Properties of Pleural Mesothelial Cells in Idiopathic Pulmonary Fibrosis and Cryptogenic Organizing Pneumonia.

BACKGROUND: Profibrotic properties of pleural mesothelial cells may play an important role in the fibrosis activity in idiopathic pulmonary fibrosis (IPF). The purpose of this study was to compare the expression of pleural mesothelial cell markers in IPF and cryptogenic organizing pneumonia (COP), with an assumption that increased expression implies increase in fibrosis. METHODS: Twenty IPF lung samples were stained by immunohistochemistry for the pleural mesothelial cell markers: leucine rich repeat neuronal 4 (LRRN4), uroplakin 3B, CC-chemokine ligand 18, and laminin-5. Nine COP lung samples were used as controls. A semi-quantitative analysis was performed to compare markers expression in IPF and COP. RESULTS: LRRN4 expression was found in epithelial lining cells along the honeycombing and fibroblastic foci in IPF, but not in the fibrotic interstitial lesion and airspace filling fibrous tufts in COP. We found a significant decrease in baseline forced vital capacity when LRRN4 expression was increased in honeycombing epithelial cells and fibroblastic foci. CONCLUSION: LRRN4 expression patterns in IPF are distinct from those in COP. Our findings suggest that mesothelial cell profibrotic property may be an important player in IPF pathogenesis and may be a clue in the irreversibility of fibrosis in IPF.

Impact of ACE2 and TMPRSS2 Expression in Patients With Preeclampsia.

BACKGROUND/AIM: As maternal morbidity and mortality during pregnancy have increased during the COVID-19 pandemic, studies on pregnancy-related complications from SARS-CoV-2 infection are being actively conducted. Considering that pregnant women with COVID-19 may develop a preeclampsia (PE)-like syndrome, it is necessary to differentiate it from PE because true PE can result in an unfavorable perinatal outcome during a hasty delivery. MATERIALS AND METHODS: We investigated the protein expression of transmembrane serine protease 2 (TMPRSS2) and angiotensin-converting enzyme 2 (ACE2) in placental samples from 42 normotensive (n=9) and PE (n=33) patients without SARS-CoV-2 infection. We isolated placental trophoblast cells from normotensive and PE patients without evidence of SARS-CoV-2 infection to determine the mRNA and protein expression levels of TMPRSS2 and ACE2. RESULTS: High ACE2 cytoplasmic expression in extravillous trophoblasts (EVTs) was correlated with lower fibrin deposition (p=0.017). In comparison with high nuclear TMPRSS2 expression, low nuclearTMPRSS2 expression in endothelial cells (ECs) was positively correlated with PE (p=0.005), significantly higher systolic blood pressure (p=0.006), and higher urine protein-to-creatinine ratio (p=0.022). In contrast, high cytoplasmic TMPRSS2 expression in fibroblasts (FBs) was correlated with higher urine protein-to-creatinine ratio (p=0.018). Trophoblast cells extracted from PE placental tissue showed lower mRNA levels for both ACE2 and TMPRSS2. CONCLUSION: TMPRSS2 nuclear expression in ECs and cytoplasmic expression in FBs of the placenta may be related to a trophoblast-independent PE mechanism, and TMPRSS2 could be a new biomarker to discriminate actual PE from a PE-like syndrome associated with COVID-19.

Factors Associated with the Severity of Pregnancy-Related Hypertensive Disorder: Significance of Clinical, Laboratory, and Histopathological Features.

The purpose of this paper is to evaluate the association of maternal clinical and laboratory features and placental histopathological changes with disease severity in pregnancy-related hypertensive disorders. From January 2021 to December 2021, clinical and laboratory data at the time of delivery and histopathological features of the placenta were collected from pregnant women with pregnancy-related hypertensive disorders at a single institution. The women were classified according to the pregnancy-related hypertensive disorder clinical severity, and each variable was compared accordingly. Gestational age-matched normotensive groups were also compared. Univariate and multivariate regression analyses were used to identify factors influencing pregnancy-related hypertensive disorder severity. Fifty-eight pregnancies were analyzed. Maternal albumin levels before delivery (beta coefficient -0.83, p = 0.043) and increased placental syncytial knots (beta coefficient 0.71, p = 0.026) are important parameters that are closely related to disease severity in women with pregnancy-related hypertensive disorders. The combination of albumin, PAPP-A, total bilirubin, and eGFR levels appears to be optimal for predicting pregnancy-related hypertensive disorder severity.

