Outer Epidermis Research Articles

Topical delivery of baricitinib-impregnated nanoemulgel: a promising platform for inhibition of JAK -STAT pathway for the effective management of atopic dermatitis.

Baricitinib, an inhibitor of Janus kinase 1/2 receptors majorly involved in the dysregulation of immune responses in atopic dermatitis, is currently approved for managing atopic dermatitis in Europe. The delivery of baricitinib through oral route is associated to several adverse effects due to off-target effects. Therefore, the current study is aimed at formulation of baricitinib loaded nanoemulgel for evaluation of topical delivery potential in the treatment of atopic dermatitis. The baricitinib-loaded nanoemulsions (0.05 and 0.1% w/w) revealed an average globule size of 162.86 ± 0.37 and 173.66 ± 4.88nm respectively with narrow PDI. The optimized batch of baricitinib nanoemulsion was converted to nanoemulgel by the addition of the mixture of gel bases SEPINEO™ DERM and SEPINEO™ P 600 along with propylene glycol, resulting in pseudoplastic shear thinning behaviour. The optimized nanoemulgels have shown prominent retention of baricitinib in the skin along with permeation. The skin distribution study of coumarin-6 loaded nanoemulgel demonstrated high fluorescence in the epidermal layer. The western blot analysis revealed significant inhibition of phosphorylated signal transducers and activators of transcriptions 1 (##p < 0.01) and 3 (#p < 0.05) by application of 0.05 and 0.1% baricitinib nanoemulgel. The baricitinib nanoemulgels have shown anti-inflammatory activity by significantly reducing expressions of various inflammatory markers. Histopathological analysis of skin tissues treated with baricitinib nanoemulgel has demonstrated a marked reduction in acanthosis, hyperkeratosis, and intact outer epidermis. These results supported the potential role of baricitinib-loaded nanoemulgel in reducing the inflammation and disease severity associated with atopic dermatitis.

Effect of Egg Washing and Hen Age on Cuticle Quality and Bacterial Adherence in Table Eggs

The cuticle covering the outer surface of an eggshell functions as both a physical and chemical barrier against invading microorganisms. Contamination of eggs by microbial pathogens progresses in four stages: bacterial attachment to the egg surface, penetration through the cuticle and eggshell, multiplication within the underlying membranes, and the final stage of contaminating the egg contents. Therefore, it is important to study bacterial count at the first point of contact, i.e., on the surface of the eggs. In this study, we have evaluated the impact of differences in cuticle quality (due to egg washing and hen age) on bacterial load. We compared bacterial adherence on the eggshell surface of white eggs which were either washed (graded) or unwashed (ungraded), collected from Lohmann laying hens of different ages: early (24–28 weeks), mid-lay (44–48 weeks), and late (66–70 weeks). We aimed to determine the impact of hen age and egg washing on differences in cuticle quality and bacterial adherence. Our results indicate that hen age (up to 70 weeks) and commercial egg washing do not significantly impact bacterial adherence on eggshell surfaces. We have developed a novel method using green fluorescent protein (GFP)-expressing Salmonella typhimurium to estimate adherence of bacteria to the eggshell surface, with independent measurement of autofluorescence to quantitate cuticle deposition. S. typhimurium were localized, adhering to cracks visible on the outer cuticle in ungraded eggs, indicating that egg-associated pathogens usually enter the egg interior either through respiratory pores in eggshells or through shell micro-cracks. The results of this study can be utilized to optimize innovative methods for predictive microbiology in order to achieve egg safety.

