Alternative Strategies Research Articles

Attract-and-kill for managing Popillia japonica (Coleoptera: Scarabaeidae) abundance and leaf injury in commercial vineyards.

Popillia japonica Newman (Coleoptera: Scarabaeidae) feeding negatively impacts many plant species, including grapes, potentially reducing fruit quality and yield. Chemical control, representing the current grower standard, relies on frequent broadcast applications of broad-spectrum insecticides, with alternative management strategies mostly lacking. Attract-and-kill (A&K) is a behavioral management strategy that combines semiochemical attractants and a killing agent on a substrate. This study assessed the impact of A&K on (i) the number of P. japonica adults and (ii) the percent of P. japonica feeding injury on grape foliage compared to the grower standard in commercial vineyards. This 2-year study was conducted at 3 commercial vineyards with 4 paired plots consisting of a grower standard control and an A&K treatment. The A&K treatment consisted of commercial lures, each placed on outside-edge grapevines, and weekly applications of carbaryl on the plants holding lures. The grower standard received broadcast insecticide applications at the grower's discretion. The A&K treatment experienced similar numbers of P. japonica adults and similar proportions of leaf injury compared to the grower standard. The use of A&K reduced by 96% the crop area treated with insecticides compared to the grower standard. The area treated by A&K was at the edge of the vineyards, where more leaf injury occurred regardless of treatment. A&K is a targeted approach that was effective at managing P. japonica and reducing chemical inputs on a small scale. It has the potential to be scaled up and refined to provide growers with a new management strategy.

A multifunctional guanosine-based carbon dots for dead microbial imaging and synergistic broad-spectrum antimicrobial therapy

Pathogenic microbial infection poses serious threats to human lives and antibiotic abuse leads to bacterial resistance. The development of alternative antibacterial agents and strategies with reduced resistance risk is urgently demanded. Heteroatom doping is a feasible method for adjusting the electronic and optical properties of carbon dots (CDs) to improve their bioimaging performance and light-mediated therapeutic efficiency. Herein, we synthesized a series of guanosine-derived CDs from guanosine (G), guanosine monophosphate (GMP), guanosine diphosphate (GDP), and guanosine triphosphate (GTP) as respective precursors. G-CDs, prepared by guanosine with strong surface negative charge and suitable size, can successfully differentiate and stain dead microbial cells (bacteria and fungi). G-CDs with a high N content exhibited the highest antibacterial activity in vitro and in vivo among the four guanosine-derived CDs, owing to triple synergistic antibacterial actions - the intrinsic antibacterial mechanism, bacterial photodynamic and photothermal inactivation against bacteria and fungi. In this paper, we present a promising new method to prepare CDs using natural biomolecules from living systems as precursors for multifunctional dead microbial cells imaging and broad-spectrum theranostic applications.

The Perceval Sutureless Bioprosthetic Aortic Valve: Evolution of Surgical Valve Technology.

The surgical treatment of aortic stenosis continues to evolve, and sutureless aortic valve replacement (SUAVR) is an emerging technology. With the Perceval S (Corcym, London, UK) as the only true sutureless valve on the market, the objective of this review is to analyze the current literature on Perceval S. Focusing on valve design and deployment as well as applications of the technology for challenging pathology, clinical outcomes are assessed, including a comparison with transcatheter AVR (TAVR). PubMed and MEDLINE were searched by 3 authors for studies analyzing SUAVR from inception to May 19, 2023. SUAVR facilitates minimally invasive surgery and offers an alternative strategy for patients with small aortic annuli. It also has a time-saving advantage for patients who require complex operations. SUAVR results in excellent long-term morbidity, mortality, durability, and hemodynamic function. In comparison with conventional surgical AVR (SAVR), SUAVR does have a greater risk of postoperative pacemaker implantation; however, increasing user experience and refinements in implantation technique have contributed to reductions in this outcome. SUAVR results in morbidity and mortality that is similar to rapid-deployment AVR. Midterm outcomes are superior to TAVR; however, further robust investigation into all of these comparisons is ultimately necessary. SUAVR bridges the gap in technology between SAVR and TAVR. The application of this exciting technology will undoubtedly grow in the coming years, during which additional investigation is paramount to optimize preoperative planning, valve deployment, and reintervention strategies.

