FURIN, IFNL4, and TLR2 gene polymorphisms in relation to COVID-19 severity: a case-control study in Egyptian patients.

BACKGROUND AND AIM: A wide range of clinical manifestations and outcomes, including liver injury, have been reported in COVID-19 patients. We investigated the association of three substantial gene polymorphisms (FURIN, IFNL4, and TLR2) with COVID-19 disease susceptibility and severity to help predict prognosis. METHODS: 150 adult COVID-19-assured cases were categorized as follows: 78 patients with a non-severe presentation, 39 patients with severe disease, and 33 critically ill patients. In addition, 74 healthy controls were included. Clinical and laboratory evaluations were carried out, including complete and differential blood counts, D-dimer, lactate dehydrogenase (LDH), C-reactive protein (CRP), procalcitonin, ferritin, interleukin-6 (Il-6), and liver and kidney functions. FURIN (rs6226), IFNL4 (rs12979860), and TLR2 (rs3804099) genotyping allelic discrimination assays were conducted using real-time PCR. RESULTS: The FURIN, IFNL4, and TLR2 genotypes and their alleles differed significantly between COVID-19 patients and controls, as well as between patients with severe or critical illness and those with a non-severe presentation. According to a multivariable regression analysis, FURIN (C/T + T/T) and TLR2 (T/C + C/C) mutants were associated with COVID-19 susceptibility, with odds ratios of 3.293 and 2.839, respectively. FURIN C/C and IFNL4 T/T mutants were significantly linked to severe and critical illnesses. Multivariate regression analysis showed that FURIN (G/C + C/C) genotypes and IFNL4 T/T homozygosity were independent risk factors associated with increased mortality. CONCLUSION: FURIN, IFNL4, and TLR2 gene variants are associated with the risk of COVID-19 occurrence as well as increased severity and poor outcomes in Egyptian patients.

Stem cells derived exosomes as biological nano carriers for VCR sulfate for treating breast cancer stem cells.

Due to vincristine sulfate's (VCR sulfate) toxicity and non-specific targeting, which might adversely damage healthy cells, its clinical application is restricted. In this study, we loaded VCR sulfate on exosomes generated from mesenchymal stem cells (MSCs) to enhance its targeted distribution. Exosomes are able to deliver molecules to specific cells and tissues and have therapeutic potential. In this study, we isolated exosomes from MSCs, and using probe-sonication approach loaded them with VCR sulfate. Using SRB assay, the cytotoxicity of VCR sulfate-Exo was assessed in T47D breast cancer cells, and the results were contrasted with those of free VCR sulfate. Then We labeled markers (CD44+/CD24-) in the cell line to assess the targeting effectiveness of VCR sulfate-Exo using flow cytometry. Our results showed that the cytotoxicity of VCR sulfate-Exo was nearly the same as that of VCR sulfate. Flow cytometry analysis revealed that VRC sulfate-Exo was more effectively targeted to MSCs than free VCR sulfate. Our study shows that loading VCR sulfate to MSCs-derived exosomes can improve their targeted delivery and lessen their side effects. Additional research is required to determine VCR sulfate-Exo's in vivo effectiveness and safety and improve the loading and delivery strategies.

Regulation of cardiomyocyte t-tubule structure by preload and afterload: Roles in cardiac compensation and decompensation.

