Manufacturer's Instructions Research Articles

First report of brown spot disease caused by Neoscytalidium dimidiatum on Disocatus ackermannii in Guangxi of China.

Disocatus ackermannii, commonly referred to as Orchid Cactus, is a striking succulent belonging to the Cactaceae family. Its unique appearance and captivating characteristics make it a sought-after addition to gardens and courtyards beautification. In June 2023, 20-30% of D. ackermannii in a flower nursery in Baise city (23°88'07.85″N, 106°56'98.96″E), Guangxi province of China showed brown spot disease. Symptoms were minute chlorotic depressed spots with a red fleck at the center, which turned to sunken, circular, light brown to dark brown lesions. Some lesions, surrounded by a yellow halo, led to cladode decay. In some cases, the diseased tissue dried out and appeared thin, producing conidia in the diseased area. For pathogen isolation, six cladodes were surface disinfected with 5% NaClO for 2 min followed by 70% ethanol for 30 s, rinsed three times with sterile water, then cultivated on potato dextrose agar (PDA) plates of 28 °C for 2 to 4 days. Emerging fungal colonies were purified by hyphal tipping. The colonies from the plates were initially white with powdery disarticulating aerial mycelium transitioned to an olive-green to greyish color and finally turned dark-gray to black after 9 days. The arthroconidia were ellipsoid to ovoid, zero to one septum, hyaline to dark brown and were 9.6 (4.2-15.4) × 4.5 (2.4-10.7) μm (n = 100). The morphological characteristics resembled those of N. dimidiatum (Crous et al. 2006; Phillips et al. 2013). Two isolates BSLJ4-1 and BSLJ5-1 were selected for DNA extraction following the manufacturer's instructions of Plant Genomic DNA Kit. Results from the BLASTn analysis of the ITS (GenBank accession nos. PP868502，PP860525), EF1-α (PP868386, PP868387), β-tubulin (PP868384, PP868385) genes of the isolates BSLJ4-1 and BSLJ5-1 showed higher than 99% identities with the epitype N. dimidiatum CBS 251.49 (ITS: KF531819; EF1-α: KF531797; β-tubulin: KF531799). The sequences were converted to FASTA files, assembled, and the maximum likelihood tree was constructed using IQ-TREE with the best model HKY+F+G4. The tree was evaluated from 1000 bootstrap replications. The result showed the isolates were related to N. dimidiatum with 100 bootstrap value supported. Spore suspensions (1 × 107 conidia/mL) were sprayed onto young cladodes of D. ackermannii. Three plants per isolate, sterile water was used as the control. Two weeks after inoculation, circular or subcircular faded green, brown sunken spots appeared on the leaves, surrounded by yellow halo even soft rotting developed but no infections observed in the water control. N. dimidiatum was successfully reisolated from inoculate-induced brown spot and subsequently identified based on its morphological characteristics and ITS sequence to fulfill the Koch's postulates. N. dimidiatum is a primary pitaya canker causative agent (Chuang et al. 2012; Lan et al. 2012; Sanahuja et al. 2016), however, in our observations, the characteristic hard brown scabs typically associated with N. dimidiatum infections on pitaya were not present on D. ackermannii. This is the first report of N. dimidiatum causing brown spot disease on D. ackermannii in China, suggesting a broader host range of N. dimidiatum within the Cactaceae family. Further studies are needed to understand the pathogen potential impact on other species and develop disease management strategies.

Clinical Performance of MAGLUMI Diagnostic Tests for the Automated Detection of Dengue Virus.

The screening and diagnosis of dengue virus infection play a crucial role in controlling the epidemic of dengue fever, highlighting the urgent need for a highly sensitive, simple, and rapid laboratory testing method. This study aims to assess the clinical performance of MAGLUMI Denv NS1 in detecting dengue virus NS1 antigen. A retrospective study was conducted to assess the sensitivity and specificity of MAGLUMI Denv NS1 using residual samples. Dengue-confirmed and excluded samples, validated by qPCR, were subjected to testing with MAGLUMI Denv NS1 in accordance with the manufacturer's instructions. The linear range, endogenous interference, and cross-reactivity of MAGLUMI Denv NS1 were verified, and a consistency analysis with commercial comparator products was carried out. The diagnostic specificity of MAGLUMI Denv NS1 is 98.41% (62/63), and the sensitivity is 98.32% (117/119). It effectively detects various serotypes of dengue virus, with no observed endogenous interference or cross-reactivity. Additionally, the consistency of NS1, IgM, and IgG tests on the MAGLUMI platform compared to commercial comparator reagents reaches 85.71%, 99.25%, and 98.97%, respectively. The MAGLUMI Denv NS1 represents a highly sensitive laboratory testing method capable of enhancing the diagnostic accuracy and efficiency of dengue virus infection detection.

Effect of mechanical instrumentation on titanium implant surface properties.

