Distinct Types Research Articles

Prevalence of Bacterial Urinary Tract Infections and Antimicrobial Sensitivity Among Pregnant Women in Anbar Governorate.

Background: Escherichia coli (E. coli) is recognized to be a prominent source of uropathogens in women worldwide. Urinary tract infection in pregnant women continues to cause clinical issues, which is a significant load on clinicians. Objective: The study aims to investigate the frequency of bacterial urinary tract infections among pregnant women and assess the susceptibility of these pathogens. Method: Urine samples 65 were collected from pregnant women in Anbar Governorate, were identified utilizing colony characteristics the organisms' gram stain response, biochemical assays Susceptibility testing aligns to the standard techniques defined by the Clinical and Laboratory Standard Institute guideline. Results: In this study, 95% (60 out of 65) of urinary tract infections (UTIs) were culture-positive. Among the identified bacteria, six distinct types were observed. The most prevalent Gram-negative isolate was Escherichia coli, accounting for 88.3% of cases, followed by Proteus mirabilis at 1.6%. Gram-positive bacteria were also identified, with Staphylococcus aureus representing 6.6% and Streptococcus at 3.3% of cases. E. coli showed resistance to ciprofloxacin (50%), norfloxacin (19%), ceftriaxone (90%), ampicillin (89%), meropenem (10%), and nitrofurantoin (60%). Staphylococcus aureus exhibited resistance to ciprofloxacin (73%), norfloxacin (35%), ceftriaxone (50%), ampicillin (71%), meropenem (8%), and nitrofurantoin (45%). Proteus mirabilis displayed resistance to ciprofloxacin (46%), norfloxacin (16%), ceftriaxone (62%), ampicillin (90%), meropenem (25%), and nitrofurantoin (33%). Streptococcus showed resistance to ciprofloxacin (43%), norfloxacin (54%), ceftriaxone (37%), ampicillin (88%), meropenem (30%), and nitrofurantoin (87%). Conclusion: Majority UTIs were caused by Gram-negative bacteria, especially E. coli. Resistance to ampicillin, nitrofurantoin, and ciprofloxacin was common, while norfloxacin and meropenem showed sensitivity. Regular urine culture during pregnancy is vital to lower UTI risk and antibiotic resistance.

Novel hierarchical bioinspired cellular structures with enhanced energy absorption under uniaxial compression

Biological thin-walled cellular structures, such as hierarchical structures of bone, exhibit unique internal structures that offer lightweight characteristics and high energy absorption capabilities. The compact bone shell can protect against local damage during high-impact events, while the porous cancellous bone is essential in absorbing energy. This research proposed five novel bioinspired cellular structures inspired by the well-arranged structure of femur bones. These structures comprise four distinct cell types: hexagonal honeycomb, re-entrant, hybrid, and hierarchical hybrid cells. A comprehensive numerical model, validated with experimental data, was employed to assess the performance of these structures under uniaxial compression. Some key characteristics were revealed, including peak elastic load, plateau load, energy absorption capacity, Poisson's ratio, and the effect of hierarchical cell size. The results demonstrated that the novel bioinspired structures surpassed the energy absorption performance of traditional designs, such as hexagonal design, re-entrant, and trabecular-bone-inspired structures. This enhanced performance was due to the progressive buckling and collapse mechanisms, showing promising implications for future engineering applications, particularly where energy absorption is of paramount importance.

Whole genome sequence-based molecular characterization of blood isolates of carbapenem-resistant Enterobacter cloacae complex from ICU patients in Kolkata, India, during 2017-2022: emergence of phylogenetically heterogeneous Enterobacter hormaechei subsp. xiangfangensis.

