Purification of andibenin and a chromanone analogue from rhizospheric Aspergillus flavus and soil-borne Penicillium notatum exhibiting cytotoxic and antibacterial properties.

The discovery of bioactive compounds from fungal natural sources holds immense potential for the development of novel therapeutics. The present study investigates the extracts of soil-borne Penicillium notatum and rhizosphere-inhabiting Aspergillus flavus for their antibacterial, antifungal, and cytotoxic potential. Additionally, two compounds were purified using chromatographic and spectroscopic techniques. The results demonstrated that the ethyl acetate fraction of A. flavus exhibited prominent cytotoxic activity against Artemia salina, whereas the ethyl acetate fraction of P. notatum displayed promising antibacterial potential. At dose concentrations of 10, 100, and 1000 µg mL-1, the ethyl acetate fraction of A. flavus showed mortality percentages of 7.6%, 66.4%, and 90%, respectively. The ethyl acetate fraction of P. notatum extract exhibited significant antibacterial activity, forming inhibition zones measuring 41, 38, 34, 34, and 30 mm against B. subtilis, S. flexneri, E. coli, K. pneumoniae, and S. aureus, respectively, at 1000 µg mL-1. At this concentration, inhibition zones of 28, 27, and 15 mm were recorded for P. vulgaris, S. typhi, and X. oryzae. Using bioassay-guided approach, one compound each was purified from the fungal extracts. The initial purification involved mass spectroscopic analysis, followed by structural elucidation using 500 MHz nuclear magnetic resonance (NMR) spectroscopy. Compound 1, derived from A. flavus, was identified as ethyl 2-hydroxy-5,6-dimethyl-4-oxocyclohex-2-ene-1-carboxylate, with a mass of 212, whereas compound 2, isolated from P. notatum, was identified as 3-amino-2-(cyclopenta-2,4-dien-1-ylamino)-8-methoxy-4H-chromen-4-one, with an exact mass of 270. Based on bioassay results, compound 1 was subjected to brine shrimp lethality assay and compound 2 was tested for its antibacterial potential. Compound 1 exhibited 30% lethality against brine shrimp larvae at a concentration of 100 µg mL-1, whereas at 1000 µg mL-1 the mortality increased to 70%. Compound 2 displayed notable antibacterial potential, forming inhibition zones of 30, 24, 19, and 12 mm against S. aureus, E. coli, B. subtilis, and S. flexneri, respectively. In comparison, the standard antibiotic tetracycline produced inhibition zones of 18, 18, 15, and 10 mm against the respective bacterial strains at the same concentration.

Fractional epidemic model of coronavirus disease with vaccination and crowding effects.

Most of the countries in the world are affected by the coronavirus epidemic that put people in danger, with many infected cases and deaths. The crowding factor plays a significant role in the transmission of coronavirus disease. On the other hand, the vaccines of the covid-19 played a decisive role in the control of coronavirus infection. In this paper, a fractional order epidemic model (SIVR) of coronavirus disease is proposed by considering the effects of crowding and vaccination because the transmission of this infection is highly influenced by these two factors. The nonlinear incidence rate with the inclusion of these effects is a better approach to understand and analyse the dynamics of the model. The positivity and boundedness of the fractional order model is ensured by applying some standard results of Mittag Leffler function and Laplace transformation. The equilibrium points are described analytically. The existence and uniqueness of the non-integer order model is also confirmed by using results of the fixed-point theory. Stability analysis is carried out for the system at both the steady states by using Jacobian matrix theory, Routh-Hurwitz criterion and Volterra-type Lyapunov functions. Basic reproductive number is calculated by using next generation matrix. It is verified that disease-free equilibrium is locally asymptotically stable if R 0 < 1 and endemic equilibrium is locally asymptotically stable if R 0 > 1 . Moreover, the disease-free equilibrium is globally asymptotically stable if R 0 < 1 and endemic equilibrium is globally asymptotically stable if R 0 > 1 . The non-standard finite difference (NSFD) scheme is developed to approximate the solutions of the system. The simulated graphs are presented to show the key features of the NSFD approach. It is proved that non-standard finite difference approach preserves the positivity and boundedness properties of model. The simulated graphs show that the implementation of control strategies reduced the infected population and increase the recovered population. The impact of fractional order parameter α is described by the graphical templates. The future trends of the virus transmission are predicted under some control measures. The current work will be a value addition in the literature. The article is closed by some useful concluding remarks.

