Spatiotemporal characteristics of tissue derived small extracellular vesicles is associated with tumor relapse and anti-PD-1 response.

Small extracellular vesicles (sEVs) residing at tumor tissues are valuable specimens for biopsy. Tumor heterogeneity is common across all cancer types, but the heterogeneity of tumor tissue-derived sEVs (Ti-sEVs) is undefined. This study aims to discover the spatial distributions of Ti-sEVs in oral squamous cell carcinoma (OSCC) tissues and explore how these vesicle distributions affect the patients' prognosis. Multi-regional sampling enabled us to uncover that Ti-sEVs' accumulation at peritumoral sites correlates with a higher disease-free survival rate, and conversely, sparse peritumoral Ti-sEVs tend to forecast a higher risk of relapse. Of those relapsed patients, Ti-sEVs strongly bind to extracellular matrix and subsequently degrade it for allowing themselves enter the bloodstream rather than staying in situ. In advanced OSCC patients, the quantity and spatial distribution of Ti-sEVs prior to anti-PD-1 treatment, as well as the temporal variance of Ti-sEVs before and after immunotherapy, strongly map the clinical response and can help to distinguish the patients with shrinking tumors from those with growing tumors. Our work elucidates the correlation of spatiotemporal features of Ti-sEVs with patients' therapeutic outcomes and exhibit the potential for using Ti-sEVs as a predictor to forecast prognosis and screen the responders to anti-PD-1 therapy.

GBA1 as a risk gene for osteoporosis in the specific populations and its role in the development of Gaucher disease.

BACKGROUND: Osteoporosis and its primary complication, fragility fractures, contribute to substantial global morbidity and mortality. Gaucher disease (GD) is caused by glucocerebrosidase (GBA1) deficiency, leading to skeletal complications. This study aimed to investigate the impact of the GBA1 gene on osteoporosis progression in GD patients and the specific populations. METHODS: We selected 8115 patients with osteoporosis (T-score ≤ - 2.5) and 55,942 healthy individuals (T-score > - 1) from a clinical database (N = 95,223). Monocytes from GD patients were evaluated in relation to endoplasmic reticulum (ER) stress, inflammasome activation, and osteoclastogenesis. An in vitro model of GD patient's cells treated with adeno-associated virus 9 (AAV9)-GBA1 to assess GBA1 enzyme activity, chitotriosidase activity, ER stress, and osteoclast differentiation. Longitudinal dual-energy X-ray absorptiometry (DXA) data tracking bone density in patients with Gaucher disease (GD) undergoing enzyme replacement therapy (ERT) over an extended period. RESULTS: The GBA1 gene variant rs11264345 was significantly associated [P < 0.002, Odds Ratio (OR) = 1.06] with an increased risk of bone disease. Upregulation of Calnexin, NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) and Apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC) was positively associated with osteoclastogenesis in patients with GD. In vitro AAV9-GBA1 treatment of GD patient cells led to enhanced GBA1 enzyme activity, reduced chitotriosidase activity, diminished ER stress, and decreased osteoclast differentiation. Long-term bone density data suggests that initiating ERT earlier in GD leads to greater improvements in bone density. CONCLUSIONS: Elevated ER stress and inflammasome activation are indicative of osteoporosis development, suggesting the need for clinical monitoring of patients with GD. Furthermore, disease-associated variant in the GBA1 gene may constitute a risk factor predisposing specific populations to osteoporosis.

Antigen-induced chimeric antigen receptor multimerization amplifies on-tumor cytotoxicity.

Ligand-induced receptor dimerization or oligomerization is a widespread mechanism for ensuring communication specificity, safeguarding receptor activation, and facilitating amplification of signal transduction across the cellular membrane. However, cell-surface antigen-induced multimerization (dubbed AIM herein) has not yet been consciously leveraged in chimeric antigen receptor (CAR) engineering for enriching T cell-based therapies. We co-developed ciltacabtagene autoleucel (cilta-cel), whose CAR incorporates two B-cell maturation antigen (BCMA)-targeted nanobodies in tandem, for treating multiple myeloma. Here we elucidated a structural and functional model in which BCMA-induced cilta-cel CAR multimerization amplifies myeloma-targeted T cell-mediated cytotoxicity. Crystallographic analysis of BCMA-nanobody complexes revealed atomic details of antigen-antibody hetero-multimerization whilst analytical ultracentrifugation and small-angle X-ray scattering characterized interdependent BCMA apposition and CAR juxtaposition in solution. BCMA-induced nanobody CAR multimerization enhanced cytotoxicity, alongside elevated immune synapse formation and cytotoxicity-mediating cytokine release, towards myeloma-derived cells. Our results provide a framework for contemplating the AIM approach in designing next-generation CARs.