Blood-borne infections caused by the carbapenem-resistant Enterobacter cloacae complex (CR-ECC) are major public threats with respect to the challenges encountered during treatment. This study describes the whole genome sequencing-based molecular characteristics of blood isolates (n = 70) of CR-ECC from patients admitted to the intensive care unit of tertiary care hospitals in Kolkata, India, during 2017-2022 with respect to species identification, antimicrobial resistance (AMR) profiling, mechanism of drug resistance, and molecular subtypes. Vitek2 MALDI and species-specific PCR identified Enterobacter hormaechei subsp. xiangfangensis (47.14%) as the emerging CR-ECC subspecies in Kolkata. The predominating carbapenemase and extended-spectrum β-lactamase genes found were blaNDM-1 (51.42%) and blaCTX-M-15 (27%), respectively. Besides, blaNDM-4, blaNDM-5, blaNDM-7, blaCMH-3, blaSFO-1, blaOXA-181, blaOXA-232, blaKPC-3, and blaDHA-7 genes were also detected, which were not previously reported from India. A multitude of Class 1 integrons (including In180, In4874, In4887, and In4888, which were novel) and plasmid replicon types (IncFIB, IncFII, IncX3, IncHI1-HI2, IncC, and IncR) involved in AMR dissemination were identified. Reverse transcription-PCR and western blot revealed that carbapenem resistance in non-carbapenemase-producing CR-ECC isolates was contributed by elevated levels of ampC, overexpression of acrAB, and loss of ompF. A total of 30 distinct sequence types (STs) were ascertained by multi-locus sequence typing; of which, ST2011, ST2018, ST2055, ST2721, and ST2722 were novel STs. Pulsed-field gel electrophoresis analysis showed heterogeneity (69 pulsotypes with a similarity coefficient of 48.40%) among the circulating isolates, suggesting multiple reservoirs of infections in humans. Phylogenetically and genetically diverse CR-ECC with multiple AMR mechanisms mandates close monitoring of nosocomial infections caused by these isolates to forestall the transmission and dissemination of AMR.IMPORTANCEThe emergence and extensive dissemination of the carbapenem-resistant Enterobacter cloacae complex (CR-ECC) have positioned it as a critical nosocomial global pathogen. The dearth of a comprehensive molecular study pertaining to CR-ECC necessitated this study, which is the first of its kind from India. Characterization of blood isolates of CR-ECC over the last 6 years revealed Enterobacter hormaechei subsp. xiangfangensis as the most prevalent subsp., exhibiting resistance to almost all antibiotics currently in use and harboring diverse transmissible carbapenemase genes. Besides the predominating blaNDM-1 and blaCTX-M-15, we document diverse carbapenemase and AmpC genes, such as blaNDM-4, blaNDM-7, blaOXA-181, blaOXA-232, blaKPC-3, blaCMH-3, blaSFO-1, and blaDHA-7, in CR-ECC, which were not previously reported from India. Furthermore, novel integrons and sequence types were identified. Our findings emphasize the need for strengthened vigilance for molecular epidemiological surveillance of CR-ECC due to the presence of epidemic clones with a phylogenetically diverse and wide array of antimicrobial resistance genes in vulnerable populations.

Double cross-linked graphene oxide hydrogel for promoting healing of diabetic ulcers.

This study explores the synthesis and characterization of a novel double cross-linked hydrogel composed of polyvinyl alcohol (PVA), sodium alginate (SA), graphene oxide (GO), and glutathione (GSH), henceforth referred to as PVA/SA/GO/GSH. This innovative hydrogel system incorporates two distinct types of cross-linking networks and is meticulously engineered to exhibit sensitivity to high glucose and/or reactive oxygen species (ROS) environments. A sequential approach was adopted in the hydrogel formation. The initial phase involved the absorption of GSH onto GO, which was subsequently functionalized with boric acid and polyethylene glycol derivatives via a bio-orthogonal click reaction. This stage constituted the formation of the first chemically cross-linked network. Subsequently, freeze-thaw cycles were utilized to induce a secondary cross-linking process involving PVA and SA, thereby forming the second physically cross-linked network. The resultant PVA/SA/GO/GSH hydrogel retained the advantageous hydrogel properties such as superior water retention capacity and elasticity, and additionally exhibited the ability to responsively release GSH under changes in glucose concentration and/or ROS levels. This feature finds particular relevance in the therapeutic management of diabetic ulcers. Preliminary in vitro evaluation affirmed the hydrogel's biocompatibility and its potential to promote cell migration, inhibit apoptosis, and exhibit antibacterial properties. Further in vivo studies demonstrated that the PVA/SA/GO/GSH hydrogel could facilitate the healing of diabetic ulcer sites by mitigating oxidative stress and regulating glucose levels. Thus, the developed PVA/SA/GO/GSH hydrogel emerges as a promising candidate for diabetic ulcer treatment, owing to its specific bio-responsive traits and therapeutic efficacy.