Significance of HER2 and VEGFR2 in Early-stage Endometrial Cancer.

BACKGROUND/AIM: The majority of patients with early-stage endometrial cancer (EC) have a good prognosis, but recurrence does occur despite diagnosis at an early stage. There is a growing need for early diagnosis of EC and novel treatment options. MATERIALS AND METHODS: Human epidermal growth factor receptor 2 (HER2) and vascular endothelial growth factor receptor 2 (VEGFR) expression in microarrays of patient EC tissue was examined in association with clinicopathological data. HER2 and VEGFR2 expression in Ishikawa cells and differences in migration and proliferation of cells with HER2-knockdown were evaluated. RESULTS: Higher expression of VEGFR2 was associated with lower International Federation of Obstetrics and Gynecology grades (p=0.044). The positive correlation between HER2 and VEFR2 expression was statistically significant in T1 stage (p=0.002) and International Federation of Obstetrics and Gynecology grade 1 tumors (p=0.004). Wound-healing assays revealed that HER2 loss in Ishikawa cells reduced confluence compared to that of control cells. CONCLUSION: The association of VEGFR2 and HER2 expression in early EC was elucidated. This study shows that the measurement of VEGFR2 expression may be useful in the preoperative assessment of EC and suggests the possibility of anti-HER2 therapy for EC.

RF Transceiver for the Multi-Mode Radar Applications.

In this work, a multi-mode radar transceiver supporting pulse, FMCW and CW modes was designed as an integrated circuit. The radars mainly detect the targets move by using the Doppler frequency which is significantly affected by flicker noise of the receiver from several Hz to several kHz. Due to this flicker noise, the long-range detection performance of the radars is greatly reduced, and the accuracy of range to the target and velocity is also deteriorated. Therefore, we propose a transmitter that suppresses LO leakage in consideration of long-range detection, target distance, velocity, and noise figure. We also propose a receiver structure that suppresses DC offset due to image signal and LO leakage. The design was conducted with TSMC 65 nm CMOS process, and the designed and fabricated circuit consumes a current of 265 mA at 1.2 V supply voltage. The proposed transmitter confirms the LO leakage suppression of 37 dB at 24 GHz. The proposed receiver improves the noise figure by about 20 dB at 100 Hz by applying a double conversion architecture and an image rejection, and it illustrates a DC rejection of 30 dB. Afterwards, the operation of the pulse, FMCW, and CW modes of the designed radar in integrated circuit was confirmed through experiment using a test PCB.

Prognostic role of macrophage migration inhibitory factor expression in patients with squamous cell carcinoma of the lung.

BACKGROUND: Macrophage migration inhibitory factor (MIF) has been shown to play an important role in the inflammatory and immune response in squamous cell carcinoma (SCC). Recent studies have reported that MIF is involved in the tumorigenesis and overexpressed in various cancers. In this study, we assessed the prognostic role of MIF expression in SCC of the lung, and demonstrated the effect of knockdown of MIF on the migration in lung SCC cell lines. METHODS: The relationship between MIF expression and clinicopathological parameters and the prognostic role of MIF expression were evaluated with immunohistochemical staining in 96 patients with SCC of the lung. The expression of MIF mRNA and protein was analyzed by semi-quantitative polymerase chain reaction and Western blot in lung SCC cell. The effect of knockdown of MIF was assessed by wound healing assay. RESULTS: The high percentage of MIF-positive cells was significantly associated with lymph node metastasis (P = 0.004), and was a poor prognostic factor of disease-free survival (DFS) (hazard ratio [HR]: 3.125; 95% confidence interval [CI], 1.628-5.998; P = 0.001) and disease-specific survival (DSS) (HR: 2.303; 95% CI, 1.172-4.525; P = 0.016). Moreover, Kaplan-Meier analysis showed that SCC patients with a high percentage of MIF-positive cells had a significantly lower DFS (P = 0.001) and DSS (P = 0.014) than those with a low percentage. Furthermore, wound healing assay revealed that knockdown of MIF resulted in decreased cellular migration. CONCLUSION: MIF is closely associated with tumor progression and could be a prognostic factor in SCC of the lung.

