Miracle-dz/UniFAR-FTUs huggingface dataset:
Antioxidant metallothionein alleviates endoplasmic reticulum stress-induced myocardial apoptosis and contractile dysfunction

Abstract Aims. Endoplasmic reticulum (ER) stress exerts myocardial oxidative stress, apoptosis, and contractile anomalies, although the precise interplay between ER stress and apoptosis remains elusive. This study was designed to examine the impact of the cysteine-rich free radical scavenger metallothionein on ER stress-induced myocardial contractile defect and underlying mechanisms. Methods and results. Wild-type friendly virus B and transgenic mice with cardiac-specific overexpression of metallothionein were challenged with the ER stress inducer tunicamycin (1 mg/kg, intraperitoneal, 48 h) prior to the assessment of myocardial function, oxidative stress, and apoptosis. Our results revealed that tunicamycin promoted cardiac remodeling (enlarged left ventricular end systolic/diastolic diameters with little changes in left ventricular wall thickness), suppressed fractional shortening and cardiomyocyte contractile function, elevated resting Ca2+, decreased stimulated Ca2+ release, prolonged intracellular Ca2+ clearance, and downregulated sarco(endo)plasmic reticulum Ca2+-ATPase levels, the effects of which were negated by metallothionein. Treatment with tunicamycin caused cardiomyocyte mitochondrial injury, as evidenced by decreased mitochondrial membrane potential (∆Ѱm, assessed by JC-1 staining), the effect of which was negated by the antioxidant. Moreover, tunicamycin challenge dramatically facilitated myocardial apoptosis as manifested by increased Bax, caspase 9, and caspase 12 protein levels, as well as elevated caspase 3 activity. Interestingly, metallothionein transgene significantly alleviated tunicamycin-induced myocardial apoptosis. Conclusion. Taken together, our data favor a beneficial effect of metallothionein against ER stress-induced cardiac dysfunction possibly associated with attenuation of myocardial apoptosis.
Timing of Twinning, X-Inactivation and Sex Proportion at Birth

Abstract Twins are not a homogeneous group. According to zygosity and chorionicity essential differences exist. The lower sex proportion at birth is due to the monozygotic twins, especially the monoamnionic variety. X-inactivation patterns in monozygotic twin girls are totally symmetrical in monoamnionic pairs, almost symmetrical in monochorionic diamnionic pairs and can be very asymmetrical in the dichorionic variety.
Comparability analysis of anti-CD20 commercial (rituximab) and RNAi-mediated fucosylated antibodies by two LC-MS approaches

In developing biosimilar or biobetter products, comparability to the reference product is required to claim similar integrity or intended purpose. In this work, an anti-CD20 monoclonal antibody developed using RNA interference to decrease core fucosylation (RNAi-mediated) was comprehensively characterized by LC-MS and compared with the commercially-available anti-CD20 rituximab (MabThera®). As anticipated, < 30% core fucose was found within the RNAi-produced molecule (compared with > 90% in rituximab), and the reduction in fucose resulting in a significant improvement in FcγRΙΙΙa binding and antibody-dependent cell-mediated cytotoxicity. Two mutations, S258Y (fully mutated) and F174I/L (partially mutated), however, were detected in the production of the RNAi-mediated molecule. An alternative LC-MS approach using dimethyl labeling (i.e., 2CH2 for rituximab and 2CD2 for the RNAi-mediated molecule) was developed to additionally compare the two mAbs and confirm the full sequence with the two mutation sites. Furthermore, disulfide linkages were found to be the same for the two antibodies, with a small portion of unpaired cysteines in both products. Disulfides were correctly linked if the samples were prepared at low pH (i.e., enzymatic digestion by pepsin at pH 2); however, trace amounts of scrambling were found by trypsin digestion at pH 6.8, and this scrambling increased significantly at pH 8. Typical modifications, such as pyro-Glu formation at the N-terminus, K clipping at the C-terminus, oxidation at Met, and deamidation at Asn, were also detected with no significant differences between the two products. Using the LC-MS approaches for the comparability study, product integrity with critical structure information was revealed for confirmation of intended purpose (core fucosylation), identification of critical parameters (e.g., sample pH), and correction as needed (amino acid mutation).
The in vitro resistance of IgG2 to proteolytic attack concurs with a comparative paucity of autoantibodies against peptide analogs of the IgG2 hinge

