Intrapatient variation in PD-L1 expression and tumor mutational burden and the impact on outcomes to immune checkpoint inhibitor therapy in patients with non-small cell lung cancer.

BACKGROUND: PD-L1 tumor proportion score (TPS) and tumor mutational burden (TMB) are key predictive biomarkers for immune checkpoint inhibitors (ICI) efficacy in non-small cell lung cancer (NSCLC). Data on their variation across multiple samples are limited. METHODS: Patients with NSCLC and multiple PD-L1 TPS and/or TMB assessments were included. Clinicopathologic and genomic data were analyzed according to PD-L1 and TMB variation. RESULTS: In total, 402 PD-L1 sample pairs and 413 TMB sample pairs were included. Concordance between pairs was moderate for PD-L1 (ρ=0.53, P<0.0001) and high for TMB (ρ=0.80, P<0.0001). Shorter time between biopsies correlated with higher concordance in PD-L1, but not in TMB. Major increases (ΔTPS≥+50%) and decreases (ΔTPS≤-50%) in PD-L1 were observed in 9.7% and 8.0% of cases, respectively. PD-L1, but not TMB, decreased with intervening ICI (P=0.02). Acquired copy number loss of CD274, PDCD1LG2, and JAK2 were associated with major decrease in PD-L1 (q<0.05). Among patients with multiple PD-L1 assessments before ICI, cases where all samples had a PD-L1 ≥1%, compared to cases with at least one sample with PD-L1 <1% and another with PD-L1 ≥1%, achieved improved objective response rate and progression-free survival (PFS). Among patients with at least one PD-L1 <1% and one ≥1% before ICI, cases where the most proximal sample was PD-L1 ≥1% had longer median PFS compared to cases where the most proximal PD-L1 was <1%. Among patients with multiple TMB assessments before ICI, patients with a TMB ≥10 mut/Mb based on the most recent assessment, as compared to those with a TMB <10 mut/Mb, achieved improved PFS and OS to ICI; instead, no differences were observed when patients were categorized using the oldest TMB assessment. CONCLUSION: Despite intrapatient concordance in PD-L1 and TMB, variation in these biomarkers can influence ICI outcomes, warranting consideration for reassessment prior to ICI initiation when feasible.

Rearing and Shipping of Uranotaenia lowii, a Frog-Biting Mosquito.

Many studies on mosquito biology rely on laboratory-reared colonies, emphasizing the need for standardized protocols to investigate critical aspects such as disease biology, mosquito behavior, and vector control methods. While much knowledge is derived from anthropophilic species from genera like Anopheles, Aedes, and Culex, there is a growing interest in studying mosquitoes that feed on non-human hosts. This interest stems from the desire to gain a deeper understanding of the evolution of diverse host range use and host specificity. However, there is currently a limited number of comprehensive protocols for studying such species. Considering this gap, we present a protocol for rearing Uranotaenia lowii, a mosquito species specialized in feeding on anuran amphibians by eavesdropping on host-emitted sound cues. Additionally, we provide instructions for successfully shipping live specimens to promote research on this species and similar ones. This protocol helps fill the current gap in comprehensive guidelines for rearing and maintaining colonies of anuran host-biting mosquitoes. It serves as a valuable resource for researchers seeking to establish colonies of mosquito species from the Uranotaeniini tribe. Ultimately, this protocol may facilitate research on the evolutionary ecology of Culicidae, as this family has recently been proposed to have originated from a frog-feeding ancestor. Key features • Rearing and maintenance of colonies of non-human host-biting mosquitoes that feed on frogs using host-emitted acoustic cues. • Provides shipping guidelines aimed to enhance the establishment of colonies by new research groups and specimen exchanges between labs.

Odontogenic Keratocyst With Moderate Epithelial Dysplasia: A Rare Entity.