Analysis of the role of the rice metallothionein gene OsMT2b in grain size regulation

Seed size is one of the three main characteristics determining rice yield. Clarification of the mechanisms regulating seed size in rice has implications for improving rice yield. Although several genes have been reported to regulate seed size, most of the reports are fragmentary. The role of metallothioneins (MTs) in regulating seed size remains unknown. Here, we found that OsMT2b was expressed in both spikelets and developing seeds. OsMT2b-overexpression lines had large and heavy seeds, and RNAi (RNA interference) lines had small and light seeds. Scanning electron microscopy (SEM) observations revealed that OsMT2b regulated spikelet hull size by affecting cell expansion in the outer epidermis. Histological analysis indicated that OsMT2b affected the number of cells in the cross-section of spikelet hulls, which affected seed size. The fresh weight of seeds was consistently higher in OsMT2b-overexpression lines than in seeds of the wild-type (WT) and RNAi lines from 6 DAP (days after pollination) until maturity, indicating that OsMT2b affected seed filling. Reverse transcription-quantitative PCR (RT-qPCR) analyses revealed that OsMT2b regulates the expression of reactive oxygen species scavenging-related genes involved in seed size regulation. In conclusion, our results indicated that OsMT2b positively regulates seed size, which provides a novel approach for regulating seed size with genetic engineering technology.

Энтомопатогенные грибы как средство подавления численности рыжего таракана

The use of mycoinsecticides can be an alternative to reduce chemical pollution in human dwellings. The use of Beauveria bassiana and Metarhizium anisopliae is gradually increasing, reaching commercial scale in such countries as Brazil, China, and Mexico. At the same time, significant progress has been made in understanding the molecular aspects of pathogenesis, allowing the selection of highly effective fungal strains. Differences in the susceptibility of adults and nymphs of the German cockroach Blattella germanica L. to entomopathogenic fungi was shown. The possibility of enhancing the pathogenicity of various types of fungi when integrated with insecticides in sublethal doses is considered. The German cockroach has evolved many physical barriers, behavioral traits, and physiological mechanisms that prevent infection by various entomopathogens. These barriers include the outer cuticle, inner cuticle, digestive tract and peritrophic membrane, and immunological barriers. Fungal conidia entering the cockroach’s digestive tract was inactivated by a combination of antimicrobial compounds produced by the cockroach and commensal microorganisms, intestinal pH and digestive enzymes. The microbiota of Bl.?germanica may play a critical role in protecting cockroaches from fungal invasion and colonization. Keywords: insecticide resistance, Blattella germanica, Beauveria bassiana, Metarhizium anisopliae, mycoinsecticides, entomopathogenic fungi.

Genome-wide identification of crustacyanin and function analysis of one isoform high-expression in carapace from Neocaridina denticulata sinensis

Lipocalin proteins transport hydrophobic molecules, including apolipoprotein D, retinol-binding protein, and crustacyanin (CRCN). CRCN can combine with astaxanthin to cause a bathochromic shift in the emission spectrum of astaxanthin from red to blue. Therefore, CRCN influences the colors and patterns of crustaceans, which are important for various biological functions such as camouflage, reproduction, and communication. For aquatic organisms, body color is economically important and can be indicative of habitat water quality. In this study, thirteen CRCN genes (NdCRCNs) were first discovered in Neocaridina denticulata sinensis, contradicting prior findings of a few isoform genes in a species. The expression pattern of NdCRCNs in tissues showed that the expression of one CRCN isoform gene, named NdCRCN-30, was the highest in the carapace. In situ hybridization (ISH) analysis revealed that NdCRCN-30 was predominantly distributed in the outer epidermis of shrimp. Interference of NdCRCN-30 could cause a change in the color of the carapace. RNA-seq was performed after knockdown with the NdCRCN-30, and differential gene enrichment analysis revealed that this gene is primarily associated with antioxidant function, pigmentation, and molting. Overall, our results will provide new insights into the biological function of the CRCN and genetic breeding for changing body color in economic crustaceans.

