Preparation of a membrane-sealed cell for studying catalyst nanoparticles in flowing gas with high vacuum x-ray photoelectron spectrometer.

Here a sealing-style x-ray photoelectron spectroscopy study of the surface of a 1.0 wt. %Ni/TiO2 nanoparticle catalyst in a flowing mixture of CO and O2 at 1 bar was performed with a graphene membrane-sealed Si3N4 window-based miniature cell. We report the details on how a commercial Si3N4 window is modified before assembling a graphene membrane, how single-layer graphene membranes are transferred from their metal supports to the modified Si3N4 window, how a modified Si3N4 window covered with a double-layer graphene membrane is assembled onto a blank cell cap, how a nanoparticle catalyst is introduced to the cell cap and then the cell cap is installed onto a cell body to form a complete reaction cell, and how a complete cell is interfaced with a high vacuum chamber of an XPS system before an XPS study of 1.0 wt. %Ni/TiO2 catalyst surface in a flowing mixture for 0.2 bar CO and 0.8 bar O2 is performed. How the characterization of a catalyst using this type of graphene membrane-sealed Si3N4 window-based miniature cell is relevant to the finding of the actual surface chemistry of a catalyst during catalysis is discussed.

Genomic characterization of Aeromonas spp. isolates from striped catfish with motile Aeromonas septicemia and human bloodstream infections in Vietnam.

Aeromonas spp. are commonly found in the aquatic environment and have been responsible for motile Aeromonas septicemia (MAS) in striped catfish, resulting in significant economic loss. These organisms also cause a range of opportunistic infections in humans with compromised immune systems. Here, we conducted a genomic investigation of 87 Aeromonas isolates derived from diseased catfish, healthy catfish and environmental water in catfish farms affected by MAS outbreaks in eight provinces in Mekong Delta (years: 2012-2022), together with 25 isolates from humans with bloodstream infections (years: 2010-2020). Genomics-based typing method precisely delineated Aeromonas species while traditional methods such as aerA PCR and MALDI-TOF were unable identify A. dhakensis. A. dhakensis was found to be more prevalent than A. hydrophila in both diseased catfish and human infections. A. dhakensis sequence type (ST) 656 followed by A. hydrophila ST251 were the predominant virulent species-lineages in diseased catfish (43.7 and 20.7 %, respectively), while diverse STs were found in humans with bloodstream infections. There was evidence of widespread transmission of ST656 and ST251 on striped catfish in the Mekong Delta region. ST656 and ST251 isolates carried a significantly higher number of acquired antimicrobial resistance (AMR) genes and virulence factors in comparison to other STs. They, however, exhibited several distinctions in key virulence factors (i.e. lack of type IV pili and enterotoxin ast in A. dhakensis), AMR genes (i.e. presence of imiH carbapenemase in A. dhakensis), and accessory gene content. To uncover potential conserved proteins of Aeromonas spp. for vaccine development, pangenome analysis has unveiled 2202 core genes between ST656 and ST251, of which 78 proteins were in either outer membrane or extracellular proteins. Our study represents one of the first genomic investigations of the species distribution, genetic landscape, and epidemiology of Aeromonas in diseased catfish and human infections in Vietnam. The emergence of antimicrobial resistant and virulent A. dhakensis strains underscores the needs of enhanced genomic surveillance and strengthening vaccine research and development in preventing Aeromonas diseases in catfish and humans, and the search for potential vaccine candidates could focus on Aeromonas core genes encoded for membrane and secreted proteins.

Synthesis of magnetic MFe2O4 (M = Ni, Co, Zn, Fe) supported on porous carbons derived from Bidens pilosa weed and their adsorptive comparison of toxic dyes.

Bidens pilosa is classified as an invasive plant and has become a problematic weed to many agricultural crops. This species strongly germinates, grows and reproduces and competing for nutrients with local plants. To lessen the influence of Bidens pilosa, therefore, converting this harmful species into carbon materials as adsorbents in harm-to-wealth and valorization strategies is required. Here, we synthesized a series of magnetic composites based on MFe2O4 (M = Ni, Co, Zn, Fe) supported on porous carbon (MFOAC) derived from Bidens pilosa by a facile hydrothermal method. The Bidens pilosa carbon was initially activated by condensed H3PO4 to increase the surface chemistry. We observed that porous carbon loaded NiFe2O4 (NFOAC) reached the highest surface area (795.7 m2 g-1), followed by CoFe2O4/AC (449.1 m2 g-1), Fe3O4/AC (426.1 m2 g-1), ZnFe2O4/AC (409.5 m2 g-1). Morphological results showed nanoparticles were well-dispersed on the surface of carbon. RhB, MO, and MR dyes were used as adsorbate to test the adsorption by MFOAC. Effect of time (0-360 min), concentration (5-50 mg L-1), dosage (0.05-0.2 g L-1), and pH (3-9) on dyes adsorption onto MFOAC was investigated. It was found that NFOAC obtained the highest maximum adsorption capacity against dyes, RhB (107.96 mg g-1) < MO (148.05 mg g-1) < MR (153.1 mg g-1). Several mechanisms such as H bonding, π-π stacking, cation-π interaction, and electrostatic interaction were suggested. With sufficient stability and capacity, NFOAC can be used as potential adsorbent for real water treatment systems.

