Microscopic Visualization Research Articles

Expanding the toolbox for microfluidic-based in situ membrane characterization via microscopy

Polyamide thin film composite membranes are the commercial standard for aqueous nanofiltration and reverse osmosis. Establishing their synthesis-structure-performance relationships (SSPs), needed for rational membrane design, is hampered by the small scale and high reaction rate of interfacial polymerization (IP). Microfluidic devices, compatible with microscopic real-time visualization of IP and performance testing of the formed film, are interesting within this respect. In this study, a new microfluidic design and operational protocol for in situ characterization of IP is developed. Difficulties encountered with microfluidics and coping strategies are highlighted. The outcome of the optimization study proves that a parylene-coated PDMS-glass chip comprising a channel lay-out with 4 inlets, 2 outlets, a channel height of 20 μm, and a reaction channel length ≤50 μm is most compatible with IP and performance testing. Varying synthesis conditions show changing film morphology and water flux in line with trends for dip-coated membranes. Addition of NaHCO3 and ethyl acetate induce morphological features and increase water flux. Increasing TMC concentrations decrease water flux until an excess is generated. By combining the developed protocol and microfluidic device with an online measurement technique to probe film formation dynamics, such as fluorescence microscopy, SSPs can be derived in the future.

Waste biomass from hypersaline potash mining byproducts: Detection and visualization of Cu(II) and Cr(VI) on Croceicoccus sp. FTI14 biosorbent

Microorganisms in hypersaline potash mining byproducts and their potential environmental applications have not been extensively reported. This study reports the diverse waste-impacted microbial communities (archaea and bacteria) adapted to extreme salinity (>10–25%). Of these, halotolerant Croceicoccus sp. FTI14 was investigated as a biosorbent for removing dissolved Cu(II) and Cr(VI) from synthetic Cu(II) and Cr(VI)-contaminated DI, groundwater and saline groundwater (0.55 M ionic strength). FTI14 biomass was oven-dried, finely ground, and investigated in batch biosorption experiments. At initial metal concentrations of 40 mg/L, FTI14 removed 40 ± 0.7% (16.3 ± 0.5 mg/g) and 19 ± 0.1% (7.8 ± 0.1 mg/g) of the Cu(II) from deionized water and saline groundwater, respectively, while only 22.9 ± 0.7% (9.6 ± 0.2 mg/g) and 2.1 ± 0.6% (1.0 ± 0.3 mg/g) Cr(VI) removal was achieved. Cu(II) uptake (mg/g) exceeded Cr(VI) uptake by a factor of 1.7–7.8. Langmuir and Freundlich models were applied on FTI14 biosorption isotherm data. The Freundlich model showed a better fit for both Cu(II) and Cr(VI), as indicated by the AIC values compared with evidence ratios. Synchrotron-based scanning transmission X-ray microscopy (STXM) visualizations of the biosorbent showed a mixture of whole cells and indistinct biomass and a spatial association between metals and biomass. Metal exposure alters the amide functional groups peak in Fourier transform Infra-red (FTIR) spectra, suggesting its role in sorption process. Thus, this study indicates culturable halotolerant microorganisms from hypersaline potash mining byproducts and its potential as biosorbent applications for metal removal from impacted groundwater.

Effect of green Fe2O3 nanoparticles in controlling Fusarium fruit rot disease of loquat in Pakistan.

The subtropical fruit known as the loquat is prized for both its flavour and its health benefits. The perishable nature of loquat makes it vulnerable to several biotic and abiotic stressors. During the previous growing season (March-April 2021), loquat in Islamabad showed signs of fruit rot. Loquat fruits bearing fruit rot symptoms were collected, and the pathogen that was causing the disease isolated and identified using its morphology, microscopic visualisation, and rRNA sequence. The pathogen that was isolated was identified as Fusarium oxysporum. Green synthesized metallic iron oxide nanoparticles (Fe2O3 NPs) were employed to treat fruit rot disease. Iron oxide nanoparticles were synthesized using a leaf extract of the Calotropis procera. Characterization of NPs was performed by different modern techniques. Fourier transform infrared spectroscopy (FTIR) determined the existence of stabilizing and reducing compounds like phenol, carbonyl compounds, and nitro compounds, on the surface of Fe2O3 NPs. X-ray diffraction (XRD) explained the crystalline nature and average size (~49 nm) of Fe2O3 NPs. Energy dispersive X-ray (EDX) exhibited Fe and O peaks, and scanning electron microscopy (SEM) confirmed the smaller size and spherical shape of Fe2O3 NPs. Following both in vitro and in vivo approaches, the antifungal potential of Fe2O3 NPs was determined, at different concentrations. The results of both in vitro and in vivo analyses depicted that the maximum fungal growth inhibition was observed at concentration of 1.0 mg/mL of Fe2O3 NPs. Successful mycelial growth inhibition and significantly reduced disease incidence suggest the future application of Fe2O3 NPs as bio fungicides to control fruit rot disease of loquat.