We present a rare case of odontogenic keratocyst (OKC) with moderate epithelial dysplasia in a 47-year-old male patient. He presented with a history of pain and swelling on the right side of his face for the past three months. The radiograph revealed multi-locular radiolucency involving the ramus, coronoid process, and condylar process of the right mandible. We have discussed the cytology, incisional biopsy, radiological investigations, surgical management, reconstruction, excisional biopsy report, and follow-up of this rare entity.

The L1014F Knockdown Resistance Mutation Is Not a Strong Correlate of Phenotypic Resistance to Pyrethroids in Florida Populations of Culex quinquefasciatus.

Culex quinquefasciatus is an important target for vector control because of its ability to transmit pathogens that cause disease. Most populations are resistant to pyrethroids and often to organophosphates, the two most common classes of active ingredients used by public health agencies. A knockdown resistance (kdr) mutation, resulting in an amino acid change from a leucine to phenylalanine in the voltage gated sodium channel, is one mechanism contributing to the pyrethroid resistant phenotype. Enzymatic resistance has also been shown to play a very important role. Recent studies have shown strong resistance in populations even when kdr is relatively low, which indicates that factors other than kdr may be larger contributors to resistance. In this study, we examined, on a statewide scale (over 70 populations), the strength of the correlation between resistance in the CDC bottle bioassay and the kdr genotypes and allele frequencies. Spearman correlation analysis showed only moderate (-0.51) or weak (-0.29) correlation between the kdr genotype and permethrin or deltamethrin resistance, respectively. The frequency of the kdr allele was an even weaker correlate than genotype. These results indicate that assessing kdr in populations of Culex quinquefasciatus is not a good surrogate for phenotypic resistance testing.

Aedes aegypti Knockdown Resistance Mutations and Dengue Virus Infection in Haiti.

Haiti is home to approximately 11 million people and has a high incidence of vector-borne disease, including more than 70,000 cases of dengue per year. Vector control is difficult in Haiti and adulticide spray of malathion is the main method of control employed during the outbreak of disease although pyrethroids are used in both bed net campaigns and in widely available aerosol cans for personal use. However, limited pathogen or insecticide resistance surveillance data are available for making operational decisions. In this study, we assessed Aedes aegypti from serial surveillance collections from 3 locations for the presence of dengue virus serotypes 1-3 (DENV1-3) by polymerase chain reaction and assessed, by melt curve analysis, samples from 10 locations in 2 departments for the presence of two mutations (V1016I and F1534C), that in combination, are linked to strong pyrethroid insecticide resistance. Only one of the 32 tested pools was positive for the presence of dengue virus. The two knockdown resistance (kdr) mutations were present in all locations. The 1016I mutation frequency varied from 0.29 to 0.91 and was in all sites lower than the 0.58-1.00 frequency of the 1534C mutation. We also observed that the genotype homozygous for both mutations (IICC), which has been linked to strong pyrethroid resistance, varied from 13 to 86% in each population. Notably, 3 locations - Ti Cousin and Christianville in Ouest department and Camp Coq in Nord department had more than 30% of the tested population without the presence of kdr mutations. These results indicate that the kdr markers of pyrethroid resistance are present in Haiti, at high frequency in several locations and, based on previous studies linking kdr genotypes and phenotypic resistance, that operational interventions with pyrethroids are not likely to be as effective as expected.

Insecticide resistance of Miami-Dade Culex quinquefasciatus populations and initial field efficacy of a new resistance-breaking adulticide formulation.

