Application of spatial transcriptomics analysis using the Visium system for the mouse nasal cavity after intranasal vaccination.

Intranasal vaccines that elicit mucosal immunity are deemed effective against respiratory tract infections such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but their ability to induce humoral immunity characterized by immunoglobulin A (IgA) and IgG production is low. It has been reported that vaccination with a mixture of a viscous base carboxyvinyl polymer (CVP) and viral antigens induced robust systemic and mucosal immune responses. In this study, we analyzed the behavior of immunocompetent cells in the nasal cavity over time by spatial transcriptome profiling induced immediately after antigen vaccination using CVP. We established a method for performing spatial transcriptomics using the Visium system in the mouse nasal cavity and analyzed gene expression profiles within the nasal cavity after intranasal vaccination. Glycoprotein 2 (Gp2)-, SRY-box transcription factor 8 (Sox8)-, or Spi-B transcription factor (Spib)-expressing cells were increased in the nasal passage (NP) region at 3-6 hr after SARS-CoV-2 spike protein and CVP (S-CVP) vaccination. The results suggested that microfold (M) cells are activated within a short period of time (3-6 hr). Subsequent cluster analysis of cells in the nasal cavity showed an increase in Cluster 9 at 3-6 hr after intranasal vaccination with the S-CVP. We found that Il6 in Cluster 9 had the highest log2 fold values within the NP at 3-6 hr. A search for gene expression patterns similar to that of Il6 revealed that the log2 fold values of Edn2, Ccl20, and Hk2 also increased in the nasal cavity after 3-6 hr. The results showed that the early response of immune cells occurred immediately after intranasal vaccination. In this study, we identified changes in gene expression that contribute to the activation of M cells and immunocompetent cells after intranasal vaccination of mice with antigen-CVP using a time-series analysis of spatial transcriptomics data. The results facilitated the identification of the cell types that are activated during the initial induction of nasal mucosal immunity.

Presence of Staphylococcus spp. carriers of the mecA gene in the nasal cavity of piglets in the nursery phase.

The presence of Staphylococcus spp. resistant to methicillin in the nasal cavity of swine has been previously reported. Considering the possible occurrence of bacterial resistance and presence of resistance genes in intensive swine breeding and the known transmissibility and dispersion potential of such genes, this study aimed to investigate the prevalence of resistance to different antibiotics and the presence of the mecA resistance gene in Staphylococcus spp. from piglets recently housed in a nursery. For this, 60 nasal swabs were collected from piglets at the time of their housing in the nursery, and then Staphylococcus spp. were isolated and identified in coagulase-positive (CoPS) and coagulase-negative (CoNS) isolates. These isolates were subjected to the disk-diffusion test to evaluate the bacterial resistance profile and then subjected to molecular identification of Staphylococcus aureus and analyses of the mecA gene through polymerase chain reaction. Of the 60 samples collected, 60 Staphylococcus spp. were isolated, of which 38 (63.33%) were classified as CoNS and 22 (36.67%) as CoPS. Of these, ten (45.45%) were identified as Staphylococcus aureus. The resistance profile of these isolates showed high resistance to different antibiotics, with 100% of the isolates resistant to chloramphenicol, clindamycin, and erythromycin, 98.33% resistant to doxycycline, 95% resistant to oxacillin, and 85% resistant to cefoxitin. Regarding the mecA gene, 27 (45%) samples were positive for the presence of this gene, and three (11.11%) were phenotypically sensitive to oxacillin and cefoxitin. This finding highlights the importance of researching the phenotypic profile of resistance to different antimicrobials and resistance genes in the different phases of pig rearing to identify the real risk of these isolates from a One Health perspective. The present study revealed the presence of samples resistant to different antibiotics in recently weaned production animal that had not been markedly exposed to antimicrobials as growth promoters or even as prophylactics. This information highlights the need for more research on the possible sharing of bacteria between sows and piglets, the environmental pressure within production environments, and the exposure of handlers during their transport, especially considering the community, hospital, and political importance of the presence of circulating resistant strains.