A Rare Case of Infantile Otalgia and Otorrhagia due to Alive Leech Bite

Abstract Pediatric ear infestation with leeches is a rare event; however, it can lead to serious complications. Leeches usually make their way into the human body through bathing with or drinking of contaminated water from nonhygienic sources and swimming in infested water. To our knowledge, this is the first report of leech removal from the external auditory canal in an infant. We described the case of a 3-month-old infant who presented to our institution with sudden left otalgia, otorrhagia, poor oral intake, and prolonged crying for 4 days, culminating in a single epileptic attack, drowsiness, and electrolyte disturbances. Microscopic visualization of the left ear revealed minimal oozing of blood with clots obscuring the ear canal and a live motile 3 cm leech deeply seated near the tympanic membrane. Extraction was performed. The bloody discharge ceased completely and all symptoms were relieved. Leech infestation is typically considered after other potential etiologies in infants with sudden otorrhagia and otalgia living in an endemic region. It should be considered a hidden cause of pediatric discomfort. Detailed history taking and meticulous ear examinations are vital for the prompt management to avoid unnecessary computed tomography imaging and radiation, unhelpful interventions, and delayed treatment. A diagnosis was reached and treatment performed by careful inspection and leech removal, respectively.

Investigation of the plugging capacity and enhanced oil recovery of flexible particle three-phase foam

In the later stage of water flooding in reservoirs, the oil production decreases. The stability of three-phase foam system is strong, enhancing oil displacement effects of ultrahigh water cut reservoirs. In this paper, the flow state of the foam system was studied by using a microscopic visualization model. Second, sandpack flooding experiments were performed to analyze the optimal plugging conditions of three-phase foam through resistance factors. Finally, single and parallel sandpack flooding experiments were carried out to analyze plugging effect and production conditions under different injection systems and permeability ratios. The research shows that three-phase foam is more stable and capable of plugging. The stable residual resistance factor after injection of the three-phase foam system is 172.75. When the foam and particles are injected simultaneously, the gas–liquid ratio is 1:1, the formation permeability is 5526.5 mD, and the plugging effect of three-phase foam is the best. The oil recovery increases by 24.9% after the injection of three-phase foam system. The three-phase foam system combines Jamin effect of foam and dynamic plugging characteristics of flexible particles. The highest oil recovery factor of 80.87% is achieved at a permeability ratio of 9.99. Three-phase foam flooding has broad application prospects.

Microscopic Distribution and Development Strategy of Residual Oil in Tight Sandstone

Pore and permeability are distributed discontinuously and unevenly in the dominant sedimentary model of the lateral accretion body inside the meandering river point bar of the Fuyang reservoir of the Yushulin oilfield. Based on the water flooding experiments of field core samples, the influence of pore permeability conditions on residual oil distribution type and water cutting rate was studied by using the microscopic visualization technology enabled through a photolithographically fabricated glass model. It is found that the residual oil in samples shows five discontinuous types, which are cluster, columnar, oil droplet, membrane, and blind end. In the stages with low, medium, and high water cutting rates, the proportion of clustered residual oil in the samples with different permeability is high, reflecting the situation that it is difficult for injected water to spread widely in tight oil reservoirs. With the decrease of permeability, the proportion of membrane and blind end residual oil gradually increases, which indicates that the thin pore throat can produce large restrictions on residual oil, resulting in residual oil enrichment. At the same time, the water flooding experiment was carried out by changing the displacement direction and periodic water injection. It was found that changing the displacement direction was beneficial to the recovery of residual oil in the thin pore throat and avoided the dominant seepage of injected water in the big pore throat, and the recovery rate was increased by more than 2.14%. Periodic water injection, which was conducive to adjusting the displacement pressure difference, reduced the constraining force of the throat on residual oil and increased the recovery rate by more than 3.98%. The actual well area with closed coring wells and dynamic production data is preferred for the application of experimental research results. Changing displacement direction and periodic water injection increased the residual oil recovery by more than 3%.

