Light Microscopy Research Articles

Systematic analysis of ocular features and responses of cultured spotted wolffish (Anarhichas minor).

A better understanding of unique anatomical and functional features of the visual systems of teleost fish could provide key knowledge on how these systems influence the health and survival of these animals in both wild and culture environments. We took a systematic approach to assess some of the visual systems of spotted wolffish (Anarhichas minor), a species of increasing importance in North Atlantic aquaculture initiatives. The lumpfish (Cyclopterus lumpus) was included in these studies in a comparative manner to provide reference. Histology, light and electron microscopy were used to study the spatial distribution and occurrence of cone photoreceptor cells and the nature of the retinal tissues, while immunohistochemistry was used to explore the expression patterns of two photoreceptor markers, XAP-1 and XAP-2, in both species. A marine bacterial infection paradigm in lumpfish was used to assess how host-pathogen responses might impact the expression of these photoreceptor markers in these animals. We define a basic photoreceptor mosaic and present an ultrastructural to macroscopic geographical configuration of the retinal pigment tissues in both animals. Photoreceptor markers XAP-1 and XAP-2 have novel distribution patterns in spotted wolffish and lumpfish retinas, and exogenous pathogenic influences can affect the normal expression pattern of XAP-1 in lumpfish. Live tank-side ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT) revealed that normal cultured spotted wolffish display novel variations in the shape of the retinal tissue. These two complementary imaging findings suggest that spotted wolffish harbour unique ocular features not yet described in marine teleosts and that visual function might involve specific retinal tissue shape dynamics in these animals. Finally, extensive endogenous biofluorescence is present in the retinal tissues of both animals, which raises questions about how these animals might use retinal tissue in novel ways for visual perception and/or communication. This work advances fundamental knowledge on the visual systems of two economically important but now threatened North Atlantic teleosts and provides a basic foundation for further research on the visual systems of these animals in health versus disease settings. This work could also be useful for understanding and optimizing the health and welfare of lumpfish and spotted wolffish in aquaculture towards a one health or integrative perspective.

Pseudo-NRBC in the Mindray BC-6800Plus analyzer: A clue for diagnostic anticipation of fungemia. Experimental and preliminary clinical reports.

Candidemia can be a significant cause of death in immunosuppressed or debilitated patients particularly. Abnormalities of the instrumental cytograms of some hematological analyzers, such as Mindray BC-6800Plus, can be related to circulating Candida. We studied the possible diagnostic usefulness of this information. A fungal bloodstream infection has been simulated by adding aliquots of Candida albicans, Candida parapsilosis, and Candida glabrata to 75 leftovers and anonymized peripheral blood samples. Cytographic abnormalities like those of experimental samples were used to select patients with possible fungemia. The microscopic review of peripheral blood smears constituted the confirmatory method. In all experimental samples, the various Candida types caused pseudo-NRBC and morphological abnormalities of WNB and DIFF cytograms. Circulating blastospores, free or engulfed by neutrophils, were the microscopic findings in the peripheral blood smears. In the clinical verification, 72 patients were recruited based on the presence of an evocative cluster in the WNB and DIFF cytograms. The microscopic review of 39 out of 72 samples was positive for NRBC. According to blood cultures, light microscopy revealed fungal forms of several Candida or non-Candida types in the remaining 33 samples. Nine of these cases were not yet known to suffer from bloodstream infection. Although further confirmatory clinical studies are required for these diagnostic abilities, the BC 6800Plus cytographic abnormalities related to fungemia have proven helpful in rapidly monitoring persistent fungemia in already diagnosed patients. In unknown or undiagnosed cases, they could be the trigger point for the subsequent diagnostic-therapeutic pathway.

Effects of oil heat treatment on poplar wood properties: A pilot scale study

Pilot-scale oil heat treatment was conducted to enhance the quality of poplar wood obtained from a fast-growing plantation. Three target temperatures, namely 180, 190, and 200 ˚C, were chosen for the oil heat treatment process. Clear, small specimens were cut from the lumber, following the prescribed standards for assessing wood quality. Light microscopy revealed the ruptures and deformation of the wood cell wall as the treatment temperature increased. The volumetric swelling of the treated specimen decreased proportionally with the increase in treatment temperature. The impact of oil heat treatment on the mechanical properties of wood varied depending on the specific type of mechanical strength and processing temperature. The modulus of rupture, toughness, and hardness of the treated specimens drastically decreased at 200°C. However, the specimens treated at 200°C exhibited much lower mass loss during the fungal decay test. The results of the analytical test showed an increase in the crystalline cellulose content and thermal stability of the treated wood, due to the thermal degradation of hemicelluloses and amorphous regions of cellulose. Overall, it is concluded that oil heat treatment of poplar wood in the optimal temperature is an efficient approach to upgrade its quality with minimal reduction in mechanical strength.