Mechanical load is a potent regulator of cardiac structure and function. Although high workload during heart failure is associated with disruption of cardiomyocyte t-tubules and Ca2+ homeostasis, it remains unclear whether changes in preload and afterload may promote adaptive t-tubule remodelling. We examined this issue by first investigating isolated effects of stepwise increases in load in cultured rat papillary muscles. Both preload and afterload increases produced a biphasic response, with the highest t-tubule densities observed at moderate loads, whereas excessively low and high loads resulted in low t-tubule levels. To determine the baseline position of the heart on this bell-shaped curve, mice were subjected to mildly elevated preload or afterload (1 week of aortic shunt or banding). Both interventions resulted in compensated cardiac function linked to increased t-tubule density, consistent with ascension up the rising limb of the curve. Similar t-tubule proliferation was observed in human patients with moderately increased preload or afterload (mitral valve regurgitation, aortic stenosis). T-tubule growth was associated with larger Ca2+ transients, linked to upregulation of L-type Ca2+ channels, Na+-Ca2+ exchanger, mechanosensors and regulators of t-tubule structure. By contrast, marked elevation of cardiac load in rodents and patients advanced the heart down the declining limb of the t-tubule-load relationship. This bell-shaped relationship was lost in the absence of electrical stimulation, indicating a key role of systolic stress in controlling t-tubule plasticity. In conclusion, modest augmentation of workload promotes compensatory increases in t-tubule density and Ca2+ cycling, whereas this adaptation is reversed in overloaded hearts during heart failure progression. KEY POINTS: Excised papillary muscle experiments demonstrated a bell-shaped relationship between cardiomyocyte t-tubule density and workload (preload or afterload), which was only present when muscles were electrically stimulated. The in vivo heart at baseline is positioned on the rising phase of this curve because moderate increases in preload (mice with brief aortic shunt surgery, patients with mitral valve regurgitation) resulted in t-tubule growth. Moderate increases in afterload (mice and patients with mild aortic banding/stenosis) similarly increased t-tubule density. T-tubule proliferation was associated with larger Ca2+ transients, with upregulation of the L-type Ca2+ channel, Na+-Ca2+ exchanger, mechanosensors and regulators of t-tubule structure. By contrast, marked elevation of cardiac load in rodents and patients placed the heart on the declining phase of the t-tubule-load relationship, promoting heart failure progression. The dependence of t-tubule structure on preload and afterload thus enables both compensatory and maladaptive remodelling, in rodents and humans.

Prevalence of Medication Associated with QTc Prolongation Used Among Critically Ill Patients.

Background: Acquired prolonged corrected QT (QTc) interval can lead to life-threatening Torsade de Pointes (TdP) arrhythmia. Multiple risk factors including medications, comorbidities, and electrolyte imbalances contribute significantly to acquired manifestations of the QTc prolongation. Critically ill patients are particularly more vulnerable to TdP due to complex medical conditions, aging, and polypharmacy. Objective: This study aimed to assess the prevalence of TdP-associated medication prescribing, identify risk factors for QTc prolongation and TdP, and determine primary predictors of high TdP medication usage in critically ill patients in Jordan. Methods: We conducted a retrospective cross-sectional analysis of electronic medical records for patients from King Abdullah University Hospital who were admitted to Intensive Care Unit (ICU) between (July 2012-July 2022). We collected data on patients' demographics, clinical characteristics, comorbidities, laboratory results, and prescribed medications. Medications were categorized into three TdP risk levels according to CredibleMeds® assessment tool. Data were analyzed using descriptive statistics and a binary logistic regression model. Results: Of the 13,300 patients (58.2% male, median age 62 years). Prescribing prevalence for medications with known TdP risk was 19%, possible risk (24.7%), conditional risk (21.6%), and confirmed conditional risk (8.3%). Common comorbidities included hypertension (40.9%), diabetes (33.3%), and cancer (15.4%). Drugs with known TdP risk included citalopram, amiodarone, clarithromycin, and ciprofloxacin. A binary regression model revealed that as age increased, the odds of TdP associated medication prescribing decreased (OR = 0.989, p < 0.001), while patients on more than five medications had higher odds (OR = 4.281, p < 0.001). Conclusion: The study identified a notable prevalence of prescribing for medications with QTc prolongation/TdP risk in critically ill patients. Healthcare providers in the ICU should exercise caution to minimize the inadvertent prescription of TdP associated medications especially among older patients and those with polypharmacy.

Automated blood glucose regulation for nonlinear model of type-1 diabetic patient under uncertainties: GWOCS type-2 fuzzy approach.