Evaluation of a medical after-care intervention among deselected elite male Academy football players: a pilot study.

Objective: This pilot study aimed to evaluate the perception and satisfaction of deselected male Academy professional footballers towards a medical after-care intervention. Methods: A quasiexperimental study design, with deselected players (aged≥18 years) at a single Premier League Academy during the 2022/2023 season, were invited to participate. The intervention included individualised health recommendations, key medical information and signposting to key support resources. Participants' perceptions and satisfaction were assessed through an electronic survey. Descriptive analyses (mean, SD, frequency and/or range) were performed for all variables. Results: Twelve out of 15 eligible participants (80% response rate) provided informed consent and completed the survey (mean age: 19.5 years). All (100%) of the participants were satisfied with receiving the medical information. Ten out of 12 (83.3%) participants agreed that all Academy players should receive this medical intervention on deselection. Nine (75%) players felt more prepared for the next steps in their careers due to the medical information shared with them. Conclusion: Deselected male Academy footballers expressed high satisfaction with an individualised medical intervention which shared key health information and signposted them to important resources (eg, mental health). Future studies across multiple clubs should explore the broader impact of this intervention among deselected male and female Academy footballers. Football clubs should consider integrating a medical after-care process for deselected players as part of routine care.

Unveiling the Potential of Ambient Air Annealing for Highly Efficient Inorganic CsPbI3 Perovskite Solar Cells.

Here, we report a detailed surface analysis of dry- and ambient air-annealed CsPbI3 films and their subsequent modified interfaces in perovskite solar cells. We revealed that annealing in ambient air does not adversely affect the optoelectronic properties of the semiconducting film; instead, ambient air-annealed samples undergo a surface modification, causing an enhancement of band bending, as determined by hard X-ray photoelectron spectroscopy measurements. We observe interface charge carrier dynamics changes, improving the charge carrier extraction in CsPbI3 perovskite solar cells. Optical spectroscopic measurements show that trap state density is decreased due to ambient air annealing. As a result, air-annealed CsPbI3-based n-i-p structure devices achieved a 19.8% power conversion efficiency with a 1.23 V open circuit voltage.

Phosphate-Solubilizing Bacillus sp. Modulate Soil Exoenzyme Activities and Improve Wheat Growth.

Phosphorus (P) is a vital mineral nutrient in agriculture and its deficiency results in reduced growth, yield, and grain quality in cereals. Much of the applied P in agriculture becomes fixed in soils, limiting its accessibility to plants. Thus, investigating sustainable strategies to release fixed P resources and enhance plant uptake is crucial. This study explored how plant-associated bacteria employ phosphate solubilizing mechanisms to improve P availability. The growth patterns of four bacterial strains, namely Bacillus subtilis ZE15 and ZR3, along with Bacillus megaterium ZE32 and ZR19, were examined in Pikovskaya's broth culture with and without the addition of insoluble phosphorus (P). In the absence of P amendment, most strains reached a stationary growth phase by the fourth day. However, their responses diverged when exposed to P-amended media. Particularly, ZE15 demonstrated the highest P solubilization capability, achieving up to 130 µg mL-1 solubilization in vitro. All strains produced organic acids in Pikovskaya's broth culture. A comparison of the influence of Ca3(PO4)2 revealed significantly greater organic acid quantities in the presence of insoluble P. Notably, strain ZE15 exhibited the highest phosphate esterase activity (3.65 nmol g-1 dry matter), while strain ZE32 showed the highest ß-D glucosidase activity (2.81 nmol g-1 dry matter) in the presence of insoluble P. The ability of Bacillus species to solubilize P in combination with increased exoenzyme activity in the rhizosphere could be used in future studies to support P uptake through enhanced solubilization and mineralization.