Identification of long-chain acyl-CoA synthetase gene family reveals that SlLACS1 is essential for cuticular wax biosynthesis in tomato

Long-chain acyl-CoA synthetases (LACSs), belonging to the acyl-activating enzyme superfamily, play crucial roles in lipid biosynthesis and fatty acid catabolism. Here, we identified 11 LACS genes in the tomato reference genome, and these genes were clustered into six subfamilies. Gene structure and conserved motif analyses indicated that LACSs from the same subfamily shared conserved gene and protein structures. Expression analysis revealed that SlLACS1 was highly expressed in the outer epidermis of tomato fruits and leaves. Subcellular localization assay results showed that SlLACS1 was located in the endoplasmic reticulum. Compared with wild-type plants, the wax content on leaves and fruits decreased by 22.5–34.2 % in SlLACS1 knockout lines, confirming that SlLACS1 was involved in wax biosynthesis in both leaves and fruits. Water loss, chlorophyll extraction, water-deficit, and toluidine blue assays suggested that cuticle permeability was elevated in SlLACS1 knockout lines, resulting in reduction in both drought stress resistance and fruit shelf-life. Overall, our analysis of the LACSs in tomato, coupled with investigations of SlLACS1 function, yielded a deeper understanding of the evolutionary patterns of LACS members and revealed the involvement of SlLACS1 in wax accumulation contribute to drought resistance and extended fruit shelf-life in tomato.

Vitamin D Supplementation in Congenital Ichthyosis: A Case Series.

Ichthyosis is a group of genetic keratinization disorders characterized by excessive scaling that is associated with hyperproliferative epidermis and/or cellular retention. Whereas normal outer epidermis thickness is 25 μm, it can be 10-fold greater in patients with ichthyosis. As a result, photoactivation of 7-dehydrocholesterol is impaired, causing systemic vitamin D deficiency.In this case series, 25 patients with congenital ichthyosis with vitamin D deficiency (<10 ng/mL) were supplemented with 60,000 IU of vitamin D3 for 10 days followed by daily allowance of 400 to 600 IU of vitamin D3 and 40 mg/kg per day of elemental calcium. The authors assessed improvement in cutaneous scaling and body and tested patients' blood and urine samples at day 1, day 10, 1 month, and 3 months. They also documented patients' Dermatology Life Quality Index score before and after treatment.All patients had normal vitamin D levels; supplementation was discontinued for two patients who reached a level of 100 ng/mL within 10 days. Subjective improvement of symptoms (dryness of the skin, allergic rhinitis, tightness of the skin, and scaling) was observed by both the provider and the patients. There was remarkable improvement in symptoms of severe ichthyosis such as lamellar ichthyosis (tightness of the skin and scaling). Marked improvement in Dermatology Life Quality Index score was also noted.This case series demonstrated remarkable symptomatic relief with vitamin D supplementation in patients with congenital ichthyosis; however, additional research should be conducted with larger sample sizes to support these findings.

The structure and proteomic analysis of byssus in Pteria penguin: Insights into byssus evolution and formation

Byssus is a unique external structure in sessile bivalves and is critical for settlement and metamorphosis. However, little is known about the stout byssus in Pteria penguin. We explored the byssus structure and proteins using scanning electron microscopy and proteomics, respectively. The results revealed that P. penguin byssus has a dense and highly aligned fiber inner core, and the outer cuticle contains protein granules embedded in the protein matrix. Proteomic analysis revealed 31 proteins in the byssus, among which 15 differentially expressed proteins were mainly enriched in the EGF/EGF-like and laminin EGF-like domains. Foot proteins were enriched in the EF-hand, immunoglobulin, and fibronectin domains. All these domains can participate in protein-protein and/or protein-metal interactions in the extracellular matrix (ECM), which, together with the seven types of ECM proteins detected in the byssus, supports the hypothesis that the byssus is derived from the ECM. We also found that in vitro acellular structures of the byssus and the shell shared commonalities in their formation processes. These results are useful for further understanding byssus evolution and the characterization of byssus-related proteins. SignificanceThis manuscript investigates the structure and the origin of Pteria penguin byssus, given that byssus is vital to provide critical protection for reproduction and even against environmental stresses that affect survival. However, there is rare research on byssus protein composition. Hence, though scanning electron microscopy and proteomic analysis, we discovered that P. penguin byssus possesses the dense and highly aligned fiber inner core, and the outer cuticle has protein granules embedded in the protein matrix. Proteomic analysis showed that there were 31 proteins in the byssus, among which 15 proteins were mainly enriched in the EGF/EGF-like and laminin EGF-like domains. Foot proteins closely related to byssus formation were enriched in EF hand, immunoglobulin, and fibronectin domains. These domains are able to participate in protein-protein and/or protein-metal interactions in the extracellular matrix (ECM), which together with the seven types of ECM proteins detected in byssus support the hypothesis that byssus derive from the ECM. We also found in vitro acellular structures the byssus and the shell share commonalities in their formation processes. These results were useful for further understanding the byssus evolution and the characterization of the byssus-related proteins.