Analysis of the mechanical behavior of an alternative sand for foundry molding

The exploration and beneficiation of iron ore result in a substantial generation of waste, predominantly directed towards tailings dams, heightening environmental concerns. Faced with this environmental challenge, numerous studies have been devoted to seeking sustainable solutions, aiming at the utilization of waste in various sectors of the economy. In this context, this study focuses on the technical feasibility investigation of using sandy iron ore waste as molding sand in foundry processes. Foundry molding sand, crucial in casting, must meet specific requirements and exhibit particular characteristics to effectively fulfill its function. To address this issue, test specimens were fabricated using three distinct types of sands: Standard Sand, Sand 01 (without magnetic separation), and Sand 02 (subjected to magnetic separation), combined with two binders (bentonite and sodium silicate). Subsequently, the test specimens underwent mechanical tests for tension, flexure, and compression, with the aim of evaluating their ability to withstand applied forces. The results of these tests revealed that Sand 02, when combined with sodium silicate, demonstrated satisfactory performance, in accordance with established standards and studies for obtaining sands intended for molding in foundry processes.

Association of pulmonary hypertension with the outcome in patients undergoing edge-to-edge mitral valve repair

ObjectivesThe association of pulmonary hypertension (PH) with the outcome after mitral transcatheter edge-to-edge repair (M-TEER) focusing on the new ESC/ERS guidelines definition for PH.BackgroundPH is frequently found in patients with...

Interplay of network topologies in aviation delay propagation: A complex network and machine learning analysis

In this study, the fundamental characteristics of flight delay propagation and the key factors influencing such propagation are investigated. Three distinct types of networks were constructed: an aviation network, a traffic flow network, and a delay propagation network. Employing complex network theory, an analysis of the fundamental topological attributes of each network was conducted, exploring the interrelations among these attributes. Findings reveal a notable resemblance between the network topology attributes of the delay propagation network and the traffic flow network. The delay propagation network overall adheres to scale-free network attributes, with a few dominant major airports playing a pivotal role in the delay propagation process. Furthermore, it was uncovered that smaller airports are more susceptible to the influence of delay propagation than their larger counterparts. Delays, in a general sense, exhibit a pronounced tendency to aggregate and spread extensively within the realm of smaller airports. Regarding delay propagation, airport traffic emerges as the primary factor precipitating this phenomenon, with a feature importance score reaching 0.889. An escalation in airport traffic significantly amplifies the extent of delay propagation, yet concurrently, this escalation is not incessant but stabilizes beyond a certain threshold.

Ecosystem Service Value Assessment of the Yellow River Delta Based on Satellite Remote Sensing Data

The Yellow River Delta (YRD) stands as a globally significant wetland, playing a pivotal role in sustaining regional ecosystem stability and offering crucial ecosystem services to humanity. However, anthropogenic activities, particularly resource development, unavoidably disrupt the ecosystem, leading to the degradation of these vital services. Utilizing satellite remote sensing data, the InVEST model, and energy analysis, this study introduces the concept of ‘emergy’ as an ‘intermediate variable’ to investigate the spatiotemporal changes in the ecosystem service value of the YRD. Five distinct types of ecosystem services are selected for quantitative assessment and analysis of the YRD’s spatiotemporal evolution from 1990 to 2020. Results indicate a 63.7% decline in the total value of ecosystem services from 1990 to 2010, followed by a 16.5% increase from 2010 to 2020. The study also unveils spatial shifts in high- and low-value areas of ecosystem services and attributes these changes to rapid urbanization and alterations in land use and cover. The assessment of ecosystem service values concretizes the intangible ecosystem service functions of natural resources. This lays the foundation for establishing a mechanism that combines positive incentives and reverse pressure to achieve the economic valuation of ecosystem service.