SARS-CoV-2 requires acidic pH to infect cells.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cell entry starts with membrane attachment and ends with spike (S) protein-catalyzed membrane fusion depending on two cleavage steps, namely, one usually by furin in producing cells and the second by TMPRSS2 on target cells. Endosomal cathepsins can carry out both. Using real-time three-dimensional single-virion tracking, we show that fusion and genome penetration require virion exposure to an acidic milieu of pH 6.2 to 6.8, even when furin and TMPRSS2 cleavages have occurred. We detect the sequential steps of S1-fragment dissociation, fusion, and content release from the cell surface in TMPRRS2-overexpressing cells only when exposed to acidic pH. We define a key role of an acidic environment for successful infection, found in endosomal compartments and at the surface of TMPRSS2-expressing cells in the acidic milieu of the nasal cavity.

The prognostic value of S-100 protein and Ki-67 index in olfactory neuroblastoma.

OBJECTIVE: To evaluate the prognostic value of S-100 protein and Ki-67 labeling index in olfactory neuroblastomas. METHODS: A retrospective study was conducted on a cohort of 85 patients with olfactory neuroblastomas. The immunohistochemical expression of S-100 and Ki-67 was assessed, and the predictive value of S-100 and Ki-67 was further evaluated. The optimal cutoff value of Ki-67 labeling index was determined using time-dependent receiver operating characteristic curve analysis. Overall survival and progression-free survival were assessed using the Kaplan-Meier method. RESULTS: A cut-off Ki-67 labeling index value of 67.5% was determined for prognosis in patients with olfactory neuroblastomas. There was a significant correlation between Ki-67 expression and cervical lymph node metastasis (P = 0.049). Compared with S-100 (+), S-100 (-) was associated with a higher rate of lymph node metastasis and a higher level of Ki-67 (P = 0.007, < 0.001, respectively), as well as an advanced Kadish stage (P = 0.037). Survival analyses showed that patients with S-100 (+) had better 5-year overall survival than those with S-100 (-) (P = 0.028), and patients with both S-100 (+) and Ki-67 (<67.5%) had superior 5-year overall survival compared with all the other patients (P = 0.0225). CONCLUSION: Our findings suggest that S-100 combined with Ki-67 labeling index are reliable prognostic factors in patients with olfactory neuroblastomas.

Physical Obstruction of Nasal Cavities With Subsequent Asphyxia, Causes Lethality of Rats in an Acute Inhalation Study With Hydrophobic HMDZ Surface-Treated Synthetic Amorphous Silica (SAS).

The aim of the present study was to understand the mechanism of lethality associated with high dose inhalation of a low-density hydrophobic surface-treated SAS observed in some acute inhalation studies. It was demonstrated that physical obstruction of the upper respiratory tract (nasal cavities) caused the effects observed. Hydrophobic surface-treated SAS was inhaled (flow-past, nose-only) by six Wistar rats (three males and three females) in an acute toxicity study at a concentration of ~500 mg/m3 for an intended 4-hr exposure. Under the conditions of the test set-up, the concentration applied was found to be the highest that can be delivered to the test animal port without significant alteration of the aerosol size distribution over time. None of the test- material-exposed animals survived the planned observation time of 4 h; three animals died between 2 34 h after starting exposure and cessation of exposure at 3 14 h, two died after transfer to their cages and the remaining animal was sacrificed due to its poor condition and welfare considerations. Histology accomplished by energy dispersive X-ray (EDX) analysis demonstrated that test material particles agglomerated and formed a gel-like substrate that ultimately blocked the upper respiratory airways, which proved fatal for the rat as an obligatory nose breather. This observation is in line with the findings reported by Hofmann et al. showing a correlation between lethality and hydrophobicity determined by contact angle measurement. The aerosol characterizations associated with this study are provided in detail by Wessely et al.