To assess the impact of mechanical decontamination using rotary brushes on the surface topography, elemental composition, roughness, and wettability of titanium implant surfaces. Four commercially available rotary brushes were used: Labrida BioClean Brush® (LB), i-Brush1 (IB), NiTiBrush Nano (NiTiB), and Peri-implantitis Brush (PIB). Seventy-five titanium discs with sandblasted, large-grit, acid-etched (SLA) surfaces were randomly assigned to five groups (n = 15): LB, IB, NiTiB, PIB, and a control group. Each disc was treated for 60 seconds with the respective rotary brush according to the manufacturer's instructions. Surface morphology was analysed using Scanning Electron Microscopy (SEM), surface elemental composition with Energy Dispersive X-ray (EDX), surface roughness via optical profilometry, and wettability with a droplet shape analyser. SEI analysis revealed morphological changes, including scratches, flattening, and loose titanium particles in the IB, PIB, and NiTiB groups, whereas the LB group preserved the original surface morphology. SEM-EDX analysis showed that LB, PIB, and NiTiB groups closely match the control elemental composition. However, IB groups showed significantly different composition. Surface roughness values in the IB, PIB, and NiTiB groups differed significantly from the control (p < 0.05), whereas the LB group had comparable roughness values (p > 0.05). Contact angle measurements indicated enhanced wettability in IB, PIB, and NiTiB groups (p < 0.05), while the LB group exhibited values comparable to the control (p > 0.05). Mechanical decontamination of implant surfaces utilising rotary brushes can alter implant surface properties.

Efficacy of cleaning, disinfection, and sterilization modalities for addressing infectious drug-resistant fungi: a review.

This is a timely and important review that focuses on the appropriateness of established cleaning, disinfection, and sterilization methods to safely and effectively address infectious fungal drug-resistant pathogens that can potentially contaminate reusable medical devices used in healthcare environment in order to mitigate the risk of patient infection. The release of the World Health Organization (WHO) fungal priority pathogen list (FPPL) in 2022 highlighted the public health crisis of antimicrobial resistance (AMR) in clinically relevant fungal species. Contamination of medical devices with drug-resistant fungal pathogens (including those on the FPPL) in healthcare is a rare event that is more likely to occur due to cross-transmission arising from lapses in hand hygiene practices. Established disinfection and sterilization methods decontaminate fungal pathogens on single-use and reusable medical devices; however, there are assumptions that reusable devices destined for semi-critical use are appropriately cleaned and do not harbour biofilms that may undermine the ability to effectively decontamination these type devices in healthcare. International standards dictate that manufacturer's instructions for use must provide appropriate guidance to healthcare facilities to meet safe reprocessing expectations that include addressing drug-resistant fungal pathogens. Increased environmental monitoring and vigilance surrounding fungal pathogens in healthcare is advised, including adherence to hand hygiene/aseptic practices and appropriate cleaning encompassing the simplification of reusable device features for 'ease-of-reach'. There are emereging opportunities to promote a more integrated multiactor hub approach to addressing these sophisticated challenges, including future use of artificial intelligence and machine learning for improved diagnostics, monitoring/surveillance (such as healthcare and wastewater-based epidemiology), sterility assurance, and device design. There is a knowledge gap surrounding the occurrence and potential persistence of drug-resistant fungal pathogens harboured in biofilms, particularly for ascertaining efficacy of high-level disinfection for semi-critical use devices.

Comparison of physical, mechanical, and optical properties between thermoplastic materials and 3-dimensional printing resins for orthodontic clear retainers.

This study investigated the physical, mechanical, and optical properties of 3-dimensional (3D) printing resins compared with thermoplastic materials to evaluate their suitability for the fabrication of orthodontic clear retainers. Samples were prepared from thermoplastic sheets (Duran [Scheu-Dental GmbH, Iserlohn, Germany] and Zendura [Bay Materials LLC, Fremont, Calif]) and biocompatible 3D-printing resins (Dental LT Clear V2 [Formlabs Inc, Somerville, Mass] and OrthoFlex [Nextdent BV, Soesterberg, The Netherlands]) according to the manufacturer's instructions. The materials were characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry, and water sorption tests. Mechanical properties were assessed by tensile tests and hardness under 3 different conditions: dry, wet (24-hour water immersion), and aged (thermocyled for 10,000 cycles). Surface characteristics were qualitatively and quantitatively evaluated by scanning electron microscopy and 3D confocal imaging, respectively. Optical properties were assessed by ultraviolet-visible spectroscopy and color stability tests by immersion into various staining solutions. The mechanical properties of the 3D-printing resins were more markedly altered in different testing conditions (dry, wet, and aged) than in thermoplastic materials. The surface roughness, transparency, and color stability of 3D-printing resins are significantly inferior, especially NextDent OrthoFlex. The evaluated 3D-printing resins are more brittle and less ductile compared with the thermoplastic materials. The 3D-printing resins also do not meet the clinical thresholds of surface roughness and optical properties for the fabrication of orthodontic clear retainers. Further postprocessing of the 3D-printing resins may be required to improve these properties.

