Assessing sequence heterogeneity in Chlorellaceae DNA barcode markers for phylogenetic inference.

Phylogenetic inference is an important approach that allows the recovery of the evolutionary history and the origin of the Chlorellaceae species. Despite the species' potential for biofuel feedstock production, their high phenotypic plasticity and similar morphological structures among the species have muddled the taxonomy and identification of the Chlorellaceae species. This study aimed to decipher Chlorellaceae DNA barcode marker heterogeneity by examining the sequence divergence and genomic properties of 18S rRNA, ITS (ITS1-5.8S rRNA-ITS2-28S rRNA), and rbcL from 655 orthologous sequences of 64 species across 31 genera in the Chlorellaceae family. The study assessed the distinct evolutionary properties of the DNA markers that may have caused the discordance between individual trees in the phylogenetic inference using the Robinson-Foulds distance and the Shimodaira-Hasegawa test. Our findings suggest that using the supermatrix approach improves the congruency between trees by reducing stochastic error and increasing the confidence of the inferred Chlorellaceae phylogenetic tree. This study also found that the phylogenies inferred through the supermatrix approach might not always be well supported by all markers. The study highlights that assessing sequence heterogeneity prior to the phylogenetic inference could allow the approach to accommodate sequence evolutionary properties and support species identification from the most congruent phylogeny, which can better represent the evolution of Chlorellaceae species.

Effects of a combined progressive resistance training and functional electrical stimulation-evoked cycling exercise on lower limb muscle strength of individuals with incomplete spinal cord injury: A randomized controlled study.

Objectives: This study was conducted to investigate the effects of combined progressive resistance training (PRT) and functional electrical stimulation-evoked leg cycling exercise (FES-LCE) on isometric peak torque and muscle volume in individuals with incomplete spinal cord injury. Patients and methods: In the single-blind, randomized controlled trial performed between April 2015 and August 2016, 28 participants were randomized between two exercise interventions (FES-LCE+PRT and FES-LCE alone), and training was conducted over 12 weeks. The isometric muscle peak torque and muscle volume for both lower limbs were measured at the baseline and after 6 and 12 weeks. Linear mixed-model analysis of variance was performed to test the effects of FES-LCE+PRT versus FES-LCE on each outcome measure over time via an intention-to-treat analysis. Results: Twenty-three participants (18 males, 5 females; mean age: 33.4±9.7 years; range 21 to 50 years) completed study (10 in the FES-LCE+PRT group, and 13 in the FES-LCE group). The 12-week pre-and posttraining change for left hamstrings' muscle peak torque in the FES-LCE+PRT group (mean difference=4.5±7.9 Nm, 45% change, p<0.05) was consistently higher than that in the FES-LCE group (mean difference=2.4±10.3 Nm, 4% change; p<0.018). The improvement in the right quadriceps muscle's peak torque of the FES-LCE+PRT group (mean difference=19±7.6 Nm, 31% change, p<0.05) was more significant compared to the FES-LCE group. The left muscle volume showed a remarkable increase after 12 weeks in the FES-LCE+PRT group (mean difference=0.3±9.3 L, 7% change, p<0.05). Conclusion: The combination of PRT and FES-LCE was better in improving lower limb muscle strength and volume in chronic incomplete individuals with spinal cord injury.

Challenges in Implementing the WHO Hospital Readiness Checklist for the COVID-19 Pandemic in Indonesian Hospitals: A Qualitative Study.

Purpose: To assist hospitals in preparing for a surge of patients during the COVID-19 pandemic, the World Health Organization (WHO) published an assessment tool called the rapid hospital readiness checklist. This checklist has been used by numerous countries, including Indonesia. However, several technical issues were discovered, primarily as a result of the manual recording of the checklist on a spreadsheet. This study aimed to identify challenges related to the hospital readiness checklist that was implemented in Indonesia. Materials and Methods: This qualitative study used focus group discussions to collect data. The study was conducted in East Java, Indonesia, in October 2021, with the participation of nine organizations from the provinces of East Java and Bali. Data were thematically analyzed, and the findings were presented in a narrative format. Results: Hospital participants had experience in filling out a hospital readiness checklist every 3 months. Some challenges faced by the hospital was the need to manually enter data into the checklist, lack of coordination and communication, there were various perceptions in hospitals as there was no technical guide in completing the checklist, absence of feedback, and the data returning empty due to filling errors. Additionally, City X health office also identified challenges in monitoring and evaluating the hospital checklists. Conclusion: This study, which included both hospitals and the health office, described the challenges encountered in the assessment of the hospital readiness checklist. Both hospitals and DHO experienced some challenges with the current system. The checklist has the potential to evolve into a public reporting to improve efficiency and faster decision-making.

Implementation of Computerized Physician Order Entry in Primary Care: A Scoping Review.