Epithelial morphometric alterations and mucosecretory responses in the nasal cavity of mice chronically exposed to hydrothermal emissions.

Air pollutants (either of natural or anthropogenic origin) represent a considerable environmental risk to human health by affecting the respiratory system and causing respiratory disorders. In this study, we investigate the effects of chronic exposure to hydrothermal emissions on the nasal cavity of mice since it is the first and the most exposed region of the respiratory system. This study, carried in S. Miguel Island, Azores-Portugal, used Mus musculus as a bioindicator species. Mice were captured in an area with non-eruptive active volcanism (Furnas Village) and another area without volcanism (Rabo de Peixe, reference site). The hydrothermal emissions present at Furnas Village are characterized by the continuous release of several gases (CO2, H2S, 222Rn) along with metals (e.g. Hg, Cd, Zn, Al) and particulate matter into the environment. We test the hypothesis whether chronic exposure to this specific type of pollution causes epithelial morphometric, mucosecretory and neuronal alterations on the nasal cavity. Thickness measurements were taken in the squamous, respiratory and olfactory epithelia. The relative density of cell types (basal, support and neurons) was also assessed in the olfactory epithelium and the mucosecretory activity was determined in the lateral nasal glands, Bowman's gland and goblet cells. Mice chronically exposed to hydrothermal emissions presented thinner olfactory epithelia and lesser mucous production, which could result in loss of olfactory capabilities as well as a decrease in the protective function provided by the mucous to the lower respiratory tract. For the first time, it is demonstrated that, in mice, this specific type of non-eruptive active volcanism causes epithelial and mucosecretory alterations, leading to the loss of olfactory capabilities.

From laboratory to clinic: Translation of extracellular vesicle based cancer biomarkers.

The past decade has witnessed a rapid growth in the field of extracellular vesicle (EV) based biomarkers for the diagnosis and monitoring of cancer. Several studies have reported novel EV based biomarkers, but the technical and clinical validation phase has been hampered by general challenges common to biomedical research field as well as specific challenges inherent to the nanoparticle field. This has led to more common failures than success stories in the biomarker discovery pipeline. As a result, more attention must be focused on the process of biomarker discovery, verification, and validation to allow for translation and application of novel EV based research to patient care. Herein, we briefly discuss the hurdles and potential solutions in EV biomarker discovery and verification and validation, and clinical translation.

An improved method of nasal swab analysis for assessing alpha internal radioactive contaminants.

When responding to nuclear and radiological emergencies, rapid and on-site detections of possible internal radioactive contaminants are required for early dose estimation and medical triage. Nasal swab analysis is an effective method to provide valuable information for early and fast estimates of alpha radionuclide inhalation intakes and resultant doses. In this study, to improve the quality of nasal swab measurements, a specialised double-detector alpha counter was designed. Various parameters including swab materials, sample pre-preparation, angle-dependence and vacuum dependence were investigated to optimise the reliability and convenience of the nasal swab method. An improved procedure of direct nasal swab measurement was eventually established, which could be used to obtain early data for initial dose assessment during the first response of nuclear and radiological emergencies.

[Computational investigation of Artemisia pollen deposition in realistic nasal cavities of residents in northwest China].