Optimization of DNA Extraction for ITS/U4U3 analysis of Rhipsalis baccifera

Genetic analysis of plants relies on high yields of pure DNA. For Rhipsalis baccifera this represents a great challenge since its plant tissue can accumulate large amounts of mucilage, polysaccharides, polyphenols and secondary metabolites, which co-purify with amplifiable DNA. These contaminating compounds lead to a poor yield and prevent acces to PCR-based analysis. These contaminating compounds lead to a poor DNA yield and prevent access to PCR-based analysis. A number of factors, including choice of plant tissue, tissue preparation, and modifications of the extraction buffer, can impact on DNA extraction process. In this study, four different DNA extraction procedures were tested aiming to develop a simple protocol based on CTAB buffer. The obtained results showed that the use of the outer cuticle of old lyophilized tissue allowed reliable results in R. baccifera plants with a good purity ranging from 1.6 to 1.8 and high DNA yield yield ? 500 ng ?L-1.

Reproductive biology of Centaurea kilaea (Asteraceae, Cardueae) – an endemic species from Türkiye

In this study, the embryology of Centaurea kilaea Boiss., a species endemic to Türkiye, was examined using light microscopy. The anthers of C. kilaea are tetrasporangiate; the anther wall development is dicotyledonous; and the tapetum is amoeboid. The meiotic division of the microspore mother cells is regular, and when the pollen grains are thrown from the anthers, they are three-celled. The ovary of C. kilaea is inferior, bicarpellary, syncarpous, and unilocular, which is characteristic of the Asteraceae family. It carries only a single ovule with basal placentation. The ovule is anatropous, unitegmic, and tenuinucellate. The megaspore mother cell undergoes meiotic division, giving rise to a linear tetrad of megaspores. The chalazal megaspore remains functional, and the other three megaspores degenerate rapidly. The functional megaspore undergoes three mitotic divisions in succession. As a result, a Polygonum-type embryo sac, with eight nuclei and seven cells, is formed. The antipodal cells persist until the first divisions of the zygote. In the mature embryo sac stage, the integument consists of the endothelium, peri-endothelial region, parenchymatous cells, and outer epidermis, from the inside out. Endosperm development is initially free nuclear, becoming cellular in the globular embryo stage. Embryo development is of the asterad type. The mature seed does not contain endosperm, but the endothelium persists.

Spatial multi-omics of human skin reveals KRAS and inflammatory responses to spaceflight

Spaceflight can change metabolic, immunological, and biological homeostasis and cause skin rashes and irritation, yet the molecular basis remains unclear. To investigate the impact of short-duration spaceflight on the skin, we conducted skin biopsies on the Inspiration4 crew members before (L-44) and after (R + 1) flight. Leveraging multi-omics assays including GeoMx™ Digital Spatial Profiler, single-cell RNA/ATAC-seq, and metagenomics/metatranscriptomics, we assessed spatial gene expressions and associated microbial and immune changes across 95 skin regions in four compartments: outer epidermis, inner epidermis, outer dermis, and vasculature. Post-flight samples showed significant up-regulation of genes related to inflammation and KRAS signaling across all skin regions. These spaceflight-associated changes mapped to specific cellular responses, including altered interferon responses, DNA damage, epithelial barrier disruptions, T-cell migration, and hindered regeneration were located primarily in outer tissue compartments. We also linked epithelial disruption to microbial shifts in skin swab and immune cell activity to PBMC single-cell data from the same crew and timepoints. Our findings present the inaugural collection and examination of astronaut skin, offering insights for future space missions and response countermeasures.