A Study on the Materials Used in Ancient Wooden Architectural Paintings at DaZhong Gate in Confucius Temple, Qufu, Shandong, China.

This study analyzes the pigments and binders used in the painted wooden structure of DaZhong Gate in the Confucius Temple in Qufu, Shandong Province, China. Five samples were collected from the building and analyzed using techniques such as polarized light microscopy (PLM), energy-dispersive X-ray spectroscopy (EDX), micro-Raman spectroscopy (m-RS), and Fourier-transform infrared spectroscopy (FT-IR). The findings reveal that the red, yellow, green, and blue pigments are identified as lead red, lead chromate yellow, emerald green, and ultramarine, respectively. The white pigment is determined to be a combination of chalk and lead white or anglesite. Considering the production period of the yellow and green pigments, it is inferred that architectural paintings underwent restoration or repainting during the late Qing Dynasty. The analysis of the binder in the pigment using pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) reveals that the binder employed is a protein-based glue. Additionally, the detected presence of Heat-bodied tung oil suggests a potential connection to traditional Chinese painting techniques on wooden surfaces. This discovery not only contributes to the historical research of the Confucius Temple but also provides crucial data for the conservation and restoration efforts of this culturally significant heritage site.

Structural colour in the brown algal genus Sporochnus (Sporochnales, Phaeophyceae)

ABSTRACT The colour produced by an object’s microstructure rather than by pigments is generally called structural colour. Various macroalgae have some mechanisms of structural colouration. The presence of structural colour in the brown algae is well known in the Dictyotales and Fucales but has recently been reported in the Sporochnales where Sporochnus dotyi has structures in the terminal assimilatory filaments (trichothallic hairs) that produce a pronounced green iridescence. Based on observations by light microscopy, the structural colour of this species was caused by iridescent bodies, 10–15 µm in diameter, within the cells of assimilatory filaments. In contrast, another Sporochnus species known as S. radiciformis in Japan (S. radiciformis auct. japon.) also contains iridescent bodies of 6–10 µm diameter in the assimilatory filaments, but no significant structural colour was observed. In epi-illumination observations using red, green and blue LED light sources, iridescent bodies of S. dotyi showed stronger reflection of green compared with red and blue, whereas the reflections were similar among those colours in S. radiciformis. Transmission electron microscopy (TEM) using a rapid freezing method revealed that homogeneous globules 130–160 nm in diameter were tightly packed inside the iridescent bodies of S. dotyi, whereas the sizes of the globules were markedly irregular in iridescent bodies of S. radiciformis. Therefore, we conclude that the structural colouration of S. dotyi is caused by the interference and diffraction of incident light caused by the crystal lattice-like structure of tightly packed and regularly arranged globules of homogeneous size in the iridescent bodies, which is similar to the mechanism causing iridescence in opal. A similar opal-like structure of iridescent bodies causing structural colouration was recently reported in Ericaria selaginoides, but that of S. dotyi lacked the dynamic shift of the globules shown in E. selaginoides that resulted in a change of the wavelength of reflected light.

Content and tissue expression of Collagen I, Collagen IV, and Laminin in the Extracellular Matrix in Prostate Adenocarcinoma.

Prostate cancer is one of the most common neoplasm in the male population. It is not known why some tumors become more aggressive than others. Although most studies show changes in the expression of cell adhesion molecules and the extracellular matrix correlated with the Gleason score, no study has objectively measured the tissue content of these molecules. This study aims to measure the content and tissue expression of collagen type I and IV and laminin in the extracellular matrix of patients with prostate adenocarcinoma and correlate these findings with the Gleason score and clinical characteristics. Forty-one patients who underwent radical prostate surgery at the Urology Department of a reference Hospital in Brazil between January 2015 and December 2020 were studied. The tissue protein content was estimated under light microscopy at a final magnification of 200 × . The mean collagen I score in prostate adenocarcinoma tissue samples was 7.16 ± 1.03 pixels/field. The mean type IV collagen score was 3.44 ± 0.61 pixels/field. The mean laminin score was 5.19 ± 0.79 pixels/field. The total Gleason score was correlated with both collagen and laminin. All the correlations were negative, which shows that the higher the collagen/laminin expression was, the lower the total Gleason score (p-value < 0,05). According to the Pearson correlation analysis, age has no statistical relationship with collagen and laminin content. PSA, in turn, showed a correlation only with laminin, but r = -0.378 (p = 0.015). Among the associated diseases and lifestyle habits, there is only statistical significance in the comparison of alcoholism for collagen I. For collagen IV and laminin, no statistical significance was obtained with the clinical variables analyzed.