Sporadic outbreaks of human cases of West Nile virus (WNV), primarily vectored by Culex quinquefasciatus Say in suburban and urban areas, have been reported since introduction of the virus into Florida in 2001. Miami-Dade County, Florida is part of one of the largest metropolitan areas in the United States, supports Cx. quinquefasciatus year-round, and recently experienced over 60 human cases of WNV during one outbreak. To facilitate more effective integrated vector management and public health protection, we used the Centers for Disease Control and Prevention (CDC) bottle bioassay method to evaluate the susceptibility of adult Cx. quinquefasciatus collected from 29 locations throughout Miami-Dade County to pyrethroid and organophosphate adulticide active ingredients (AIs) used by Miami-Dade County Mosquito Control. We also determined the frequency of the 1014 knockdown resistance (kdr) mutation for Cx. quinquefasciatus from a subset of 17 locations. We detected resistance to two pyrethroid AIs in all tested locations (permethrin: 27 locations, deltamethrin: 28 locations). The 1014F allele was widely distributed throughout all 17 locations sampled; however, 29.4% of these locations lacked 1014F homozygotes even though phenotypic pyrethroid resistance was present. Organophosphate resistance was more variable; 20.7% of the locations tested were susceptible to malathion, and 33.3% of the populations were susceptible to naled. We subsequently conducted a field trial of ReMoa Tri, a recently approved multiple AI adulticide formulation labelled for resistant mosquitoes, against a mixed location field population of Miami-Dade Cx. quinquefasciatus. Average 24-hr mortality was 65.1 ± 7.2% and 48-hr mortality increased to 85.3 ± 9.1%, indicating good control of these resistant Cx. quinquefasciatus. This current study shows that insecticide resistance is common in local Cx. quinquefasciatus but effective options are available to maintain control during active disease transmission in Miami-Dade County.

Unusual Presentation of Primary Hyper-IgE-Related Salivary Gland Disease in a 13-Year-Old Male.

Adenomas are common pathologies of the salivary glands that are often associated with the major salivary glands and occur in the fourth to sixth decades of life. They are seldom seen in the pediatric age group and rarely in the minor salivary glands. Autoimmune sialadenosis of the minor salivary glands is a new phenomenon that has seldom been reported in the literature, with as few as three cases. Histopathological examination of the excised specimen is the definitive diagnosis, and these lesions have to be differentiated from adenomas and low-grade malignancies of the minor salivary glands. Management strategies of these lesions are extremely variable, ranging from wait-and-watch principle to the use of immunosuppressants and excision of the gland. This case report discusses the etiopathogenesis of the autoimmune sialadenosis and the management strategies.

Comparison of the Efficacy of Midazolam and Dexmedetomidine As Sedative Agents in Third Molar Surgery.

Introduction Minor dental and oral surgical procedures have been made comfortable with the rise in the use of daycare sedatives. Of these sedatives, midazolam is deemed a common sedative used for minor oral surgical procedures. Newer and safer sedatives such as dexmedetomidine have certain properties that may prove more efficient in oral surgical procedures. Third molar surgery is one of the most common minor oral surgical procedures performed in dentistry. Thus, this study aims to compare the efficacy of midazolam and dexmedetomidine as sedative agents in third molar surgery. Materials and methods Sixty young adult patients free from other comorbidities were included in the study with ages ranging between 18 and 50 years. The samples were matched for the difficulty of impacted teeth and randomly distributed among the groups. Groups were administered the respective sedative drugs midazolam and dexmedetomidine and their effects were observed through the Observer's Assessment of Alertness/Sedation scale. The intraoperative vitals and sedation effects were checked every 15 minutes. Statistical analysis was done using IBM SPSS Statistics for Windows, Version 22 (Released 2013; IBM Corp., Armonk, New York, United States). Independent samples t-test and analysis of variance were the statistical tests employed to analyze the obtained data with p<0.05 considered as statistically significant. Results The depth of sedation has been both subjectively and objectively assessed and had no significant difference among the groups. The intra-operative heart rate assessment proved a more efficient reduction of pulse rate in the dexmedetomidine group as compared with the midazolam group. However, it was not statistically significant (p=0.121). The mean diastolic blood pressure showed a statistically significant difference between the groups with dexmedetomidine having lower blood diastolic pressure compared to midazolam (p=0.004). Quick arousal was witnessed in the dexmedetomidine group. Conclusion It can be concluded from the study that both dexmedetomidine and midazolam were equally effective as sedative agents for third molar surgery. However, the nature of cardio-protection, anti-sialagogue, and analgesic properties of dexmedetomidine can prove helpful, especially in minor oral surgical procedures like third molar surgery and it is recommended.