Prognostic impact of cortactin in patients with hypopharyngeal cancer and its role for tegafur-uracil maintenance after adjuvant chemoradiotherapy.

The prognosis of patients with hypopharyngeal cancer (HPC) remains poor. Our study aims to investigate the prognostic impact of cortactin in patients with HPC and its role for tegafur-uracil (UFUR) maintenance after adjuvant chemoradiotherapy (CRT). Patients who were diagnosed to have HPC and underwent laryngopharyngectomy followed by adjuvant CRT were enrolled into our study. Immunohistochemical staining was performed for cortactin evaluation. Kaplan-Meier curves were depicted for recurrence-free survival (RFS) and overall survival (OS). A total of 157 patients were enrolled into our study. After stratified by cortactin, 53 patients were cortactin (+) and 104 patients were cortactin (-). The median RFS was 86.7 months in cortactin (-) and 10.2 months in cortactin (+) (P < 0.001). The median OS was 93.4 months in cortactin (-) and 16.9 months in cortactin (+) (P < 0.001). Patients were further classified according to UFUR maintenance or not after adjuvant CRT. In cortactin (+) patients, the median RFS and OS were 13.6 months versus 7.0 months (P = 0.006) and 24.0 months versus 10.0 months (P < 0.001) in UFUR (+) and UFUR (-), respectively. In cortactin (-) patients, the median RFS and OS were 96.0 months versus 72.2 months (P = 0.262) and 98.5 months versus 105.0 months (P = 0.665) in UFUR (+) and UFUR (-), respectively. Cortactin has a significantly impact in HPC patients. UFUR maintenance provided survival benefits in patients with cortactin (+) after adjuvant CRT.

Elucidating the Cancer Phenotype in Turner Syndrome: A 20-Year Observational Cohort Study.

BACKGROUND/AIM: Turner syndrome confers increased cancer susceptibility; however, large-scale epidemiological evidence is lacking. This study aimed to analyze the incidence and prevalence of various malignancies in patients with Turner syndrome over 20 years of age to inform screening strategies. PATIENTS AND METHODS: We performed a retrospective cohort analysis of 11,502 patients with Turner syndrome from 2000 to 2020 utilizing the TriNetX research network database. The outcomes encompassed the incidence and prevalence of 20 cancers. Stratified analyses were used to evaluate variations in age, sex, and race. RESULTS: Key findings demonstrated markedly elevated risks of breast (1.7%), colon (1.0%), renal (0.4%), gonadoblastoma (0.4%), and other cancers. Significant demographic variations were observed in the incidence of cancers, such as gonadoblastoma, renal, and colon cancer. CONCLUSION: This large real-world study offers novel insights into the spectrum of cancer risk across adulthood in Turner syndrome. Our findings elucidate Turner syndrome's complex cancer phenotype to inform clinical decision-making, prognostication, and tailored screening strategies to ultimately advance patient care.

Incidence of Pulmonary and Respiratory Conditions in Gaucher Disease from 2000 to 2020: A Multi-institutional Cohort Study.

BACKGROUND/AIM: Gaucher disease (GD) is a rare lysosomal storage disorder that can involve the lungs and pulmonary vasculature. The long-term effects of GD on respiratory health remain unclear due to limited data on the natural history of this disease. We analyzed electronic health records for 11,004 patients with GD over 10-20 years to determine the incidence of pulmonary hypertension (PH), lung disease, and other respiratory comorbidities and better understand disease course to guide management. PATIENTS AND METHODS: We conducted a retrospective cohort study using the TriNetX research database of 130 million international patients. The incidence of primary/secondary PH, pulmonary heart disease, interstitial/obstructive/restrictive lung disease, pulmonary hemorrhage, and pulmonary embolism was assessed in patients with GD from 2000-2020. RESULTS: Incidence rates of all conditions assessed increased from 10 to 20 years of follow-up. Excess risk of PH, lung disease, and pulmonary hemorrhage was significantly higher in GD patients after 20 versus 10 years. CONCLUSION: Extended follow-up in GD is associated with substantially higher risks of PH, lung disease and other respiratory comorbidities, highlighting the need for close monitoring and early intervention to mitigate long-term pulmonary decline. Improved understanding of mechanisms driving respiratory deterioration can support the development of novel treatments to optimize outcomes in this population at high risk of pulmonary morbidity and mortality.