Inhibition of spinal BRD4 alleviates pyroptosis and M1 microglia polarization via STING-IRF3 pathway in morphine-tolerant rats

BackgroundMorphine tolerance has been a challenging medical issue. Neuroinflammation is considered as a critical mechanism for the development of morphine tolerance. Bromodomain-containing protein 4 (BRD4), a key regulator in cell damage and inflammation, participates in the development of chronic pain. However, whether BRD4 is involved in morphine tolerance and the underlying mechanisms remain unknown. MethodsThe morphine-tolerant rat model was established by intrathecal administration of morphine twice daily for 7 days. Behavior test was assessed by a tail-flick latency test. The roles of BRD4, pyroptosis, microglia polarization and related signaling pathways in morphine tolerance were elucidated by Western blot, real-time quantitative polymerase chain reaction, and immunofluorescence. ResultsRepeated morphine administration upregulated BRD4 level, induced pyroptosis, and promoted microglia M1-polarization in spinal cord, accompanied by the release of proinflammatory cytokines, such as TNF-α and IL-1β. JQ-1, a BRD4 antagonist, alleviated the development of morphine tolerance, diminished pyroptosis and induced the switch of microglia from M1 to M2 phenotype. Mechanistically, stimulator of interferon gene (STING)- interferon regulatory factor 3 (IRF3) pathway was activated and the protective effect of JQ-1 against morphine tolerance was at least partially mediated by inhibition of STING-IRF3 pathway. ConclusionThis study demonstrated for the first time that spinal BRD4 contributes to pyroptosis and switch of microglia polarization via STING-IRF3 signaling pathway during the development of morphine tolerance, which extend the understanding of the neuroinflammation mechanism of morphine tolerance and provide an alternative strategy for the precaution against of this medical condition.

Evaluating the Genetic Diversity of the Plasmodium vivax siap2 Locus: A Promising Candidate for an Effective Malaria Vaccine?

Malaria is the deadliest parasitic disease in the world. Traditional control measures have become less effective; hence, there is a need to explore alternative strategies, such as antimalarial vaccines. However, designing an anti-Plasmodium vivax vaccine is considered a challenge due to the complex parasite biology and the antigens´ high genetic diversity. Recently, the sporozoite invasion-associated protein 2 (SIAP2) has been suggested as a potential antigen to be considered in vaccine design due to its significance during hepatocyte invasion. However, its use may be limited by the incomplete understanding of gene/protein diversity. Here, the genetic diversity of pvsiap2 using P. vivax DNA samples from Colombia was assessed. Through PCR amplification and sequencing, we compared the Colombian sequences with available worldwide sequences, revealing that pvsiap2 displays low genetic diversity. Molecular evolutionary analyses showed that pvsiap2 appears to be influenced by directional selection. Moreover, the haplotypes found differ by a few mutational steps and several of them were shared between different geographical areas. On the other hand, several conserved regions within PvSIAP2 were predicted as potential B-cell or T-cell epitopes. Considering these characteristics and its role in hepatocyte invasion, the PvSIAP2 protein emerges as a promising antigen to be considered in a multi-antigen-multi-stage (multivalent) fully effective vaccine against P. vivax malaria.