Expression, activity, and consequences of biochemical inhibition of α- and ß-glucosidases in different life stages of Culex quinquefasciatus.

Mosquitoes have a wide range of digestive enzymes that enable them to utilize requisite blood and sugar meals for survival and reproduction. Sugar meals, typically derived from plant sources, are critical to maintain energy in both male and female mosquitoes, whereas blood meals are taken only by females to complete oogenesis. Enzymes involved in sugar digestion have been the subject of study for decades but have been limited to a relatively narrow range of mosquito species. The southern house mosquito, Culex quinquefasciatus, is of public health importance and seldom considered in these types of studies outside of topics related to Bacillus sphaericus, a biocontrol agent that requires interaction with a specific gut-associated α-glucosidase. Here we sought to describe the nature of α-glucosidases and unexplored ß-glucosidases that may aid Cx. quinquefasciatus larvae in acquiring nutrients from cellulosic sources in their aquatic habitats. Consistent with our hypothesis, we found both α- and ß-glucosidase activity in larvae. Interestingly, ß-glucosidase activity all but disappeared at the pupal stage and remained low in adults, while α-glucosidase activity remained in the pupal stage and then exceeded larval activity by approximately 1.5-fold. The expression patterns of the putative α- and ß-glucosidase genes chosen did not consistently align with observed enzyme activities. When the α-glucosidase inhibitor acarbose was administered to adults, mortality was seen especially in males but also in females after two days of exposure and key energetic storage molecules, glycogen and lipids, were significantly lower than controls. In contrast, administering the ß-glucosidase inhibitor conduritol ß-epoxide to larvae did not produce mortality even at the highest soluble concentration. Here we provide insights into the importance of α- and ß-glucosidases on the survival of Cx. quinquefasciatus in their three mobile life stages.

Larvicidal constituents from Poncirus trifoliata root extracts.

In the search for effective and environmentally friendly mosquito control agents, we have examined natural sources, such as microbes and plants, and the synthetic analogs of natural products. These plants and microbes have evolved in their ecological niches to produce defensive compounds against other competing organisms in their surroundings such as microbes, plants, and insects as a means to enhance their survival. Thus, some of these plants and microbes have bioactive compounds with insecticidal, fungicidal, and phytotoxic activities. In our previous research, we successfully isolated bioactive constituents from natural sources. We have carried out synthetic modifications and total synthesis of marginally active isolated compounds to achieve significantly higher active compounds. We have focused on plants in the Rutaceae family as the members of this family are known to possess bioactive compounds with algicidal, antifungal, insecticidal, and fungicidal activities. In this article, we report the isolation and structure elucidation of mosquito larvicidal constituents from Poncirus trifoliata (Rutaceae) root extract.

A Survey on the COVID-19 Awareness and Vaccination Acceptance/Hesitance Among the Transgender Population in Various Districts of Tamilnadu.

Introduction: Coronavirus Disease of 2019 (COVID-19) has been deemed as one of the biggest global health crisis. The pandemic has caused a landslide in governance, economics, health care systems, and the daily lifestyle of man for the past 2 years. One of the chief weapons installed by various governments worldwide was the rapid development of an efficient vaccine to combat the disease. This study aims to identify the awareness of COVID 19 and seeks to find the challenges faced by the community in vaccination. This study hypothesized the presence of a multifaceted issue among the transgender populations' hesitance to vaccination. The results from the study could be exploited by the policy makers of the government to provide policies that best alleviate the hesitance among the transgender population. Material and Methods: This study is a cross sectional questionnaire based study. A total of 121 transgender people above the age of 18 years who were voluntarily willing for the study were included in the study, while excluding the male and female participants and those transgender participants who were unwilling for the study. Results and Discussion: A 100% response regarding COVID-19 awareness from the 121 participants showed exemplary reach to the population. With 76% of individuals being vaccinated, the acceptance of the vaccine and reach of the vaccine was high. COVID-19 awareness has reached the people from all walks of life. With little tweaks to the current policies, India could achieve absolute vaccination against the pandemic.