Influence of the shortened warming protocol on human blastocyst viability: an in vitro experimental study

Research QuestionDoes the shortened warming protocol impact the cell viability and outgrowth competence of human vitrified blastocysts warmed with or without fatty acids (FAs)? DesignIn this study, 326 discarded vitrified human blastocysts donated for research by consenting couples were used. The blastocysts were randomly allocated to five groups depending on the warming solutions, protocols, and recovery culture media: the control-conventional, control-shortened, FA-conventional, FA-shortened, and FA-shortened/recovery culture with FA (RF) groups. The blastocysts were warmed with or without FAs following the manufacturer's instruction (conventional method) or using the shortened method, in which blastocysts were immersed in a thawing solution for 1 min and then cultured in the recovery medium for 2 h. The embryo volume recovery, cell viability, intracytoplasmic lipid droplets, and outgrowth competence were evaluated. ResultsThe degree of blastocyst volume recovery was significantly higher after shortened warming than after conventional warming. Cell membrane collapse was observed during the shortened warming. The blastocyst survival rate and expansion status after the recovery culture were comparable among the five groups. However, the proportion of necrotic cells was increased in the control-shortened, FA-shortened, and FA-shortened/RF groups. The adhesion rates were comparable among all the groups; however, the outgrowth area was significantly higher in the FA-conventional group than in the other groups. ConclusionsThe efficacy of the shortened warming protocol depended on the warming solutions used. The shortened protocol effectively improves the daily workflow; however, each laboratory should examine its impact thoroughly before introducing it to prioritise clinical outcomes.

Magnetic Carbon Bead-Based Concentration Method for SARS-CoV-2 Detection in Wastewater.

Wastewater surveillance for pathogens is important to monitor disease trends within communities and maintain public health; thus, a quick and reliable protocol is needed to quantify pathogens present in wastewater. In this study, a method using a commercially available magnetic carbon bead-based kit, i.e., the Carbon Prep (C.prep) method (Life Magnetics), was employed to detect and quantify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as well as bacteriophage Phi6 and pepper mild mottle virus (PMMoV) in wastewater samples. The performance of this method was evaluated by modifying several steps and comparing it with the polyethylene glycol (PEG) precipitation method to demonstrate its applicability to virus detection in wastewater. The protocol of the C.prep method, based on the manufacturer's instructions, could not detect SARS-CoV-2 RNA, while the optimized protocol could detect it in the tested samples at concentrations that were not significantly different from those obtained using the PEG precipitation method. However, the optimized C.prep method performed more poorly in recovering Phi6 and detecting PMMoV than the PEG precipitation method. The results of this study indicated that the full workflow of the C.prep method was not sufficient to detect the target viruses in wastewater and that an additional RNA extraction step was needed to increase its detection sensitivity.

Microtomographic Analysis of the Biomechanical Performance of Thermally Treated Instruments in Flattened and Curved Root Canals.

This study aimed to evaluate, using micro-computed tomography (microCT), the impact of various mechanized systems on the geometry of the root canal system (RCS) and dentin thickness in mandibular first molars. The hypothesis proposed that different systems would produce significant variations in RCS preparation. Mesial and distal canals of mandibular molars were selected and divided into five groups (n=10) based on the system used: 2Shape (25.06), Reciproc Blue (25.08), ProTaper Ultimate (25.08), ProTaper Next (25.06), and Univy New (25.06). Biomechanical preparation was performed following each manufacturer's instructions. MicroCT scans were conducted before and after preparation to assess three-dimensional parameters (volume, surface area, SMI, and prepared root canal walls) and two-dimensional parameters (area, perimeter, diameters, roundness, and transportation). ProTaper Ultimate exhibited the greatest volume increase in mesial canals (1.73 ± 0.19), while ProTaper Next demonstrated the highest increase in distal canals (1.95 ± 0.25). Furthermore, ProTaper Ultimate resulted in the greatest dentin removal and the largest post-preparation area. Reciproc Blue achieved the best results for roundess and had the lower unprepared walls (15.3 ± 6.97). All systems produced comparable outcomes, with ProTaper Ultimate yielding the greatest volume increase and dentin removal, indicating a less conservative preparation. Reciproc Blue provided optimal roundness and minimized unprepared root canal wall surfaces.

Gasdermin D (GSDM D) as a Potential Diagnostic Biomarker in Bladder Cancer: New Perspectives in Detection.