Development and anatomy of petals with specialized nectar holder and pollen container in Fumarioideae (Papaveraceae).

Petal developmental characteristics in Fumarioideae were similar at early stages, and the specialized nectar holder/pollen container formed by the outer/inner petals. The micro-morphology of these two structures, however, shows diversity in seven species. Elaborate petals have been modified to form different types, including petal lobes, ridges, protuberances, and spurs, each with specialized functions. Nectar holder and pollen container presumably have a function in plant-pollinator interactions. In Fumarioideae, four elaborate petals of the disymmetric/zygomorphic flower present architecture forming the "nectar holder" and "pollen container" structure at the bottom and top separately. In the present study, the petals of seven species in Fumarioideae were investigated by scanning electron microscopy, light microscope, and transmission electron microscopes. The results show that petal development could divided into six stages: initiation, enlargement, adaxial/abaxial differentiation, elaborate specializations (sacs, spurs, and lobes formed), extension, and maturation, while the specialized "nectar holder" and "pollen container" structures mainly formed in stage 4. "Nectar holder" is developed from the shallow sac/spur differentiated at the base of the outer petal, eventually forming a multi-organized complex structure, together with staminal nectaries (1-2) with individual sizes. A semi-closed ellipsoidal "pollen container" is developed from the apical part of the 3-lobed inner petals fused by middle lobes and attain different sizes. The adaxial epidermis cells are specialized, with more distinct punctate/dense columnar protrusions or wavy cuticles presented on obviously thickening cell walls. In addition, a large and well-developed cavity appears between the inner and outer epidermis of the petals. As an exception, Hypecoum erectum middle lobes present stamen mimicry. Elaborate petal structure is crucial for comprehending the petal diversity in Fumarioideae and provides more evidence for further exploration of the reproductive study in Papaveraceae.

Genetic and Molecular Regulation of Cotton Fiber Initiation and Elongation

Cotton fiber, a crucial and sustainable resource for global textile production, undergoes a complex five-stage developmental process, encompassing initiation, elongation, transition, secondary cell wall biosynthesis, and maturation. These elongated single-cell fibers originate from the outer ovule epidermis. The development of cotton fibers involves intricate changes in gene expression and physiological processes, resulting in a nearly pure cellulose product that is vital for the global cotton industry. Decoding the genes associated with fiber development enhances our understanding of cotton fiber mechanisms and facilitates the cultivation of varieties with enhanced quality. In recent decades, advanced omics approaches, including genomics, transcriptomics, and proteomics, have played a pivotal role in identifying the genes and gene products linked to cotton fiber development, including the MYB transcription factor family, which coordinates cotton fiber development. Molecular studies have revealed the transcription factors, like MYB, WRKY, Homeodomain Leucine Zipper (HD-ZIP), and basic helix–loop–helix (bHLH), influencing fiber initiation and elongation. The intricate interplay of phytohormones, like auxin, gibberellic acid (GA), brassinosteroids (BRs), jasmonic acid (JA), ethylene, abscisic acid (ABA), and cytokinin, is explored, providing a comprehensive perspective on the shaping of cotton fibers. Numerous candidate genes and cellular processes affecting various aspects of fiber development hold promise for genetic engineering or marker-assisted breeding to improve fiber quality. This review presents a comprehensive overview of key achievements in cotton molecular biology, with a specific emphasis on recent advancements in understanding the transcription factors and phytohormones involved in cotton fiber initiation and elongation.

Fluid-Solid Interfacial Properties and Drag-Reducing Characterization of the Flexible Conical Microstructured Film Inspired by the Streamlined Body Surface of the Pufferfish.