Optimization of hydrothermal synthesis conditions of Bidens pilosa-derived NiFe2O4@AC for dye adsorption using response surface methodology and Box-Behnken design.

The presence of stable and hazardous organic dyes in industrial effluents poses significant risks to both public health and the environment. Activated carbons and biochars are widely used adsorbents for removal of these pollutants, but they often have several disadvantages such as poor recoverability and inseparability from water in the post-adsorption process. Incorporating a magnetic component into activated carbons can address these drawbacks. This study aims to optimizing the production of NiFe2O4-loaded activated carbon (NiFe2O4@AC) derived from a Bidens pilosa biomass source through a hydrothermal method for the adsorption of Rhodamine B (RhB), methyl orange (MO), and methyl red (MR) dyes. Response surface methodology (RSM) and Box-Behnken design (BBD) were applied to analyze the key synthesis factors such as NiFe2O4 loading percentage (10-50%), hydrothermal temperature (120-180 °C), and reaction time (6-18 h). The optimized condition was found at a NiFe2O4 loading of 19.93%, a temperature of 135.55 °C, and a reaction time of 16.54 h. The optimum NiFe2O4@AC demonstrated excellent sorption efficiencies of higher than 92.98-97.10% against all three dyes. This adsorbent was characterized, exhibiting a well-developed porous structure with a high surface area of 973.5 m2 g-1. Kinetic and isotherm were studied with the best fit of pseudo-second-order, and Freundlich or Temkin. Qmax values were determined to be 204.07, 266.16, and 177.70 mg g-1 for RhB, MO, and MR, respectively. By selecting HCl as an elution, NiFe2O4@AC could be efficiently reused for at least 4 cycles. Thus, the Bidens pilosa-derived NiFe2O4@AC can be a promising material for effective and recyclable removal of dye pollutants from wastewater.

Dysregulation of the WNK4-SPAK/OSR1 pathway has a minor effect on baseline NKCC2 phosphorylation.

The with-no-lysine kinase 4 (WNK4)-sterile 20/SPS-1-related proline/alanine-rich kinase (SPAK)/oxidative stress-responsive kinase 1 (OSR1) pathway mediates activating phosphorylation of the furosemide-sensitive Na+-K+-2Cl- cotransporter (NKCC2) and the thiazide-sensitive NaCl cotransporter (NCC). The commonly used pT96/pT101-pNKCC2 antibody cross-reacts with pT53-NCC in mice on the C57BL/6 background due to a five amino acid deletion. We generated a new C57BL/6-specific pNKCC2 antibody (anti-pT96-NKCC2) and tested the hypothesis that the WNK4-SPAK/OSR1 pathway strongly regulates the phosphorylation of NCC but not NKCC2. In C57BL/6 mice, anti-pT96-NKCC2 detected pNKCC2 and did not cross-react with NCC. Abundances of pT96-NKCC2 and pT53-NCC were evaluated in Wnk4-/-, Osr1-/-, Spak-/-, and Osr1-/-/Spak-/- mice and in several models of the disease familial hyperkalemic hypertension (FHHt) in which the CUL3-KLHL3 ubiquitin ligase complex that promotes WNK4 degradation is dysregulated (Cul3+/-/Δ9, Klhl3-/-, and Klhl3R528H/R528H). All mice were on the C57BL/6 background. In Wnk4-/- mice, pT53-NCC was almost absent but pT96-NKCC2 was only slightly lower. pT53-NCC was almost absent in Spak-/- and Osr1-/-/Spak-/- mice, but pT96-NKCC2 abundance did not differ from controls. pT96-NKCC2/total NKCC2 was slightly lower in Osr1-/- and Osr1-/-/Spak-/- mice. WNK4 expression colocalized not only with NCC but also with NKCC2 in Klhl3-/- mice, but pT96-NKCC2 abundance was unchanged. Consistent with this, furosemide-induced urinary Na+ excretion following thiazide treatment was similar between Klhl3-/- and controls. pT96-NKCC2 abundance was also unchanged in the other FHHt mouse models. Our data show that disruption of the WNK4-SPAK/OSR1 pathway only mildly affects NKCC2 phosphorylation, suggesting a role for other kinases in NKCC2 activation. In FHHt models NKCC2 phosphorylation is unchanged despite higher WNK4 abundance, explaining the thiazide sensitivity of FHHt.NEW & NOTEWORTHY The renal cation cotransporters NCC and NKCC2 are activated following phosphorylation mediated by the WNK4-SPAK/OSR1 pathway. While disruption of this pathway strongly affects NCC activity, effects on NKCC2 activity are unclear since the commonly used phospho-NKCC2 antibody was recently reported to cross-react with phospho-NCC in mice on the C57BL/6 background. Using a new phospho-NKCC2 antibody specific for C57BL/6, we show that inhibition or activation of the WNK4-SPAK/OSR1 pathway in mice only mildly affects NKCC2 phosphorylation.