Balancing risks and rewards of alternate strategies in the seaward extent, duration and timing of fjord use in contemporary anadromy of brown trout (Salmo trutta)

BackgroundAnadromy comprises a successful life-cycle adaptation for salmonids, with marine migration providing improved feeding opportunities and thus improved growth. These rewards are balanced against costs from increased energy expenditure and mortality risk. Anthropogenic-induced environmental changes that reduce benefits and/or increase costs of migration e.g., aquaculture and hydropower, may therefore result in adaptations disfavouring anadromy. We tagged brown trout (Salmo trutta) smolts (N = 175) and veteran migrants (N = 342), from five adjacent riverine populations located in Sognefjorden, the longest Norwegian fjord-system supporting anadromous brown trout populations (209 km). Over four years, 138 acoustic telemetry receivers were deployed to track migrations of tagged individuals from freshwater and throughout Sognefjorden. Detected movements were used to fit migration models and multi-state mark-recapture models of survival and movement for each life-stage. Seaward migration distance was modelled to examine the fitness consequences from alternate migration strategies, with these models used to simulate the extent of fjord-use by individuals and accompanying growth, fecundity and survival consequences. We compared these findings with mark-recapture data collected prior to aquaculture and hydropower development.ResultsThe telemetry data revealed that the outermost-fjord region was utilised by all populations albeit by few individuals. However, historical recaptures were located at a greater distance from the river mouth (87.7 ± 70.3 km), when compared to maximum migration distances of present-day counterparts (58.6 ± 54.9 km). River of origin influenced observed migratory behaviour and differential survival was estimated for each population and life-stage. The simulations based on telemetry-data models revealed a 30% and 23% difference in survival among populations for smolts and veteran migrants, respectively. At the individual-level, a long-distance migration strategy was rewarded with enhanced fecundity. However, the main contribution to population-level fecundity was overwhelmingly derived from middle-distance migrants, due to higher mortality rates and limited numbers of long-distant migrants.ConclusionsWe conclude that present-day anadromy is precarious, but potential risk varies considerably between life-stages and populations, even within a single fjord system. Our findings suggest that selection for extended migration is under pressure, we therefore stress the importance of monitoring and management actions to secure genetic variation pertinent to preserve fitness gains of anadromy.

Marketing Mix to Increase Sales of Wedding Package Resort Buyout at Alila Villas Uluwatu

Purpose: The purpose of this research is to determine strengths, weaknesses, opportunities and threats of an effort to increase sales of wedding package resort buyout, to find out marketing mix strategy that should be used by Sales & Marketing Department to increase sales of wedding package resort buyout. Research methods: Data collection methods that used were participation observation, interview, documentation, and questionnaire. The analysis technique that used are qualitative descriptive analysis, quantitative descriptive analysis, Internal Factor Analysis Summary (IFAS) matrix, External Factor Analysis Summary (EFAS) matrix, Internal-External (IE) matrix, and SWOT analysis. Results and discussion: The result of the research show that there are internal and external factors that influence the increase in upselling which carried out by Sales & Marketing Department at Alila Villas Uluwatu. Based on SWOT analysis score of IFAS was 2.88 and score EFAS was 2.86 that made the position of Alila Villas Uluwatu in current condition in cell 5, named the strategy of concentration through horizontal integration or stability (no change in profit). It means that this company is in a moderate attractive industry, the strategy that used has the goal relatively more defensive, namely to avoid losing sales and losing profits. Based on the SWOT analysis, alternative strategies that can be used to increase the sales of wedding package resort buyout is obtained by finding new markets, adding wedding inclusion on the wedding package, improving the payment regulation, making new building for attendees and equipment store, and making competitor list that shows if hotel is competed to the competitors. Implication: The advice given to the management of Alila Villas Uluwatu is to add new markets that are potential guests who hold wedding resort buyout such as India and Singapore, add more inclusions to the wedding package in order to make the guests interested, and improve the payment regulations to make the wedding operational runs better. Keywords: Marketing mix, IFAS matrix, EFAS matrix.