Given the challenges in accurately replicating the surface of the pufferfish, this study employed three-dimensional (3D) printing to create a model based on inverse modeling. The morphology of the pufferfish exhibits a streamlined configuration, characterized by a gradual widening from the anterior oral region to the central ocular area, followed by a progressive narrowing from the midabdominal region toward the caudal extremity. The RNG k-ε turbulence simulation results demonstrate that the streamlined body surface of the pufferfish diminishes differential pressure resistance. This enhancement promotes laminar flow formation, delays fluid separation, minimizes turbulence-induced vortices, and reduces frictional resistance. Moreover, the pufferfish's supple and uneven outer epidermis was simplified into a flexible, nonsmooth planar film to conduct fluid-solid coupling simulations. These revealed that the pufferfish's unique skin can absorb turbulent energy and minimize momentum transfer between the fluid and the solid film, lowering the fluid resistance during swimming. In summary, The high-efficiency swimming capacity of pufferfish stems not only from their streamlined body surface but also significantly from the unique structural characteristics and mechanical properties of their flexible skin. This research provides critical theoretical underpinnings for the design of functional bionic surfaces aimed at drag reduction.

TESTING THE ACTIVITY AND FORMULATION OF NATURAL HAND SOAP BASED ON NATURAL SURFACTANTS OF LERAK FRUIT (Sapindus rarak DC.) AGAINST Staphylococcus aureus

Lerak (Sapindus rarak DC.) is a fruit belonging to the Sapindaceae family that usually grows in tropical forests. Lerak fruit flesh contains the main active compound, saponin. Saponin is a natural surfactant with foaming and emulsifying properties. The use of synthetic surfactants in the manufacture of hand soap sold on the market can have negative impacts, such as irritation and denaturation of proteins in the outer epidermis layer in humans if exposed too often. This study aimed to determine whether lerak can be formulated into natural hand soap as a natural surfactant and determine its antibacterial activity against Staphylococcus aureus bacteria. Lerak fruit solutions were prepared using a maceration method. The resulting solution was then formulated into hand soap at concentrations of 10%, 30%, and 50%. The evaluation of the natural hand soap preparation from the lerak fruit included organoleptic evaluation, homogeneity, pH, viscosity, foam height, and antibacterial activity against Staphylococcus aureus bacteria using the disc diffusion method. Based on the research results, Lerak Fruit (Sapindus rarak DC.) can be formulated as a natural surfactant in hand soap at concentrations of 0%, 10%, 30%, and 50%. Based on the evaluation of the physical properties in the foam height test, F1, F2, and F3 met the requirements, whereas F0 did not meet the requirements in the foam stability test because it did not contain lerak fruit extract, so no foam formed ...

Morphology, anatomy, and quantitative determination of corosolic acid in Lagerstroemia calyculata Kurz in the Southeast region, Vietnam

Objectives: To describe the morphological characteristics, anatomy (stem, leaves), and quantify the corosolic acid in the leaves of Lagerstroemia calyculata Kurz collected in the Southeast region, Vietnam. Subjects and methods: Using the comparative morphological method to determine the scientific name of species. Studying the microscopic characteristics of Lagerstroemia calyculataKurz according to the research method of N. Ba (2009) [1] and quantifying the content of corosolic acid in the leaves according to the United States Pharmacopeia 2017 (USP 40), with adjustments to suit the conditions of the laboratory. Results: Lagerstroemia calyculata Kurz is tree, deciduous, height 30-35 m, smooth bark. Leaves lanceolate, lateral veins 9-11 pairs, hairy. Inflorescence paniculate, small white flowers. Capsules ovoid, 6-valved. Seeds are numerous, brown, and smooth. The stem has a bicollateral vascular bundle, conduction tissue arranged continuously, double phloem pillars and leaf veins exposed on both sides. The outer epidermis has multicellular stellate hairs. Scattered in soft tissue there are crystalline particles. Stomata are present only on the underside of leaves. The quantification of corosolic acid in three leaf samples of Lagerstroemia calyculata Kurz was from 0.05 to 0.12%. Conclusions: This study has detailed morphological, anatomical characteristics, and quantitative determination of corosolic acid in three leaf samples of Lagerstroemia calyculata Kurz collected in the Southeast region.