Pelvic arteriovenous malformation mimicking benign prostatic hyperplasia: A case report.

Pelvic arteriovenous malformation is a rare vascular abnormality, espescially in male patients, and is difficult to treat because of its nature supplied by multiple arterial feeders. We report a 70-year-old male patient admitted due to symptoms of benign prostatic hyperplasia. Ultrasound was performed initially, and no other abnormalities were found other than an enlarged prostate. CT scan later demonstrated a pelvic arteriovenous malformation adjacent to the prostate, with multiple arterial feeders from the right internal iliac artery. Angiography confirmed the diagnosis, and transaterial embolization was successfully done. The symptoms disappeared several days later, and the patient remained asymptompmatic during follow-up.

The exit of nanoparticles from solid tumours.

Nanoparticles enter tumours through endothelial cells, gaps or other mechanisms, but how they exit is unclear. The current paradigm states that collapsed tumour lymphatic vessels impair the exit of nanoparticles and lead to enhanced retention. Here we show that nanoparticles exit the tumour through the lymphatic vessels within or surrounding the tumour. The dominant lymphatic exit mechanism depends on the nanoparticle size. Nanoparticles that exit the tumour through the lymphatics are returned to the blood system, allowing them to recirculate and interact with the tumour in another pass. Our results enable us to define a mechanism of nanoparticle delivery to solid tumours alternative to the enhanced permeability and retention effect. We call this mechanism the active transport and retention principle. This delivery principle provides a new framework to engineer nanomedicines for cancer treatment and detection.

A new snake of the genus Dendrelaphis Boulenger, 1890 (Squamata: Colubridae) from the coastal area of southern Vietnam.

We describe a new species of Dendrelaphis from the coastal area in southern Vietnam based on morphological data and nucleotide sequences from one partial mitochondrial gene (Cytb). Dendrelaphis binhi sp. nov. is diagnosed by the following morphological characters: body scale rows 13 at neck and midbody, 9-11 before vent; vertebral scale row feebly enlarged; ventrals 154-158 in males and 161-170 in females; subcaudals 95-106 in males and 95-102 in females; one supralabial touching the eye; 18 or 19 maxillary teeth; hemipenis spinose, reaching 10th or 11th SC, with a slender papilla; cloacal plate divided; dorsum dark brown; and a white stripe along the lower flank. The new species differs from its congeners by an uncorrected p-distance in Cytb sequences of at least 8.1%.

Toward Predicting Nanoparticle Distribution in Heterogeneous Tumor Tissues.

Nanobio interaction studies have generated a significant amount of data. An important next step is to organize the data and design computational techniques to analyze the nanobio interactions. Here we developed a computational technique to correlate the nanoparticle spatial distribution within heterogeneous solid tumors. This approach led to greater than 88% predictive accuracy of nanoparticle location within a tumor tissue. This proof-of-concept study shows that tumor heterogeneity might be defined computationally by the patterns of biological structures within the tissue, enabling the identification of tumor patterns for nanoparticle accumulation.

High production of poly(3-hydroxybutyrate) in Escherichia coli using crude glycerol.