The mammalian antibody repertoire comprises immunoglobulin (Ig) molecules of multiple isotypes and subclasses with varying functional properties. Among the four subclasses of the human IgG isotype, we found that IgG2 exhibits a particular resistance to human and bacterial proteases that readily cleave the IgG1 hinge region in vitro. Autoantibodies (IgGs) that recognize points of proteolytic cleavage in the IgG1 hinge are widespread in the healthy human population, suggesting that IgG1 fragmentation and the generation of cryptic antigens for host immune surveillance commonly occur in vivo. We previously reported that autoantibodies to cleaved IgG1s can restore Fc-mediated effector functions that are lost following proteolytic cleavage of the hinge. In contrast, it was not possible to demonstrate an analogous cohort of autoantibodies to IgG2 hinge epitope analogs, and there appeared to be no functional component in human serum with the ability to reconstitute Fc effector functions to a cell-bound IgG2 fragment. Thus, the results indicate that among the IgG subclasses, human IgG2 is uniquely resistant to a number of known pathological proteases and that autoimmune recognition to potential cleavage points in the IgG2 hinge appears to be absent in human circulation.
Gastroretentive drug delivery system of acyclovir-loaded alginate mucoadhesive microspheres: Formulation and evaluation

In the present study, mucoadhesive alginate microspheres of acyclovir were prepared to prolong the gastric residence time using a simple emulsification phase separation technique. The particle size of drug-loaded formulations was measured by SEM and the particle size distribution was determined using an optical microscope and mastersizer. The release profile of acyclovir from microspheres was examined in simulated gastric fluid (SGF pH 1.2). The particles were found to be discreet and spherical with the maximum particles of an average size (70.60 ± 2.44 µm). The results indicated that the mean particle size of the microspheres increased with an increase in the concentration of polymer and decreased with increase in stirring speed. The entrapment efficiency was found to be in the range of 51.42−80.46%. The concentration of the calcium chloride (% w/v) of 10% and drug–polymer ratio of 1:4 resulted in an increase in the entrapment efficiency and the extent of drug release. The optimized alginate microspheres were found to possess good mucoadhesion (66.42 ± 1.01%). The best fit model with the highest regression coefficient values (R2) was predicted by Peppas model (0.9813). In Gamma scintigraphy analysis, the section of GIT was critically analyzed and much differentiation was present at each time point after oral administration, which revealed that the optimized formulation demonstrated gastroretention in vivo for more than 4 h, which revealed that optimized formulation could be a good choice for gastroretentive systems.
An in vitro model for Pelger-Huët anomaly

The principal human blood granulocyte (neutrophil) possesses a lobulated and deformable nucleus, important to facilitate rapid egress from blood vessels as these cells migrate to sites of bacterial or fungal infection. This unusual nuclear shape is a product of elevated levels of an integral membrane protein of the nuclear envelope lamin B receptor (LBR) and of decreased amounts of lamin A/C. In humans, a genetic deficiency of LBR produces Pelger-Huët anomaly, resulting in blood neutrophils that exhibit hypolobulated nuclei with redistributed heterochromatin. Structural changes in nuclear architecture occur during granulopoiesis within bone marrow. The exact mechanisms of this nuclear shape change and of heterochromatin redistribution remain largely unknown. As a tool to facilitate analysis of these mechanisms, a stable LBR knockdown subline of HL-60 cells was established. During in vitro granulopoiesis induced with retinoic acid, the LBR knockdown cells retain an ovoid shaped nucleus with reduced levels of lamin A/C; whereas, the parent cells develop lobulated nuclei. Macrophage forms induced in LBR knockdown cells by in vitro treatment with phorbol ester were indistinguishable from the parent cells, comparing both nuclear shape and attached cell morphology. The capability of differentiation of LBR knockdown HL-60 cells should facilitate a detailed analysis of the molecular relationship between LBR levels, granulocyte nuclear shape and heterochromatin distribution.
Development, in vitro and in vivo characterization of Eudragit RL 100 nanoparticles for improved ocular bioavailability of acetazolamide