Cellular patterning and cyto-architectural organization of the skin of electric catfish (Malapterurus electricus, Siluriformes) with a particular emphasis on its ampullary electroreceptor

The functional morphology of the skin of Malapteruridae is presumably evolved to cope with a diversified range of ambient physiological, environmental, and behavioral conditions. Herein, we firstly characterized the microstructures and intriguing patterning of the skin of twelve adult electric catfish (Malapterurus electricus, Malapteruridae) using histological, histochemical, immunofluorescent, and ELISA standard methodology. The skin comprises three sequentially–oriented layers: the epidermis, dermis, and hypodermis with a significantly increased thickness of the former. The epidermis contains four types of cells: the surface epithelial cells, mucous cells, granular cells, and club cells. We defined distinctive ampullary electroreceptors in the outer epidermis that possess flask-shaped sensory crypt containing electroreceptor cells together with vertical collagen rods. Dermis and hypodermis are composed of connective tissue; however, the former is much more coarse and dense with comparable reactivity for Masson–Goldner trichrome (MT). Placing our data in the context of the limited body of previous work, we showed subtle changes in the expression of mucin subunits together with cytoskeletal fractions of collagens, myosin, F-actin, keratins, and tubulins. Taken as a whole, our results convincingly showed that the skin of M. electricus shares some structural similarities to other Siluriformes, however, it has some functional modifications that are implicated in protection, defense, and foraging behavior.

Larvicidal potential, antimicrobial properties and molecular docking analysis of Egyptian Mint (Mentha rotundifolia) against Culex pipiens L. (Diptera: Culicidae) and Midgut-borne Staphylococcus aureus

Mosquitoes prefer stagnant areas near hospitals to live and easily spread pathogenic bacteria. Our current study aims to isolate multidrug-resistant (MDR) Staphylococcus aureus isolates from midguts of Mosquito Culex pipiens and study the potential of mint as a biocontrol strategy against C. pipiens larvae and their midgut-borne S. aureus. Samples of the third and fourth larval instars of C. pipiens were collected from water ponds around three Cairo hospitals. Ciprofloxacin, gentamycin and tetracycline, as well as various concentrations of mint leaf extract (MLE) were tested for antibiotic susceptibility. Sixty-five isolates were obtained and showed antibiotic resistance to tetracycline, gentamycin, ciprofloxacin, and undiluted MLE with resistant percentages (%) of 27.69, 30.76, 17.46, and 23.08%, respectively. Undiluted MLE inhibited 61.53% of the multidrug S. aureus isolates, whereas it couldn't inhibit any of these isolates at dilutions less than 50 μg/mL. The MIC of MLE was ≤ 700 µg/mL, while the MIC of the antibiotics ranged from 0.25 to 5.0 µg/mL for the three antibiotics. The most inhibited S. aureus isolate was identified by 16SrRNA sequencing approach and registered in GenBank as S. aureus MICBURN with gene accession number OQ766965. MLE killed all larval stages after 72 h of exposure, with mortality (%) reaching 93.33 and 100% causing external hair loss, breakage of the outer cuticle epithelial layer of the abdomen, and larvae shrinkage. Histopathology of treated larvae showed destruction of all midgut cells and organelles. Gas chromatography (GC) of MLE revealed that menthol extract (35.92%) was the largest active ingredient, followed by menthone (19.85%), D-Carvone (15.46%), Pulegone (5.0579%). Docking analysis confirmed that alpha guanine and cadinol had the highest binding affinity to both predicted active sites of Culex pipiens acetylcholinesterase. As a result, alpha-guanine and cadinol might have a role as acetylcholinesterase inhibitors.