Tools of the Trade: Image Analysis Programs for Confocal Laser-Scanning Microscopy Studies of Biofilms and Considerations for Their Use by Experimental Researchers.

Confocal laser-scanning microscopy (CLSM) is the bedrock of the microscopic visualization of biofilms. Previous applications of CLSM in biofilm studies have largely focused on observations of bacterial or fungal elements of biofilms, often seen as aggregates or mats of cells. However, the field of biofilm research is moving beyond qualitative observations alone, toward the quantitative analysis of the structural and functional features of biofilms, across clinical, environmental, and laboratory conditions. In recent times, several image analysis programs have been developed to extract and quantify biofilm properties from confocal micrographs. These tools not only vary in their scope and relevance to the specific biofilm features under study but also with respect to the user interface, compatibility with operating systems, and raw image requirements. Understanding these considerations is important when selecting tools for quantitative biofilm analysis, including at the initial experimental stages of image acquisition. In this review, we provide an overview of image analysis programs for confocal micrographs of biofilms, with a focus on tool selection and image acquisition parameters that are relevant for experimental researchers to ensure reliability and compatibility with downstream image processing.

Antibacterial Activity and Mechanisms of TroHepc2-22, a Derived Peptide of Hepcidin2 from Golden Pompano (Trachinotus ovatus).

Hepcidin, a cysteine-rich antimicrobial peptide, has a highly conserved gene structure in teleosts, and it plays an essential role in host immune response against various pathogenic bacteria. Nonetheless, few studies on the antibacterial mechanism of hepcidin in golden pompano (Trachinotus ovatus) have been reported. In this study, we synthesized a derived peptide, TroHepc2-22, from the mature peptide of T. ovatus hepcidin2. Our results showed that TroHepc2-22 has superior antibacterial abilities against both Gram-negative (Vibrio harveyi and Edwardsiella piscicida) and Gram-positive (Staphylococcus aureus and Streptococcus agalactiae) bacteria. Based on the results of a bacterial membrane depolarization assay and propidium iodide (PI) staining assay in vitro, TroHepc2-22 displayed antimicrobial activity by inducing the bacterial membrane depolarization and changing the bacterial membrane permeability. Scanning electron microscopy (SEM) visualization illustrated that TroHepc2-22 brought about membrane rupturing and the leakage of the cytoplasm for the bacteria. In addition, TroHepc2-22 was verified to have hydrolytic activity on bacterial genomic DNA in view of the results of the gel retardation assay. In terms of the in vivo assay, the bacterial loads of V. harveyi in the tested immune tissues (liver, spleen, and head kidney) were significantly reduced in T. ovatus, revealing that TroHepc2-22 significantly enhanced the resistance against V. harveyi infection. Furthermore, the expressions of immune-related genes, including tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin 1-β (IL-1β), IL-6, Toll-like receptor 1 (TLR1), and myeloid differentiation factor 88 (MyD88) were significantly increased, indicating that TroHepc2-22 might regulate inflammatory cytokines and activate immune-related signaling pathways. To summarize, TroHepc2-22 possesses appreciable antimicrobial activity and plays a vital role in resisting bacterial infection. The observation of our present study unveils the excellent application prospect of hepcidin as a substitute for antibiotics to resist pathogenic microorganisms in teleosts.