Regulating blood glucose level (BGL) for type-1 diabetic patient (T1DP) accurately is very important issue, an uncontrolled BGL outside the standard safe range between 70 and 180 mg/dl results in dire consequences for health and can significantly increase the chance of death. So the purpose of this study is to design an optimized controller that infuses appropriate amounts of exogenous insulin into the blood stream of T1DP proportional to the amount of obtained glucose from food. The nonlinear extended Bergman minimal model is used to present glucose-insulin physiological system, an interval type-2 fuzzy logic controller (IT2FLC) is utilized to infuse the proper amount of exogenous insulin. Superiority of IT2FLC in minimizing the effect of uncertainties in the system depends primarily on the best choice of footprint of uncertainty (FOU) of IT2FLC. So a comparison includes four different optimization methods for tuning FOU including hybrid grey wolf optimizer-cuckoo search (GWOCS) and fuzzy logic controller (FLC) method is constructed to select the best controller approach. The effectiveness of the proposed controller was evaluated under six different scenarios of T1DP using Matlab/Simulink platform. A 24-h scenario close to real for 100 virtual T1DPs subjected to parametric uncertainty, uncertain meal disturbance and random initial condition showed that IT2FLC accurately regulate BGL for all T1DPs within the standard safe range. The results indicated that IT2FLC using GWOCS can prevent side effect of treatment with blood-sugar-lowering medication. Also stability analysis for the system indicated that the system operates within the stability region of nonlinear system.

Long-term health complications of chemical weapon exposure: a study on Halabja chemical attack survivors (Iraqi Kurds).

OBJECTIVE: In 1988, the Iraqi government used a range of chemical weapons (CWs) against the Iraqi Kurds of Halabja. Here, we aim to investigate the long-term health consequences in exposed survivors as they are not sufficiently studied. MATERIALS AND METHODS: This was a retrospective study conducted from November 2019 to May 2020 assessing the health status of all exposed Halabja chemical attack survivors compared to non-exposed people from the same area. RESULTS AND DISCUSSION: Two hundred thirty survivors and 240 non-exposed participants were enrolled in this study, with control participants matched to age, gender, and occupation. Among the survivors, females were more prevalent. The respiratory system was the most common single exposure route (83, 36.1%), with 138 (60%) of the survivors being exposed by multiple routes. The vast majority (88.7%) of survivors had activities of daily living (ADL) impairment. There was female predominance in mild and moderate cases, with more males in severe cases (p < 0.01). Respiratory and cardiac diseases were significantly more common in the survivors compared to the controls (p < 0.001). Survivors with multiple CW exposure routes had significantly higher rates of ADL impairment (p < 0.001) and cardiac disease, respiratory diseases, and miscarriage (p < 0.01), than those with a single exposure route. CONCLUSION: In this study comparing CW survivors with a local control population, a single, high-dose exposure to CWs was associated with significant increases in chronic respiratory and cardiac conditions, in addition to high rates of ADL impairment. Similar studies are needed in other, more recent CW survivor cohorts.

Adherence and utilization of short-term antibiotics: Randomized controlled study.

Enhancing adherence to medication has the potential to improve clinical outcomes and decrease healthcare cost. The role of clinical pharmacist-led education on adherence to short-term antibiotic has never been investigated in Jordan. This study aimed to evaluate the impact of an educational intervention on antibiotic short-term adherence and to assess the antibiotic utilization pattern. A prospective, single blinded, randomized controlled study was conducted in a tertiary referral hospital in Jordan. Adult patients diagnosed with acute infection and prescribed a short-term antibiotic course (< 30 day) were included in the study. Recruited patients were randomly allocated into control and intervention groups. Pharmaceutical education about the correct use of antibiotic/s was provided to the intervention group. The results showed that penicillins were the most prescribed antibiotics (38.7%) followed by fluoroquinolones (23.9%) and cephalosporines (20.9%). Patients in the intervention group were more likely to be adherent to the prescribed antibiotics compared to control group (OR = 1.445, 95CI% = 1.029-2.030, p = 0.033). Employed patients, less frequent administration of antibiotic, and searching information related to the prescribed antibiotics were factors associated with better adherence to short-term antibiotic (p<0.05). The most common reasons for non-adherence were feeling better and forgetfulness to take medication. These findings highlighted that pharmacist-led educational intervention significantly enhance adherence to prescribed short-term antibiotics which is a major drive to control antibiotic resistance. Initiatives should be adopted to include patient education as a regular element in the medication dispensing process. Clinical trial registration: The trial is registered at ClinicalTrials.gov (identifier: NCT05293977).