Traditional methods of bladder cancer (BC) diagnosis include clinical examination, imaging, urine tests, cystoscopy, and biopsy. Due to the complexity of detection, diagnostic markers of bladder cancer measured in blood are still being sought. The pathogenesis of BC is complex and depends on many factors. However, the available literature data suggest that gasdermin D (GSDM D) has an influence in the pathogenesis of cancers. This study is the first that assesses the significance and diagnostic utility of serum GSDM D in bladder cancer. A total of 80 serum samples were obtained and analysed from healthy volunteers (C) and bladder cancer patients. The cancer patients were further classified into two groups, low-grade (LG) and high-grade (HG) bladder cancer, according to the TNM classification. The serum levels of GSDM D, CEA, and CA19-9 were assessed following the manufacturer's instructions using immunoassay techniques. The concentrations of GSDM D were nearly twice as high in the serum of BC patients compared to controls. Additionally, the median of GSDM D concentration was notably elevated in LG and HG bladder cancer than in C. In the total study group, the GSDM D concentration correlated with CRP and CEA levels. The diagnostic sensitivity of GSDM D was higher than that of CEA, but slightly lower in comparison to CA19-9. Moreover, the negative predictive value of GSDM D was the highest among all tested markers. The highest positive predictive value and diagnostic accuracy were observed for CEA, while the accuracy of GSDM D was comparable. GSDM D had an AUC value of 0.741, which was similar to the AUC value of CEA. GSDM D in serum appears to be a valuable diagnostic biomarker, especially when its measurement is used in conjunction with other markers such as CEA. Its high sensitivity makes it effective for the early detection of bladder cancer.

Long-Term Outcomes and Late Complications of Thoracic Endovascular Aortic Repair Using the GORE TAG or Conformable GORE TAG.

Previous clinical studies have reported on the short- and mid-term prognoses of thoracic endovascular aortic repair (TEVAR) using the GORE thoracic aortic graft (TAG) or conformable GORE TAG. We evaluated the long-term prognoses of patients who underwent TEVAR using the GORE TAG or conformable GORE TAG in a real-world setting. This retrospective observational study analyzed the data of all consecutive patients aged 20 years or older who underwent TEVAR using either the GORE TAG or the conformable GORE TAG at a single center. The surgical criteria did not deviate from global standards. In principle, TEVAR was performed with the TAG or conformable thoracic aortic graft (CTAG) according to the manufacturer's instructions. The primary outcomes of this study were the 10-year overall survival rate and the rate of freedom from aorta-related deaths at 10 years. Between January 2011 and December 2021, 331 patients underwent TEVAR, of whom 175 consecutive patients treated with TEVAR using either the GORE TAG or the conformable GORE TAG were enrolled. The mean age of the patients was 72.1 ± 9.3 years. In addition, the aneurysm and chronic dissection were 61.7% and 38.3%, respectively. Of all patients, 157 had a proximal landing zone ≥3, and 18 had a zone <3. The mean follow-up time was 46.5 ± 33.1 months. The 30-day mortality rate was 2.86%. The 3-, 5-, 7-, and 10-year overall survival rates were 76.9 ± 3.4%, 63.4 ± 4.1%, 55.9 ± 4.9%, and 42.4 ± 8.4%, respectively. The rate of freedom from aorta-related deaths at 3, 5, 7, and 9 years was 97.4 ± 1.3%, 95.0 ± 2.1%, 95.0 ± 2.1%, and 95.0 ± 2.1%, respectively. Late complications occurred in 23 (13.1%) patients. Endovascular re-intervention was performed because of endoleak, false lumen enlargement, or stent graft-induced new entry tear in 14 patients. Three patients required open conversion, and 6 were observed without re-intervention. Thoracic endovascular aortic repair using the GORE TAG or conformable GORE TAG is a safe and effective procedure for the treatment of thoracic aortic aneurysms and aortic dissection. Thoracic endovascular aortic repair using the GORE thoracic aortic graft (TAG) or conformable GORE TAG is a safe and effective procedure for the treatment of thoracic aortic aneurysms and aortic dissection. The 30-day mortality rate was 2.86%. During 10 years of follow-up, the overall survival rate was low; however, the risk of aorta-related deaths was also low. No significant differences in freedom from late complications and re-intervention are seen between the TAG and conformable thoracic aortic graft (CTAG) groups. Further investigations into the treatment of late complications are required.

The Impact of Pre-Treatment with Desensitizing Agents on the Effectiveness of In-Office Bleaching: An In Vitro Study.

In-office teeth bleaching is an esthetic dental procedure performed to whiten teeth. A desensitizing procedure often requires prior in-office bleaching to relieve tooth hypersensitivity. The objective of this study was to evaluate the bleaching efficiency of teeth specimens pre-treated with different desensitizing agents by analyzing the color parameters. A total of 25 bovine specimens were stained and divided into five groups (n = 5). The specimens were pre-treated with GLUMA, ICON, bonding, or fluoride varnish according to the manufacturer's instructions. Subsequently, the samples underwent three consecutive 20 min bleaching sessions using an in-office bleaching product. Color values in the CIE L*, a*, and b* color space were initially recorded, both before any treatment and after each bleaching session. The color difference (ΔE) was then computed. The data were analyzed using SPSS 24.0 software. The color alteration after pre-treatment was analyzed using a one-way ANOVA test, and the color alteration in each bleaching session was compared using a repeated-measure ANOVA test. A one-way ANOVA analysis showed a statistically significant difference in the bleaching efficiency of teeth specimens pre-treated with different desensitizers (p < 0.001). The repeated-measure ANOVA analysis showed no statistically significant difference in additional bleaching sessions (p = 0.133). The ICON and bonding pre-treatment showed the color alteration of teeth specimens before the bleaching procedure. Although the different desensitizer pre-treatments showed significant differences in bleaching efficiency, the differences in color parameters of the teeth specimens at the end of three bleaching sessions were statistically insignificant. Desensitizer pre-treatment influenced the degree of color change during multiple bleaching sessions; however, the outcome of bleaching was not affected by desensitizer pre-treatment.