Improved synthesis and evaluation of preclinical pharmacodynamic parameters of a new monocyclic ß-lactam DPI-2016.

Monocyclic ß-lactams are stable to a number of ß-lactamases and are the focus of researchers for the development of antibacterial drugs, particularly against Enterobacterales. We recently synthesized and reported the bactericidal activity of diverse series of aztreonam appended with amidine moieties as siderophores. One of the derivatives exhibiting the highest MIC value in vitro was selected for further preclinical studies. The compound DPI-2016 was reassessed for its synthetic routes and methods that were improved to find the maximum final yields aimed at large-scale synthesis. In addition, the results of the pharmacological studies were determined with reference to aztreonam. It has been found that the compound DPI-2016 showed comparable or slightly improved ADMET as well as pharmacokinetic parameters to aztreonam. It is estimated that the compound could be a potential lead for further clinical evaluation.

Application of relative importance metrics for CMIP6 models selection in projecting basin-scale rainfall over Johor River basin, Malaysia.

The most recent set of General Circulation Models (GCMs) derived from the Coupled Model Intercomparison Project Phase 6 (CMIP6) was used in this work to analyse the spatiotemporal patterns of future rainfall distribution across the Johor River Basin (JRB) in Malaysia. A group of 23 GCMs were chosen for comparative assessment in simulating basin-scale rainfall based on daily rainfall from the historical period of the Climate Hazards Group InfraRed Precipitation with Station Data (CHIRPS). The methodological novelty of this study lies in the application of relative importance metrics (RIM) to rank and select historical GCM simulations for reproducing rainfall at 109 CHIRPS grid points within the JRB. In order to choose the top GCMs, the rankings given by RIM were aggregated using the compromise programming index (CPI) and Jenks optimised classification (JOC). It was found that ACCESS-ESM1-5 and CMCC-ESM2 were ranked the highest in most of the grid. The final GCM was then bias-corrected using the linear scaling method before being ensemble based on the Bayesian model averaging (BMA) technique. The spatiotemporal assessment of the ensemble model for the different months over the near-future period 2021-2060 and far-future period 2061-2100 was compared with those under Shared Socioeconomic Pathways (SSPs), namely, SSP1-2.6, SSP2-4.5, SSP3-7.0, and SSP5-8.5. Heterogeneous changes in rainfall were projected across the JRB, with both increasing and decreasing trends. In the near-future and far-future scenarios, higher rainfall was projected for December, indicating an elevated risk of flooding during the end of the North East monsoon (NEM). Conversely, August showed a decreasing trend in rainfall, implying an increasing risk of severe drought. The findings of this study provide valuable insights for effective water resource management and climate change adaptation in the region.

Genetic Diversity, Evolutionary Dynamics, and Ongoing Spread of Pedilanthus Leaf Curl Virus.