Age at puberty in Obispo Creole lambs from La Montaña of Guerrero, Mexico

Objective. Determine the age and body weight at the onset of puberty in Criollo Obispo lambs from a sheep herd. Materials and methods. The present study was carried out ex situ (18⁰ 15’ 52’’ LN). 10 lambs born during the spring were used, which were detected at 3 m of age. The detachment of the urethral process was verified at the age of 5.0 to 5.5 months of age. A semen sample was taken every two weeks with the help of an electro-ejaculator. Sperm concentration was evaluated with a digital photometer, while light microscopy was used for progressive motility and sperm morphology. The Kaplan-Meier procedure was used to estimate the risk or survival time function.to determine the days of onset of puberty. A male is considered to have entered puberty when the ejaculate registers at least 150 million sperm per mL, with a minimum of 50% abnormal cells and a motility greater than 30%. Results. It was observed that the average age of onset of puberty was 211.4±25.09 d, with a body weight of 19.3±1.27 kg. Conclusions. Criollo Obispo lambs begin puberty at 7 m of age and with a body weight of 19.3±1.27 kg.

Remission induced by renal protective therapy in nephrotic syndrome with thin basement membrane in an older patient: a case report

BackgroundAdult nephrotic syndrome is a well-known kidney disease that causes heavy proteinuria, hypoalbuminemia, hypercholesterolemia, edema, and hypertension. The treatment varies according to its underlying cause but often faces medication resistance or adverse drug effects.Case presentationA Japanese woman in her 80s presented with nephrotic syndrome after a 3 year latent period of urinary protein and occult blood. She did not have any secondary causes of nephrotic syndrome. Renal biopsy revealed thin glomerular basement membrane, partial foot process fusion on electron microscopy with minor glomerular change on light microscopy, and slight coarse immunoglobulin M deposition in the mesangium on immunofluorescence microscopy, which was inconsistent with any other glomerular diseases. Without steroid treatment, she dramatically remitted from proteinuria after the administration of the renal protective agents enalapril, ezetimibe, rosuvastatin, and dapagliflozin. Recurrence after 8 months of follow-up subsided with the administration of additional doses of the agents.ConclusionsThis case illustrated the novel outcomes of combining medical treatment without steroid use for nephrotic syndrome with thin glomerular basement membrane disease. At the time of writing this report, the patient’s renal function was stable and she was free of edema, although moderate proteinuria and occult hematuria persisted. The final diagnosis was uncertain because of the lack of genetic investigation; however, the response to the aforementioned medical treatment suggests the effectiveness of the supportive therapy.

Histological Disruptive Effects of Tau-Fluvalinate in Zebrafish (Danio rerio) Testis

Due to the high use of pesticides, undesirable effects are observed in the environment and toxic effects occur in terrestrial and aquatic organisms via pesticide bioaccumulation. Although bioaccumulation levels are low, especially in aquatic ecosystems, toxicological effects on aquatic organisms have been observed. Synthetic pyrethroids are pesticides produced for domestic and agricultural pests and tau-fluvalinate is also a broad-spectrum synthetic pyrethroid. Our study investigated the histopathological effects of zebrafish exposed to tau-fluvalinate at two different doses (8µg/L,16µg/L) on testicular tissue. After 5 days of exposure, testicular tissues were dissected and routine histological methods were applied. Tissues were investigated under a light microscope after they were stained with hematoxylin &amp; eosin. As a result of tau-fluvalinate exposure, deterioration in seminiferous tubule morphology, interstitial fibrosis, vascular congestion, karyorrhexis and hypertrophic spermatogenic cells were detected. Tau-fluvalinate exposure was found to induce apoptosis and cause germ cell deterioration.