UHPLC-MS/MS standardized extract of Vernonia amygdalina leaf inhibits CYP2C9 and CYP3A4 activities in hepatic cells of control and streptozotocin-induced diabetic rats.

Vernonia amygdalina Del. is a perennial tropical shrub from Asteraceae. The fresh leaf of V.amygdalina is consumed as a vegetable due to its medicinal and nutritional properties. The present study focused on the quantification of bioactive compounds, luteolin-7-O-glucoside, luteolin-7-O-glucuronide, and 1,5-O-dicaffeoylquinic acid from aqueous leaf extract of V.amygdalina. The study also aims to investigate the effects of the aqueous leaf extract of V.amygdalina on cytochrome P450 2C9 (CYP2C9), and cytochrome P450 3A4 (CYP3A4) in hepatic cells of control and diabetic rats. The quantification of the bioactive compounds was conducted using ultra-high-performance liquid chromatography multiple reactions monitoring tandem mass spectrometry (UHPLC-MS/MS-MRM) technique. The effect of the extract on CYP2C9 and CYP3A4 activities was determined using a fluorometric screening kit according to the manufacturer's instructions. The three bioactive compounds were detected and quantified in the aqueous leaf extract. Results showed that the content of luteolin-7-O-glucuronide (47 μg/mg) was the highest followed by luteolin-7-O-glucoside (3.5 μg/mg) and 1,5-O-dicaffeoylquinic acid (1.07 μg/mg). The extract showed an inhibitory effect on CYP3A4 and CYP2C9 enzyme activities in control and diabetic rats. The UHPLC-MS/MS-MRM method is sensitive and reliable for the quality control of V.amygdalina leaf extract. The inhibitory effect of the extract suggests that concomitant use of V.amygdalina leaf preparations with conventional drugs metabolized and eliminated from the body by CYP3A4 and CYP2C9 enzymes may lead to possible interaction.

COMPARISON OF SOME MECHANICAL PROPERTIES OF ADDING ZrO2 NANOPARTICLES TO MICROWAVE-CURED ACRYLIC AND HOT-CURED DENTURE BASE ACRYLIC MATERIALS

Background: The mechanical properties of denture-base acrylic resin have declined over the past decade, it is imperative to take immediate action to investigate alternative possibilities, such as employing metal oxide nanoparticles, which have shown considerable promise in diverse applications in the fields of nanobiotechnology, specifically in biological and health care fields. Aims: Exploring the effect of incorporating zirconium oxide (ZrO2) nanoparticles into microwave-cured acrylic and hot-cured acrylic denture base materials, on the tensile strength, impact strength and structural chemical changes. Methods: A total of 66 samples were utilized. The manufacturer's instructions have been followed for the production of specimens for each product. The impact of addition 0%, 3% and 5% zirconium oxide (ZrO2) nanoparticles were examined. Results: Addition of 3% and 5% ZrO2 nanoparticles improved the tensile and impact strengths with no chemical alterations changes with the addition of ZrO2 NPs, with 3% ZrO2 showing a greater enhancement than 5%. Conclusions: Incorporating 3% ZrO2 nanoparticles into both microwave-cured and hot-cured acrylic denture base materials enhances tensile and impact strengths more effectively than 5%, with no chemical changes observed.

INSTALASI PEMBUMIAN DAN INSTALASI LISTRIK MUSHOLA AL MUTTAQIIN TEMBESI KIBING-BATU AJI BATAM

The condition of the electrical installation system in people's homes and especially places of pray (Mushola) in Tembesi Kalurahan Kibing, Batu Aji District, Batam City, most people still do not understand about electrical installations and there are still many things that do not comply with general regulations regarding electrical installations, apart from There are still many electrical installation connections carried out by the community themselves which do not comply with the provisions. Community service by providing counseling and going directly to see the location that is the target for carrying out service activities, namely the Al Muttaqiin prayer room. By holding community service (PKM), residents of Kibing Village will understand how to install electricity properly, correctly and safely. The equipment selected for installation in an electrical installation must meet applicable standards and must be suitable for its environment. Installation of equipment must comply with the provisions in PUIL, and match the equipment manufacturer's instructions. Electrical installations must be regularly inspected and tested for misuse, damage or poor installation, including loose connections. To protect against leakage currents, grounding and ELCB can be installed.In order to achieve an increase in public knowledge and understanding about the importance of electrical hazards, this activity needs to be sustainable

First report of potato pink rot caused by Phytophthora erythroseptica in Colorado.

