL-89 was prepared and charged with a payload of sildenafil (generating Sil@nanoMIL-89). Sildenafil release was measured by Enzyme-Linked Immunosorbent Assay (ELISA), and its effect on cell viability and dilator function in mouse aorta were assessed. Results showed that Sil@nanoMIL-89 released sildenafil over 6 h, followed by a more sustained release over 72 h. Sil@nanoMIL-89 showed no significant toxicity in human blood outgrowth endothelial cells for concentrations up to100µg/ml; however, it reduced the viability of the human pulmonary artery smooth muscle cells (HPASMCs) at concentrations > 3 µg/ml without inducing cellular cytotoxicity. Finally, Sil@nanoMIL-89 induced vasodilation of mouse aorta after a lag phase of 2-4 h. To our knowledge, this study represents the first demonstration of a novel nanoformulation displaying delayed drug release corresponding to vasodilator activity. Further pharmacological assessment of our nanoformulation, including in PAH models, is required and constitutes the subject of ongoing investigations.

Factors Associated with the Perception of Risk and Knowledge of Contracting the SARS-Cov-2 among Adults in Bangladesh: Analysis of Online Surveys.

This study investigated the perception and awareness of risk among adult participants in Bangladesh about Coronavirus Disease 2019 (COVID-19). During the lockdown era in Bangladesh at two different time points, from 26-31 March 2020 (early lockdown) and 11-16 May 2020 (late lockdown), two self-administered online surveys were conducted on 1005 respondents (322 and 683 participants, respectively) via social media. To examine risk perception and knowledge-related factors towards COVID-19, univariate and multiple linear regression models were employed. Scores of mean knowledge (8.4 vs. 8.1, p = 0.022) and perception of risk (11.2 vs. 10.6, p < 0.001) differed significantly between early and late lockdown. There was a significant decrease in perceived risk scores for contracting SARS-Cov-2 [ß = -0.85, 95%CI: -1.31, -0.39], while knowledge about SARS-Cov-2 decreased insignificantly [ß = -0.22, 95%CI: -0.46, 0.03] in late lockdown compared with early lockdown period. Self-quarantine was a common factor linked to increased perceived risks and knowledge of SARS-Cov-2 during the lockdown period. Any effort to increase public awareness and comprehension of SARS-Cov-2 in Bangladesh will then offer preference to males, who did not practice self-quarantine and are less worried about the propagation of this kind of virus.

Internalization of Metal-Organic Framework Nanoparticles in Human Vascular Cells: Implications for Cardiovascular Disease Therapy.

Abstract: Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality worldwide. Alteration of endothelial cells and the underlying vasculature plays a central role in the pathogenesis of various CVDs. The application of nanoscale materials such as nanoparticles in biomedicine has opened new horizons in the treatment of CVDs. We have previously shown that the iron metal-organic framework nanoparticle, Materials Institut Lavoisier-89 (nanoMIL-89) represents a viable vehicle for future drug delivery of pulmonary arterial hypertension. In this study, we have assessed the cellular uptake of nanoMIL-89 in pulmonary artery endothelial and smooth muscle cells using microscopy imaging techniques. We also tested the cellular responses to nanoMIL-89 using molecular and cellular assays. Microscopic images showed cellular internalization of nanoMIL-89, packaging into endocytic vesicles, and passing to daughter cells during mitosis. Moreover, nanoMIL-89 showed anti-inflammatory activity without any significant cytotoxicity. Our results indicate that nanoMIL-89 formulation may offer promising therapeutic opportunities and set forth a new prototype for drug delivery not only in CVDs, but also for other diseases yet incurable, such as diabetes and cancer.

In silico and in vivo models for Qatari-specific classical homocystinuria as basis for development of novel therapies.

Homocystinuria is a rare inborn error of methionine metabolism caused by cystathionine ß-synthase (CBS) deficiency. The prevalence of homocystinuria in Qatar is 1:1,800 births, mainly due to a founder Qatari missense mutation, c.1006C>T; p.R336C (p.Arg336Cys). We characterized the structure-function relationship of the p.R336C-mutant protein and investigated the effect of different chemical chaperones to restore p.R336C-CBS activity using three models: in silico, ΔCBS yeast, and CRISPR/Cas9 p.R336C knock-in HEK293T and HepG2 cell lines. Protein modeling suggested that the p.R336C induces severe conformational and structural changes, perhaps influencing CBS activity. Wild-type CBS, but not the p.R336C mutant, was able to restore the yeast growth in ΔCBS-deficient yeast in a complementation assay. The p.R336C knock-in HEK293T and HepG2 cells decreased the level of CBS expression and reduced its structural stability; however, treatment of the p.R336C knock-in HEK293T cells with betaine, a chemical chaperone, restored the stability and tetrameric conformation of CBS, but not its activity. Collectively, these results indicate that the p.R336C mutation has a deleterious effect on CBS structure, stability, and activity, and using the chemical chaperones approach for treatment could be ineffective in restoring p.R336C CBS activity.

Chemical and biological assessment of metal organic frameworks (MOFs) in pulmonary cells and in an acute in vivo model: relevance to pulmonary arterial hypertension therapy.

Pulmonary arterial hypertension (PAH) is a progressive and debilitating condition. Despite promoting vasodilation, current drugs have a therapeutic window within which they are limited by systemic side effects. Nanomedicine uses nanoparticles to improve drug delivery and/or reduce side effects. We hypothesize that this approach could be used to deliver PAH drugs avoiding the systemic circulation. Here we report the use of iron metal organic framework (MOF) MIL-89 and PEGylated MIL-89 (MIL-89 PEG) as suitable carriers for PAH drugs. We assessed their effects on viability and inflammatory responses in a wide range of lung cells including endothelial cells grown from blood of donors with/without PAH. Both MOFs conformed to the predicted structures with MIL-89 PEG being more stable at room temperature. At concentrations up to 10 or 30 µg/mL, toxicity was only seen in pulmonary artery smooth muscle cells where both MOFs reduced cell viability and CXCL8 release. In endothelial cells from both control donors and PAH patients, both preparations inhibited the release of CXCL8 and endothelin-1 and in macrophages inhibited inducible nitric oxide synthase activity. Finally, MIL-89 was well-tolerated and accumulated in the rat lungs when given in vivo. Thus, the prototypes MIL-89 and MIL-89 PEG with core capacity suitable to accommodate PAH drugs are relatively non-toxic and may have the added advantage of being anti-inflammatory and reducing the release of endothelin-1. These data are consistent with the idea that these materials may not only be useful as drug carriers in PAH but also offer some therapeutic benefit in their own right.