Macrocheles muscaedomesticae (Acari: Macrochelidae) associated with Stomoxys calcitrans (Diptera: Muscidae) in the municipality of Sabanalarga, Antioquia

Objective: To identify mites in Diptera in a wooded area of Sabanalarga, Department of Antioquia, Colombia. Materials and methods: Two "stable flies" (Stomoxys calcitrans) were collected in a corral used temporarily to move cattle located in a wooded area of the municipality of Sabanalarga, Antioquia-Colombia. The flies were stored in Eppendorf tubes containing 70% ethyl alcohol for further processing. In the laboratory, the flies were identified, and then the mites they carried on their ventral area were removed and counted. Subsequently, the mites were stored in 70% ethyl alcohol to be rinsed on slides and identified under a light microscope. Results: A total of 15 specimens of a mite of the order Mesostigmata, family Macrochelidae of the species Macrocheles muscaedomesticae, parasitizing the flies S. calcitrans, were identified. Conclusions: This is the first report on the parasitic association between phoretic mites Macrocheles muscaedomesticae and stable flies Stomoxys calcitrans in Sabanalarga, Antioquia-Colombia.

Determination of prevalence, risk factors and symptoms of urinary schistosomiasis among school children in Kaduna South Local Government Area, Kaduna State

Schistosomiasis negatively impacts children’s health. Children expose themselves daily to schistosome infections due to lack of awareness and knowledge about the disease or the danger of infested water bodies. This study was aimed at determining the prevalence, risk factors and symptoms of urinary schistosomiasis among school children in Kaduna South Local Government Area, Kaduna State, Nigeria. Four hundred (400) consented school children submitted 10mL urine each and responded to structured questionnaires. Sediment of each urine sample was examined by light microscopy, after centrifugation at 3000 rpm for 5 min. Overall prevalence of urinary schistosomiasis was 4.5%. Both male and female children were equally infected (4.5% each). Older children between 13-17 years old were more infected (5.0%) and more at risk (OR = 2.982) than younger children (8-12 years old) who had 1.7% of the infection. No infection was recorded among children who had awareness about the disease. Water-based activities that served as significant risk factors for the infection included swimming in river (8.7%, P=0.016, OR= 3.064), fishing (12.2%, P=0.005, OR=3.942) and washing of clothes in river (17.4%, P=0.002, OR=5.459). Significant symptoms of urinary schistosomiasis identified were frequent urination (12.5%, P=0.023, OR=3.612), abdominal pain (10.0%, P=0.045, OR=2.880), and terminal haematuria (13.6%, P=0.002, OR=4.526). Infected individuals were more at risk of experiencing pains during urination (7.4%, OR=2.047). Unawareness promotes the spread of schistosomiasis among Nigerian children; their continuous exposure to infested water poses a greater risk. Creation of awareness amongst children will help to control their play habits in unsafe water bodies.

Distinct ultrastructural phenotypes of glial and neuronal alpha-synuclein inclusions in multiple system atrophy.

Multiple System Atrophy is characterized pathologically by the accumulation of alpha-synuclein (aSyn) into glial cytoplasmic inclusions (GCIs). The mechanism underlying the formation of GCIs is not well understood. In this study, correlative light and electron microscopy was employed to investigate aSyn pathology in the substantia nigra and putamen of post-mortem multiple system atrophy brain donors. Three distinct types of aSyn immuno-positive inclusions were identified in oligodendrocytes, neurons and dark cells presumed to be dark microglia. Oligodendrocytes contained fibrillar GCIs that were consistently enriched with lysosomes and peroxisomes, supporting the involvement of the autophagy pathway in aSyn aggregation in multiple system atrophy. Neuronal cytoplasmic inclusions exhibited ultrastructural heterogeneity resembling both fibrillar and membranous inclusions, linking multiple systems atrophy and Parkinson's disease. The novel aSyn pathology identified in the dark cells, displayed GCI-like fibrils or non-GCI-like ultrastructures suggesting various stages of aSyn accumulation in these cells. The observation of GCI-like fibrils within dark cells suggests these cells may be an important contributor to the origin or spread of pathological aSyn in multiple system atrophy. Our results suggest a complex interplay between multiple cell types that may underlie the formation of aSyn pathology in multiple system atrophy brain and highlight the need for further investigation into cell-specific disease pathologies in multiple system atrophy.