Retinoic acid receptor activation reprograms senescence response and enhances anti-tumor activity of natural killer cells

Cellular senescence can exert dual effects in tumors, either suppressing or promoting tumor progression. The senescence-associated secretory phenotype (SASP), released by senescent cells, plays a crucial role in this dichotomy. Consequently, the clinical challenge lies in developing therapies that safely enhance senescence in cancer, favoring tumor-suppressive SASP factors over tumor-promoting ones. Here, we identify the retinoic-acid-receptor (RAR) agonist adapalene as an effective pro-senescence compound in prostate cancer (PCa). Reactivation of RARs triggers a robust senescence response and a tumor-suppressive SASP. In preclinical mouse models of PCa, the combination of adapalene and docetaxel promotes a tumor-suppressive SASP that enhances natural killer (NK) cell-mediated tumor clearance more effectively than either agent alone. This approach increases the efficacy of the allogenic infusion of human NK cells in mice injected with human PCa cells, suggesting an alternative therapeutic strategy to stimulate the anti-tumor immune response in "immunologically cold" tumors.

Do risk scores improve use of faecal immunochemical testing for haemoglobin in symptomatic patients in primary care?

Faecal immunochemical testing (FIT) is used in the detection of colorectal cancer (CRC). FIT is invariably used at a single faecal haemoglobin (f-Hb) concentration threshold. The aim of this observational study was to explore risk scoring models (RSMs) with f-Hb and other risk factors for CRC in symptomatic patients attending primary care, potentially speeding diagnosis and saving endoscopy resources. Records of patients completing FIT were linked with The Scottish Cancer Registry and with other databases with symptoms, full blood count and demographic variables, and randomized into derivation and validation cohorts. Stepwise multivariable logistic regression created RSMs assessed in the validation cohort. Of 18 805 unique patients, 9374 and 9431 were in the derivation and validation cohorts, respectively: f-Hb, male sex, increasing age, iron deficiency anaemia and raised systemic immune inflammation index created the final RSM. A risk score threshold of ≥2.363, generating the same number of colonoscopies as a f-Hb threshold of ≥10 μg Hb/g gave improved sensitivity for CRC in both cohorts. A RSM which excluded f-Hb was used to investigate the effect of raising the f-Hb threshold from ≥10 to ≥20 μg Hb/g in those with a low risk score. This approach would have generated 234 fewer colonoscopies but missed four CRCs. The RSM conferred no significant benefit to patients with very low f-Hb and CRC. Alternative strategies combining FIT with other variables may be more appropriate for safety-netting of symptomatic patients. Further work to develop and investigate the value of RSM for significant bowel disease other than CRC may also be beneficial.

Electrolyte Regulation in Stabilizing the Interface of a Cobalt-Free Layered Cathode for 4.8 V High-Voltage Lithium-Ion Batteries.

The cobalt-free layered oxide cathode of LiNi0.65Mn0.35O2 is promising for high-energy-density lithium-ion batteries (LIBs). However, under high-voltage conditions, severe side reactions between the Co-free cathode and electrolyte, as well as grain boundary cracks and pulverization of particles, hinder its practical applications. Herein, an electrolyte regulation strategy is proposed by adding fluoroethylene carbonate (FEC) and LiPO2F2 as electrolyte additives in carbonate-based electrolytes to address the above issues. As a result, a homogeneous and dense organic-inorganic hybrid cathode electrolyte interface (CEI) film is formed on the cathode surface. The CEI film consists of C-F, LiF, Li2CO3, and LixPOyFz species, which is proven to be highly conductive and effective in suppressing structure damage and alleviating the interfacial reactions between the cathode and electrolyte. With such a CEI film, the interfacial stability of the Co-free cathode and the high-voltage cycling performance of Li||LiNi0.65Mn0.35O2 are greatly improved. A reversible capacity of 155.1 mAh g-1 and a capacity retention of 81.3% over 150 cycles are attained at a 4.8 V charge cutoff voltage with the tamed electrolyte, whereas the cell without the additives only retains 76.1% capacity retention. Therefore, our work demonstrates the synergistic effect of FEC and LiPO2F2 in stabilizing the interface of Co-free cathode materials and provides an alternative strategy for the electrolyte design of high-voltage LIBs.