Structures and bonding properties of lithium polysulfide clusters LiSn-/0 (n = 3-5) and Li2S4-/0: size-selected anion photoelectron spectroscopy and theoretical calculations.

The structures and bonding properties of several lithium polysulfide clusters LiSn-/0 (n = 3-5) and Li2S4-/0 were investigated by size-selected anion photoelectron spectroscopy coupled with quantum chemistry calculations. The vertical detachment energies of LiS3-, LiS4-, and LiS5- were estimated to be 2.17 ± 0.08, 3.30 ± 0.08 and 3.66 ± 0.08 eV, respectively, and that of Li2S4- was estimated to be 3.21 ± 0.08 eV. It is found that LiS3- and LiS3 have planar quadrilateral structures, and LiS4- and LiS4 have distorted five-membered ring structures. LiS5- has a distorted six-membered ring structure while neutral LiS5 has a book-shaped structure. The lowest-lying structure of Li2S4- can be viewed as a S2 unit connecting to the Li-Li edge of a Li2S2 tetrahedron. The lowest-lying structure of neutral Li2S4 can be viewed as a S2 unit connecting to the S atoms of a Li2S2 quadrilateral. The natural population analysis (NPA) and electron localization function (ELF) analyses show that the excess electron of LiSn- is mainly localized over the sulfur chains, especially on the S atoms interacting with Li, thus, the most stable structures of LiSn- can be regarded as a Li+ cation interacting with a Sn2- dianion. The results may be useful for understanding the formation of lithium polysulfides in lithium sulfur batteries.

Reducing Medication Errors in Children's Hospitals.

OBJECTIVES: Knowledge of the prevalence and characteristics of medication errors in pediatric and neonatal patients is limited. This study aimed to evaluate the incidence and medication error characteristics in a pediatric hospital over 5 years and to determine whether serial error prevention programs to optimize a computerized physician order entry (CPOE) system reduce error incidence. METHODS: We retrospectively reviewed medication errors documented between January 2015 and December 2019. RESULTS: A total of 2,591,596 prescriptions were checked, and 255 errors were identified. Wrong dose prescriptions constituted the most common errors (56.9%). Medications with the highest rate of errors were antibiotics/antiviral drugs (36.9%). Oral route medications comprised the highest portion (60.8%), followed by intravenous ones (28.6%). The most common stage for medication errors was physician ordering (93.3%). Junior residents were responsible for most errors (45.9%). Most errors occurred in the pediatric ward (53.7%). In total, 221 (86.7%) errors were near misses. Only 4 errors (1.6%) were considered significant and required active monitoring or intervention. Type of error, stage of error, staff composition, and severity level of errors were significantly related to the number of errors in different years. There was a statistically significant decrease in errors per 100,000 prescriptions across different years after optimizing the CPOE system. CONCLUSIONS: The incidence of medication errors decreased with extensive use of the CPOE system. Continuous application of the CPOE optimization program can effectively reduce medication errors. Further incorporation of pediatric-specific decision-making and support tools and error prevention measures into CPOE systems is needed.

[Characteristic Analysis of nirS Denitrifying Bacterial Community in Lijiahe Reservoir During Stratification].