Assessing pyrethroid resistance status in the Culex pipiens complex (Diptera: Culicidae) from the northwest suburbs of Chicago, Illinois using Cox regression of bottle bioassays and other detection tools.

Culex pipiens complex is an important vector of epizootic and zoonotic pathogens, including West Nile virus. Chicago, Illinois and its suburbs have suffered high incidence of human West Nile virus infections in the past. This makes abatement programs in and around the Chicago area an essential service. The control of Cx. pipiens is often complicated by rapidly evolving resistance to pyrethroids, which are the most widely used chemical class in US mosquito abatement programs. The present study assessed Sumithrin® resistance in Cx. pipiens collected from five locations around Cook County, Illinois, neighboring the city limits of Chicago. According to CDC guidelines, samples from all five locations demonstrated some resistance to Sumithrin®. When assessed with Anvil®, a formulated product made of Sumithrin® synergized with piperonyl butoxide, susceptibility was rescued in mosquitoes from three out of the five locations, suggesting involvement of mixed-function oxidases and/or carboxylesterases in Sumithrin® resistance at these locations. Not all locations had susceptibility rescued by Anvil®, but these locations had relatively low knockdown resistance allele frequencies, suggesting that mechanisms other than knockdown resistance may be involved. Enzyme activities did not reveal any marked trends that could be related back to mortality in the bottle bioassays, which highlights the need for multiple types of assays to infer enzymatic involvement in resistance. Future directions in pyrethroid resistance management in Chicago area Cx. pipiens are discussed.

Insecticidal and Attractant Activities of Magnolia citrata Leaf Essential Oil against Two Major Pests from Diptera: Aedes aegypti (Culicidae) and Ceratitis capitata (Tephritidae).

In this study, Magnolia citrata Noot and Chalermglin (Magnoliaceae) essential oil (MCEO) was evaluated for insecticidal activity against the yellow fever mosquito Aedes aegypti and attractant activity for the Mediterranean fruit fly Ceratitis capitata. The leaves of Magnolia citrata (Gioi chanh) were collected from northwestern Vietnam, and the water-distilled MCEO was analyzed by gas-chromatography and mass spectrometry (GC-MS). The major constituents of MCEO were identified as linalool 19%, geranial 16%, citronellal 14%, neral 14%, and sabinene 12%. MCEO showed 100% mortality at 1 µg/µL against 1st instar larvae of Ae. aegypti (Orlando strain, ORL), and the oil exhibited 54% (ORL) and 68% (Puerto Rico strain) mortality at 5 µg/mosquito against Ae. aegypti adult females. Initial screens showed that MCEO had weak insecticidal activity compared to the positive control permethrin. In bioassays with sterile male C. capitata, MCEO exhibited moderately strong attraction, comparable to that observed with a positive control, Tetradenia riparia essential oil (TREO). Herein, the insecticidal and attractant activities of MCEO are reported for the first time.

Insecticidal Activity and Free Radical Scavenging Properties of Isolated Phytoconstituents from the Saudi Plant Nuxia oppositifolia (Hochst.).