Optimized type-2 fuzzy controller based on IoMT for stabilizing the glucose level in type-1 diabetic patients.

Due to advancements in existing Internet of Medical Things (IoMT) systems and devices, the blood glucose level (BGL) for type-1 diabetic patients (T1DPs) is effectively and continually monitored and controlled by Artificial Pancreas. Because the regulation of BGL is a very complex process, many efforts have been conducted to design a powerful and effective controller for the exogenous insulin infusion system. The main objective of this study is to propose an optimized interval type-2 fuzzy (IT2F) based controller of artificial pancreas for regulation BGL of T1DP based on IoMT. The proposed controller should avoid the risk of hyperglycemia and hypoglycemia situations that T1DP faces during the infusion of exogenous insulin. The main contribution of this work is using meta-heuristic method called grey wolf optimizer (GWO) to tune the footprint of uncertainty for IT2F's membership functions to inject the proper dose of insulin under different conditions. The nonlinear extended Bergman minimal model (EBMM) with uncertainty is used to represent the blood glucose regulation and represent the dynamics of meal disturbance in T1DP. The effectiveness and the performance of the proposed controller are investigated using MATLAB/Simulink platform. Simulation results show that the proposed controller can avoid both severe hypoglycemia and hyperglycemia for nominal parameters of the model, in addition to model under the presence of both parametric uncertainty and uncertain meal disturbance.

A suspension of inactivated bacteria used for vaccination against recurrent urinary tract infections increases the phagocytic activity of murine macrophages.

Background: Urinary tract infections are a major cause of the consumption of antibiotics in humans. Methods: We studied the effect of a vaccine (StroVac®, containing inactivated bacteria and used to prevent recurrent urinary tract infections) licensed in Germany on the release of pro-inflammatory cytokines and the phagocytosis of Escherichia (E.) coli in primary murine macrophages and the macrophage cell line J774A.1. Results: StroVac® increased the release of the cytokines TNF-α, IL-6, IL-12/23 p40, and IL-1ß and stimulated the phagocytosis of E. coli in a dose-dependent manner. This effect was independent of LPS as shown by the use of macrophages isolated from LPS-resistant C3H/HeJ mice. At concentrations up to 30 mg/l it was not toxic to bacteria or eukaryotic cells. Conclusion: StroVac® does not only act via the adaptive but also by stimulating the innate immune system. This stimulation may help to build trained innate immunity against bacterial pathogens involved in recurrent urinary tract infections.

Exploiting fungi in bioremediation for cleaning-up emerging pollutants in aquatic ecosystems.

Aquatic pollution negatively affects water bodies, marine ecosystems, public health, and economy. Restoration of contaminated habitats has attracted global interest since protecting the health of marine ecosystems is crucial. Bioremediation is a cost-effective and eco-friendly way of transforming hazardous, resistant contaminants into environmentally benign products using diverse biological treatments. Because of their robust morphology and broad metabolic capabilities, fungi play an important role in bioremediation. This review summarizes the features employed by aquatic fungi for detoxification and subsequent bioremediation of different toxic and recalcitrant compounds in aquatic ecosystems. It also details how mycoremediation may convert chemically-suspended matters, microbial, nutritional, and oxygen-depleting aquatic contaminants into ecologically less hazardous products using multiple modes of action. Mycoremediation can also be considered in future research studies on aquatic, including marine, ecosystems as a possible tool for sustainable management, providing a foundation for selecting and utilizing fungi either independently or in microbial consortia.

Students' experiences and attitudes toward a newly developed simulation-based pharmaceutical care clinical rotation: A pre-post study design.