Pink rot, caused by the oomycete Phytophthora erythroseptica, is a serious disease that may cause substantial losses to potato growers. In 2023, high infection rates of pink rot were noticed in the San Luis Valley (SLV), Colorado around harvest time and in storage with several growers losing complete storage pins to pink rot. In September 2023, tubers with pink rot symptoms were collected from two grower fields of Russet Norkotah (~2-3% infection) and Canela Russet (~5% infection) near the San Luis Valley Research Center at harvest time. Infected tubers showed dark discoloration on the surface at the stem end. Upon cutting, affected tissue developed salmon pink color about 20-30 min later (Fig. 1). DNA was extracted from two symptomatic tubers from each cultivar and healthy tubers using DNeasy PowerSoil Pro Kit (Qiagen, MD, USA) according to the manufacturer's instructions. The conventional PCR and qPCR assays developed by Cullen et al. (2007) were used to detect P. erythroseptica. In the conventional PCR, the primer pair Pery2F1/R1 produced expected bands of 135bp from symptomatic tubers but not from the healthy tubers. The symptomatic tubers were positive in the qPCR using the primer-probe set 99F, 177R, and 133T (Cullen et al., 2007). P. erythroseptica was isolated by culturing specimens of infected tissues on PARP selective media (Jeffers and Martin, 1986). A week later colonies of white color grew on the media. These colonies were confirmed to be of P. erythroseptica by PCR as mentioned above. The colonies of two isolates were transferred to water agar media, then three days later a mycelium tip was excised and cultured on PDA media for single isolation. On PDA media the colonies showed chrysanthemum pattern. Four days later DNA was extracted from the two isolates, RN296-1 and Canela-2, and tested positive for P. erythroseptica by the above-mentioned PCR. PDA agar plugs of two isolates were used to inoculate surface sterilized potato tubers. The isolate SLV-2023-Canela2 was inoculated on tubers of Russet Norkotah and Reveille Russet, 10 each, while10 tubers of Russet Norkotah were inoculated with the isolate SLV-2023-RN296-1. Ten tubers of Russet Norkotah were inoculated by sterilized plugs of PDA for negative controls. Tubers were left at room temperature for a week. All tubers of Russet Norkotah inoculated with SLV-2023-Canela2 and RN296-4 developed pink rot symptoms and tested positive for P. erythroseptica by PCR, while only 9 tubers of Reveille Russet inoculated with SLV-2023-Canela2 developed symptoms and were positive to P. erythroseptica by PCR. No symptoms were observed on the mock inoculated tubers and no P. erythroseptica was detected by the PCR. P. erythroseptica was isolated again from Russet Norkotah tubers inoculated with RN296-1 on PDA media and tested positive for P. erythroseptica by PCR one week later. To further confirm the identity, the ITS region was amplified using the primers ITS-1 and ITS-4 (White et al., 1990) from DNA extracted from inoculated Russet Norkotah tubers with SLV-2023-Canela2 isolate. PCR product was purified using QIAquick Gel Extraction Kit (Qiagen, MD, USA)and cloned using TOPO TA Cloning Kit for Sequencing (Invitrogen, MD, USA). Four clones were submitted for sequencing. The 846 nt sequence of the ITS was submitted to the GenBank (accession number PP587391) shared 99.76% identity with the P. erythroseptica strain BBA 62683 (KJ755119). P. erythroseptica was identified previously using morphological tools from tubers with symptoms resembling pink rot collected in the SLV (Tyler et al., 2002). However, in that report neither Koch's Postulates, nor molecular identification were conducted, hence, the current study presents the first molecular report of pink rot in the SLV, Colorado. Rapidly changing climate and increased temperature around harvest time may lead to an increase in the incidence of pink rot, hence effective management strategies need to be developed and implemented.

Trends in Bacterial Distribution and Antimicrobial Resistance in Intensive Care Units of Hubei Province, China: A Four-year Surveillance Study (2020-2023).