Abstract Glaucoma is characterized by increased intra ocular pressure (IOP) which results in blindness if left untreated. Acetazolamide (ACZ) is used to treat glaucoma since long back. Since it is a Class IV drug [According to Biopharmaceutics Classification System (BCS)], so its topical delivery results in poor ocular bioavailability. Objective of the present study is to increase the topical ocular bioavailability and to sustain the release of drug for longer time. ACZ-loaded Eudragit® RL 100 nanoparticle suspension (ACZ-E-NPs) was prepared by the nanoprecipitation method. Ratio of organic to aqueous phase and composition of organic phase were altered to get the best formulation. Formulations prepared with acetone and methanol as organic phase were smallest in size. EE was in the range of 57.8% to 68.5%. According to drug release study almost all the formulations released 80% of drug in 8 h duration. The kinetics of drug release showed that the drug release pattern followed Higuchi’s model (highest R2 values) and further it was fitted to the Korsemeyer–Peppas model, which showed the release was as per Fickian diffusion. IOP lowering effects of plain drug solution and ACZ-E-NPs were compared in adult male albino rabbits with a Riester Tonometer. The data revealed that the ACZ-E-NPs lower the IOP for longer time and of higher magnitude also. The difference was significant (p < 0.001). Short-term stability study showed that none of the formulations was having remarked difference in their physicochemical properties after 6 months of storage at various temperatures.
The Use of Midazolam in Paediatric Dentistry: A Review of the Literature

Evidence Acquisition: To conduct this review of the literature, Pub-Med and Scopus were searched using the following terms: midazolam, oral, transmucosal, intravenous, intramuscular, rectal, premedication, paediatric dentistry, general anaesthesia. Fourty-five published papers relating to the use of midazolam in paediatric dentistry were shortlisted out of an initial 124 publications and assessed according to the “critical appraisal skills programme” criteria. Randomized controlled trials, non- randomized studies, clinical trials and case reports were reviewed with regard to midazolam's indications, contraindications, drug interactions and various administration routes when used as a single sedative agents. Research and reviews based on the single-drug use of midazolam in children with adequate sample size as well as clinical guidelines were included. However, those conducted on adult populations, using poly-pharmacy for sedation and clinical investigations with flaws or non-significant results were excluded. Conclusions: Midazolam could be used as a safe and effective drug for conscious sedation, general anaesthetic premedication and the treatment of seizures during dental treatment. However, further research on paediatric patients would be beneficial.
Directed assembly of cell-laden hydrogels for engineering functional tissues

Tissue engineering aims to develop functionalized tissues for organ replacement or restoration. Biodegradable scaffolds have been used in tissue engineering to support cell growth and maintain mechanical and biological properties of tissue constructs. Ideally cells on these scaffolds adhere, proliferate, and deposit matrix at a rate that is consistent with scaffold degradation. However, the cellular rearrangement within these scaffolds often does not recapitulate the architecture of the native tissues. Directed assembly of tissue-like structures is an attractive alternative to scaffold-based approach for tissue engineering which potentially can build tissue constructs with biomimetic architecture and function. In directed assembly, shape-controlled microstructures are fabricated in which organized structures of different cell types can be used as tissue building blocks. To fabricate tissue building blocks, hydrogels are commonly used as biomaterials for cell encapsulation to mimic the matrix in vivo. The hydrogel-based tissue building blocks can be arranged in pre-defined architectures by various directed tissue assembly techniques. In this paper, recent advances in directed assembly-based tissue engineering are summarized as an emerging alternative to meet challenges associated with scaffold-based tissue engineering and future directions are addressed.
Separate mechanisms act concurrently to shed and release the prion protein from the cell