Poly(3-hydroxybutyrate) (PHB) is a prominent bio-plastic and recognized as the potential replacement of petroleum-derived plastics. To make PHB cost-effective, the production scheme based on crude glycerol was developed using Escherichia coli. The heterogeneous synthesis pathway of PHB was introduced into the E. coli strain capable of efficiently utilizing glycerol. The central metabolism that links to the synthesis of acetyl-CoA and NADPH was further reprogrammed to improve the PHB production. Key genes were targeted for manipulation, involving those in glycolysis, the pentose phosphate pathway, and the tricarboxylic cycle. As a result, the engineered strain gained a 22-fold increase in the PHB titer. Finally, the fed-batch fermentation was conducted with the producer strain to give the PHB titer, content, and productivity reaching 36.3 ± 3.0 g/L, 66.5 ± 2.8%, and 1.2 ± 0.1 g/L/h, respectively. The PHB yield on crude glycerol accounts for 0.3 g/g. The result indicates that the technology platform as developed is promising for the production of bio-plastics.

Pan-cancer whole-genome comparison of primary and metastatic solid tumours.

Metastatic cancer remains an almost inevitably lethal disease1-3. A better understanding of disease progression and response to therapies therefore remains of utmost importance. Here we characterize the genomic differences between early-stage untreated primary tumours and late-stage treated metastatic tumours using a harmonized pan-cancer analysis (or reanalysis) of two unpaired primary4 and metastatic5 cohorts of 7,108 whole-genome-sequenced tumours. Metastatic tumours in general have a lower intratumour heterogeneity and a conserved karyotype, displaying only a modest increase in mutations, although frequencies of structural variants are elevated overall. Furthermore, highly variable tumour-specific contributions of mutational footprints of endogenous (for example, SBS1 and APOBEC) and exogenous mutational processes (for example, platinum treatment) are present. The majority of cancer types had either moderate genomic differences (for example, lung adenocarcinoma) or highly consistent genomic portraits (for example, ovarian serous carcinoma) when comparing early-stage and late-stage disease. Breast, prostate, thyroid and kidney renal clear cell carcinomas and pancreatic neuroendocrine tumours are clear exceptions to the rule, displaying an extensive transformation of their genomic landscape in advanced stages. Exposure to treatment further scars the tumour genome and introduces an evolutionary bottleneck that selects for known therapy-resistant drivers in approximately half of treated patients. Our data showcase the potential of pan-cancer whole-genome analysis to identify distinctive features of late-stage tumours and provide a valuable resource to further investigate the biological basis of cancer and resistance to therapies.

In situ and operando study of catalysts during high-temperature high-pressure catalysis in a fixed-bed plug flow reactor with x-ray absorption spectroscopy.

Numerous important catalytic reactions, such as Fischer-Tropsch synthesis (FTS), are performed under harsh conditions in terms of high temperature of a catalyst in a mixture of reactants at a high pressure. There has been a lack of an intrinsic correlation between a catalytic performance and its corresponding catalyst structure due to the unavailable information on the authentic structure of the catalyst during catalysis under a high-temperature high-pressure (HTHP) condition. Here, we report in situ/operando studies of Co catalysts during catalysis under HTHP conditions using x-ray absorption spectroscopy (XAS). A high-temperature high-pressure catalysis-XAS (HTHP Catalysis-XAS) system using a thin, small quartz or beryllium tube as the reactor was built for in situ/operando characterization of high-energy absorption edges of 4d transition metals or low-energy absorption edges of 3d/4d transition metals under high-temperature high-pressure conditions, respectively. This reactor can be used for HTHP catalysis performed at a temperature of up to 550 °C and a gas pressure of up to 60 bars for uncovering the chemical states and coordination environments of metal atoms of these catalysts during HTHP catalysis. The capability of collecting XAS data during HTHP catalysis was confirmed through tests at 400oC in the mixture of 20 bar mixture of reactants at beamline endstation. The operando studies of Ru catalyst particles under Fischer-Tropsch catalytic conditions with extended x-ray absorption fine structure spectroscopy revealed a restructuring of the Ru catalyst at 250 °C in the mixture of 6 bars CO and 12 bars H2 during FTS (30 ml/min), which was not observed at 300 °C in 1 bar H2 (20 ml/min). This observation suggests new chemistry for metal catalysts under HTHP condition inaccessible due to a lack of applicable characterizations. These tests confirmed the function of this HTHP Catalysis-XAS system for in situ/operando characterizations of catalysts during HTHP catalysis.

Insights into the synthesis strategies of plant-derived cyclotides.