Purpose: This study aims to evaluate Doctor of Pharmacy (PharmD) students' experience with the newly developed simulation-based pharmaceutical care (PC) rotation by evaluating their knowledge and attitudes towards PC before and after the rotation. Methods: A self-administered questionnaire was distributed to sixth year PharmD students enrolled in the clinical training rotation "Comprehensive Pharmaceutical Care" during the 2020/2021 academic semesters at Jordan University of Science and Technology's (JUST) Faculty of Pharmacy. Questionnaires were distributed before and after completing four experiential training weeks and consisted of three sections. The first section collected students' demographic details while the second and third sections evaluated students' knowledge about, and attitudes toward PC, respectively. Descriptive statistics were used to describe and compare changes in students' knowledge and attitudes pre-and post-rotation. Results: A total of 106 valid questionnaires were completed with a response rate of 99.07%. The rates of correct answers increased after the rotation with median total knowledge score increasing from 8 to 10 (out of 13, P value < 0.001). Significant improvements in students' understanding of aspects relating to the concept and process of PC, and the role of clinical pharmacist in PC provision, were shown post the simulation-based clinical rotation. Similarly, their attitudes toward performing PC were either improved or emphasized. In contrast results also revealed that specific aspects of the rotation require further refinement, such as the comprehensiveness of the PC process and responsibilities in providing PC. Conclusions: PharmD students' understanding and attitudes toward PC were either improved or emphasized after the simulation-based PC rotation. This study highlights the value of simulation as a unique instructional technique that can assist educators to develop PC competencies for pharmacy students.

DNA Methylation Analysis Identifies Novel Epigenetic Loci in Dilated Murine Heart upon Exposure to Volume Overload.

Left ventricular (LV) dilatation, a prominent risk factor for heart failure (HF), precedes functional deterioration and is used to stratify patients at risk for arrhythmias and cardiac mortality. Aberrant DNA methylation contributes to maladaptive cardiac remodeling and HF progression following pressure overload and ischemic cardiac insults. However, no study has examined cardiac DNA methylation upon exposure to volume overload (VO) despite being relatively common among HF patients. We carried out global methylome analysis of LV harvested at a decompensated HF stage following exposure to VO induced by aortocaval shunt. VO resulted in pathological cardiac remodeling, characterized by massive LV dilatation and contractile dysfunction at 16 weeks after shunt. Although methylated DNA was not markedly altered globally, 25 differentially methylated promoter regions (DMRs) were identified in shunt vs. sham hearts (20 hypermethylated and 5 hypomethylated regions). The validated hypermethylated loci in Junctophilin-2 (Jph2), Signal peptidase complex subunit 3 (Spcs3), Vesicle-associated membrane protein-associated protein B (Vapb), and Inositol polyphosphate multikinase (Ipmk) were associated with the respective downregulated expression and were consistently observed in dilated LV early after shunt at 1 week after shunt, before functional deterioration starts to manifest. These hypermethylated loci were also detected peripherally in the blood of the shunt mice. Altogether, we have identified conserved DMRs that could be novel epigenetic biomarkers in dilated LV upon VO exposure.

Metabolomic Profiling in Patients with Different Hemodynamic Subtypes of Severe Aortic Valve Stenosis.

Severe aortic stenosis (AS) is a common pathological condition in an ageing population imposing significant morbidity and mortality. Based on distinct hemodynamic features, i.e., ejection fraction (EF), transvalvular gradient and stroke volume, four different AS subtypes can be distinguished: (i) normal EF and high gradient, (ii) reduced EF and high gradient, (iii) reduced EF and low gradient, and (iv) normal EF and low gradient. These subtypes differ with respect to pathophysiological mechanisms, cardiac remodeling, and prognosis. However, little is known about metabolic changes in these different hemodynamic conditions of AS. Thus, we carried out metabolomic analyses in serum samples of 40 AS patients (n = 10 per subtype) and 10 healthy blood donors (controls) using ultrahigh-performance liquid chromatography-tandem mass spectroscopy. A total of 1293 biochemicals could be identified. Principal component analysis revealed different metabolic profiles in all of the subgroups of AS (All-AS) vs. controls. Out of the determined biochemicals, 48% (n = 620) were altered in All-AS vs. controls (p < 0.05). In this regard, levels of various acylcarnitines (e.g., myristoylcarnitine, fold-change 1.85, p < 0.05), ketone bodies (e.g., 3-hydroxybutyrate, fold-change 11.14, p < 0.05) as well as sugar metabolites (e.g., glucose, fold-change 1.22, p < 0.05) were predominantly increased, whereas amino acids (e.g., leucine, fold-change 0.8, p < 0.05) were mainly reduced in All-AS. Interestingly, these changes appeared to be consistent amongst all AS subtypes. Distinct differences between AS subtypes were found for metabolites belonging to hemoglobin metabolism, diacylglycerols, and dihydrosphingomyelins. These findings indicate that relevant changes in substrate utilization appear to be consistent for different hemodynamic subtypes of AS and may therefore reflect common mechanisms during AS-induced heart failure. Additionally, distinct metabolites could be identified to significantly differ between certain AS subtypes. Future studies need to define their pathophysiological implications.