Chromatographic purification of the alcoholic extract from the aerial parts of the Saudi plant Nuxia oppositifolia (Hochst.), Benth., resulted in five isolated phenolic compounds. Two flavones, hispidulin (1) and jaceosidin (2), and the phenylethanoid glycosides, verbascoside (3), isoverbascoside (4), and conandroside (5), were identified and their chemical structures were determined by spectroscopic analyses. The insecticidal activity of compounds 1 and 2, in addition to 11 compounds isolated in a previous research (6-16), was evaluated against the Yellow Fever mosquito, Aedes aegypti. Four compounds displayed adulticidal activity with LD50 values of 2-2.3 µg/mosquito. Free radical scavenging properties of the plant extracts and compounds (1-5) were evaluated by measuring the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate radical cation (ABTS•+) scavenging activity. All compounds exhibited notable activity, compared with the positive control, l-Ascorbic acid. This study suggests that N. oppositifolia could be a promising source of secondary metabolites, some with lethal adulticidal effect against Ae. aegypti.

Molecular Characterization of Bartonella Species Discovered in Ectoparasites Collected from Domestic Animals, Cuzco, Peru.

Rickettsiae and bartonellae are Gram-negative bacteria that can cause zoonotic and human diseases and are vectored by hematophagous arthropods. In the Americas, rickettsioses and bartonelloses have reemerged as significant public health threats. Bartonella species have been identified as causing zoonotic infections responsible for a variety of clinical syndromes in humans and animals. The aim of this study was to investigate the distribution, prevalence, and molecular heterogeneity of Rickettsia spp. and Bartonella spp. among ectoparasites collected from domestic animals in 14 farming communities in the Andes Mountains of Cuzco, Peru. A total of 222 domestic animals representing 8 different species (sheep, donkeys, goats, cattle, pigs, llamas, guinea pigs, and horses) were sampled. Nine species of ectoparasites (n = 1,697) collected from 122 animals were identified resulting in 1,657 chewing lice, 39 ticks, and 1 flea. DNA was individually extracted from a random sample of 600 (35.4%) considering variability of ectoparasite species, hosts, and sample location elevation. All 600 samples were negative for rickettsial DNA by a genus-specific molecular assay. A subset of 173 (28.8%) samples were selected based on variability of arthropods species, host, and location for Bartonella testing. Ninety-one (52.6%) of these samples including Melophagus ovinus (90/110) and Bovicola bovis (1/7) were positive for Bartonella by a genus-specific molecular assay. Five Bartonella genes of seven DNA samples from M. ovinus were analyzed by the multilocus sequence typing for characterization. We identified five identical Bartonella melophagi specimens and two specimens with Bartonella species related to B. melophagi from the seven M. ovinus. The Bartonella agents detected were widely distributed and frequent in multiple studied locations.

Spatial variation in the frequency of knockdown resistance genotypes in Florida Aedes aegypti populations.

BACKGROUND: The development of insecticide resistance in disease-vectoring mosquito species can lead to vector control failure and disease resurgence. However, insecticide applications remain an essential public health intervention. In Florida, insecticide resistance in Aedes aegypti, an anthropophilic mosquito species capable of transmitting dengue, chikungunya, and Zika virus, is a major concern. Understanding the location, scale, and driving factors of insecticide resistance can enhance the ability of vector control organizations to target populations effectively. METHODS: We used previously collected data on frequencies of mutations that confer resistance to commonly used pyrethroid insecticides in Ae. aegypti specimens from 62 sites distributed across 18 counties in Florida. To determine the scale of clustering for the most resistant variant, we used a Ripley's K function. We also used a spatial scanning statistic technique to identify locations of clusters where higher than expected frequencies of susceptible or resistant mosquitoes occurred. We then tested for associations between landscape, demographic, and insecticide-use factors using a beta regression modelling approach and evaluated the effect of spatial lag and spatial error terms on overall explanatory power of these models. RESULTS: The scale at which maximum clustering of the most resistant variant occurs is approximately 20 kilometers. We identified statistically significant clusters of genotypes associated with resistance in several coastal cities, although some of these clusters were near significant clusters of susceptible mosquitoes, indicating selection pressures vary at the local scale. Vegetation density, distance from roads, and pyrethroid-use by vector control districts were consistently significant predictors of knockdown resistance genotype frequency in the top-performing beta regression models, although pyrethroid use surprisingly had a negatively associated with resistance. The incorporation of spatial lags resulted in improvements to the fit and explanatory power of the models, indicating an underlying diffusion process likely explains some of the spatial patterns observed. CONCLUSIONS: The genetic mutations that confer resistance to pyrethroids in Ae. aegypti mosquitoes in Florida exhibit spatial autocorrelation and patterns that can be partially explained by landscape and insecticide-use factors. Further work at local scales should be able to identify the mechanisms by which these variables influence selection for alleles associated with resistance.