Cyclotides are plant peptides characterized with a head-to-tail cyclized backbone and three interlocking disulfide bonds, known as a cyclic cysteine knot. Despite the variations in cyclotides peptide sequences, this core structure is conserved, underlying their most useful feature: stability against thermal and chemical breakdown. Cyclotides are the only natural peptides known to date that are orally bioavailable and able to cross cell membranes. Cyclotides also display bioactivities that have been exploited and expanded to develop as potential therapeutic reagents for a wide range of conditions (e.g., HIV, inflammatory conditions, multiple sclerosis, etc.). As such, in vitro production of cyclotides is of the utmost importance since it could assist further research on this peptide class, specifically the structure-activity relationship and its mechanism of action. The information obtained could be utilized to assist drug development and optimization. Here, we discuss several strategies for the synthesis of cyclotides using both chemical and biological routes.

Cysteine-rich peptides: From bioactivity to bioinsecticide applications.

Greater levels of insect resistance and constraints on the use of current pesticides have recently led to increased crop losses in agricultural production. Further, the health and environmental impacts of pesticides now restrict their application. Biologics based on peptides are gaining popularity as efficient crop protection agents with low environmental toxicity. Cysteine-rich peptides (whether originated from venoms or plant defense substances) are chemically stable and effective as insecticides in agricultural applications. Cysteine-rich peptides fulfill the stability and efficacy requirements for commercial uses and provide an environmentally benign alternative to small-molecule insecticides. In this article, cysteine-rich insecticidal peptide classes identified from plants and venoms will be highlighted, focusing on their structural stability, bioactivity and production.

A critical review on the bio-mediated green synthesis and multiple applications of magnesium oxide nanoparticles.

Nowadays, advancements in nanotechnology have efficiently solved many global problems, such as environmental pollution, climate change, and infectious diseases. Nano-scaled materials have played a central role in this evolution. Chemical synthesis of nanomaterials, however, required hazardous chemicals, unsafe, eco-unfriendly, and cost-ineffective, calling for green synthesis methods. Here, we review the green synthesis of MgO nanoparticles and their applications in biochemical, environmental remediation, catalysis, and energy production. Green MgO nanoparticles can be safely produced using biomolecules extracted from plants, fungus, bacteria, algae, and lichens. They exhibited fascinating and unique properties in morphology, surface area, particle size, and stabilization. Green MgO nanoparticles served as excellent antimicrobial agents, adsorbents, colorimetric sensors, and had enormous potential in biomedical therapies against cancers, oxidants, diseases, and the sensing detection of dopamine. In addition, green MgO nanoparticles are of great interests in plant pathogens, phytoremediation, plant cell and organ culture, and seed germination in the agricultural sector. This review also highlighted recent advances in using green MgO nanoparticles as nanocatalysts, nano-fertilizers, and nano-pesticides. Thanks to many emerging applications, green MgO nanoparticles can become a promising platform for future studies.

Green synthesis of ZnFe2O4@ZnO nanocomposites using Chrysanthemum spp. floral waste for photocatalytic dye degradation.

The occurrence of textile dyeing wastewater discharged into the environment has been recently increasing, resulting in harmful effects on living organisms and human health. The use of green nanoparticles for water decontamination has received much attention. Floral waste can be extracted with the release of natural compounds, which act as reducing and stabilizing agents during the biosynthesis of nanoparticles. Herein, we report the utilization of Chrysanthemum spp. floral waste extract to synthesize green ZnFe2O4@ZnO (ZFOZx) nanocomposites for the photocatalytic degradation of Congo red under solar light irradiation. The various molar ratio of ZnFe2O4 (0-50%) was incorporated into ZnO nanoparticles. The surface area of green ZFOZx nanocomposites was found to increase (7.41-42.66 m2 g-1) while their band gap energy decreased from 1.98 eV to 1.92 eV. Moreover, the results exhibited the highest Congo red dye degradation efficiency of 94.85% at a concentration of 5.0 mg L-1, and a catalyst dosage of 0.33 g L-1. The •O2- reactive species played a vital role in the photocatalytic degradation of Congo red dye. Green ZFOZ3 nanocomposites had good recyclability with at least three cycles, and an excellent stability. The germination results showed that wastewater treated by ZFOZ3 was safe enough for bean seed germination. We expect that this work contributes significantly to developing novel green bio-based nanomaterials for environmental remediation as well as reducing the harm caused by flower wastes.

Atomic-Scale Mechanism of Platinum Catalyst Restructuring under a Pressure of Reactant Gas.