Parameters Variation and Flow Characteristics When CO2 Displacing Brine in Four Micromodels Simulating Carbon Sequestration in Saline Aquifers

Two-phase flow of CO2/brine in porous media is critical to the capacity and safety of carbon sequestration into the brine aquifer. In order to provide valuable information and important theoretical basis for site selection and CO2 injection, the microscopic visualization technology was employed in this study to conduct displacement experiments of CO2/brine at the pore scale. Four micromodels with different sizes and structures, five injection rates of CO2, and six salinities of brine were used to study the effects of micromodel’s structure and displacement pattern on two-phase flow. Several parameters including the differential pressure, contact angle, permeability, velocity field, and force field were obtained by experimental measurement, image post-processing, and theoretical analysis, and then, these parameters’ variation was investigated. Phenomena such as thin film, corner flow, and Haines jump were also found during the displacement. Although brine could be completely displaced by CO2 in the capillary duct, the backflow of the wetting phase would occur at a low injection rate. Phenomena different from the theoretical analysis also occurred in pore doublet models: some brine was residual in the homogeneous pore doublet model at a low injection rate, while the heterogeneous pore doublet model was fully occupied by CO2 at a high injection rate. These phenomena are very useful for two-phase flow, and multiple factors need to be comprehensively considered to determine the operating conditions of CO2 storage into the brine aquifer.

Molecular mechanisms of regulation by a β-alanine-responsive Lrp-type transcription factor from Acidianus hospitalis.

The leucine-responsive regulatory protein (Lrp) family of transcriptional regulators is widespread among prokaryotes and especially well-represented in archaea. It harbors members with diverse functional mechanisms and physiological roles, often linked to the regulation of amino acid metabolism. BarR is an Lrp-type regulator that is conserved in thermoacidophilic Thermoprotei belonging to the order Sulfolobales and is responsive to the non-proteinogenic amino acid β-alanine. In this work, we unravel molecular mechanisms of the Acidianus hospitalis BarR homolog, Ah-BarR. Using a heterologous reporter gene system in Escherichia coli, we demonstrate that Ah-BarR is a dual-function transcription regulator that is capable of repressing transcription of its own gene and activating transcription of an aminotransferase gene, which is divergently transcribed from a common intergenic region. Atomic force microscopy (AFM) visualization reveals a conformation in which the intergenic region appears wrapped around an octameric Ah-BarR protein. β-alanine causes small conformational changes without affecting the oligomeric state of the protein, resulting in a relief of regulation while the regulator remains bound to the DNA. This regulatory and ligand response is different from the orthologous regulators in Sulfolobus acidocaldarius and Sulfurisphaera tokodaii, which is possibly explained by a distinct binding site organization and/or by the presence of an additional C-terminal tail in Ah-BarR. By performing site-directed mutagenesis, this tail is shown to be involved in ligand-binding response.

Host switching by an ambrosia beetle fungal mutualist: Mycangial colonization of indigenous beetles by the invasive laurel wilt fungal pathogen.

Ambrosia beetles require their fungal symbiotic partner as their cultivated (farmed) food source in tree galleries. While most fungal-beetle partners do not kill the host trees they inhabit, since their introduction (invasion) into the United states around ~2002, the invasive beetle Xyleborus glabratus has vectored its mutualist partner (but plant pathogenic) fungus, Harringtonia lauricola, resulting in the deaths of over 300 million trees. Concerningly, indigenous beetles have been caught bearing H. lauricola. Here, we show colonization of the mycangia of the indigenous X. affinis ambrosia beetle by H. lauricola. Mycangial colonization occurred within 1 h of feeding, with similar levels seen for H. lauricola as found for the native X. affinis-R. arxii fungal partner. Fungal mycangial occupancy was stable over time and after removal of the fungal source, but showed rapid turnover when additional fungal cells were available. Microscopic visualization revealed two pre-oral mycangial pouches of ~100-200 × 25-50 μm/each, with narrow entry channels of 25-50 × 3-10 μm. Fungi within the mycangia underwent a dimorphic transition from filamentous/blastospore growth to yeast-like budding with alterations to membrane structures. These data identify the characteristics of ambrosia beetle mycangial colonization, implicating turnover as a mechanism for host switching of H. lauricola to other ambrosia beetle species.