Monitoring of Thermoacoustic Combustion Instability via Recurrence Quantification Analysis and Optimized Deep Belief Network

Thermoacoustic oscillation is indeed a phenomenon characterized by the symmetric coupling of thermal and acoustic waves. This paper introduces a novel approach for monitoring and predicting thermoacoustic combustion instability using a combination of recurrence quantification analysis (RQA) and an optimized deep belief network (DBN). Six samples of combustion state data were collected using two distinct types of burners to facilitate the training and validation of GA-DBN. The proposed methodology leverages RQA to extract intricate patterns and dynamic features from time series data representing combustion behavior. By quantifying the recurrence plot of specific patterns, the analysis provides valuable insights into the underlying thermoacoustic dynamics. Among three different feature extraction methods, RQA stands out remarkably in performance. These RQA-derived features serve as input to a carefully tuned DBN, which is trained to learn the complex relationships within the combustion process. The classification accuracy of deep belief network optimized by genetic algorithm (GA-DBN) reached an impressive 99.8%. Subsequent multiple comparisons were conducted between GA-DBN, DBN, and support vector machine (SVM), revealing that GA-DBN consistently demonstrated satisfactory classification results. This method holds significant importance in monitoring intricate combustion states.

Improving acid-stressed anaerobic digestion processes with biochar - towards a combined biomass and carbon management system

Abstract Interest in biochar as an additive to enhance anaerobic digestion (AD) has grown in the context of biomass cascading use and the 2050 net-zero goal. However, few studies have investigated the effects of biochar on AD from a biochar production perspective, including biomass feedstocks and pyrolysis temperatures. To valorise biomass and better understand the mechanisms and environmental implications of using biochar in AD, we investigated the effects of distinct biochar types on AD under acid stress-induced process inhibition using batch tests. The results demonstrated that biochar can mitigate acid stress and enhance the methane production rate. The kinetic rate constant of methane production is positively related to the buffer capacity of the tested biochars (R2 = 0.88). The choice of feedstocks is a crucial factor (P = 0.003), particularly the best-performing biochars derived from raw grass silage. In contrast, the pyrolysis temperature effect was less significant (P = 0.18). Furthermore, the analysis of biochar indicates that the alkali (K) and alkaline earth (Ca, Mg) metals contained in biochar may be one of the important factors contributing to buffer capacity (R2 = 0.82 to 0.86). Hence, buffer capacity is a crucial quality criteria when evaluating biochar for AD applications. Raw grass silage biochars are promising for acid stress mitigation due to their high buffer capacity, while carbon-rich woody biochars have high CO2 sequestration potential. A compromise between mitigating acid stress and sequestering carbon is the use of pre-treated grass biochar. Overall, the use of biochar-enriched digestate offers a potential way to close material loops and complete the biomass-to-biochar value chain.

Evaluating AI-Generated Emails: A Comparative Efficiency Analysis

This study investigates the efficiency of large language models (LLMs) in producing routine, negative, and persuasive business emails for educational purposes within the context of Business Writing. Specifically, it compares the outputs generated by four widely-used LLMs (ChatGPT 3.5, Llama 2, Bing Chat, and Bard) when presented with identical email scenarios. These generated emails are evaluated using an elaborate rubric, allowing for a systematic assessment of LLMs' performance across three distinct email types. The results of the study show that the output with the same prompt varies greatly despite the rather formulaic nature of business emails. For instance, some LLMs struggle with following the requested structure and maintaining consistency in tone, while others have issues with unity and conciseness. The findings of this research hold implications for teaching business writing (rubrics, task instructions, in-class implementation), as well as for the integration of AI in professional communication at large.