Human Amniotic MSC Response in LPS-Stimulated Ascites from Patients with Cirrhosis: FOXO1 Gene and Th17 Activation in Enhanced Antibacterial Activation.

Spontaneous bacterial peritonitis (SBP) is a severe complication in patients with decompensated liver cirrhosis and is commonly treated with broad spectrum antibiotics. However, the rise of antibiotic resistance requires alternative therapeutic strategies. As recently shown, human amnion-derived mesenchymal stem cells (hA-MSCs) are able, in vitro, to promote bacterial clearance and modulate the immune and inflammatory response in SBP. Our results highlight the upregulation of FOXO1, CXCL5, CXCL6, CCL20, and MAPK13 in hA-MSCs as well as the promotion of bacterial clearance, prompting a shift in the immune response toward a Th17 lymphocyte phenotype after 72 h treatment. In this study, we used an in vitro SBP model and employed omics techniques (next-generation sequencing) to investigate the mechanisms by which hA-MSCs modify the crosstalk between immune cells in LPS-stimulated ascitic fluid. We also validated the data obtained via qRT-PCR, cytofluorimetric analysis, and Luminex assay. These findings provide further support to the hope of using hA-MSCs for the prevention and treatment of infective diseases, such as SBP, offering a viable alternative to antibiotic therapy.

The interplay of local adaptation and gene flow may lead to the formation of supergenes.

Supergenes are genetic architectures resulting in the segregation of alternative combinations of alleles underlying complex phenotypes. The co-segregation of alleles at linked loci is often facilitated by polymorphic chromosomal rearrangements suppressing recombination locally. Supergenes are involved in many complex polymorphisms, including sexual, colour or behavioural polymorphisms in numerous plants, fungi, mammals, fish, and insects. Despite a long history of empirical and theoretical research, the formation of supergenes remains poorly understood. Here, using a two-island population genetic model, we explore how gene flow and the evolution of overdominant chromosomal inversions may jointly lead to the formation of supergenes. We show that the evolution of inversions in differentiated populations, both under disruptive selection, leads to an increase in frequency of poorly adapted, immigrant haplotypes. Indeed, rare allelic combinations, such as immigrant haplotypes, are more frequently reshuffled by recombination than common allelic combinations, and therefore benefit from the recombination suppression generated by inversions. When an inversion capturing a locally adapted haplotype spreads but is associated with a fitness cost hampering its fixation (e.g. a recessive mutation load), the maintenance of a non-inverted haplotype in the population is enhanced; under certain conditions, the immigrant haplotype persists alongside the inverted local haplotype, while the standard local haplotype disappears. This establishes a stable, local polymorphism with two non-recombining haplotypes encoding alternative adaptive strategies, that is, a supergene. These results bring new light to the importance of local adaptation, overdominance, and gene flow in the formation of supergenes and inversion polymorphisms in general.

Enhanced bone regeneration in rat calvarial defects through BMP2 release from engineered poly(ethylene glycol) hydrogels

The clinical standard therapy for large bone defects, typically addressed through autograft or allograft donor tissue, faces significant limitations. Tissue engineering offers a promising alternative strategy for the regeneration of substantial bone lesions. In this study, we harnessed poly(ethylene glycol) (PEG)-based hydrogels, optimizing critical parameters including stiffness, incorporation of arginine-glycine-aspartic acid (RGD) cell adhesion motifs, degradability, and the release of BMP2 to promote bone formation. In vitro we demonstrated that human bone marrow derived stromal cell (hBMSC) proliferation and spreading strongly correlates with hydrogel stiffness and adhesion to RGD peptide motifs. Moreover, the incorporation of the osteogenic growth factor BMP2 into the hydrogels enabled sustained release, effectively inducing bone regeneration in encapsulated progenitor cells. When used in vivo to treat calvarial defects in rats, we showed that hydrogels of low and intermediate stiffness optimally facilitated cell migration, proliferation, and differentiation promoting the efficient repair of bone defects. Our comprehensive in vitro and in vivo findings collectively suggest that the developed hydrogels hold significant promise for clinical translation for bone repair and regeneration by delivering sustained and controlled stimuli from active signaling molecules.