The synergism of improving interfacial properties between betaine and crude oil for enhanced oil recovery

Betaine, an emerging surfactant for enhanced oil recovery (EOR), has garnered increased attention and recognition. This study focuses on linear alkylbenzene sulfonate betaine (ASB) as the research subject, and different crude oils were utilized to prepare two types of oil–water formulations with high interfacial film strengths but differed interfacial tensions. The interfacial properties were characterized and explored using interfacial dilational rheology, droplet squeeze coalescence, interfacial tension, emulsification, microscopic visualization, and other measuring methods. The actual emphasis in interfacial film properties of betaine as a standalone chemical EOR formulation was clarified. The results indicated that betaine had generally high interfacial film strength with selected crude oils, at this time, the slight difference in film strength had a negligible effect on enhancing the recovery. The main contributing factor was the difference in interfacial tension between the oil and water. Ultra-low interfacial tension could increase the stripping efficiency of oil in pore, reduce the energy required to emulsification or oil droplets initiation, and ensure that droplets were displaced and collected in smaller sizes, thereby avoiding large oil droplets trapped in the pore throat and unable to pass. Making good use of the advantages of improving sweep efficiency brought by high interfacial film strength, at the same time giving the system ultra-low oil–water interfacial tension, ensuring the start-up and migration of porous crude oil, is the core construction principle of betaine surfactant applied to EOR.

Down-regulation of β-lactam antibiotics resistance and biofilm formation by Staphylococcus epidermidis is associated with isookanin.

Biofilm formation is the major pathogenicity of Staphylococcus epidermidis (S. epidermidis), which enhances bacterial resistance to antibiotics. Isookanin has potential inhibitory activity on biofilm. The inhibiting mechanisms of isookanin against biofilm formation through surface hydrophobicity assay, exopolysaccharides, eDNA, gene expression analysis, microscopic visualization, and molecular docking were explored. Additionally, the combination of isookanin and β-lactam antibiotics were evaluated by the broth micro-checkerboard assay. The results showed that isookanin could decrease the biofilm formation of S. epidermidis by ≥85% at 250 μg/mL. The exopolysaccharides, eDNA and surface hydrophobicity were reduced after treatment with isookanin. Microscopic visualization analysis showed that there were fewer bacteria on the surface of the microscopic coverslip and the bacterial cell membrane was damaged after treatment with isookanin. The down-regulation of icaB and up-regulation of icaR were observed after treatment with isookanin. Additionally, the RNAIII gene was significantly up-regulated (p < 0.0001) at the mRNA level. Molecular docking showed that isookanin could bind to biofilm-related proteins. This indicated that isookanin can affect biofilm formation at the initial attachment phase and the aggregation phase. The FICI index showed that the combination of isookanin and β-lactam antibiotics were synergistic and could reduce doses of antibiotics by inhibiting biofilm formation. This study improved the antibiotic susceptibility of S. epidermidis through inhibition of the biofilm formation, and provided a guidance for the treatment of antibiotic resistance caused by biofilm.

Dynamic real-time optical microscopy of oral mucosal lesions using confocal laser endomicroscopy.

Confocal laser endomicroscopy (CLE) is a novel non-invasive point-of-care optical biopsy technology that enables real-time in vivo microscopic visualisation of cellular and tissue architecture. In this study, we assessed the diagnostic accuracy of a hand-held fluorescence single-fibre distal-scanning CLE (fsdCLE) platform for diagnosing oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC). Forty-seven patients presenting with 63 distinct oral mucosal lesions were subjected to optical biopsy using a miniaturised fsdCLE system (ViewnVivo®, Optiscan Imaging Ltd) and topical exogenous acriflavine hydrochloride contrast agent before undergoing tissue biopsy and histopathological consensus review by four pathologists. CLE images were captured in vivo in real-time during clinical examination and assessed on-the-fly for the presence of cellular and architectural features of OED/OSCC offering an instantaneous diagnosis. Predicted optical diagnoses were compared to definitive consensus tissue histopathology. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy were calculated for the presence/absence of dysplasia/malignancy on optical biopsy. Percentage agreement, Fleiss' kappa, and intraclass correlation coefficient (ICC) were calculated for each assessment stage during the consensus histopathology process. Diagnostic accuracy was extremely high at 88.9%. Other metrics were sensitivity 86.8%, specificity 92%, PPV 94.3% and NPV 82.1%. One hundred percent of carcinoma cases were detected accurately using CLE in the clinic. fsdCLE is a highly accurate, easy-to-use, rapid and slide-free point-of-care in vivo optical technology for diagnosing OED/OSCC and discriminating between dysplastic and non-dysplastic pathology. It demonstrates near-perfect agreement with traditional consensus histopathology without the need for physical tissue biopsy.