What American children's museum mission statements say about Lifelong Competencies

AbstractChildren's museums are a distinct type of museum—one particularly poised to engage and support the development of Lifelong Competencies that will help children live into their own successful and fulfilling futures. Understudied in the children's museum, which one(s) they prioritize is largely unknown, and, thus, so is their optimization. This work draws upon the important communication tool of institutional mission statements. Frequency analysis of the terms in 321 American children's museum mission statements (Study A) identified that children's museums prioritize the Lifelong Competencies of creativity and curiosity. Thematic analysis of the 119 mission statements that included these terms (Study B) indicated that children's museums' intended practice for achieving creativity and curiosity is through experiences, environments, and play with a focus on hands‐on and interactive elements, family‐friendly features, eliciting a positive affect, and experiences that are educational and innovative. Furthermore, creativity and curiosity are primarily valued as mediators toward learning.

Asymptotic profiles of a spatial vector‐borne disease model with Fokker–Planck‐type diffusion

AbstractThis paper is concerned with a spatially heterogeneous vector‐borne disease model that follows the Fokker–Planck‐type diffusion law. One of the significant features in our model is that Fokker–Planck‐type diffusion is used to characterize individual movement, which poses new challenges to theoretical analysis. We derive for the first time the variational characterization of basic reproduction ratio for the model under certain conditions and investigate its asymptotic profiles with respect to the diffusion rates. Furthermore, via overcoming the difficulty of the associated elliptic eigenvalue problem, the asymptotic behaviors of endemic equilibrium for the model are discussed. Our results imply that whether rapid or slow movement of susceptible and infected individuals are conducive to disease control depends on the degree of disease risk in the habitat. Numerically, we verify the theoretical results and detect that Fokker–Planck‐type diffusion may amplify the scale of disease infection, which in turn increases the complexity of disease transmission by comparing the impacts of distinct dispersal types on disease dynamics.

Identification of a Gene Signature That Predicts Dependence upon YAP/TAZ-TEAD.

Targeted therapies are effective cancer treatments when accompanied by accurate diagnostic tests that can help identify patients that will respond to those therapies. The YAP/TAZ-TEAD axis is activated and plays a causal role in several cancer types, and TEAD inhibitors are currently in early-phase clinical trials in cancer patients. However, a lack of a reliable way to identify tumors with YAP/TAZ-TEAD activation for most cancer types makes it difficult to determine which tumors will be susceptible to TEAD inhibitors. Here, we used a combination of RNA-seq and bioinformatic analysis of metastatic melanoma cells to develop a YAP/TAZ gene signature. We found that the genes in this signature are TEAD-dependent in several melanoma cell lines, and that their expression strongly correlates with YAP/TAZ activation in human melanomas. Using DepMap dependency data, we found that this YAP/TAZ signature was predictive of melanoma cell dependence upon YAP/TAZ or TEADs. Importantly, this was not limited to melanoma because this signature was also predictive when tested on a panel of over 1000 cancer cell lines representing numerous distinct cancer types. Our results suggest that YAP/TAZ gene signatures like ours may be effective tools to predict tumor cell dependence upon YAP/TAZ-TEAD, and thus potentially provide a means to identify patients likely to benefit from TEAD inhibitors.

Infrared diode laser enhances human periodontal ligament stem cells behaviour on titanium dental implants