Application of a recombinant novel trypsin from Trichinella spiralis for serodiagnosis of trichinellosis

BackgroundThe excretory/secretory (ES) antigen of Trichinella spiralis muscle larvae (ML) is currently the most widely used diagnostic antigen to detect T. spiralis infection. However, this antigen has certain drawbacks, such as a complicated ES antigen preparation process and lower sensitivity during the early phase of infection. The aim of this study was to investigate the features of a novel T. spiralis trypsin (TsTryp) and evaluate its potential diagnostic value for trichinellosis.MethodsThe TsTryp gene was cloned and recombinant TsTryp (rTsTryp) expressed. Western blotting and an enzyme-linked immunosorbent assay (ELISA) were performed to confirm the antigenicity of rTsTryp. The expression pattern and distribution signature of TsTryp at various life-cycle stages of T. spiralis were analyzed by quantitative PCR, western blotting and the immunofluorescence test. An ELISA with rTsTryp and ML ES antigens was used to detect immunoglobulins G and M (IgG, IgM) in serum samples of infected mice, swine and humans. The seropositive results were further confirmed by western blot with rTsTryp and ML ES antigens.ResultsTsTryp expression was observed in diverse T. spiralis life-cycle phases, with particularly high expression in the early developmental phase (intestinal infectious larvae and adults), with distribution observed mainly at the nematode outer cuticle and stichosome. rTsTryp was identified by T. spiralis-infected mouse sera and anti-rTsTryp sera. Natural TsTryp protease was detected in somatic soluble and ES antigens of the nematode. In mice infected with 200 T. spiralis ML, serum-specific IgG was first detected by rTsTryp-ELISA at 8 days post-infection (dpi), reaching 100% positivity at 12 dpi, and first detected by ES-ELISA at 10 dpi, reaching 100% positivity at 14 dpi. Specific IgG was detected by rTsTryp 2 days earlier than by ES antigens. When specific IgG was determined in serum samples from trichinellosis patients, the sensitivity of rTsTryp-ELISA and ES antigens-ELISA was 98.1% (51/52 samples) and 94.2% (49/52 samples), respectively (P = 0.308), but the specificity of rTsTryp was significantly higher than that of ES antigens (98.7% vs. 95.4%; P = 0.030). Additionally, rTsTryp conferred a lower cross-reaction, with only three serum samples in total testing positive from 11 clonorchiasis, 20 cysticercosis and 24 echinococcosis patients (1 sample from each patient group).ConclusionsTsTryp was shown to be an early and highly expressed antigen at intestinal T. spiralis stages, indicating that rTsTryp represents a valuable diagnostic antigen for the serodiagnosis of early Trichinella infection.Graphical

Microchemistry of Single Hair Strands Below and Above the Scalp: Impacts of External Contamination on Cuticle and Cortex Layers.

External contamination of hair is the most significant challenge to it becoming an accepted matrix for monitoring endogenous metal exposure and nutritional deficiency. Here we use laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to quantify elemental concentrations in hair strands below and above the scalp in the cuticle and cortex layers to determine the extent of external contamination in a reference population. Evidence of hair strand contamination occurred for barium, calcium, iron, magnesium, and strontium in both the outer cuticle and the inner cortex layers, with increasing concentrations from root to tip. Aluminum, boron, copper, lead, and manganese showed significant contamination in the cuticle layer only, suggesting some protection of the inner cortex. Phosphorus and potassium decreased outside the scalp suggesting loss by washing, while chromium, mercury, selenium, sodium, titanium, and zinc showed no evidence of loss or external contamination above the scalp. The results clearly show that for most elements, hair chemistry above the scalp is unreliable for use in interpretation of endogenous exposures or deficiencies, and that the below-scalp portion provides a more accurate monitoring tool. This is the first paper to provide a reference range of elemental hair chemistry that is not impacted by the external environment.