Heterogeneous catalysis is key for chemical transformations. Understanding how catalysts' active sites dynamically evolve at the atomic scale under reaction conditions is a prerequisite for accurately determining catalytic mechanisms and predictably developing catalysts. We combine in situ time-dependent scanning tunneling microscopy observations and machine-learning-accelerated first-principles atomistic simulations to uncover the mechanism of restructuring of Pt catalysts under a pressure of carbon monoxide (CO). We show that a high CO coverage at a Pt step edge triggers the formation of atomic protrusions of low-coordination Pt atoms, which then detach from the step edge to create sub-nano-islands on the terraces, where under-coordinated sites are stabilized by the CO adsorbates. The fast and accurate machine-learning potential is key to enabling the exploration of tens of thousands of configurations for the CO-covered restructuring catalyst. These studies open an avenue to achieve an atomic-scale understanding of the structural dynamics of more complex metal nanoparticle catalysts under reaction conditions.

A new species of the genus Rhabdophis Fitzinger, 1843 (Squamata: Colubridae) from the Hoang Lien range, northwest Vietnam.

We describe a new species of the Natricinae genus Rhabdophis Fitzinger, 1843 from the Hoang Lien range, northwest Vietnam. The new species is distinct from all congeneric species on the grounds of morphometric and molecular data. The new species is most similar to Rhabdophis leonardi in terms of morphology but can be distinguished from it based on differences in maxillary tooth count, scalation, and genetic data. A sequence on GenBank from a Rhabdophis specimen collected in Honghe, Yunnan, China was identical to the species we describe, and it is likely that the new species is not restricted to Vietnam. As a priority, future work should focus on determining the distribution of this species, as well as understanding population and life history traits such as reproductive rate.

Amplification of olfactory signals by Anoctamin 9 is important for mammalian olfaction.

Sensing smells of foods, prey, or predators determines animal survival. Olfactory sensory neurons in the olfactory epithelium (OE) detect odorants, where cAMP and Ca2+ play a significant role in transducing odorant inputs to electrical activity. Here we show Anoctamin 9, a cation channel activated by cAMP/PKA pathway, is expressed in the OE and amplifies olfactory signals. Ano9-deficient mice had reduced olfactory behavioral sensitivity, electro-olfactogram signals, and neural activity in the olfactory bulb. In line with the difference in olfaction between birds and other vertebrates, chick ANO9 failed to respond to odorants, whereas chick CNGA2, a major transduction channel, showed greater responses to cAMP. Thus, we concluded that the signal amplification by ANO9 is important for mammalian olfactory transduction.

NOR-1 (NR4A3) immunostaining on cytologic preparations for the preoperative diagnosis of acinic cell carcinoma of the salivary gland.

INTRODUCTION: Acinic cell carcinoma of the salivary gland (ACC-SG) is characterized by recurrent rearrangements in the nuclear receptor subfamily 4 group A member 3 (NR4A3). Immunostaining using an antibody targeting this rearrangement, neuron-derived orphan receptor 1 (NOR-1), has been recently studied on surgical specimens and cell block material of fine-needle aspirates for the diagnosis of ACC-SG. Our goal was to evaluate whether NOR-1 immunostaining could reliably be performed on destained cytologic preparations. MATERIALS AND METHODS: This was a retrospective multi-institutional study. Immunostaining with the NOR-1 antibody (sc-393902 [H-7], Santa Cruz Biotechnology Inc.) was performed at a titer of 1:30 on destained cytologic preparations. ACC-SG cases (n = 17) were represented by twelve cases with alcohol-fixed preparations (n = 12), including direct smears and SurePath preparations, as well as 5 cases with air-dried preparations (n = 5). These were compared to 27 mimicker lesions (n = 27): normal acini (4), chronic sialadenitis (3), oncocytoma (2), pleomorphic adenoma (6), Warthin tumor (8), mucoepidermoid carcinoma (1), secretory carcinoma (2), and salivary duct carcinoma (1). RESULTS: The positivity of NOR-1 in ACC-SG cases was 100% on destained alcohol-fixed preparations (12/12) and 60% on air-dried preparations (3/5). All 27 mimicker lesions were negative for NOR-1 (0/27). Evaluation of 2 ACC-SG cases with both types of cytologic preparations showed that NOR-1 was positive on the alcohol-fixed slides but negative on the air-dried slides. CONCLUSIONS: NOR-1 immunostaining can reliably be performed on alcohol-fixed direct smears and liquid-based preparations for the diagnosis of ACC-SG. Air-dried preparations show a lower positivity rate and may be less suitable for diagnostic immunostaining.