An ex vivo system for investigation of Plasmodium berghei invasion of the salivary gland of Anopheles stephensi mosquitoes.

Plasmodium sporozoites associated with the midgut and in the hemolymph of mosquitoes differ from sporozoites in the secretory cavities and ducts of the insects' salivary glands in their transcriptome, proteome, motility, and infectivity. Using an ex vivo Anopheles stephensi salivary gland culture system incorporating simple microfluidics and transgenic Plasmodium berghei with the fluorescent protein gene mCherry under the transcriptional control of the Pbuis4 promoter whose expression served as a proxy for parasite maturation, we observed rapid parasite maturation in the absence of salivary gland invasion. While in vivo Pbuis4::mCherry expression was only detectable in sporozoites within the salivary glands (mature parasites) as expected, the simple exposure of P. berghei sporozoites to dissected salivary glands led to rapid parasite maturation as indicated by mCherry expression. These results suggest that previous efforts to develop ex vivo and in vitro systems for investigating sporozoite interactions with mosquito salivary glands have likely been unsuccessful in part because the maturation of sporozoites leads to a loss in the ability to invade salivary glands.

A case study of a real-time internet of things system for site-specific potato crop management in El-Salhia Area-Egypt.

The site-specific management is the technology that considers the natural variability within the same field of factors related to crop growth to improve its management practices such that the agricultural treatments are varied for field's small production zones saving resources and environment, and improving crop quality and size. Since site-specific decisions are not far from the Fourth Industrial Revolution and the concept of processes automation, this work addresses improving the process of spatial variability analysis and thus supporting management decisions by developing a system-entitled EGYPADS-based on the Internet of Things and its enabling technologies. EGYPADS automates data collection, zones delineation according to their land suitability evaluation, and maps generation. The paper addresses a case study of potato crop in a specific area in Egypt, El-Salhia, in which eighty-five sites were chosen as main dataset for the modeling process during different stages of crop growth. Three management zones were recognized of the selected field based on the differentiation in their land suitability characteristics, representing about 5%, 65%, and 30% of the total area, respectively. The structure, screens, and services of EGYPADS are described in this paper. EGYPADS offered services include: management zones delineation using absolute and virtual coordinates, Land Suitability Assessment (LSA), data entry from field in real-time as well as from excel sheets, saving maps in suitable format for variable rate application, real-time and historical data processing, centralized management, and flexible formulation of events and related actions. The implementation of EGYPADS was verified. The system dynamically produces non-contiguous isobands, each representing a specific range of parameter values, and can be properly exported for use by other programs or smart machinery. It was proven that EGYPADS supports more than one land with different geometry, area, location, and number of nodes. EGYPADS was compared with the traditional LSA method, and was found to produce similar management zones.

Hemodynamic stress-induced cardiac remodelling is not modulated by ablation of phosphodiesterase 4D interacting protein.