To explore the composition of the nirS denitrifying bacterial community during stratification in spring(March to May) in a drinking water reservoir and its relationship with water quality, the water quality and relative abundance and structure of the denitrifying bacterial community were analyzed using in-situ monitoring coupled with Illumina high-throughput sequencing technology in the Lijiahe Reservoir. The results showed that:① through high-throughput sequencing, 4 phyla and 13 genera were identified. The dominant bacterial phylum was Proteobacteria, and its relative abundance was between 52.5% and 70.6%. The overall trend of the relative abundance of Proteobacteria decreased on the time scale (P<0.05), and its relative abundance in the surface and middle layers was higher than that of the bottom layer on the spatial scale (P<0.05). There was no difference in the proportion of Proteobacteria between the surface and middle layers (P>0.05), and the abundance of its bottom layer was relatively stable; eight genera of bacteria with denitrification function were identified, among which the dominant bacterial genera (relative abundance>1%) were Dechloromonas and Pseudomonas. The relative abundance of Dechloromonas showed a trend of first decreasing and then increasing on the time scale, whereas the relative abundance of Pseudomonas showed a trend of increasing first and then decreasing on the time scale. There were no differences on the spatial scale between these two genera (P>0.05); the changes in bacterial diversity and abundance were basically similar, with a trend of first increasing and then decreasing on the time scale. The highest diversity and abundance of the bacterial community gradually increased with increasing depth on the spatial scale. ② ρ(TN) of the reservoir during stratification was 2.35-2.91 mg·L-1, and the nitrogen pollution was more serious. In March and April, ρ(TN) on the vertical scale was basically similar and showed a decreasing trend. In May, the content of total nitrogen was higher than that in March and April, and the highest value of total nitrogen content occurred in the surface layer. ③ Redundancy analysis showed that water temperature, dissolved oxygen, nitrate, and ammonia nitrogen were the main driving factors, and ammonia nitrogen showed a significantly negative correlation with Dechlormonas. In summary, the study of nirS-type denitrification communities and related influencing factors will contribute to analyzing the characteristics of denitrifying bacterial community changes in a micro-polluted drinking water reservoir and provide a theoretical research basis for the biological remediation of nitrogen pollution in such reservoirs in the future.

[Mechanism of Algal Community Dynamics Driven by the Seasonal Water Bacterial Community in a Stratified Drinking Water Reservoir].

Phytoplankton and bacteria are crucial components of aquatic food webs, playing critical roles in the structure and function of freshwater ecosystems. However, there are few studies on how the algal and bacterial communities interact and respond to changing environmental conditions in the water reservoirs. Thus, the ecological interaction relationship between the temporal succession of the phytoplankton community and the bacterial community was investigated using 16S rDNA high-throughput sequencing and a co-occurrence network in the Lijiahe Reservoir. The results showed that Bacillariophyta and Chlorophyta were also dominant taxa in the phytoplankton community. In August, Cyanobacteria replaced Bacillariophyta as the second-most dominant taxa, with an average relative abundance of 30.13%. DNA sequencing showed that Proteobacteria, Actinobacteria, and Bacteroidetes dominated throughout the year. Proteobacteria reached a maximum relative abundance of 71.68% in July. Acidobacteria and Deinococcus-Thermus, which were rare taxa, reached maximum relative abundances of 10.20% and 5.56%, respectively. The co-occurrence network showed that the association between algae and bacteria was mainly positive, indicating that the interaction between them may be dominated by mutualism. As a keystone taxa, Methylotenera was significantly and positively related to Chlorella. Scenedesmus was also a keystone taxa and was significantly and negatively correlated with various bacteria, such as Methylobacter, Solitalea, and Rhodoferax. An RDA analysis showed that the succession of algal and bacterial communities was significantly regulated by water temperature, pH, and conductivity, and the environmental factors explained 93.1% and 90% of the variation in the algal community and bacterial community, respectively. The results will provide a scientific basis for exploring the micro-ecological driving mechanism of the interaction between algae and bacteria in deep drinking water reservoir ecosystems.

[Advances in Microbe-Derived Taste and Odor Compounds in Typical Aquatic Ecosystems].

Olfactory microorganisms mainly include actinomycetes, algae, and myxobacteria, and are widely found in typical water ecosystems such as water source reservoirs, lakes, streams, and oceans. Biologically-derived taste and odor compounds can cause severe off-flavor problems at trace concentrations, threatening the safety of water supplies. Physical and chemical methods have been implemented, but they need to be improved and optimized. Microbial methods show attractive prospects due to their environmentally friendly characteristics. This article reviews the literature regarding the types, biological sources, detection methods, control measures, influencing factors, and odor-producing mechanisms of microbial methods. Future research directions and fields of investigation are discussed in order to promote the related research of taste and odor compounds and ensure the safety of urban water supplies.