Pedilanthus leaf curl virus (PeLCV) is a monopartite begomovirus (family Geminiviridae) discovered just a few decades ago. Since then, it has become a widely encountered virus, with reports from ca. 25 plant species across Pakistan and India, indicative of its notable evolutionary success. Viruses mutate at such a swift rate that their ecological and evolutionary behaviors are inextricably linked, and all of these behaviors are imprinted on their genomes as genetic diversity. So, all these imprints can be mapped by computational methods. This study was designed to map the sequence variation dynamics, genetic heterogeneity, regional diversity, phylogeny, and recombination events imprinted on the PeLCV genome. Phylogenetic and network analysis grouped the full-length genome sequences of 52 PeLCV isolates into 7 major clades, displaying some regional delineation but lacking host-specific demarcation. The progenitor of PeLCV was found to have originated in Multan, Pakistan, in 1977, from where it spread concurrently to India and various regions of Pakistan. A high proportion of recombination events, distributed unevenly throughout the genome and involving both inter- and intraspecies recombinants, were inferred. The findings of this study highlight that the PeLCV population is expanding under a high degree of genetic diversity (π = 0.073%), a high rate of mean nucleotide substitution (1.54 × 10-3), demographic selection, and a high rate of recombination. This sets PeLCV apart as a distinctive begomovirus among other begomoviruses. These factors could further exacerbate the PeLCV divergence and adaptation to new hosts. The insights of this study that pinpoint the emergence of PeLCV are outlined.

Ethno-Dentistry of Medicinal Plants Used in North Waziristan, Pakistan.

OBJECTIVE: This study aimed at recording therapeutic plant species used by inhabitants to treat dental disorders in the district of North Waziristan, Pakistan. The indigenous people of the studied area are dependent on medicinal plants for their basic health care needs including dental care. METHODS: Ethnomedicinal data were collected using a semi-structured questionnaires, and in addition 130 local informants were interviewed. The collected data were evaluated using various quantitative indices, including use value (UV), relative frequency of citation (RFC), fidelity level (FL%), and Jaccard Index (JI). RESULTS: A total of 69 plants belonging to 48 plant families used in dental disorders were identified. The Lamiaceae was the leading family that shared 7 species, followed by Solanaceae (4 spp).The dominant life form used was herbs (47.83%), folowed by leaves (43.90%) in preparing remedies for different dental disorders. Decoction was the most common mode of preparation (34.21%), followed by pastes (21.05%). The highest RFC (0.36) was reported for Bergenia ciliata, followed by Salvadora oleoides (0.35). The majority of the plants (36 spp) were utilised as herbal medicine to treat toothache, followed by 13 species for periodontal (gum) infections, 11 species used for teeth cleaning, and 9 species for halitosis (bad breath). CONCLUSIONS: This study is the first-ever record of ethnomedicinal applications for the treatment of dental diseases from Pakistan. Some of the forgeoing hebal medications should be further evalauted for the development of pahrmaceutical bio-products for the treatment of dental disorders.

Astrocyte and L-lactate in the anterior cingulate cortex modulate schema memory and neuronal mitochondrial biogenesis.

Astrocyte-derived L-lactate was shown to confer beneficial effects on synaptic plasticity and cognitive functions. However, how astrocytic Gi signaling in the anterior cingulate cortex (ACC) modulates L-lactate levels and schema memory is not clear. Here, using chemogenetic approach and well-established behavioral paradigm, we demonstrate that astrocytic Gi pathway activation in the ACC causes significant impairments in flavor-place paired associates (PAs) learning, schema formation, and PA memory retrieval in rats. It also impairs new PA learning even if a prior associative schema exists. These impairments are mediated by decreased L-lactate in the ACC due to astrocytic Gi activation. Concurrent exogenous L-lactate administration bilaterally into the ACC rescues these impairments. Furthermore, we show that the impaired schema memory formation is associated with a decreased neuronal mitochondrial biogenesis caused by decreased L-lactate level in the ACC upon astrocytic Gi activation. Our study also reveals that L-lactate-mediated mitochondrial biogenesis is dependent on monocarboxylate transporter 2 (MCT2) and NMDA receptor activity - discovering a previously unrecognized signaling role of L-lactate. These findings expand our understanding of the role of astrocytes and L-lactate in the brain functions.

Tamoxifen-Loaded Eudragit Nanoparticles: Quality by Design Approach for Optimization of Nanoparticles as Delivery System.