Adrenergic stimulation in the heart activates the protein kinase A (PKA), which phosphorylates key proteins involved in intracellular Ca2+ handling. PKA is held in proximity to its substrates by protein scaffolds, the A kinase anchoring proteins (AKAPs). We have previously identified the transcript of phosphodiesterase 4D interacting protein (Pde4dip; also known as myomegalin), one of the sarcomeric AKAPs, as being differentially expressed following hemodynamic overload, a condition inducing hyperadrenergic state in the heart. Here, we addressed whether PDE4DIP is involved in the adverse cardiac remodelling following hemodynamic stress. Homozygous Pde4dip knockout (KO) mice, generated by CRISPR-Cas9 technology, and wild-type (WT) littermates were exposed to aortocaval shunt (shunt) or transthoracic aortic constriction (TAC) to induce hemodynamic volume overload (VO) or pressure overload (PO), respectively. The mortality, cardiac structure, function and pathological cardiac remodelling were followed up after hemodynamic injuries. The PDE4DIP protein level was markedly downregulated in volume-overloaded- but upregulated in pressure-overloaded-WT hearts. Following shunt or TAC, mortality rates were comparably increased in both genotypes. Twelve weeks after shunt or TAC, Pde4dip-KO animals showed a similar degree of cardiac hypertrophy, dilatation and dysfunction as WT mice. Cardiomyocyte hypertrophy, myocardial fibrosis, reactivation of cardiac stress genes and downregulation of ATPase, Ca2+ transporting, cardiac muscle, slow twitch 2 transcript did not differ between WT and Pde4dip-KO hearts following shunt or TAC. In summary, despite a differential expression of PDE4DIP protein in remodelled WT hearts, Pde4dip deficiency does not modulate adverse cardiac remodelling after hemodynamic VO or PO.

Heat shock protein A4 ablation leads to skeletal muscle myopathy associated with dysregulated autophagy and induced apoptosis.

BACKGROUND: Molecular chaperones assist protein folding, facilitate degradation of misfolded polypeptides, and thereby maintain protein homeostasis. Impaired chaperone activity leads to defective protein quality control that is implicated in multiple skeletal muscle diseases. The heat shock protein A4 (HSPA4) acts as a co-chaperone for HSP70. Previously, we showed that Hspa4 deletion causes impaired protein homeostasis in the heart. However, its functional role in skeletal muscle has not been explored. METHODS: We performed a comparative phenotypic and biochemical analyses of Hspa4 knockout (KO) mice with wild-type (WT) littermates. RESULTS: HSPA4 is markedly upregulated in regenerating WT muscle in vivo, and in differentiated myoblasts in vitro. Hspa4-KO mice are marked by growth retardation and increased variability in body weight, accompanied by 35% mortality rates during the peri-weaning period. The surviving Hspa4-KO mice experienced progressive skeletal muscle myopathy, characterized by increased number of muscle fibers with centralized nuclei, heterogeneous myofiber size distribution, inflammatory cell infiltrates and upregulation of embryonic and perinatal myosin heavy chain transcripts. Hspa4-KO muscles demonstrated an accumulation of autophagosome-associated proteins including microtubule associated protein1 light chain 3-II (LC3-II) and p62/sequestosome accompanied by increased number of TUNEL-positive nuclei. CONCLUSIONS: Our findings underscore the indispensable role of HSPA4 in maintenance of muscle integrity through contribution in skeletal muscle autophagy and apoptosis, which might provide a novel therapeutic strategy for skeletal muscle morbidities.

Different activation of MAPKs and Akt/GSK3ß after preload vs. afterload elevation.

AIMS: Pressure overload (PO) and volume overload (VO) lead to concentric or eccentric hypertrophy. Previously, we could show that activation of signalling cascades differ in in vivo mouse models. Activation of these signal cascades could either be induced by intrinsic load sensing or neuro-endocrine substances like catecholamines or the renin-angiotensin-aldosterone system. METHODS AND RESULTS: We therefore analysed the activation of classical cardiac signal pathways [mitogen-activated protein kinases (MAPKs) (ERK, p38, and JNK) and Akt-GSK3ß] in in vitro of mechanical overload (ejecting heart model, rabbit and human isolated muscle strips). Selective elevation of preload in vitro increased AKT and GSK3ß phosphorylation after 15 min in isolated rabbit muscles strips (AKT 49%, GSK3ß 26%, P < 0.05) and in mouse ejecting hearts (AKT 51%, GSK49%, P < 0.05), whereas phosphorylation of MAPKs was not influenced by increased preload. Selective elevation of afterload revealed an increase in ERK phosphorylation in the ejecting heart (43%, P < 0.05), but not in AKT, GSK3ß, and the other MAPKs. Elevation of preload and afterload in the ejecting heart induced a significant phosphorylation of ERK (95%, P < 0.001) and showed a moderate increased AKT (P = 0.14) and GSK3ß (P = 0.21) phosphorylation, which did not reach significance. Preload and afterload elevation in muscles strips from human failing hearts showed neither AKT nor ERK phosphorylation changes. CONCLUSIONS: Our data show that preload activates the AKT-GSK3ß and afterload the ERK pathway in vitro, indicating an intrinsic mechanism independent of endocrine signalling.