Objective: To investigate the deposition rate of Artemisia pollen in different nasal cavity regions and its influence factors in residents of northwest China. Methods: Thirty healthy adults from northwest China were enrolled. The computational fluid dynamics (CFD) and discrete phase model (DPM) were used for numerical simulation of nasal structures. The pollen deposition fraction in each anatomical part was counted and the effects of pollen density and breathing rate on deposition were analyzed. SPSS 19.0 software was used for statistical analysis. Results: The hottest deposition parts of Artemisia pollen were nasal septum (30.70%±12.27%), vestibule (27.45%±8.21%), middle turbinate area (13.59%±8.98%) and nasopharynx (7.14%±5.90%). When the inspiratory flow rate increased to 30 L/min, the deposition rates of pollen in nasal vestibule and nasal septum were significantly higher than that at the rate of 15 L/min (43.20%±11.14% vs 27.45%±8.21%, 51.48%±9.77% vs 30.70%±12.27%, t value was -8.126,-5.264, respectively, all P<0.05), which indicated that with the increase of the inspiratory flow rate, the deposition hotspot moved forward. Compared with the wet Artemisia pollen, the deposition rate of the dry pollen in nasal vestibule and nasal septum decreased significantly (16.55%±4.33% vs 27.45%±8.21%, 7.09%±3.69% vs 30.70%±12.27%, t value was 8.669, 9.173, respectively, all P<0.05). The escape rate at outlet increased from 17.00%±9.57% to 43.48%±13.43% (t=-9.282, P<0.05). Conclusions: The deposition of Artemisia pollen in nasal cavity is highly concentrated. The inhalation velocity and the dry-wet degree of pollen are the main determinants of the deposition site.

The pH Value as a Factor Modifying Bacterial Colonization of Sinonasal Mucosa in Healthy Persons.

OBJECTIVE: The aim of the present study is to determine and compare the range of pH value in nasal and sinus cavities in vivo regarding the presence of bacteria colonizing sinonasal mucosa among healthy subjects. METHODS: The nasal pH value measurement using a portable pH meter (Dx-pH System, Restech) and the microbiological culture swab were taken from beneath the middle turbinate and in the sinus cavity in 39 healthy subjects during maxillary bone corrective osteotomy with the Le Fort I technique. RESULTS: The mean pH value (independently of sex, P = .441) in the healthy sinus cavity was statistically higher than in the nasal middle meatus: 7.96 (SD ± 0.29) versus 7.83 (SD ± 0.30) (P = .032). Forty-eight strains of bacteria were cultured from sinus maxillaries cavities-aerobic 36.8%, aerobic and anaerobic 52.6%, anaerobic only 10.5%-and 23 strains from the nasal meatus-aerobic 25%, aerobic and anaerobic 75%. A statistically significant correlation was found between the type and location of 8 microorganisms, especially Propionibacterium acnes, identified only in the sinus cavities. CONCLUSIONS: Differences in the pH value between the middle nasal meatus and the maxillary sinus are characteristic of healthy subjects and could be associated with the diverse bacterial flora. The role of bacteria Propionibacterium acnes seems to be crucial for the pH range and sinus flora in healthy subjects.

Clinicopathologic and Molecular Features of a Series of 41 Biphenotypic Sinonasal Sarcomas Expanding Their Molecular Spectrum.

Biphenotypic sinonasal sarcoma (BSNS) is a locally aggressive tumor occurring in the sinonasal region. It harbors both myogenic and neural differentiation and is characterized by PAX3 rearrangement with MAML3 as the most frequent fusion partner, but the partner of PAX3 remains unidentified in a subset of cases. About 70 cases have been reported so far. In this study, we report a series of 41 cases with clinical, pathologic, and molecular description. Twenty-five (61%) patients were female individuals, and the median age was 49 years. Tumors arose predominantly in the nasal cavity and ethmoidal sinuses. Local recurrences occurred in 8 cases of the 25 (32%). Histologic features were characteristic of BSNS, with 5 cases showing focal rhabdomyoblastic differentiation. Immunohistochemistry showed a constant positivity of S100 protein and PAX3 and negativity of SOX10. MyoD1 was focally positive in 91% of cases, whereas only 20% were positive for myogenin. Molecular analysis showed a PAX3-MAML3 transcript in 37 cases (90%). RNA sequencing was performed in the 4 negative cases for PAX3-MAML3 fusion, and it showed that 1 case harbored a PAX3-FOXO1 fusion, as previously described in the literature, and 2 novel fusions: PAX3-WWTR1 fusion in 2 cases and PAX3-NCOA2 fusion in 1 case. RNA sequencing results were confirmed by fluorescence in situ hybridization, reverse transcription-polymerase chain reaction, and Sanger sequencing. The PAX3-NCOA2-positive case showed focal rhabdomyoblastic differentiation. In conclusion, we report 2 novel fusions (PAX3-WWTR1 and PAX3-NCOA2) in BSNS and show that MyoD1 is more sensitive than myogenin for demonstrating myogenic differentiation in this tumor.