Nanoparticles have numerous applications as drug carriers in drug delivery. The aim of the study was to produce tamoxifen nanoparticles with a defined size and higher encapsulation for efficient tissue uptake with controlled drug release. The quality by design approach was utilized to produce tamoxifen-loaded Eudragit nanoparticles by identifying the significant process variables using the nanoprecipitation method. The process variables (amount of drug, polymer, and surfactant) were altered to analyze the influence on particle size (PS), % encapsulation efficiency (EE). The results showed that the drug and polymer individually as well as collectively have an impact on PS, while the surfactant has no impact on the PS. The %EE was influenced by the surfactant individually and in interaction with the drug. The linear regression model was endorsed to fit the data showing high R2 values (PS, 0.9146, %EE, 0.9070) and low p values (PS, 0.0004, EE, 0.0005). The PS and EE were confirmed to be 178 nm and 90%, respectively. The nanoparticles were of spherical shape, as confirmed by SEM and TEM. The FTIR confirmed the absence of any incompatibility among the ingredients. The TGA confirmed that the NPs were thermally stable. The in vitro release predicted that the drug release followed Higuchi model.

Interface Modification for Energy Level Alignment and Charge Extraction in CsPbI3 Perovskite Solar Cells.

In perovskite solar cells (PSCs) energy level alignment and charge extraction at the interfaces are the essential factors directly affecting the device performance. In this work, we present a modified interface between all-inorganic CsPbI3 perovskite and its hole-selective contact (spiro-OMeTAD), realized by the dipole molecule trioctylphosphine oxide (TOPO), to align the energy levels. On a passivated perovskite film, with n-octylammonium iodide (OAI), we created an upward surface band-bending at the interface by TOPO treatment. This improved interface by the dipole molecule induces a better energy level alignment and enhances the charge extraction of holes from the perovskite layer to the hole transport material. Consequently, a Voc of 1.2 V and a high-power conversion efficiency (PCE) of over 19% were achieved for inorganic CsPbI3 perovskite solar cells. Further, to demonstrate the effect of the TOPO dipole molecule, we present a layer-by-layer charge extraction study by a transient surface photovoltage (trSPV) technique accomplished by a charge transport simulation.

Purification and structure elucidation of Penicillium chrysogenum derived antifungal compound with potential anti-Candida property: in silico and in vitro evidence.

Following preliminary bioactivity testing, the fungal strain identified as Penicillium crysogenum was cultured in a modified Czapec Yeast Broth medium (CYB) for the production of antifungal compounds. Several chromatographic techniques including HPLC were used to purify the fungal metabolites from the crude extract. The mass determination of the purified compound was performed using Water's LCMS system while the structure of the compound was elucidated using 400 and 500 Varian NMR machines. The chemical name of the purified compound is (2 R, 4S) -2, 4-dimethyl-4-((E)-2-((3S, 4S)-2, 4, 5-trihydroxy-3-methoxy-4-phenyl-1, 2, 3, 4-tetrahydroquinolin-6-yl) vinyl) cyclohexanone with the chemical formula C26H31NO5 and exact mass of 437.2. Molecular docking predicted compound docking score with dihydrofolate reductase enzyme and lanosterol 14α-demethylase enzyme as -8.1 kcal/mol and -9.8 kcal/mol respectively. Further, the compounds showed stable binding mode with the enzymes and reported robust binding energies. After insilico analysis, the compound with mass 437 was tested for its antifungal potential in vitro against two pathogenic yeast species (i.e. Candida albicans and Candida glaberata) using the agar tube diffusion method. Using sterile di-methyl sulfoxide (DMSO) the compound was prepared in four dose concentrations (100, 250, 500, 1000 µg mL-1) and mixed with autoclaved semisolid Potato Dextrose Agar (PDA) medium in screw-capped test tubes labelled with the corresponding dose concentration. The fungal strains were inoculated on this medium and linear growth inhibition of the fungal strains was calculated using fluconazole as the control drug. The results from in vitro experiments were encouraging as at concentrations of 500 and 1000 µg mL-1 the compound inhibited the growth of C. albicans by 17% and 38% while 19% and 41% inhibition were recorded against C. glaberata. The compound showed antifungal activity in silico and in vitro against both the Candida species and can act as a potent antifungal candidate in the future upon further investigation.Communicated by Ramaswamy H. Sarma.