Low level laser treatment (LLLT) is known for its photobiostimulatory and photobiomodulatory characteristics, which stimulate cell proliferation, increase cellular metabolism, and improve cellular regeneration. The objective of the present research was to assess the possible influence of infrared diode laser irradiation on the behaviour, attachment, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) seeded on different types of dental implants. Two distinct types of implants, one subjected to laser surface treatment and the other treated with acid etching, were longitudinally divided into two halves and submerged in six wells culture plates. Both implants were subjected to infrared diode laser treatment, and subsequently, the morphology and attachment of cells were examined using scanning electron microscopy (SEM) after 14 and 21 days. The behaviour of (hPDLSCs) towards two types of implants, when exposed to osteogenic medium and low-level laser therapy (LLLT), was assessed using quantitative real-time polymerase chain reaction to measure the expression of stemness markers and osteogenic markers. The scanning electron microscopy (SEM) demonstrated that the application of infrared diode laser irradiation substantially improved the attachment of cells to both types of implants. The stemness gene markers were significantly down regulated in cells seeded on both surfaces when challenged with osteogenic media in relation to control. At 14 days, early osteogenic markers, were upregulated, while late osteogenic markers, were downregulated in both challenged groups. At the 21-day mark, hPDLSCs seeded on an acid-etched implant exhibited increased expression of all osteogenic markers in response to stimulation with osteogenic media and infra-red diode laser, in contrast to hPDLSCs seeded on a laser surface treated implant under the same conditions. Finally, the findings of our research revealed that when subjected to infrared diode laser, human periodontal ligament stem cells cultured on both types of implants demonstrated improved cellular attachment and differentiation. This suggested that infrared diode laser enhanced the activity of the cells surrounding the implants. Hence, the use of infrared diode laser could be pivotal in improving and expediting the clinical osseointegration process around dental implants.

On chip all-optical distinguishing of independently placed distinct types of single Rayleigh particle

In order to determine whether a particle is plasmonic, dielectric, or chiral, different complex processes and chemicals are applied in lab setups and pharmaceutical industries. Sorting or categorizing a particle based on distinct optical forces can be a novel technique. When a beam of light interacts with a particle, it usually pushes the particle in the direction of the light's propagation. Counterintuitively, it can also pull the particle toward the light beam or move it toward a lateral direction. As far as we know, to date, no comprehensive report exists regarding a single optical arrangement capable of inducing entirely distinct behaviors of force for three disparate types of independently placed single Rayleigh particle. This study introduces an all-optical technique aimed at effectively sorting nanoscale Rayleigh-sized objects employing a plasmonic substrate, when each distinct type of single particle is placed over the substrate independently. Unfortunately, this proposed technique does not work for the cluster or mixture of distinct particles. In our proposed configuration, a simple linearly polarized plane wave is incident onto the plasmonic substrate, thereby engendering completely different responses from three different types of nanoparticles: Gold (plasmonic), SiO2 (dielectric), and Chiral particles. We conducted individual tests for our setup using linearly polarized plane waves at angles of 30-degree, 45-degree, and 60-degree individually. Consistent results were obtained across all angles. In each of the three distinct setups involving the aforementioned particle, a dielectric Rayleigh particle experiences an optical pulling force, a plasmonic Rayleigh particle experiences an optical pushing force, and a chiral Rayleigh particle encounters an optical lateral force. These distinctive force behaviors manifest as a result of the intricate interplay between the material properties of the nanoparticles and the characteristics of the plane-polarized beam, encompassing aspects such as plasmonic response, chirality, and refractive index. Moreover, this technique presents an environmentally sustainable and economically viable alternative to the utilization of expensive and potentially hazardous chemicals in nanoparticle sorting processes within industrial domains.

Peripheral Blood Non-Coding RNA as Biomarker for Schizophrenia: A Review.