Ultra high frequency ultrasound enables real-time visualization of blood supply from chorioallantoic membrane to human autosomal dominant polycystic kidney tissue

Ultra high frequency (UHF) ultrasound enables the visualization of very small structures that cannot be detected by conventional ultrasound. The utilization of UHF imaging as a new imaging technique for the 3D-in-vivo chorioallantoic membrane (CAM) model can facilitate new insights into tissue perfusion and survival. Therefore, human renal cystic tissue was grafted onto the CAM and examined using UHF ultrasound imaging. Due to the unprecedented resolution of UHF ultrasound, it was possible to visualize microvessels, their development, and the formation of anastomoses. This enabled the observation of anastomoses between human and chicken vessels only 12 h after transplantation. These observations were validated by 3D reconstructions from a light sheet microscopy image stack, indocyanine green angiography, and histological analysis. Contrary to the assumption that the nutrient supply of the human cystic tissue and the gas exchange happens through diffusion from CAM vessels, this study shows that the vasculature of the human cystic tissue is directly connected to the blood vessels of the CAM and perfusion is established within a short period. Therefore, this in-vivo model combined with UHF imaging appears to be the ideal platform for studying the effects of intravenously applied therapeutics to inhibit renal cyst growth.

Prevalence of urinary schistosomiasis among school children in Giwa and Markafi Local Government Areas, Kaduna State, Nigeria

Urinary schistosomiasis is a neglected tropical disease that is affecting the health of many children, especially in Nigeria. Awareness about schistosomiasis in some communities is low; hence more children undertake activities in unsafe bodies of water. This study determined the prevalence, socio-demography, risk factors and symptoms of urinary schistosomiasis among school children in Giwa and Makarfi LGAs. A total of 200 school children were enrolled in this study and each of them submitted 10 mL urine sample. Each urine sample was concentrated by centrifugation and the sediment was examined using a light compound microscope. Data collected were analyzed using IBM SPSS Version 23 at 95% confidence interval. Overall prevalence of urinary schistosomiasis in this study was 19(9.5%). Spatial prevalence was 10.0% and 9.0% in Giwa and Makarfi LGAs respectively. There was no infection among the females (0.0%), but 13.5% of infection occurred among the males (P=0.003, OR =1.156). School children within the age-group of 18–19 years old had highest infection of 20.0%, followed by 12.1% and 5.6% among those within age-groups of 14-15 and 12-13 years old respectively. The infection was more occurring among children in junior secondary school (10.4%) than among those in senior secondary school (7.7%). Significant risk factors were irrigation farming, swimming, fishing and washing of clothes in rivers (P&lt;0.05, OR &gt;1). Significant symptoms of the disease among them included painful urination and terminal haematuria (P&lt;0.05, OR &gt;1). However, infected individuals mostly encountered frequent urination and abdominal pains (OR&gt;1). Urinary schistosomiasis prevails in the study area. It is important to create widespread awareness and treatment intervention.

A new Actinella species (Eunotiaceae, Bacillariophyta) from the Yangambi Biosphere Reserve, Democratic Republic of the Congo

Background and aims – Actinella species from Central Africa are only rarely reported. The acid waters from many stream and small rivers in the Congo Basin form a potential environment for species of this genus. We studied several samples from the region of Yangambi to confirm whether species belonging to this genus are present. Material and methods – Samples for diatom investigation were collected in several streams and small rivers (Moni, Ngima, and Libongo), in the tropical rainforest in the Yangambi Biosphere Reserve, Tshopo Province, Democratic Republic of the Congo. The samples were prepared to obtain permanent microscope slides for light microscopy studies using standard methods and cleaned material for scanning electron microscopy. Key results – A novel Actinella was observed in the studied samples. The taxon was compared to other species such as A. disjuncta, A. lange-bertalotii, A. modesta, A. pereunotioides, and A. pseudohantzschia, although the new species morphologically resembles most A. eunotioides, a species discovered in the Amazon basin in Brazil, and A. eunotioides var. minor, described in 1966 from the Central African Republic. The taxon observed in our material differs from A. eunotioides mainly in the valve dimensions, being much smaller, and the higher density of the marginal spines. Stria density also tends to be coarser. The valve length of A. eunotioides var. minor, on the other hand, corresponds with the smallest valves of our taxon but the valves are narrower, with a higher stria density. The ventral margin is slightly convex, while in the new taxon it is slightly concave. Moreover, in the original description of A. eunotioides var. minor, the absence of spines is mentioned, while they are present on the valves in the materials we investigated. Based on the observed morphological differences, we consider the taxon recorded in the streams and small rivers in the Yangambi Biosphere Reserve distinct from A. eunotioides and its var. minor and subsequently, we describe Actinella kufferathiana as a species new to science.