Study on Flow Characteristics of Flue Gas and Steam Co-Injection for Heavy Oil Recovery

Flue gas is composed of N2 and CO2, and is often used as an auxiliary agent for oil displacement, with good results and very promising development prospects for co-injection with steam to develop heavy oil. Although research on the oil displacement mechanism of flue gas has been carried out for many years, the flow characteristics of steam under the action of flue gas have rarely been discussed. In this paper, the flow resistance and heat transfer effect of flue gas/flue gas + steam were evaluated by using a one-dimensional sandpack, a flue gas-assisted steam flooding experiment was carried out using a specially customized microscopic visualization model, and the microscopic flow characteristics in the process of the co-injection of flue gas and steam were observed and analyzed. The results showed that flue gas could improve the heat transfer effect of steam whilst accelerating the flow of steam in porous media and reducing the flow resistance of steam. Compared with pure steam, when the volume ratio of flue gas and steam was 1:2, the mobility decreased by 2.8 and the outlet temperature of the sandpack increased by 35 °C. This trend intensified with an increase in the proportion of flue gas. In the microscopic oil displacement experiments, the oil recovery and sweep efficiency of the flue gas and steam co-injection stage increased by 4.7% and 32.9%, respectively, compared with the pure steam injection stage due to the effective utilization of blocky remaining oil and corner remaining oil caused by the expansion of fluid channels, the flow of flue gas foam, and the dissolution and release of flue gas in heavy oil.

In vitro-in vivo relationship for amorphous solid dispersions using a double membrane dissolution-permeation setup

The use of amorphous solid dispersions (ASDs) is one commonly applied formulation strategy to improve the oral bioavailability of poorly water-soluble drugs by overcoming dissolution rate and/or solubility limitations. While bioavailability enhancement of ASDs is well documented, it has often been a challenge to establish a predictive model describing in vitro-in vivo relationship (IVIVR). In this study, it is hypothesized that drug absorption might be overestimated by in vitro dissolution-permeation (D/P)-setups, when drug in suspension has the possibility of directly interacting with the permeation barrier. This is supported by the overprediction of drug absorption from neat crystalline efavirenz compared to four ASDs in a D/P-setup based on the parallel artificial membrane permeability assay (PAMPA). However, linear IVIVR (R2 = 0.97) is established in a modified D/P-setup in which the addition of a hydrophilic PVDF-filter acts as a physical boundary between the donor compartment and the PAMPA-membrane. Based on microscopic visualization, the improved predictability of the modified D/P-setup is due to the avoidance of direct dissolution of drug particles in the lipid components of the PAMPA-membrane. In general, this principle might aid in providing a more reliable evaluation of formulations of poorly water-soluble drugs before initiating animal models.

Host factor TNK2 is required for influenza virus infection.

Host factors are required for Influenza virus infection and have great potential to become antiviral target. Here we demonstrate the role of TNK2 in influenza virus infection. CRISPR/Cas9 induced TNK2 deletion in A549 cells. CRISPR/Cas9-mediated deletion of TNK2. Western blotting and qPCR was used to measure the expression of TNK2 and other proteins. CRISPR/Cas9-mediated deletion of TNK2 decreased the replication of influenza virus and significantly inhibited the ex-pression of viral proteins and TNK2 inhibitors (XMD8-87 and AIM-100) reduced the expression of influenza M2, while over-expression of TNK2 weakened the resistance of TNK2-knockout cells to influenza virus infection. Furthermore, a decrease of nuclear import of IAV in the infected TNK2 mutant cells was observed in 3h post-infection. Interestingly, TNK2 deletion enhanced the colocalization of LC3 with autophagic receptor p62 and led to the attenuation of influenza virus-caused accumulation of autophagosomes in TNK2 mutant cells. Further, confocal microscopy visualization result showed that influenza viral matrix 2 (M2) was colocalized with Lamp1 in the infected TNK2 mutant cells in early infection, while almost no colocalization between M2 and Lamp1 was observed in IAV-infected wild-type cells. Moreover, TNK2 depletion also affected the trafficking of early endosome and the movement of influenza viral NP and M2. Our results identified TNK2 as a critical host factor for influenza viral M2 protein trafficking, suggesting that TNK2 will be an attractive target for the development of antivirals therapeutics.