The cellular prion protein (PrPC) is attached to the cell membrane via its glycosylphosphatidylinositol (GPI)-anchor and is constitutively shed into the extracellular space. Here, three different mechanisms are presented that concurrently shed PrPC from the cell. The fast α-cleavage released a N-terminal fragment (N1) into the medium and the extreme C-terminal cleavage shed soluble full-length (FL-S) PrP and C-terminally cleaved (C1-S) fragments outside the cell. Also, a slow exosomal release of full-length (FL) and C1-fragment (C1) was demonstrated. The three separate mechanisms acting simultaneously, but with different kinetics, have to be taken into consideration when elucidating functional roles of PrPC and also when processing of PrPC is considered as a target for intervention in prion diseases. Further, in this study it was shown that metalloprotease inhibitors affected the extreme C-terminal cleavage and shedding of PrPC. The metalloprotease inhibitors did not influence the α-cleavage or the exosomal release. Taken together, these results are important for understanding the different mechanisms acting in parallel in the shedding and cleavage of PrPC.
Quality of Care, Nurse Staffing, and Patient Outcomes

Widespread reports of declining levels of quality of care and patient safety in hospitals, an escalation in calls for legislation mandating minimum nurse staffing ratios, and growing levels of nurse burnout and a looming nursing shortage have focused attention on the working conditions that nurses face and their implications for patient outcomes. This article reports on the preliminary results of an international study on how nurse staffing levels and the nursing practice environment affect the quality of care and patient outcomes in hospitals. Surveys of staff nurses working in acute care hospitals in Pennsylvania reveal that one out of every five staff nurses reported the quality of care on their unit as fair or poor. Workload played a role in these quality assessments, but it was the consequences of workload, such as the reports of unfinished nursing at the end of the last shift and the frequency of adverse events among patients, that played a much more prominent role.
Emerging infections and bioterrorism emergencies: Where do we go from here?

Emerging infectious disease outbreaks and bioterrorism attacks warrant urgent public health and medical responses. Response plans for these events may include use of medications and vaccines for which the effects on pregnant women and fetuses are unknown. Recent experiences with outbreaks of severe acute respiratory syndrome, monkey pox and anthrax, as well as response planning for bioterrorism and pandemic influenza, illustrate the challenges of making recommendations about medical interventions for victims. Experience with bioterrorism attacks (anthrax), viroterrorism attacks (arena viruses) and emergency response preparedness (smallpox vaccination) has been gained. Understanding the physiology of the body, the factors that influence the teratogenic potential of medications and vaccines and the infection control measures that may stop an outbreak will aid planners in making recommendations for care during large-scale infectious disease emergencies. Key words: Bioterrorism, emerging infections, emergence preparedness, pathogens, viroterrorism.
Potential c-myc Antisense Oligonucleotide Carriers: PCl/PEG/PEI and PLL/PEG/PEI

Abstract: In this work, positively charged, micelle-forming polymers were synthesized and used as a model vector to deliver antisense oligodeoxynucleotide (ASODN) into melanoma cells. Polymers and polymer/ASODN complexes were characterized by DLS according to size, charge, and critical micelle concentration. Nanosize and spherical complexes were observed by AFM. Complexes did not reveal significant toxicity to melanoma cells. Antiproliferative effect of the complexes was observed by immunocytochemical staining and estimated as 56.8% with N/P:9. High amount of apoptosis and very small amount of necrosis were estimated. According to the results, these positively charged polymers forming micelle-like structures seem promising as ASODN carriers.
Influence of health literacy on outcomes using telehomecare technology: A systematic review