Failure of Permethrin-Treated Military Uniforms to Protect Against a Laboratory-Maintained Knockdown-Resistant Strain of Aedes aegypti.

Military forces and the recreational industry rely on the repellent properties of permethrin-treated fabrics and N,N-diethyl-meta-toluamide (deet)-based lotions to provide protection from disease vectors and hematophagous organisms. Concerns regarding efficacy have been raised as pyrethroid resistance becomes more common and recent publications present contradictory conclusions. In this preliminary study, consenting volunteers were exposed to pyrethroid-susceptible and pyrethroid-resistant Aedes aegypti strains while wearing sleeves of untreated or permethrin-treated army uniform fabric as well as with untreated or deet-treated exposed forearms. Deet was nearly 100% effective against both susceptible and resistant strains. However, permethrin treatment provided no significant protection against the resistant Puerto Rico strain relative to untreated control sleeves. These results confirm that pyrethroid-resistant vectors can negate the efficacy of permethrin-treated uniforms. Additional testing with resistant field strains is needed to better understand the risk to service members.

Insecticidal and repellent properties of novel trifluoromethylphenyl amides III.

The adulticidal, larvicidal, and repellent activity of 18 trifluoromethylphenyl amides (TFMPAs) was determined against Aedes aegypti mosquitoes. The compounds studied are the third generation designed from active structures of the previous two generations. N-(3,5-Bis(trifluoromethyl)phenyl)-2-chloroacetamide (8f) and N-(3,5-bis(trifluoromethyl)phenyl)-2,2,3,3,3-pentafluoropropanamide (8h) were most active against 1st stage Ae. aegypti larvae with LC50 values of 125 and 2.53 µM; for comparative purposes, the published LC50 for fipronil is 0.014 µM. Compound 8h was the most toxic against adult female Ae. aegypti with an LD50 = 2.12 nmol/mg, followed by 8f, and N-(3,5-bis(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (8g) with LD50 values of 4.27 and 4.73 nmol/mg, respectively, although these compounds were significantly less toxic than fipronil against adult female Ae. aegypti. Compounds N-(2-(trifluoromethyl)phenyl)butyramide (9c), N-(2-(trifluoromethyl)phenyl)pentanamide (9d) and N-(2-(trifluoromethyl)phenyl)hex-5-enamide (9e) were the best repellents for female Ae. aegypti, with minimum effective dosages (MEDs) of 0.026, 0.052, and 0.091 µmol/cm2, respectively, compared to DEET at 0.052 µmol/cm2. Out of 52 TFMPAs (total number of compounds from three generations of this research) compound 9c was the most active repellent along with two synthesized in our previous studies, 2-chloro-N-(3-(trifluoromethyl)phenyl)acetamide (6a) and 2,2,2-trifluoro-N-(2-(trifluoromethyl)phenyl)acetamide (4c).

Chemical composition of the essential oil and n-hexane extract of Stachys tmolea subsp. Tmolea Boiss., an endemic species of Turkey, and their mosquitocidal activity against dengue vector Aesdes aegypti.