Bovine respiratory syncytial virus-specific IgG-1 in nasal secretions of colostrum-fed neonatal calves.

In order to determine whether nasal secretions of young calves contain passively derived antibodies to bovine respiratory syncytial virus (BRSV) and if there are differences in presence and/or subclass of these antibodies between calves fed different colostrum replacement products, 17 Holstein calves were fed 150 g of IgG in either a sprayed-dried colostrum-based (CR; n = 8) or a plasma-based colostrum replacement product (PR; n = 9) within 6 h of birth. Venous blood and nasal secretions obtained before feeding and at 24 h of age were assayed for total IgG (serum) by radial immunodiffusion and for BRSV-specific total IgG, IgG-1, and IgG-2 by indirect enzyme-linked immunosorbent assay (ELISA). Calves that were fed a CR had higher concentrations of BRSV-specific IgG and IgG-1 in their serum and nasal secretions compared to calves fed product PR; calves fed the PR had higher levels of serum BRSV-specific IgG-2. The only subclass of antibodies detected in nasal secretions was IgG-1. Re-secretion of passive IgG with neutralizing activity, onto the nasal mucosa could contribute to BRSV-associated disease-sparing observed in the laboratory and in the field. Use of PR will result in lower nasal antibodies since IgG-2 is not re-secreted.

IgG-1 spécifique au virus respiratoire syncytial bovin dans les sécrétions nasales des veaux néonataux nourris au colostrum. Afin de déterminer si les sécrétions nasales des jeunes veaux contenaient des anticorps dérivés passivement envers le virus respiratoire syncytial bovin (VRS) et s'il y a des différences dans la présence et/ou la sous-catégorie de ces anticorps entre les veaux nourris avec différents produits de remplacement du colostrum, 17 veaux Holstein ont été nourris avec 150 g d'IgG soit sous forme de produit vaporisé-séché à base de colostrum (CR; n = 8) ou d'un produit de remplacement de colostrum à base de plasma (PR; n = 9) au cours des 6 premières heures après la naissance. Du sang veineux et des sécrétions nasales obtenus avant le nourrissage et à l'âge de 24 h ont été analysés pour obtenir la quantité d'IgG totale (sérum) par immunodiffusion radiale et le total des quantités d'IgG, d'IgG-1 et d'IgG-2 spécifiques au VRS par ELISA indirecte. Les veaux qui avaient été nourris d'un CR avaient des concentrations supérieures d'IgG et dIgG-1 spécifiques au VRS dans leur sérum et les sécrétions nasales comparativement aux veaux nourris de produits PR; les veaux nourris d'un PR avaient des niveaux supérieurs d'IgG-2 sérique spécifique au VRS. La seule sous-catégorie d'anticorps détectée dans les sécrétions nasales était l'IgG-1. La re-secrétion passive d'IgG avec de l'activité neutralisante sur les muqueuses nasales pourrait contribuer à l'immunité associée au VRS observée en laboratoire et sur le terrain. L'usage de PR produira des anticorps nasaux inférieurs vu que l'IgG-2 n'est pas re-secrété.(Traduit par Isabelle Vallières).

Frequent expression of somatostatin receptor 2a in olfactory neuroblastomas: a new and distinctive feature.