Lung Progenitor and Stem Cell Transplantation as a Potential Regenerative Therapy for Lung Diseases.

Chronic lung diseases are debilitating illnesses ranking among the top causes of death globally. Currently, clinically available therapeutic options capable of curing chronic lung diseases are limited to lung transplantation, which is hindered by donor organ shortage. This highlights the urgent need for alternative strategies to repair damaged lung tissues. Stem cell transplantation has emerged as a promising avenue for regenerative treatment of the lung, which involves delivery of healthy lung epithelial progenitor cells that subsequently engraft in the injured tissue and further differentiate to reconstitute the functional respiratory epithelium. These transplanted progenitor cells possess the remarkable ability to self-renew, thereby offering the potential for sustained long-term treatment effects. Notably, the transplantation of basal cells, the airway stem cells, holds the promise for rehabilitating airway injuries resulting from environmental factors or genetic conditions such as cystic fibrosis. Similarly, for diseases affecting the alveoli, alveolar type II cells have garnered interest as a viable alveolar stem cell source for restoring the lung parenchyma from genetic or environmentally induced dysfunctions. Expanding upon these advancements, the use of induced pluripotent stem cells to derive lung progenitor cells for transplantation offers advantages such as scalability and patient specificity. In this review, we comprehensively explore the progress made in lung stem cell transplantation, providing insights into the current state of the field and its future prospects.

Evaluation of breeding cost in the French maritime pine breeding program and perspectives for alternative strategies using molecular markers

The economic efficiency of conventional breeding strategies for forest trees based on biparental crosses is compared with that of alternative strategies based on pedigree reconstruction using molecular markers. Analyses of economic efficiency is based on comparisons of breeding scenarios corresponding to the same total investment. The first step is the description and cost evaluation of each basic operation, from crossing to genetic selection and clonal archive establishment. Breeding scenarios are then compared by stochastic sampling with a parametric genetic model (POPSIM), the comparison criteria in this case being genetic gain in the seed orchard for a given level of genetic diversity. Additionally, the economic gain resulting from the use of improved material is estimated for different levels of breeding investment. Our analysis shows that genotyping costs account for a much smaller proportion of total investment than phenotyping costs. We also show that, in comparisons of breeding scenarios corresponding to the same total investment, the three main breeding strategies (biparental crosses, polymix crosses, and open pollination) achieve similar genetic gains provided that sufficiently large numbers of parents are considered. These results open up promising perspectives for the wider integration of molecular markers into forest tree breeding strategies.

Preliminary report on therapeutic potential of coal-derived carbon quantum dots against SARS-CoV-2 virus

Due to the pandemic of COVID-19 and subsequent emerging of new mutant strains, there has been a worldwide hunt for therapeutic and protective agents for its inhibition. In this short communication, for the first time, we report the coal-derived carbon quantum dot (CQD) for the possible therapeutic application against SARS-CoV-2. The synthesized C1-CQD is observed to be safe towards the normal cell line at highest dose, while effectively inhibiting growth of SARS-CoV2 (>95%) with IC50 value of 5.469 μg/mL. Moreover, C1-CQD showed activity against SARS-CoV-2 infection which is comparable to known inhibitory antiviral drug i.e., Remdesivir. These novel findings indicate that coal-based CQDs have highly potent anti-viral activity and could be investigated further for developing cheap and safer alternative therapeutic strategies for inhibition of SARS-CoV-2.