Illuminating the problem of blue verditer synthesis in the early modern English period: chemical characterisation and mechanistic understanding

We present a study into early modern English production of blue verditer, an early synthetic copper-based blue pigment chemically analogous to azurite. Verditers have been identified in numerous wall and easel paintings. While initial documentation occurs in the mid 1500s and production recipes were documented by the 17th c., the synthesis was known to be unreliable. This study replicates historical and recent scientific work on blue verditer and represents a significant advance in our understanding of verditer production and its challenges. Procedures for verditer synthesis are drawn from both 17th c. documentation and 20th c. replication work. The effects of temperature, copper and carbonate sources, solution stirring, copper ion concentration, and atmospheric composition are studied in order to elucidate the mechanism of synthesis and explain its unreliability in early modern refineries. Products are characterised by polarised light microscopy, scanning electron microscopy, Raman spectroscopy, and powder X-Ray diffraction. Rouaite, a green basic copper nitrate, is for the first time confirmed as a product of the refiners’ synthesis and a precursor to blue verditer in laboratory syntheses. This result problematises the blanket identification of green verditer as basic copper carbonate and provides important clues to the mechanism for blue verditer synthesis. Solution chemistry and ion equilibria allow us to explain the route by which rouaite is first formed and then converted to blue verditer. Conditions favouring blue verditer production are also clarified further. Although it is commonly stated that low temperatures are required for blue verditer production, blue verditer is produced here at a range of ambient temperatures. The reaction is found instead to be controlled by solution equilibria and heavily favoured by high partial pressures of carbon dioxide. Alongside archival materials about refining and verditer production, these results are contextualised and explanations for the unreliability of historical synthesis are proposed.

Electromagnetic drop-on-demand (DoD) technology as an innovative platform for amorphous solid dispersion production

Production of amorphous solid dispersions (ASDs) is an effective strategy to promote the solubility and bioavailability of poorly water soluble medicinal substances. In general, ASD is manufactured using a variety of classic and modern techniques, most of which rely on either melting or solvent evaporation. This proof-of-concept study is the first ever to introduce electromagnetic drop-on-demand (DoD) technique as an alternative solvent evaporation-based method for producing ASDs. Herein 3D printing of ASDs for three drug-polymer combinations (efavirenz-Eudragit L100-55, lumefantrine-hydroxypropyl methylcellulose acetate succinate, and favipiravir-polyacrylic acid) was investigated to ascertain the reliability of this technique. Polarized light microscopy, differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), and Fourier Transform Infrared (FTIR) spectroscopy results supported the formation of ASDs for the three drugs by means of DoD 3D printing, which significantly increases the equilibrium solubility of efavirenz from 0.03 ± 0.04 µg/ml to 21.18 ± 4.20 µg/ml, and the equilibrium solubility of lumefantrine from 1.26 ± 1.60 µg/ml to 20.21 ± 6.91 µg/ml. Overall, the reported findings show how this new electromagnetic DoD technology can have a potential to become a cutting-edge 3D printing solvent-evaporation technique for on-demand and continuous manufacturing of ASDs for a variety of drugs.

A Pilot Study of Aerosolization of InfectiousMurine Norovirus in an Experimental Setup.

Human norovirus is transmitted mainly via the faecal-oral route, but norovirus disease outbreaks have been reported in which airborne transmission has been suggested as the only explanation. We used murine norovirus (MNV) as a surrogate for human norovirus to determine the aerosolization of infectious norovirus in an experimental setup. A 3-l air chamber system was used for aerosolization of MNV. Virus in solution (6 log10 TCID50/ml) was introduced into the nebulizer for generating aerosols and a RAW 264.7 cell dish without a lid was placed in the air chamber. Cell culture medium samples were taken from the dishes after the aerosol exposure time of 30 or 90min, and the dishes were placed in a 37°C, 5% CO2 incubator and inspected with a light microscope for viral cytopathic effects (CPEs). We determined both the infectious MNV TCID50 titre and used an RT-qPCR assay. During the experiments, virus infectivity remained stable for 30 and 90min in the MNV solution in the nebulizer. Infectious MNV TCID50 values/ml of 2.89 ± 0.29 and 3.20 ± 0.49 log10 were measured in the chamber in RAW 264.7 cell dish media after the 30-min and 90-min exposure, respectively. The MNV RNA loads were 6.20 ± 0.24 and 6.93 ± 1.02 log10 genome copies/ml, respectively. Later, a typical MNV CPE appeared in the aerosol-exposed RAW cell dishes. We demonstrated that MNV was aerosolized and that it remained infectious in the experimental setup used. Further studies required for understanding the behaviour of MNV in aerosols can thus be performed.