Olfactory neuroblastoma (ONB) is a malignant neuroendocrine neoplasm with a usually slow course, but with considerable recurrence rate. Many neuroendocrine tumors have shown good response to the treatment with somatostatin analogs and somatostatin radioreceptor therapy. In ONBs, there are scarce data on somatostatin-based treatment and the cellular expression of somatostatin receptors (SSTR), the prerequisite for binding and effect of somatostatin on normal and tumor cells. The aim of our study was to investigate the immunohistochemical expression of SSTR2A and SSTR5 in a cohort of 40 ONBs. In addition, tissue microarrays containing 40 high-grade sinonasal carcinomas as well as 6 sinonasal lymphomas, 3 rhabdomyosarcomas, and 3 Ewing sarcomas were evaluated. Volante system was applied for staining evaluation. Thirty cases (75%) were immunopositive for SSTR2A and 3 (7.5%) for SSTR5. Among the 30 SSTR2A-positive ONBs, 19 tumors (63.3%) scored 2+ and 11 (36.7%) scored 3+. All SSTR5-positive ONBs scored 2+. Neither sinonasal carcinomas nor sinonasal small round blue cell neoplasms expressed SSTR2A or SSTR5. The frequent expression of SSTR2A provides a rationale for radioreceptor diagnosis and therapy with SST analogs in ONBs. SSTR2A expression in ONBs is a helpful adjunct in the differential diagnosis of ONBs.

Comparative histochemical analysis of glycoconjugates in the nasal vestibule of camel and sheep.

While Corriedale sheep survive in a wide range of climates, which prevents them to specialize for one climatic condition only, dromedary camels strictly adapted to desert areas. This demands more adaptive mechanisms to hot, dry conditions in camels than in sheep. Being the entrance of the nasal cavity, nasal vestibule is subjected to various environmental stressors. A protective way is the lining epithelium which is cornified in camel, but not in sheep. Mucus nasal secretions also play a key role in the protection of underlyings. Additionally, arterio-venous anastomosis is present in the lamina propria of the nasal vestibule of camel. In the present paper, sugar residues in the nasal vestibule of camel were analyzed and compared with those of sheep using 14 types of lectins to explore the distribution of glycoconjugates that may help the function of camel nasal vestibule in desert environment. In camel, none of the lectins could label the basal cells of the vestibular epithelium, although the basal cells reacted with six lectins in sheep. In camel, LEL and RCA-120 markedly labeled the luminal surface. WGA, DBA, SBA, and VVA produced marked intensities on the luminal surface in sheep. The mucous glands reacted with six lectins: WGA, s-WGA, VVA, PNA, PHA-E, and PHA-L in camel, while all lectins used except s-WGA and PHA-E reacted in the sheep. In summary, great differences are observed in the glycoconjugate expression between camel and sheep. This suggests that these glycoconjugate are related to camel's tolerance for environmental stressors.

Mammary Analog Secretory Carcinoma of the Nasal Cavity: Characterization of 2 Cases and Their Distinction From Other Low-grade Sinonasal Adenocarcinomas.

Secretory carcinoma, originally described as mammary analog secretory carcinoma (MASC), is a low-grade salivary gland tumor characterized by a t(12;15)(p13;q25) translocation, resulting in an ETV6-NTRK3 gene fusion. Most MASCs are localized to the parotid gland and intraoral minor salivary glands. Moreover, ETV6-rearranged carcinomas with secretory features have been reported recently in the thyroid (with and without a history of radiation exposure), skin, and in very rare instances in the sinonasal tract. Here, we describe 2 cases of primary MASC in the sinonasal tract and provide a detailed clinical and histopathologic characterization of their morphology, immunohistochemical profile, and genetic background and highlight features allowing for its separation from its recently described molecular mimicker, ETV6-rearranged low-grade sinonasal adenocarcinoma.

Nasal ventilation is an important factor in evaluating the diagnostic value of nasal nitric oxide in allergic rhinitis.