PURPOSE: This scoping review aimed to assess the implementation and outcomes of computerized physician order entry (CPOE) in primary care. METHODS: A scoping review was carried out in accordance with the Joanna Briggs Institute's guidelines (JBI). The databases PubMed, CINAHL, Science Direct, and Google Scholar were all searched. The full text of each article was reviewed for eligibility after the title and abstract were evaluated. JBI data extraction were used to extract data. Donabedian's framework served as the foundation for the data discussion. RESULTS: Based on the inclusion criteria, seven studies were included. The studies' main goal in common was to analyze the outcome or impact of implementing CPOE systems in ambulatory or primary care settings. Several studies described the framework, current state of implementation, and evaluation or recommendation following CPOE system implementation. Many positive effects were felt by physicians or prescribers, pharmacists, patients, and primary care providers, with patient safety being the primary goal. CONCLUSION: Although this study discovered some issues and factors associated with CPOE implementation and adoption, such as infrastructure, workflow, level of engagement, and safety culture, CPOE has many positive outcomes for patients, physicians, and primary care. To improve CPOE adoption in healthcare, particularly primary care, more research into the structure, framework, and components of CPOE deployment is required.

Molecular and allergenic characterization of recombinant tropomyosin from mud crab Scylla olivacea.

BACKGROUND: Tropomyosin is a major allergen in crustaceans, including mud crab species, but its molecular and allergenic properties in Scylla olivacea are not well known. Thus, this study aimed to produce the recombinant tropomyosin protein from S. olivacea and subsequently investigate its IgE reactivity. METHODS AND RESULTS: The tropomyosin gene was cloned and expressed in the Escherichia coli system, followed by SDS-PAGE and immunoblotting test to identify the allergenic potential of the recombinant protein. The 855-base pair of tropomyosin gene produced was found to be 99.18% homologous to Scylla serrata. Its 284 amino acids matched the tropomyosin of crustaceans, arachnids, insects, and Klebsiella pneumoniae, ranging from 79.03 to 95.77%. The tropomyosin contained 89.44% alpha-helix folding with a tertiary structure of two-chain alpha-helical coiled-coil structures comprising a homodimer heptad chain. IPTG-induced histidine tagged-recombinant tropomyosin was purified at the size of 42 kDa and confirmed as tropomyosin using anti-tropomyosin monoclonal antibodies. The IgE binding of recombinant tropomyosin protein was reactive in 90.9% (20/22) of the sera from crab-allergic patients. CONCLUSIONS: This study has successfully produced an allergenic recombinant tropomyosin from S. olivacea. This recombinant tropomyosin may be used as a specific allergen for the diagnosis of allergy.

In vitro antibacterial activity of endophytic fungus Aspergillus flavus IBRL-C8 from Senna siamea against Gram-bacteria

Aims@#To evaluate the antibacterial efficacy of ethyl acetate extract of Aspergillus flavus IBRL-C8 against Gram-positive and Gram-negative bacteria.@*Methodology and results@#In this experiment, an endophytic fungus which identified as A. flavus IBRL-C8 was extracted using ethyl acetate and methanol, from Senna siamea, prior to in vitro antibacterial test on eight Gram-bacteria. The results were significantly more enunciated to the ethyl acetate extract since the Gram-bacteria signified 9.0 to 20.0 mm of inhibition zones on Muller Hinton Agar (MHA) during disc diffusion assay. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extract were ranged from 125-1000 µg/mL and 125-2000 µg/mL, respectively. Time-kill assay depicted the ethyl acetate extract of A. flavus IBRL-C8 exceptionally retarded methicillin-resistant Staphylococcus aureus (MRSA) and also manifested extended antibacterial activity. The maximum reduction in cell numbers occurred at 2MIC concentration (250 µg/mL) during the interval time of 16 h. The malformations noticed from microscopic observations where the transformation of structural annihilation from regular spherical morphology to non-spherical shape with an irregular surface and also disruption around the cell membrane when the MRSA treated with ethyl acetate extract of A. flavus IBRL-C8. @*Conclusion, significance and impact of study@#This study proposed the ethyl acetate extract of A. flavus IBRL-C8 as a potential antibacterial agent against MRSA infection, which can be useful in pharmaceutical application.

Inactivation of the catalytic subunit of cAMP-dependent protein kinase A causes delayed appressorium formation and reduced pathogenicity of Colletotrichum gloeosporioides.

The cyclic AMP- (cAMP-) dependent protein kinase A signaling pathway is one of the major signaling pathways responsible for regulation of the morphogenesis and pathogenesis of several pathogenic fungi. To evaluate the role of this pathway in the plant pathogenic fungus, Colletotrichum gloeosporioides, the gene encoding the catalytic subunit of cAMP-dependent protein kinase A, CgPKAC, was cloned, inactivated, and the mutant was analyzed. Analysis of the Cgpkac mutant generated via gene replacement showed that the mutants were able to form appressoria; however, their formation was delayed compared to the wild type. In addition, the mutant conidia underwent bipolar germination after appressoria formation, but no appressoria were generated from the second germ tube. The mutants also showed reduced ability to adhere to a hydrophobic surface and to degrade lipids localized in the appressoria. Based on the number of lesions produced during a pathogenicity test, the mutant's ability to cause disease in healthy mango fruits was reduced, which may be due to failure to penetrate into the fruit. These findings indicate that cAMP-dependent protein kinase A has an important role in regulating morphogenesis and is required for pathogenicity of C. gloeosporioides.