Objective: To conduct a systematic review of the literature describing the interaction between the use of telehomecare technology and level of health literacy among chronic patients. The aim of the review was both to explore whether and how level of health literacy affects the ability to use telehomecare technology and, additionally, whether and how the use of telehomecare technology influences the level of health literacy. Design: Systematic literature review. Method: Four relevant studies, reported in five papers, were identified and evaluated according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The studies include a total of 1,120 participants in the telehomecare intervention groups and 617 participants in the control groups. The following data were extracted from the four included studies: author, year, country, number of patients, type of patients, time horizon, health literacy screening tool, nature of the intervention and description of the compared alternatives, for example, usual care. Results: The studies report conflicting observations, and a synthesis of their results therefore provides no clear picture of the interaction between health literacy and the use of telehomecare technology. Conclusion: Exploring the interaction between health literacy and the use of telehomecare was not a primary objective in any of the four included studies. Further research is needed to provide a better picture of the interaction between health literacy and the use of telehomecare technology.
Self-treatment for malaria: the evidence and methodological issues.

Malaria remains an important cause of death especially in sub-Saharan Africa. Self-treatment with anti-malarial drugs is a common practice that raises important issues for policy-makers. A number of important questions concerning factors related to self-treatment adequacy of self-treatment and the role of self-treatment in malaria mortality remain unanswered. Although there are some common patterns there is considerable diversity in treatment practices even within a single country. Social science research on malaria treatment needs to move beyond description to evaluation of interventions. This will require a greater degree of methodological rigor and more attention to the generation of data that can be compared across time periods and studies. Definitions of malaria cases and the role of local disease categories in identifying cases need to be more explicit. Illnesses should be classified by severity using measures of perceived severity as well as biomedical signs of severity. Each treatment step should be considered in terms of four levels of analysis: who provided the treatment or advice what the treatment was where it was obtained and when it was taken in relationship to onset of illness. (authors)
Integration of Telemedicine Practice Into Correctional Medicine: An Evolving Standard

The application of telemedicine in corrections has grown rapidly. A comprehensive literature review was conducted to develop a status report on the value that telemedicine has added to health care delivery in the prison setting. When most state and federal correctional telemedicine projects started, operation costs were high and applications were limited. However, with technologies increasingly affordable, access to specialized consultations for prisoners expanded greatly. Further, many legal barriers to service have been resolved. The number of health complaints that result in trips outside of a secure facility has dropped significantly. Each avoided trip saves hundreds of dollars in transport and security expenses, which has been the best indicator of success. Published material supports the value of telemedicine as a highly effective tool in health care delivery without compromise to patient safety or confidentiality. In fact, telemedi-cine can increase quality of care due to interaction between specialist and caregiver, as well as greater access to specialty care.
CCA1 alternative splicing as a way of linking the circadian clock to temperature response in Arabidopsis

Most living organisms on the earth have the circadian clock to synchronize their biochemical processes and physiological activities with environmental changes to optimize their propagation and survival. CIRCADIAN CLOCK-ASSOCIATED1 (CCA1) is one of the core clock components in Arabidopsis. Notably, it is also associated with cold acclimation. However, it is largely unknown how CCA1 activity is modulated by low temperatures. We found that the CCA1 activity is self-regulated by a splice variant CCA1β and the CCA1β production is modulated by low temperatures, linking the circadian clock with cold acclimation. CCA1β competitively inhibits the activities of functional CCA1α and LATE ELONGATED HYPOCOTYL (LHY) transcription factors by forming nonfunctional CCA1α-CCA1β and LHY-CCA1β heterodimers. Consequently, CCA1β-overexpressing plants (35S:CCA1β) exhibit shortened circadian periods as observed in cca1 lhy double mutants. In addition, elongated hypocotyls and petioles and delayed flowering of CCA1α-overexpressing plants (35S:CCA1α) were rescued by coexpression of CCA1β. Interestingly, low temperatures suppress CCA1 alternative splicing and thus derepress the CCA1α activity in inducing cold tolerance. These observations indicate that a cold-responsive self-regulatory circuit of CCA1 plays a role in plant responses to low temperatures.