Biogenic synthesis of Psidium guajava‐mediated silver nanoparticles: A comprehensive antibiofilm and antivirulence study

AbstractIn this study, at sub‐MIC, the efficacy of green synthesized Psidium guajava‐mediated silver nanoparticles (Pg@AgNPs) in inhibiting quorum sensing facilitated virulence factors and suppressing biofilm formation was evaluated against clinical multidrug resistant (MDR) isolates, specifically Chromobacterium violaceum MCC‐2290 and Pseudomonas aeruginosa PAO1. The AgNPs were synthesized by adding the leaf extract to a solution of silver nitrate, and the reduction of Ag+ ions to Ag0 was confirmed by the appearance of brown color. The Pg@AgNPs were characterized using Fourier transform infrared (FTIR), UV/Vis spectroscopy, TEM, and XRD. The UV/Vis spectrum showed the characteristic absorption peak of Pg@AgNPs in the range of 350–480 nm. FTIR analysis substantiated the existence of functional groups implicated in the synthesis and stabilization of AgNPs. The XRD analysis elucidated the crystalline characteristics of the nanoparticles. The TEM images further confirmed the spherical shape and size of the nanoparticles, which ranged between 5 and 25 nm. The AgNPs demonstrated notable antibacterial efficacy against C. violaceum and P. aeruginosa. Moreover, the Pg@AgNPs effectively suppressed both biofilm formation and the production of quorum sensing‐mediated virulence factors such as pyoverdin (77.2%), pyocyanin (78.12%), and violacein (83.4%). Additionally, the Pg@AgNPs demonstrated broad‐spectrum activity against biofilm formation, showing the biofilm inhibition in C. violaceum and P. aeruginosa by 81.86% and 87.89%, respectively. Under light microscopy, it is also observed that Pg@AgNPs reduce the formation of biofilm in both tested bacteria. The present finding highlights the potential of green‐synthesized AgNPs using P. guajava leaf extract as antimicrobial agents with potential applications in combating bacterial infections and biofilm‐related diseases.

Neurokinin B Administration Induces Dose Dependent Proliferation of Seminal Vesicles in Adult Rats.

Neurokinin B; an endogenous decapeptide, mediates its reproductive physiological actions through gonadotropin releasing hormone. Despite the potential role of Neurokinin B on seminal vesicles, its effects on seminal vesicles in adult male mammals remain elusive. We aimed to investigate the potentials of variable doses of Neurokinin B, its agonist and antagonist on histomorphology and expression of NK3R on seminal vesicles, and secretory activity of seminal vesicles in adult male rats. Adult male Sprague Dawley rats (n=10 in each group) were administered intraperitoneally with Neurokinin B in three variable doses: 1 μg, 1 ηg and 10 ρg while, Senktide (Neurokinin B agonist) and SB222200 (Neurokinin B antagonist) in 1 μg doses consecutively for 12 days. After 12 days of peptide treatment, half of the animals (n=05) in each group were sacrificed while remaining half (n=05) were kept for another 12 days without any treatment to investigate treatment reversal. Seminal vesicles were dissected and excised tissue was processed for light microscopy, immunohistochemistry and estimation of seminal fructose levels. Treatment with Neurokinin B and Senktide significantly increased while SB222200 slightly decrease the seminal vesicles weight, epithelial height and seminal fructose levels as compared to control. Light microscopy revealed increased epithelial height and epithelial folding as compared to control in all Neurokinin B and Senktide treated groups while decreased in SB222200. Effects of various doses of Neurokinin B, Senktide and SB222200 on seminal vesicles weight, epithelial height, seminal fructose levels and histomorphology were reversed when rats were maintained without treatments. Immuno-expression of Neurokinin B shows no change in treatment and reversal groups. Continuous administration of Neurokinin B and Senktide effect positively while SB222200 have detrimental effects on cellular morphology, epithelial height and seminal fructose levels in seminal vesicles. Effects of peptide treatments depicted a reversal towards control group when rats were kept without any treatment.