Schizophrenia (SCZ) is a complex and heterogeneous neuropsychiatric disorder that lacks objective diagnostic indicators and the pathogenesis remain unclear. Genetic factors may exert a significant impact on the development of the condition. While obtaining brain tissue for biopsy in the course of adjuvant diagnosis of SCZ patients may not be possible, the collection of peripheral blood is more accessible and easier to implement. In recent years, the development and application of RNA sequencing technology has made seeking biomarkers of SCZ becomes more feasible. There is emerging evidence suggesting that certain non-coding RNAs (ncRNA) are distinctly different in the peripheral blood of SCZ patients and healthy controls. Although the mechanisms remain unclear, these aberrantly expressed ncRNAs may be intimately associated with the onset and development of SCZ and may be of great significance for the diagnosis and treatment of SCZ. Therefore, we reviewed the expression of distinct types of ncRNAs that have been found in the peripheral blood of SCZ patients and explored their potential application as diagnostic biomarkers of SCZ. Differentially expressed ncRNAs in the peripheral blood of SCZ patients could not only serve as potential diagnostic biomarkers and therapeutic targets for SCZ but may also have implications for advancing understanding of the molecular mechanisms underlying the development of SCZ and elucidating the complex etiology of SCZ. Early diagnostic biomarkers obtained directly from peripheral blood are of great significance for the timely diagnosis and treatment of SCZ. Our review will enhance the comprehension of molecular mechanisms of SCZ and contribute to the identification of promising ncRNAs in peripheral blood for both diagnosis and therapy of SCZ.

First report of Truncatella angustata causing leaf blight on Thuja plicata Donn ex D. Don in Canada.

Western redcedar (Thuja plicata Donn ex D. Don) is one of the most important commercial tree species in British Columbia, generates more than $1 billion in economic activity annually and about 8-10 million trees are planted in reforestation efforts (Gregory et al. 2018). It has been selected as the provincial tree of British Columbia (BC) because of its tremendous economic, ecoogical and cultural value. However, foliar diseases such as leaf blights have serious impact on redcedar growth and may cause significant loss of tree volume (Russell, 2007). Our 2014 - 2015 surveys of western redcedar forests in coastal areas of BC indicated high incidence of a distinctive type of blight. We observed the incidence of this disease on more than 80% of western redcedar (approximately 493) trees from late May to early December. Early symptoms appeared as circular to oval, brownish to black spots (2-3 mm), 1-5 spots per branch tip, scattered at the tip margins. Sequentially, the spots enlarged and developed into necrotic lesions on both young and old leaves. More than 50 symptomatic leaves from 10 different trees were collected and rinsed in distilled water then surface-sterilized with three times washing in Tween 20 (%5 solution) for 2 minutes (each time) and %70 ethanol for 30 second (3 times repeat). Tissues from under lesions were placed on MEA (Malt Extract Agar; Phyto Tech® labs-Product ID: M498) and PDA (Potato Dextrose Agar; Phyto Tech® Labs-Product ID: P772). The plates were incubated at 21°C in the dark. They developed distinct dull white to brown, cottony colonies with each black acervuli approximately 450-500µm. The isolates produced fusiform conidia with four cells. They didn't have any distinct color. The conidiophore size was approximately 23-24 x 2-3 µm with mostly hyaline to light brown color, branched and conidiogenous was hyaline and not branched and simple. The spore size was approximately 15-20μm by 7-10μm with three transverse septa and endogenous papillae with hyaline apical appendages. Next, we collected spores and replated them on fresh MEA media culture and placed back in the incubator to produce pure cultures. We studied conidia from leaves of trees mentioned above using light and electron microscopy using Hitachi S-3500N Scanning Electron Microscope (Noshad et al. 2023). After morphological study, further identification to the species level conducted using Zambounis and Wenneker's approach (Zambounis 2019; Wenneker,2017). Genomic DNA from two single-spore isolates were isolated and sequenced. Sequences of ITS (Internal Transcribed Spacer) region amplified using primers ITS1/ITS4 and sequenced. Final sequences were deposited in Genbank and published (accession numbers OP086244 and OP086251). Blast analysis of these sequences showed 99% and 99% resemblances with T. angustata sequence (Sutton 1980). To verify its pathogenicity, we performed a comprehensive pathogenicity test to fulfill Koch's postulates. We collected their distinctive spores in an aseptic environment and standardized them (5000/ml) using a haemocytometer. Then we inoculated 100 western redcedar seedlings (three years old) by injecting standardized spore suspension solution (inoculum) using ultra-fine 0.3ml, 31G, 8mm syringes (approximately 0.1ml per inoculation site). Ten positive control seedlings were inoculated with distilled water and ten negative control seedlings were not inoculated at all. All inoculated (experimental) seedlings demonstrated same symptoms (black spots and characteristic spores) after eight weeks. None of the control seedlings showed any similar symptoms. In the next stage, we isolated and cultured spores from inoculated seedlings and studied them. The identity of reisolates confirmed using DNA sequencing. We used these spores for our next set of disease screening which was successful again. We identified Truncatella angustata (Pers.) Hughes as the causal agent for shoot-tip blight (STB) on western redcedar by examining morphological and molecular characteristics of the pathogen. This is the first report of T. angustata as a primary pathogen on western redcedar in British Columbia, Canada.