[Spatial and Temporal Succession Characteristics of Aerobic Anoxygenic Photosynthesis Bacteria in a Stratified Reservoir].

Aerobic anoxygenic photosynthesis bacteria (AAPB) play a significant role in the material circulation of the hydrosphere, with diverse community structure and unique metabolic functions. To investigate the spatial and temporal succession characteristics of AAPB abundance and community structure in Jinpen Reservoir, a quantitative real-time polymerase chain reaction and Illumina MiSeq high-throughput sequencing technique targeting the pufM gene were applied. Furthermore, redundancy analysis was used to determine the influence of environmental factors on their community structure. The results showed that the AAPB abundance ranged from (6.70±0.43)×103 to (2.69±0.15)×104 copies·mL-1, with the maximum value appearing in October, and decreased with an increase in water depth. Samples were mainly classified into 19 genera (except for the unclassified genus); the most abundant AAPB genera were Bradyrhizobium sp. and Methylobacterium sp., which were affiliated to the α-Proteobacteria, and the proportion of the Bradyrhizobium sp. was highest in November, accounting for more than 60% (except 10 m). Furthermore, Rubrivivax sp., belonging to ß-Proteobacteria, was found to have a low proportion. There was a strong interaction relationship between AAPB genera. For example, Rhodobacter sp. was positively correlated with Rhodovulum sp., while Hydrogenophaga sp. was negatively correlated with Bradyrhizobium sp.. The community structure composition and distribution of AAPB were significantly different, mainly affected by temperature (T), total nitrogen (TN), NO3--N, and light intensity and comprehensively regulated by environmental factors. For instance, T, TN, and total phosphorus had a significant impact on the AAPB community structure of water samples at 0, 5, and 15 m in October, whereas light intensity, pH, DO, and chlorophyll-a were major structuring factors in the AAPB assemblages of water samples at 5 m in December. The results have guiding significance for parsing the spatial and temporal variability of AAPB abundance and diversity in stratified reservoirs, and simultaneously provide a theoretical basis for exploring the driving factors of AAPB population structure.

[Water Quality and Bacterial Population Driving Mechanism of Algae Vertical Succession in Stratified Reservoir].

Phytoplankton and bacteria are important components of the aquatic food web, and play a critical role in substance circulation and energy exchange in freshwater ecosystems. The succession of algae is closely related to the metabolism and structural succession of bacterial populations in the water column. Thus, in this study, the vertical succession characteristics of phytoplankton and bacteria community structure and their coupling with water quality were investigated during an algal bloom in the Lijiahe Reservoir using high-throughput DNA sequencing and Biolog technologies. The results showed that the Lijiahe Reservoir was in the thermal stratification stage in August, and the pH, dissolved oxygen, and NH4+-N of the water column gradually decreased with depth (P<0.001). Algal cell concentration and chlorophyll a exhibited a simultaneous trend (P<0.001), and the maximum values in the surface layer were 3363.33×104 cells·L-1 and 7.03 µg·L-1, respectively. The algal community structure was dominated by Microcystis at water depths of 0 m and 3 m, and at 6 m water depth, Cyclotella replaced Microcystis as the most dominant algae, with a relative abundance of 57.28%. Biolog analysis indicated that the outbreak of Microcystis had a significant impact on bacterial metabolic activity and its relative abundance, but the diversity of bacterial population metabolic activity varied less. A total of 1420 operational taxonomic units were found by high-throughput sequencing, belonging to 10 bacterial phyla. Of these, Actinobacteria and Proteobacteria dominated in all water layers, and their relative abundances were more than 50%. The relative abundance of Chlorobi and Planctomycetes varied significantly with water depth, reaching their maxima at a depth of 6 m with values of 10.29% and 6.78%, respectively, which were both negatively correlated with algal density (P<0.05). Firmicutes and Gemmatimonadetes were positively correlated with algal density (P<0.05). A heat map fingerprint showed that the vertical distribution of the bacterial community structure of the Lijiahe Reservoir varied significantly, and with the increase in water depth, the bacterial community was more uniformly distributed and tended to diversify. Redundancy analysis (RDA) showed that the vertical distribution of the bacterial and algal community structure was regulated by different water qualities, and the difference was significant. This study investigated the coupling mechanism of algal and bacterial communities during the algal bloom in the Lijiahe Reservoir, and the results provided a scientific basis for the investigation of the molecular microecological driving mechanism of water-source algal blooms.