To investigate the distribution characteristics of common bacteria and changes in antimicrobial resistance in intensive care unit (ICU) patients in 58 hospitals in Hubei Province from 2020-2023. The antimicrobial agents for antimicrobial susceptibility tests was selected based on the 2022 China Antimicrobial Resistance surveillance system (CARSS) technical scheme, and the specific experimental operation was based on the requirements of the CLSI M02 and M07 documents. The commercial instruments were used following the manufacturer's instructions. The interpretation of antimicrobial susceptibility test results was based on the 2023 CLSI M100 standard. There were 15 585, 19 258, 23 423 and 22 395 clinical isolates in the ICU from 2020 to 2023, respectively. Among them, gram-positive bacteria accounted for 20.5% (3190/15 585), 21.2% (4089/19 258), 21.6% (5067/23 423) and 21.6% (4 831/22 395), respectively. Gram-negative bacteria accounted for 79.5% (12 395/15 585), 78.8% (15 169/19 258), 78.4% (18 356/23 423) and 78.4% (17 564/22 395) of the bacteria, respectively. The top 5 isolates of gram-positive bacteria were Staphylococcus aureus, Enterococcus faecium, Streptococcus pneumoniae, Enterococcus faecalis, Staphylococcus epidermidis and gram-negative bacteria were Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa and Stenotrophomonas maltophil, respectively, but the proportions and rankings of the isolates in different years slightly differed. The detection rate of methicillin-resistant S. aureus (MRSA) decreased from 44.4% in 2020 to 36% in 2023, and that of methicillin-resistant coagulase-negative Staphylococcus (MRCNS) decreased from 79.8% in 2020 to 73.8% in 2022 and increased to 78.4% in 2023. The detection rates of both vancomycin-resistant E. faecium and E. faecalis were lower than 1%. The detection rate of carbapenem-resistant P. aeruginosa (CRPA) decreased from 25% in 2020 to 19.7% in 2022 and increased slightly to 20.6% in 2023. The detection rate of carbapenem-resistant A. baumannii (CRAB) decreased from 81.9% in 2020 to 79.7% in 2022 and increased to 82.9% in 2023. The detection rate of third-generation cephalosporin-resistant E. coli decreased from 59.8% in 2020 to 53.1% in 2022 and increased to 52.5% in 2023. The detection rate of fluoroquinolone-resistant E. coli decreased from 62.7% in 2020 to 50.2% in 2022 and increased slightly to 51.0% in 2023. The detection rate of carbapenem-resistant E. coli (CRECO) decreased from 3.3% in 2020 to 1.8% in 2022 and slightly increased to 2.1% in 2023. The detection rate of third-generation cephalosporin-resistant K. pneumoniae decreased from 34.3% in 2020 to 26.3% in 2022 and then increased to 32.4% in 2023. The detection rate of carbapenem-resistant K. pneumoniae (CRKPN) increased from 17.9% to 19.4% in 2020, decreased to 13.2% in 2022, and rose sharply to 20.4% in 2023. MRSA showed a continuous downwards trend from 2020 to 2023, while the detection rates of MRCNS and most multidrug-resistant gram-negative bacteria continuously decreased from 2020 to 2022 but tended to increase in 2023. Therefore, it is still necessary to strengthen the monitoring of bacterial resistance and rational application of antibiotics and actively and effectively control nosocomial infections.

Bonding performance of universal adhesive systems to enamel - Effects of the acidic composition.

To evaluate the self-etch bonding potential of universal adhesive systems with varying acidic compositions by analyzing the wettability properties, topographical change, and microshear bond strength (µSBS) to enamel. Eight universal adhesives were tested: All-Bond Universal (Bisco), Ambar Universal (FGM), Gluma Bond Universal (Kulzer), OptiBond Universal (Kerr), Peak Universal Bond (Ultradent), Prime&Bond Universal (Dentsply), Singlebond Universal (3 M ESPE), and Tetric N-Bond Universal (Ivoclar). Bovine incisors were prepared and treated with each adhesive according to the manufacturer's instructions. The following tests were carried out: contact angle to measure the adhesives' wetting ability; optical profilometry to evaluate the topographical changes obtained with adhesives' application; energy dispersive X-ray spectroscopy (EDX) to quantify the removal of inorganic elements from enamel; and µSBS to verify the bonding potential of adhesives to enamel. Statistical analyzes included ANOVA, Kruskal-Wallis, Weibull analysis, Pearson correlation, and chi square test (α=0.05). Significant differences were found in contact angles immediately after adhesive application and after 30 s, with Gluma Bond Universal and Tetric N-Bond Universal showing the lowest (the best wetting ability) and the highest values (the lowest wetting ability), respectively. EDX revealed varying degrees of Ca and P reduction, with Gluma Bond Universal resulting in the greatest Ca and P reduction from enamel (∼65 % and ∼62 % reduction, respectively. Topographical analysis indicated significant changes in roughness, mainly for Gluma Bond Universal. The µSBS mean values ranged from 14.7 MPa (Ambar Universal) to 26.8 MPa (Tetric N-Bond Universal). OptiBond Universal and Tetric N-Bond Universal exhibited the highest resin-enamel bonds. Weibull analysis was performed to better understand the bonding reliability of the tested adhesives, with OptiBond Universal demonstrating increased modulus and characteristic strength, performing as one of the most interesting compositions for enamel bonding. Adhesives based on carboxylic acids were more acidic than adhesives containing phosphate-based monomers (p = 0.033), and the removal of Ca and P elements from enamel was more intense with the use of adhesives based on carboxylic acids (p = 0.029). There was a strong relationship between the Sz values of adhesively treated enamel and the baseline contact angle (wettability) of adhesives (R² = -0.771; p = 0.025). The study highlights significant variability among universal adhesives in terms of their interaction with enamel. Adhesives based on GPDM (OptiBond Universal) and 10-MDP (Tetric N-Bond Universal) demonstrated the best bonding performance, suggesting they may be preferable in clinical settings where strong and reliable enamel bonds are critical. The presence of 10-MDP was not significantly associated with improved dental bonds. The present findings provide valuable information for dentists in selecting adhesives that optimize clinical outcomes in enamel substrate using the self-etching adhesive approach.