Co-spray dried inhalable composite powders of ciprofloxacin and alginate oligosaccharide as anti-biofilm therapy

The treatment of chronic respiratory infections caused by biofilm formation are extremely challenging owing to poor drug penetration into the complex biofilm structure and high drug resistance. Local delivery of an antibiotic together with a non-antibiotic adjuvant to the lungs could often enhance the therapeutic responses by targeting different bacterial growth pathways and minimizing drug resistance. In this study, we designed new inhalable dry powders containing ciprofloxacin (CIP) and OligoG (Oli, a low-molecular-weight alginate oligosaccharide impairing the mucoid biofilms by interacting with their cationic ions) to combat respiratory bacterial biofilm infections. The resulting powders were characterized with respect to their morphology, solid-state property, surface chemistry, moisture sorption behavior, and dissolution rate. The aerosol performance and storage stability of the dry powders were also evaluated. The results showed that inhalable dry powders composed of CIP and Oli could be readily accomplished via the wet milling and spray drying process. Upon the storage under 20 ± 2 °C/20 ± 2 % relative humidity (RH) for one month, there was no significant change in the in vitro aerosol performances of the dry powders. In contrast, the dry powders became non-inhalable following the storage at 20 ± 2 °C/53 ± 2 % RH for one month due to the hygroscopic nature of Oli, which could be largely prevented by incorporation of leucine. Collectively, this study suggests that the newly developed co-spray-dried powders composed of CIP and Oli might represent a promising and alternative treatment strategy against respiratory bacterial biofilm infections.

Effectiveness of Bilingual Project-Based Materials to Facilitate Literacy and Numeracy Teaching

The purpose of this study is to find out how well bilingual project-based learning resources may improve the literacy and numeracy abilities of primary school students. Numerous researches conducted at many levels presented the effectiveness of the project-based approach. Yet, few of them address the potential benefits of incorporating it into the curriculum, focusing on both language proficiency and mathematical competence. This experimental study utilized a nonrandomized control group design involving two classes of 6-8-year-old students. A two-way ANOVA was used to evaluate the data from 39 students that were gathered through exams and paperwork. The results demonstrated that using bilingual project-based learning resources can be an alternate strategy for improving primary school students' literacy and numeracy skills.

Pyrethroid resistance status and co-occurrence of V1016G, F1534C and S989P mutations in the Aedes aegypti population from two dengue outbreak counties along the China-Myanmar border