Cortactin as a potential predictor of second esophageal neoplasia in hypopharyngeal carcinoma.

OBJECTIVE: Hypopharyngeal carcinoma has a very poor prognosis. The high incidence of second esophageal neoplasia is one of the major causes. To establish an efficient follow-up scheme for increasing the diagnostic yield and reducing the adverse impact of second esophageal neoplasia on survival, the purpose of this study was to explore a biomarker to predict second esophageal neoplasia. METHODS: In this retrospective cohort study, consecutive tissue specimens from those patients who underwent tumor resection between September 2007 and October 2015 were collected. Gene amplification was performed by real-time PCR. The expression of cortactin was evaluated by immunohistochemistry. The predictive risk factors of developing second esophageal neoplasia and prognostic factors related to survival were analyzed. RESULTS: A total of 187 patients were included with a mean follow-up of 48months (12-118months). Second esophageal tumors were found in 53 (28.3%), including 41 (21.9%) esophageal squamous cell carcinoma and 12 severe dysplasia. The results of multivariate analyses revealed that age (OR 2.81, 95% CI 1.16-6.78), cortactin overexpression (OR 2.49, 95% CI 1.17-5.33), and stage IV versus I (OR 6.49, 95% CI 1.68-25.18) were independent predictors of second esophageal neoplasia, and second esophageal neoplasia (HR 1.78, 95% CI 1.05-3.01) was an independent predictor of overall survival. CONCLUSION: This is the first report to identify a potential biomarker for predicting second esophageal neoplasia in patients with hypopharyngeal carcinoma. In those patients with cortactin overexpression and younger age (≤60years old), close surveillance for second esophageal neoplasia is required. In addition, the real effect of cortactin overexpression on development of primary esophageal carcinoma is required to be validated in a large cohort study.

SERPINB5 Expression: Association with CCRT Response and Prognostic Value in Rectal Cancer.

Background: Due to the varying characteristics and conflicting outcomes on the overall survival of rectal cancer patients, many studies have been undertaken to determine various prognostic and predictive factors for the mainstay treatment of CCRT followed by surgery. Cancer cell motility contributes to tumor invasion, migration and eventually metastasis. However, the genes associated with cell motility (i.e., GO:0048870) have not been systemically evaluated in rectal cancers. Methods: A comparative analysis of gene expression profiles was applied to the transcriptomic dataset (GSE35452) with a focus on genes associated with cell motility (GO:0048870), where SERPINB5 was recognized as the most significantly up-regulated gene. Tumor samples from 172 primary rectal cancer patients who underwent neoadjuvant CCRT followed by surgical resection were collected. Immunohistochemistry was used to semi-quantitatively assess the expression level of SERPINB5 protein. Statistical analyses of SERPINB5 expression and various clinicopathological features as well as survival were then performed. Results: High immunoreactivity of SERPINB5 was significantly linked to pre- and post-CCRT advanced disease, lymphovascular invasion, and poor response to CCRT (all P ≤ 0.015). SERPINB5 overexpression was not only negatively associated with disease-specific survival (DSS), local recurrence-free survival (LRFS) and metastasis-free survival (MeFS) rates in univariate analyses but also was an independent prognostic factor for DSS and MeFS in rectal cancer patients (all P ≤ 0.043). Conclusion: SERPINB5 may play an important role in rectal cancer progression and response to neoadjuvant CCRT and serve as a novel prognostic factor.

A Hot-Polymer Fiber Fabry-Perot Interferometer Anemometer for Sensing Airflow.