Phage-plasmids promote recombination and emergence of phages and plasmids

Phages and plasmids are regarded as distinct types of mobile genetic elements that drive bacterial evolution by horizontal gene transfer. However, the distinction between both types is blurred by the existence of elements known as prophage-plasmids or phage-plasmids, which transfer horizontally between cells as viruses and vertically within cellular lineages as plasmids. Here, we study gene flow between the three types of elements. We show that the gene repertoire of phage-plasmids overlaps with those of phages and plasmids. By tracking recent recombination events, we find that phage-plasmids exchange genes more frequently with plasmids than with phages, and that direct gene exchange between plasmids and phages is less frequent in comparison. The results suggest that phage-plasmids can mediate gene flow between plasmids and phages, including exchange of mobile element core functions, defense systems, and antibiotic resistance. Moreover, a combination of gene transfer and gene inactivation may result in the conversion of elements. For example, gene loss turns P1-like phage-plasmids into integrative prophages or into plasmids (that are no longer phages). Remarkably, some of the latter have acquired conjugation-related functions to became mobilisable by conjugation. Thus, our work indicates that phage-plasmids can play a key role in the transfer of genes across mobile elements within their hosts, and can act as intermediates in the conversion of one type of element into another.

A Raman topography imaging method toward assisting surgical tumor resection

AbstractAchieving complete tumor resection upon initial surgical intervention can lead to better patient outcomes by making adjuvant treatments more efficacious and reducing the strain of repeat surgeries. Complete tumor resection can be difficult to confirm intraoperatively. Methods like touch preparation (TP) have been inconsistent for detecting residual malignant cell populations, and fatty specimens like breast cancer lumpectomies are too fatty to process for rapid histology. We propose a novel workflow of immunostaining and topographic surface imaging of freshly excised tissue to ensure complete resection using highly sensitive and spectrally separable surface-enhanced Raman scattering nanoparticles (SERS NPs) as the targeted contrast agent. Biomarker-targeting SERS NPs are ideal contrast agents for this application because their sensitivity enables rapid detection, and their narrow bands enable extensive intra-pixel multiplexing. The adaptive focus capabilities of an advanced Raman instrument, combined with our rotational accessory device for exposing each surface of the stained specimen to the objective lens, enable topographic mapping of complete excised specimen surfaces. A USB-controlled accessory for a Raman microscope was designed and fabricated to enable programmatic and precise angular manipulation of specimens in concert with instrument stage motions during whole-surface imaging. Specimens are affixed to the accessory on an anti-slip, sterilizable rod, and the tissue surface exposed to the instrument is adjusted on demand using a programmed rotating stepper motor. We demonstrate this topographic imaging strategy on a variety of phantoms and preclinical tissue specimens. The results show detail and texture in specimen surface topography, orientation of findings and navigability across surfaces, and extensive SERS NP multiplexing and linear quantitation capabilities under this new Raman topography imaging method. We demonstrate successful surface mapping and recognition of all 26 of our distinct SERS NP types along with effective deconvolution and localization of randomly assigned NP mixtures. Increasing NP concentrations were also quantitatively assessed and showed a linear correlation with Raman signal with an R2 coefficient of determination of 0.97. Detailed surface renderings color-encoded by unmixed SERS NP abundances show a path forward for content-rich, interactive surgical margin assessment.