Gastrointestinal tuberculosis: An autopsy-based study

BackgroundTuberculosis is a disease of great relevance since it is one of the leading causes of morbidity and mortality worldwide. Gastrointestinal tuberculosis is an unusual presentation. It is defined as the involvement of any segment of the digestive tract, associated viscera, and peritoneum. The study's main objective is to collect information from autopsies of patients diagnosed with gastrointestinal tuberculosis in a Pathology reference center in Colombia. MethodsThis is a retrospective and descriptive study of autopsy reports. A total of 4,500 autopsies were performed between January 2004 and December 2020. The inclusion criteria were authorization of a family member following local law regulations and a final autopsy diagnosis of gastrointestinal tuberculosis using microscopic visualization. ResultsForty-eight patients with gastrointestinal tuberculosis autopsies were included in our study. Most of the patients were male (n = 35, 72.9%) with a median age of 40.5 years old. Human immunodeficiency virus infection history was reported in 28 cases (58.33%). The most affected gastrointestinal tract site was the terminal ileum. Ulcers and thickened epithelium were common autopsies macroscopic findings. Tuberculosis multiorgan compromise was a relevant finding in patients with gastrointestinal tuberculosis. ConclusionsGastrointestinal tuberculosis is a disease of great importance, being its diagnosis a clinical challenge. Underdiagnosis can be reported in a high percentage of cases, so autopsy diagnosis can help reveal more accurate data about this condition.

Microscopy-Guided 3-D Reconstruction of Nanodendrites in Biosensors

Nano-dendritic structures have gained increasing popularity in electrochemical sensors. However, it is still rare to generate a 3-dimensional model in a short period of time to understand the structure-function relationship of the sensors. Here, we report the construction of a 3-dimensional model for nano-dendritic, metallic structures frequently grown on top of bioelectronics. This is achieved by merging two sources of 2-dimensional dendritic information, which includes top-view images from scanning electron microscopy and side-view visualization from Monte-Carlo simulations. The microscopy images provide the boundary conditions to tune the Monte-Carlo simulations to construct the 3-dimensional dendritic morphology. We validated the 3-dimensional model by comparing the dendritic area densities predicted via this model with those computed from microscopy images. Additionally, tuning the simulation parameters in the 3-dimensional model can be used to find the optimized dendritic density, which is an essential indicator for sensitivity enhancement. The success of this model provides a means to understand the sensitivity limits of bio-electronics through dendritic growth without the need for the time-consuming sensor fabrication and testing. Further, our SEM-guided Monte-Carlo technique provides a dendritic model with a significant resemblance to experimental images. It possesses the potential for applications in 3-dimensional morphological investigations for future biosensor design.

Antibacterial and Antibiofilm Activity of Different Species of Fabiana sp. Extract Obtained via Maceration and Ultrasound-Assisted Extraction against Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen that, under certain conditions, can induce aggravated infectious processes, mainly in immunosuppressed patients. Moreover, S. epidermidis is one of the leading causes of medical device- and implant-associated infections and is also recognized as a canonical biofilm producer. Fabiana punensis, F. densa and F. patagonica are three medicinal plants that grow in arid environments in Argentina (Altoandina, Puna, Prepuna and Monte regions). In this work, we studied the antimicrobial activity of alcoholic extracts of these plant species obtained via maceration (M) and ultrasound-assisted extraction (UAE) against S. epidermidis. In addition, the antibiofilm activity of the F. densa extract was also evaluated. It was found that the extracts obtained via M did not present differences with those obtained via UAE regarding the chemical profile. F. densa showed the lowest minimum inhibitory concentration (MIC) value (75 µg GAE/mL). At concentrations higher than the MIC, the extract induced the release of cellular constituents. At the concentration of 1/8× MIC, the extract inhibited biofilm formation by 78%, reducing metabolic activity by 67%. On the other hand, it presented a low percentage of preformed biofilm removal. In all assays, gallic acid (GA) has been used as a reference antimicrobial compound. Finally, it was shown via microscopy visualization that the extract reduces adhesion to hydrophobic and hydrophilic surfaces. Thus, F. densa extracts could potentially be used for the antibiotic treatment of infections produced by S. epidermidis or as an inhibitor agent of production biofilm, avoiding infections caused by medical devices.