Vegetation-environment interactions: plant species distribution and community assembly in mixed coniferous forests of Northwestern Himalayas.

One of the main goals of ecological studies is to disentangle the dynamics that underlie the spatiotemporal distribution of biodiversity and further functions of the ecosystem. However, due to many ecological and geopolitical reasons, many remote areas with high plant species diversity have not been assessed using newly based analytical approaches for vegetation characterization. Here, we classified and characterized different vegetation types (i.e., major plant communities) based on indicator species and on the influence of different environmental gradients in the Himalayan mixed coniferous forest, Pakistan. For that, we addressed the following questions: Does the vegetation composition of the Himalayan mixed coniferous forest correlate with climatic, topographic, geographic, and edaphic variables? Is it possible to identify plant communities through indicator species in relation to environmental gradients using multivariate approaches? Can this multivariate be helpful for conservation planning? During four consecutive years we assessed the vegetation composition and environmental variables (21 variables divided in geographic, climatic, topographic, and edaphic groups) of 156 50 m-trasects between an elevation of 2000-4000 m. Using newly based analytical approaches for community characterization, we found a total of 218 plant species clustered into four plant communities with the influence of environmental gradients. The highest index of similarity was recorded between Pinus-Cedrus-Viburnum (PCV) and Viburnum-Pinus-Abies (VPA) communities, and the highest index of dissimilarity was recorded between PCV and Abies-Juniperus-Picea (AJP) communities. Among these four communities, highest number of plant species (156 species) was recorded in PCV, maximum alpha diversity (H' = 3.68) was reported in VPA, highest Simpson index (0.961) and Pielou's evenness (0.862) were reported in VPA and AJP. The edaphic gradients (i.e., organic matter, phosphorous, pH and soil texture) and climatic factors (temperature, humidity) were the strongest environmental gradients that were responsible for structuring and hosting the diverse plant communities in mixed coniferous forest. Finally, the Himalayan mixed coniferous structure is more influenced by the spatial turnover beta-diversity process (ßsim) than by the species loss (nestedness-resultant, ßsne). Our analysis of the vegetation structure along the environmental gradient in the Himalayan mixed coniferous forest supported by sophisticated analytical approaches reveled indicator species groups, which are associated to specific microclimatic zones (i.e., vegetation communities). Within this focus, we side with the view that these results can support conservation planning and management for similar and different areas providing mitigating and preventive measures to reduce potential negative impacts, such as anthropic and climatic.

How long to wait after local infiltration anaesthesia: systematic review.

BACKGROUND: Conflicting evidence exists regarding the optimal waiting time for stable analgesic and vasoconstrictive effects after local infiltration of lidocaine with epinephrine. An objective review is needed to dispel surgical dogma. METHODS: This systematic review (PROSPERO ID: CRD42022362414) included RCTs and prospective cohort studies. Primary outcomes were (1) onset of analgesia and (2) onset of stable hypoperfusion, assessed directly, or measured indirectly using perfusion imaging. Other data extracted include waiting strategies, means of outcome assessment, anaesthetic concentrations, volume/endpoint of infiltration, and injection sites. Methodological quality was evaluated using the Cochrane risk-of-bias tool for randomized trials. Articles describing waiting strategies were critically appraised by the Joanna Briggs Institute tools. RESULTS: Twenty-four articles were analysed, comprising 1013 participants. Ten investigated analgesia onset. Their pooled mean was 2.1 min (range 0.4-9.0 min). This varied with anatomic site and targeted nerve diameter. Fourteen articles investigated onset of stable hypoperfusion. Four observed bleeding intraoperatively, finding the minimum time to hypoperfusion at 7.0 min in the eyelid skin and 25.0 min in the upper limb. The ten remaining studies used perfusion imaging, reporting a wide range of results (0.0-30.0 min) due to differences in anatomic sites and depth, resolution and artefacts. Studies using near-infrared reflectance spectroscopy and hyperspectral imaging correlated with clinical observations. Thirteen articles discussed waiting strategies, seven relating to large-volume tumescent local infiltration anaesthesia. Different waiting strategies exist for emergency, arthroscopic and cosmetic surgeries, according to the degree of hypoperfusion required. In tumescent liposuction, waiting 10.0-60.0 min is the norm. CONCLUSION: Current literature suggests that around 2 min are required for most patients to achieve complete analgesia in all sites and with all anaesthesia concentrations. Waiting around 7 min in eyelids and at least 25 min in other regions results in optimal hypoperfusion. The strategies discussed inform decisions of when and how long to wait.