This work proposes the first hot-polymer fiber Fabry-Perot interferometer (HPFFPI) anemometer for sensing airflow. The proposed HPFFPI is based on a single-mode fiber (SMF) endface that is attached to a UV-cured polymer to form an ultracompact fiber Fabry-Perot microcavity. The proposed polymer microcavity was heated using a low-cost chip resistor with a controllable dc driving power to achieve a desired polymer's steady-state temperature (T) that exceeds the T of the surrounding environment. The polymer is highly sensitive to variations of T with high repeatability. When the hot polymer was cooled by the measured flowing air, the wavelength fringes of its optical spectra shifted. The HPFFPI anemometers have been experimentally evaluated for different cavity lengths and heating power values. Experimental results demonstrate that the proposed HPFFPI responses well in terms of airflow measurement. A high sensitivity of 1.139 nm/(m/s) and a good resolution of 0.0088 m/s over the 0~2.54 m/s range of airflow were achieved with a cavity length of 10 µm and a heating power of 0.402 W.

Biochemical features of the adhesion G protein-coupled receptor CD97 related to its auto-proteolysis and HeLa cell attachment activities.

CD97 belongs to the adhesion GPCR family characterized by a long ECD linked to the 7TM via a GPCR proteolytic site (GPS) and plays important roles in modulating cell migration and invasion. CD97 (EGF1-5) is a splicing variant of CD97 that recognizes a specific ligand chondroitin sulfate on cell membranes and the extracellular matrix. The aim of this study was to elucidate the extracellular molecular basis of the CD97 EGF1-5 isoform in protein expression, auto-proteolysis and cell adhesion, including epidermal growth factor (EGF)-like domain, GPCR autoproteolysis-inducing (GAIN) domain, as well as GPS mutagenesis and N-glycosylation. Both wild-type (WT) CD97-ECD and its truncated, GPS mutated, PNGase F-deglycosylated, and N-glycosylation site mutated forms were expressed and purified. The auto-proteolysis of the proteins was analyzed with Western blotting and SDS-PAGE. Small angle X-ray scattering (SAXS) and molecular modeling were used to determine a structural profile of the properly expressed receptor. Potential N-glycosylation sites were identified using MS and were modulated with PNGase F digestion and glyco-site mutations. A flow cytometry-based HeLa cell attachment assay was used for all aforementioned CD97 variants to elucidate the molecular basis of CD97-HeLa interactions. A unique concentration-dependent GPS auto-proteolysis was observed in CD97 EGF1-5 isoform with the highest concentration (4 mg/mL) per sample was self-cleaved much faster than the lower concentration (0.1 mg/mL), supporting an intermolecular mechanism of auto-proteolysis that is distinct to the reported intramolecular mechanism for other CD97 isoforms. N-glycosylation affected the auto-proteolysis of CD97 EGF1-5 isoform in a similar way as the other previously reported CD97 isoforms. SAXS data for WT and deglycosylated CD97ECD revealed a spatula-like shape with GAIN and EGF domains constituting the body and handle, respectively. Structural modeling indicated a potential interaction between the GAIN and EGF5 domains accounting for the absence of expression of the GAIN domain itself, although EGF5-GAIN was expressed similarly in the wild-type protein. For HeLa cell adhesion, the GAIN-truncated forms showed dramatically reduced binding affinity. The PNGase F-deglycosylated and GPS mutated forms also exhibited reduced HeLa attachment compared with WT CD97. However, neither N-glycosylation mutagenesis nor auto-proteolysis inhibition caused by N-glycosylation mutagenesis affected CD97-HeLa cell interactions. A comparison of the HeLa binding affinities of PNGase F-digested, GPS-mutated and N-glycosylation-mutated CD97 samples revealed diverse findings, suggesting that the functions of CD97 ECD were complex, and various technologies for function validation should be utilized to avoid single-approach bias when investigating N-glycosylation and auto-proteolysis of CD97. A unique mechanism of concentration-dependent auto-proteolysis of the CD97 EGF1-5 isoform was characterized, suggesting an intermolecular mechanism that is distinct from that of other previously reported CD97 isoforms. The EGF5 and GAIN domains are likely associated with each other as CD97 expression and SAXS data revealed a potential interaction between the two domains. Finally, the GAIN and EGF domains are also important for CD97-HeLa adhesion, whereas N-glycosylation of the CD97 GAIN domain and GPS auto-proteolysis are not required for HeLa cell attachment.

Quiz. Correct answer to the quiz. Check your diagnosis. Clear cell papillary renal cell carcinoma.

We incidentally observed a case of clear cell papillary renal cell carcinoma of an 81-year-old woman, presenting with intermittent left flank pain. It is a recently described rare renal parenchymal tumor.