Stachys tmolea subsp. tmolea Boiss. is endemic to Turkey and is a species of the genus Stachys L. which is one of the largest genera of the family Lamiaceae with about 300 species. The aims of this study were to examine the chemical composition of the essential oil and n-hexane extract of S. tmolea subsp. tmolea as natural sources of insecticidal activity against the dengue vector, Aedes aegypti. Analysis of the essential oil by GC-FID and gas chromatography-mass spectrometry (GC-MS) systems identified hexahydrofarnesyl acetone (15%), viridiflorol (10%), hexadecanoic acid (7%) and 9-geranyl-p-cymene (6%) as major components. The volatile components of the n-hexane extract were extracted using headspace solid-phase microextraction (HS-SPME) and were analyzed using GC-MS. The principal constituents were 3,4-dimethyl decane (16%), 3-methyl-3-pentanol (15%), 2-methyl-2-pentanol (12%), 1,4-bis (1,1-dimethylethyl) benzene (12%), heptanal (10%), acetic acid (6%) and decane (4%). Bioassay of the n-hexane extract, at 5 µg/mosquito, produced 90% mortality against adult Ae. aegypti while the S. tmolea essential oil demonstrated 13% mortality. No larvicidal activity was observed both in essential oil and n-hexane extract. Further studies are needed to assess the adulticidal activity of the responsible compounds in the crude extract.

Chemical composition and antioxidant, cytotoxic, and insecticidal potential of Valeriana alliariifolia in Turkey.

Valeriana is a common plant species used for various healing purposes in folk medicine since antiquity. This study investigates the phytochemical profile, antioxidant, cytotoxic, and insecticidal activity of Valeriana alliariifolia Adams, a species that has traditionally been used in Turkey. For the analyses we prepared four root extracts of V. alliariifolia Adams using hexane (HM1), chloroform (CM1), ethanol (EM1), and water (WM1) for maceration. Additionally, two extracts were also prepared from its roots by maceration separately with ethanol (EM2) and water (WM2). One sample was prepared as a water infusion (WI), according to the procedure used in Turkish traditional medicine. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical cation scavenging activity tests showed that ethanol extracts had the strongest antioxidant activity: EM1 (IC50 - DPPH: 17.694 µg/mL; ABTS: 23.8 µg/mL) and EM2 (IC50 - DPPH: 20 µg/mL; ABTS: 21.5 µg/mL). The hexane extract, HM1, was the most cytotoxic (IC50<10 µg/mL against HepG2 and HUVEC) and EM2 strongly cytotoxic (IC50<10 µg/mL against HepG2 and IC50: 11.96 µg/mL against HUVEC). The extracts with demonstrated cytotoxic activities were further examined to check their insecticidal activity against adult female mosquito Aedes aegypti and first instar Ae. aegypti larvae. HM1 was the most effective (90±10 %), which was consistent with its cytotoxic activity. Because of the high antioxidant, cytotoxic, and insecticidal activities, we ran phytochemical analyses of the HM1, EM1, and EM2 extracts with GC-MS (for HM1) and LC-MS/MS (for EM1 and EM2). We also analysed the composition of the essential oil obtained from V. alliariifolia roots by micro-distillation in order to compare its content with HM1, which contains volatile compounds. Phytochemical analyses revealed that the major compound in HM1 was isovaleric acid (16 %) and in the essential oil 1,8-cineole (2.9 %). EM1 and EM2 contained 5-O-caffeoylquinic acid (chlorogenic acid), verbascoside (acteoside), and 3,5-dicaffeoylquinic acid as major components. In the light of our findings and available literature, we can conclude that V. alliariifolia has a good bioactive potential that could be used for different purposes, including the development of new agents for the treatment of various diseases. The difference in the content between the essential oil and HM1 was remarkable. It suggests that the variability observed in the activity of the samples was a result of composition and that, therefore, the aim of treatment should dictate which type of preparation is to be selected. An added value of our study is that it determined verbascoside and methylquercetin rutinoside for the first time in the Valeriana extracts.