Applying the consolidated framework for implementation research to evaluate the community rapid intervention service.

BACKGROUND: Developed in 2019, the Community Rapid Intervention Service (CRIS) is a community intervention service aiming to prevent hospital admissions. CRIS provides a response within two hours to patients with sub-acute medical needs in their usual place of residence. This evaluation aimed to identify challenges and facilitators to implementation of the service, with a view to informing future service development, optimising patient care and disseminating learning to other areas looking to implement similar services. METHODS: This study used the Consolidated Framework for Implementation Research (CFIR) as an evaluation framework. We conducted semi-structured interviews with local healthcare system leaders, clinicians that worked within the CRIS, and clinicians who interfaced with the CRIS. The CFIR was used to guide data collection and analysis. Two Community of Practice (CoP) meetings were held to gather stakeholders' perspectives of the evaluation. RESULTS: Three key themes were identified from the analysis of 13 interviews: contextual factors influencing implementation, service identity and navigating complexity. Contextual factors such the influence of the Covid 19 pandemic upon health services and the expansion of the CRIS were discussed by participants. The adaptability of the service was deemed both a facilitator and challenge of implementation. Ways to build-on and improve the existing CRIS model were suggested. CONCLUSION: This evaluation has shown that the CRIS may need to be redefined with clarity provided as to how the service interfaces with other urgent and planned care offered in acute, primary, community and social services. Structuring the evaluation around the CFIR was helpful in identifying facilitators and challenges that influenced the implementation of the CRIS.

Synthesis, Characterization, Acute Dermal Toxicity, Anti-inflammatory, and Wound Healing Potential of Biogenic Silver Nanoparticles in Balb C Mice.

AIM: The current study aimed to develop an economic plant-based therapeutic agent to improve the treatment strategies for diseases at the nano-scale. METHOD: In the current research, silver nanoparticles were synthesized using Trillium govanianum aqueous extract. Characterizations were done using UV-Visible spectrophotometer, X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopy. In vivo biological activities such as acute dermal toxicity, wound healing, and anti-inflammatory were done on Balb C mice. Absorbance at 295 nm corresponds to the out-of-plane quadrupole Plasmon-resonance while at 350 nm corresponds to in-plane dipole resonance. SEM images showed the morphology of TGAgNPs is not exactly spherical while XRD analysis shows that highly crystalline TGAgNPs with an average size of 27.94 nm. The FTIR spectrum represents sharp peaks of aldehyde, amide I, aromatic rings, and polysaccharides. The microscopic assessment did not find any epidermal and dermal layer abnormalities in Blab C mice when exposed to TGAgNPs during acute dermal toxicity. RESULT & DISCUSSION: Results revealed that 1000 mg/kg is not a lethal dose. In the wound healing activity, no mortality and no abnormal signs were observed when petroleum jelly, Nitrofuranose, TGaqu, and TGAgNPs-based ointments were applied. Enhanced epithelization was recorded in TGaqu and TGAgNPs treated mice (p≤0.001). The wound contraction percentage was higher in nitrofuranose-treated mice (74%) followed by TGAgNPs (71%), and TGaqu (69%) compared to vehicle-treated and open-wounded mice. The paw edema model proved the potential use of TGAgNPs and TGaqu as anti-inflammatory agents. CONCLUSION: Hence, the results proved that both TGaqu and TGAgNPs are not toxic and possessed strong anti-inflammatory and wound-healing effects due to the presence of phytochemical constituents and could be used in various drug production as a therapeutic agent.