Effects of 5 Nanosilica Concentrations and Humid Environments at 6 Different pH Levels on Fracture Toughness and Moisture Absorption of Dental Polymethyl Methacrylate Resin‎ Reinforced With Silica Nanoparticles: An Explorative Experimental Scanning Electron Microscopy Study.

No study has assessed the effects of nanosilica within polymethyl methacrylate (PMMA) resin and environmental pH on resin's fracture resistance and moisture absorption. A total of 90 specimens were divided into 30 subgroups of three, as per the ASTM D5045 standard: five groups of nanosilica percentages (0%/2%/5%/7%/10%), each ‎divided into six subgroups of pH levels (pH = 5/6/7/8/9, + "dry" control). The specimens were prepared by mixing silica nanoparticles with PMMA powder in a vacuum mixer. Then, the specimens were mixed with a diluent liquid (TEGDMA) according to the manufacturer's instructions. For each of the five weight percentages, 36 samples were produced. The 18 specimens in each group were randomly divided into six subgroups of pH levels. The specimens were kept in containers of liquid at different pH levels at room temperature for 1 week. Their before- and after-storage weights were recorded to calculate moisture absorption. The fracture resistance test was performed (ASTM D5045 standard) using the three-point bending method. Scanning electron microscopy was performed. Data were analyzed. Both nanosilica extents and pH levels significantly affected the fracture toughness with a significant interaction (p < ‎‎0.00001). All post hoc comparisons of different pH levels (except pH= 5 vs. 6) were significant (p < ‎‎0.0001). All post hoc comparisons of different nanosilica concentrations were significant (p < ‎‎0.0001). Both nanosilica extents and pH levels significantly influenced the fracture toughness with a significant interaction (p < ‎‎0.00001). All post hoc comparisons of different pH levels and also between different nanosilica concentrations were significant (p < ‎‎0.0001). The correlation between moisture absorption and fracture toughness was significant (R = -0.382, p = 0.0009). Fracture toughness decreases when placed in humid and acidic environments. Also, the samples that were placed in a humid environment suffered a brittle fracture. Increasing silica nanoparticles improved fracture toughness (becoming optimal at 5 wt% nanosilica). The objective of this study was to investigate the fracture toughness of dental samples made of PMMA reinforced with various percentages of nanosilica at various pH levels. For this purpose, dental resins with various amounts of nanosilica were placed in moist media at different pH values ranging from 5 to 9 (mimicking the normal pH range of the human mouth).

Xpert MTB/XDR assay: rapid TB drug resistance detection.

To assess the Xpert MTB/XDR assay's efficiency in promptly detecting resistance to isoniazid, fluoroquinolones, ethionamide, and second-line injectable drugs among tuberculosis (TB) patients. From August 2020 to July 2021, TB suspected patient samples were enrolled at a tertiary care center for our study. We conducted mycobacterial culture, phenotypic DST using proportion method in liquid culture at WHO-recommended concentrations, and the line probe assay (LPA). Simultaneously, the Index test, Xpert MTB/XDR, was performed following the manufacturer's instructions. Among 360 samples, 107 were excluded due to incomplete information. Resistance to isoniazid, levofloxacin and moxifloxacin was found in 45/251, 21/251 and 20/251 samples, respectively by phenotypic DST. The diagnostic accuracy of Index test, taking phenotypic DST as a reference standard, was 95.8%, 99.04%, and 99.05% for isoniazid, levofloxacin, and moxifloxacin, respectively. The Index test assay demonstrated a specificity of 99.1% for detecting SLID resistance, yielding a diagnostic accuracy of 99.2. Comparing the Index test with LPA revealed a significant enhancement in sensitivity for detecting isoniazid resistance (86.7% vs. 82.2%). The Index test exhibited promising outcomes in identifying resistance to isoniazid and fluoroquinolones, surpassing the performance of the LPA. This could be valuable for promptly initiating treatment in cases of drug-resistant tuberculosis.

Inflammatory protein levels in asthmatic bronchitis: A study in the Duhok population, Iraq.

This study aims to determine the levels of TNF-α, IGF-1, IL-6, and IL-10 protein in blood samples and their potential link to bronchitis-asthma diseases in the Iraq Duhok population, highlighting the prevalence of these long-term inflammatory diseases. Sixty blood samples were used and separated into patients (n = 43) and control (n = 17) groups. Serum samples were separated for each individual. Elisa method was used in terms of 4 different proteins investigated in blood samples with the manufacturer's instruction brand kits. This study evaluated TNF-α, IGF-1, IL-6, and IL-10 protein levels in blood samples from asthmatic bronchitis patients in Duhok. Although these levels were elevated compared to controls, the differences were not statistically significant. The differences thought to be related to the bronchitis-asthma diseases could not be demonstrated between the patient and control groups in Iraq Duhok population. Future research should explore larger sample sizes and stratified patient groups to identify potential biomarkers.