BACKGROUND: The role of nasal nitric oxide (NO) in the diagnosis of allergic rhinitis (AR) is controversial. The aim of this study was to identify factors that may affect levels of nasal NO in AR patients and evaluate the role of nasal NO in the diagnosis of AR. METHODS: Seventy-five AR patients and 31 healthy controls were enrolled in this study. AR symptom scores were assessed using the visual analog scale. Eosinophil cationic protein (ECP) was detected by enzyme-linked immunoassay, nasal NO was measured using a chemiluminescence analyzer, and nasal airway resistance (NAR) was assessed by active anterior rhinomanometry. RESULTS: Nasal obstruction score, ECP, and NAR were found to be independently associated with nasal NO. Nasal NO level in patients with nasal obstruction score <7 (mild-to-moderate obstruction) was significantly increased compared with healthy subjects (282.1 ± 122.6 vs 150.7 ± 48.4 ppb; p < 0.001), and significantly decreased in patients with nasal obstruction score ≥7 (severe obstruction) (97.2 ± 52.2 vs 150.7 ± 48.4 ppb; p < 0.001). Nasal NO and ECP in secretion were positively correlated in patients with mild-to-moderate nasal obstruction (r = 0.678), but not in patients with severe nasal obstruction (r = 0.077). In patients with NAR <0.65 Pa/cm3 /s, the correlation coefficient was highest between NO and ECP (r = 0.685). The areas under the receiver operating characteristic curve for nasal NO level were 0.878 and 0.939 in patients with nasal obstruction scores <7 and NAR <0.65 Pa/cm3 /s, respectively. CONCLUSION: Nasal patency affects nasal NO level significantly, and may reflect the severity of nasal inflammation in AR patients with mild-to-moderate nasal obstruction, but not in patients with severe nasal obstruction.

Histochemical and surface ultrastructural characteristics of the nasal cavity of laughing dove.

Ten apparently healthy, adult laughing doves were used to document detailed histological, histochemical and surface ultrastructural features of the nasal cavity and to investigate the structure-function relationship of the nasal cavity in this species. We observed that the nasal cavity of the laughing dove was composed of three main regions: nasal vestibule, respiratory and olfactory. Each region presented a characteristic epithelial lining. The epithelium varied along the nasal vestibule from keratinized stratified squamous rostrally to non-keratinized stratified squamous in the middle and stratified cuboidal in the caudal region of the nasal vestibule. The respiratory region was lined with pseudostratified columnar epithelium and was initially devoid of both goblet cells and cilia, but cilia then appeared and increased gradually in number close to the olfactory region. The caudal part of the respiratory region presented a stratified cuboidal epithelium. Strong alcianophilic, intra-epithelial mucous glands were identified, starting at the caudal region of the nasal vestibule and extended into the respiratory region. The olfactory region was lined with a pseudostratified epithelium that consisted of three different cell types: olfactory, support cells and basal cells. In conclusion, the current investigation presents new information concerning the histological, histochemical and ultrastructural features of the laughing dove's nasal cavity. Furthermore, the findings of this study may prove to be a valuable contribution to the avian histology and pathology literature.

RT-QuIC Assays for Prion Disease Detection and Diagnostics.

In coping with prion diseases, it is important to have tests that are practical enough for routine applications in medicine, agriculture, wildlife biology, and research, yet sensitive enough to detect minimal amounts of infectivity. Real-time quaking-induced conversion (RT-QuIC) assays have evolved to the point where they fulfill these criteria in applications to various human and animal prion diseases. For example, RT-QuIC assays of cerebrospinal fluid and nasal brushings allow for highly sensitive (77-97%) and specific (99-100%) identification of human sCJD patients. Recent improvements have markedly enhanced sensitivity and reduced the assay time required for many samples to a matter of hours rather than days. By combining analyses of cerebrospinal fluid and nasal brushings, diagnostic sensitivities and specificities of nearly 100% can be achieved. RT-QuIC assays are based on prion-seeded amyloid fibril formation by recombinant prion protein (rPrPSen) in multiwell plates using a Thioflavin T fluorescence readout. Here we describe our current RT-QuIC methodologies as well as technical considerations in executing, troubleshooting, and adapting the assay to new strains of prions and sample types.