BackgroundOver the past two decades, dengue fever (DF) has emerged as a significant arboviral disease in Yunnan province, China, particularly in the China-Myanmar border area. Aedes aegypti, an invasive mosquito species, plays a crucial role in transmitting the dengue virus to the local population. Insecticide-based vector control has been the primary tool employed to combat DF, but the current susceptibility status of Ae. aegypti to commonly used insecticides is unknown. Assessment of Ae. aegypti resistance to pyrethroid insecticides and an understanding of the underlying mechanisms of this resistance in the China-Myanmar border region is of significant strategic importance for effectively controlling the DF epidemic in the area.MethodsAedes aegypti larvae collected from Ruili and Gengma counties in Yunnan Province were reared to adults in the laboratory and tested for susceptibility to three pyrethroid insecticides (3.20% permethrin, 0.08% lambda-cyhalothrin and 0.20% deltamethrin) by the standard WHO susceptibility bioassay. Genotyping of mutations in the knockdown gene (kdr), namely S989P, V1016G and F1534C, that are responsible for resistance to pyrethroid insecticides was performed using allele-specific PCR methods. A possible association between the observed resistant phenotype and mutations in the voltage-gated sodium channel gene (VGSC) was also studied.ResultsAedes aegypti mosquitoes collected from the two counties and reared in the laboratory were resistant to all of the pyrethroids tested, with the exception of Ae. aegypti from Gengma County, which showed sensitivity to 0.20% deltamethrin. The mortality rate of Ae. aegypti from Ruili county exposed to 3.20% permethrin did not differ significantly from that of Ae. aegypti from Gengma County (χ2 = 0.311, P = 0.577). By contrast, the mortality rate of Ae. aegypti from Ruili County exposed to 0.08% lambda-cyhalothrin and 0.20% deltamethrin, respectively, was significantly different from that of Ae. aegypti from Gengma. There was no significant difference in the observed KDT50 of Ae. aegypti from the two counties to various insecticides. Four mutation types and 12 genotypes were detected at three kdr mutation sites. Based on results from all tested Ae. aegypti, the V1016G mutation was the most prevalent kdr mutation (100% prevalence), followed by the S989P mutation (81.6%) and the F1534C mutation (78.9%). The constituent ratio of VGSC gene mutation types was significantly different in Ae. aegypti mosquitoes from Ruili and those Gengma. The triple mutant S989P + V1016G + F1534C was observed in 274 Ae. aegypti mosquitoes (60.8%), with the most common genotype being SP + GG + FC (31.4%). The prevalence of the F1534C mutation was significantly higher in resistant Ae. aegypti from Ruili (odds ratio [OR] 7.43; 95% confidence interval [CI] 1.71–32.29; P = 0.01) and Gengma (OR 9.29; 95% CI 3.38–25.50; P = 0.00) counties than in susceptible Ae. aegypti when exposed to 3.20% permethrin and 0.08% lambda-cyhalothrin, respectively. No significant association was observed in the triple mutation genotypes with the Ae. aegypti population exposed to 3.20% permethrin and 0.20% deltamethrin resistance (P > 0.05), except for Ae. aegypti from Gengma County when exposed to 0.08% lambda-cyhalothrin (OR 2.86; 95% CI 1.20–6.81; P = 0.02).ConclusionsAedes aegypti from Ruili and Gengma counties have developed resistance to various pyrethroid insecticides. The occurrence of multiple mutant sites in VGSC strongly correlated with the high levels of resistance to pyrethroids in the Ae. aegypti populations, highlighting the need for alternative strategies to manage the spread of resistance. A region-specific control strategy for dengue vectors needs to be implemented in the future based on the status of insecticide resistance and kdr mutations.Graphical

Erythropoietin effects on cryopreserved/transplanted cat ovarian tissue: A comparison of two incubation methods

Many feline species are currently threatened with extinction. Therefore, germplasm bank establishment has become imperative. However, cryoinjury and ischemia-reperfusion injury pose significant obstacles to both cryopreservation and xenotransplantation. In this regard, erythropoietin (Epo) represents a potential alternative strategy due to its properties. This study aimed to assess the incubation of domestic cat ovarian tissue in Epo, both before and after cryopreservation, and investigate its effectiveness in promoting revascularization following xenotransplantation. Sixteen ovaries from 8 healthy cats were sliced following elective bilateral ovariohysterectomy (OHE). Subsequently, 8 fragments measuring 3 mm³ each were obtained from the cortical region of each ovary. The fragments were allocated into 3 treatment groups: Cryo group, fragments were cryopreserved, thawed and immediately transplanted; Cryo + Epo group, fragments were first cryopreserved in nitrogen, thawed, incubated in Epo (100 IU) for 2h and transplanted; and the Epo + Cryo group, in which fragments were first incubated in Epo (100 IU) for 2h, cryopreserved, thawed and immediately transplanted. The fragments were then xenotransplanted into the dorsal subcutaneous region of ovariectomized female nude mice and retrieved at 7, 14, 21, and 28 days post-transplantation. The results indicated that Epo effectively enhanced follicular survival, preservation of viability, and tissue revascularization. The Epo + Cryo group displayed better revascularization rates on D14 and D21 post-transplantation and an increase in primordial and growing follicles on D28, the Cryo + Epo group exhibited significantly more follicles on D14 and D21, with fewer degenerated follicles.