Upgrading the preparation of high-quality chitosan from Procambarus clarkii wastes over the traditional isolation of shrimp chitosan.

Crustacean waste is one of the most severe issues, posing significant environmental and health risks. This study aims to improve managing marine waste by isolating chitosan from Procambarus clarkii by devising a new methodology, incorporating technical steps, e.g., washing, decolorization and deacetylation under a reflexive condenser and dialysis purification. A comparison was made between the prepared P. clarkii chitosan and four types of shrimp chitosans: commercial, high, low, and nano. The obtained chitosan has a low molecular weight and viscosity compared to the commercial shrimp chitosan used in various applications. P. clarkii chitosan was prepared in high quality from a cheap source, as its color and quality were better than those of the commercial shrimp chitosan. The new methodology has successfully extracted chitosan from P. clarkii in a good quality and high purity, achieving 89% deacetylation, high solubility, high purity, and medium molecular weight. Analysis of the different chitosan samples with Fourier transform infrared spectroscopy (FTIR), atomic force microscopy, Raman spectrum referred indicated high similarity between the chitosan different types, regardless of its source. The 3D image of P. clarkii showed the distance between the highest and most profound points of extracted chitosan is 728.94 nm, revealing homogeneous, smooth surfaces, apparently free of pores and cracks. FTIR and Raman spectrum of P. clarkii indicated various functional groups, e.g., alcohol, amines, amides, and phenols. These active groups are responsible for about 60% of the antioxidant activity of that product. Evaluating the quality traits indicated the excellence of the chitosan prepared from P. clarkii, especially in color, viscosity, and antioxidant activity, nominating it for different food applications.

Preparation of Modified Chitosan-based Nanoparticles for Efficient Delivery of Doxorubicin and/or Cisplatin to Breast Cancer Cells.

PURPOSE: The aim is to develop a novel pH-responsive modified chitosan-based nanoparticles system for active loading of doxorubicin (DOX) and triggered intracellular release. METHODS: Nanoparticles were formed in an aqueous medium via ionic interaction between negatively charged chitosan derivative and positively charged DOX at neutral pH and then transformed in situ into cisplatin (CIS) cross-linked nanoparticles through cross-linking the formed micelles via chelation interaction between the negatively charged polymeric carrier and cisplatin. Nanoparticles were characterized in terms of particle size and zeta potential using DLS and TEM. Drug loading efficiency and encapsulation efficiency were determined based on the physio-chemical proprieties of the polymer and the amount of the cross-linking agent. In vitro release studies were performed using the dialysis method at different pHs. Finally, the cytotoxic effects of these nanoparticles were performed against the MCF-7 BrCA cell line under different pHs. RESULTS: The average particle size of polymer alone and DOX nanoparticles was 277.401 ± 13.50 nm and 290.20 ± 17.43 nm, respectively. The zeta potential was -14.6 ± 1.02 mV and -13.2 ± 0.55 mV, respectively, with a low polydispersity index. Drug loading and encapsulation deficiencies were determined, dependent on the amount of the cross-linking agent. In vitro release studies showed that the release of DOX from these nanoparticles was pH-dependent. Moreover, results showed that the cytotoxicity magnitude of DOX-loaded nanoparticles against MCF-7 BrCA cells was higher compared with free DOX. CONCLUSION: These novel pH-sensitive nanoparticles proved to be a promising Nano-drug delivery for tumor-targeted delivery of DOX.