Genetic Investigation of Consanguineous Pakistani Families Segregating Rare Spinocerebellar Disorders.

Spinocerebellar disorders are a vast group of rare neurogenetic conditions, generally characterized by overlapping clinical symptoms including progressive cerebellar ataxia, spastic paraparesis, cognitive deficiencies, skeletal/muscular and ocular abnormalities. The objective of the present study is to identify the underlying genetic causes of the rare spinocerebellar disorders in the Pakistani population. Herein, nine consanguineous families presenting different spinocerebellar phenotypes have been investigated using whole exome sequencing. Sanger sequencing was performed for segregation analysis in all the available individuals of each family. The molecular analysis of these families identified six novel pathogenic/likely pathogenic variants; ZFYVE26: c.1093del, SACS: c.1201C>T, BICD2: c.2156A>T, ALS2: c.2171-3T>G, ALS2: c.3145T>A, and B4GALNT1: c.334_335dup, and three already reported pathogenic variants; FA2H: c.159_176del, APTX: c.689T>G, and SETX: c.5308_5311del. The clinical features of all patients in each family are concurrent with the already reported cases. Hence, the current study expands the mutation spectrum of rare spinocerebellar disorders and implies the usefulness of next-generation sequencing in combination with clinical investigation for better diagnosis of these overlapping phenotypes.

qPCR Assay as a Tool for Examining Cotton Resistance to the Virus Complex Causing CLCuD: Yield Loss Inversely Correlates with Betasatellite, Not Virus, DNA Titer.

Cotton leaf curl disease (CLCuD) is a significant constraint to the economies of Pakistan and India. The disease is caused by different begomoviruses (genus Begomovirus, family Geminiviridae) in association with a disease-specific betasatellite. However, another satellite-like molecule, alphasatellite, is occasionally found associated with this disease complex. A quantitative real-time PCR assay for the virus/satellite components causing CLCuD was used to investigate the performance of selected cotton varieties in the 2014-2015 National Coordinated Varietal Trials (NCVT) in Pakistan. The DNA levels of virus and satellites in cotton plants were determined for five cotton varieties across three geographic locations and compared with seed cotton yield (SCY) as a measure of the plant performance. The highest virus titer was detected in B-10 (0.972 ng·µg-1) from Vehari and the lowest in B-3 (0.006 ng·µg-1) from Faisalabad. Likewise, the highest alphasatellite titer was found in B-1 (0.055 ng·µg-1) from Vehari and the lowest in B-1 and B-2 (0.001 ng·µg-1) from Faisalabad. The highest betasatellite titer was found in B-23 (1.156 ng·µg-1) from Faisalabad and the lowest in B-12 (0.072 ng·µg-1) from Multan. Virus/satellite DNA levels, symptoms, and SCY were found to be highly variable between the varieties and between the locations. Nevertheless, statistical analysis of the results suggested that betasatellite DNA levels, rather than virus or alphasatellite DNA levels, were the important variable in plant performance, having an inverse relationship with SCY (-0.447). This quantitative assay will be useful in breeding programs for development of virus resistant plants and varietal trials, such as the NCVT, to select suitable varieties of cotton with mild (preferably no) symptoms and low (preferably no) virus/satellite. At present, no such molecular techniques are used in resistance breeding programs or varietal trials in Pakistan.