Myoferlin silencing inhibits VEGFR2-mediated proliferation of metastatic clear cell renal cell carcinoma.

Recently, ramucirumab, a drug that targets vascular endothelial growth factor receptor (VEGFR), was clinically approved; therefore, we evaluated VEGFR2 expression and its predictive roles in tumor progression in clear cell renal cell carcinoma (CCRCC). Since we do not have many options for treating aggressive renal cell carcinoma patients, the application of anti-VEGFR2 therapy might be useful. Myoferlin (MYOF) is a 230 kDa transmembrane multi-C2-domain protein that contributes to plasma membrane repair, fusion, and endocytosis and is overexpressed in several invasive cancer cell lines, including breast, pancreas, and malignant melanoma. It forms a complex with VEGFR2 to inhibit VEGFR2 degradation. In this study, a total of 152 patients who had undergone nephrectomy for CCRCC were enrolled. Based on tissue microarray (TMA) blocks, the positive intensity and high proportion of MYOF showed a statistically significant correlation with the negative intensity (p < 0.001) and low proportion (p < 0.001) of VEGFR2, respectively. In addition, Fuhrman's nuclear grade ≥3 showed a significant correlation with VEGFR2 expression. In multivariate analysis, CCRCC patients with positive MYOF and negative VEGFR2 expression demonstrated poor clinical outcomes. We confirmed that positive MYOF expression and negative VEGFR2 expression were positively correlated in this CCRCC population. Knocking down MYOF in Caki-1 cells resulted in the downregulation of VEGFR2 at both mRNA and protein levels. Wound healing assays revealed that the loss of MYOF in Caki-1 cells decreased cell confluence compared to that in control cells. We demonstrated that MYOF influences cellular proliferation of the metastatic CCRCC cell line by regulating VEGFR2 degradation. Combined therapies targeting the MYOF and VEGFR2 pathways might be effective against metastatic CCRCC to increase patient survival.

Effect of 222-nm krypton-chloride excilamp treatment on inactivation of Escherichia coli O157:H7 and Salmonella Typhimurium on alfalfa seeds and seed germination.

We examined the control effect of a 222-nm KrCl excilamp on foodborne pathogens on alfalfa seeds and compared it with a conventional 254-nm low-pressure (LP) Hg lamp. When the 222-nm KrCl excilamp treated seeds at 87, 174 and 261 mJ/cm2, the log reductions of Escherichia coli O157:H7 (E. coli O157:H7) were 0.85, 1.77, and 2.77, respectively, and Salmonella Typhimurium (S. Typhimurium) experienced log reductions of 1.22, 2.27, and 3.04, respectively. When the 254-nm LP Hg lamp was applied at 87, 174, and 261 mJ/cm2, the log reductions of E. coli O157: H7 were 0.7, 1.16, and 1.43, respectively, and those of S. Typhimurium were 0.75, 1.15, and 1.85, respectively. Therefore, it was shown that the 222-nm KrCl excilamp was more effective than the 254-nm LP Hg lamp in reducing foodborne pathogens. The germination rate decreased to less than 80% after 261 mJ/cm2 treatment with the 254-nm LP Hg lamp, while more than 90% was maintained with 261 mJ/cm2 222-nm KrCl excilamp treatment. DNA damage assay showed that the difference in germination rate was due to DNA damage resulting from 254-nm LP Hg lamp treatment. However, 222 nm KrCl excilamp treatment did not cause DNA damage, resulting in no difference in germination rate compared to that of non-treated alfalfa seeds. Overall, these results demonstrate the utility of the 222-nm KrCl excilamp as a foodborne pathogen control intervention for the alfalfa seed industry.

A study of the transition between the non-polar and bipolar resistance switching mechanisms in the TiN/TiO2/Al memory.

Thermochemical and electronic trapping/detrapping mechanism-based resistance switching in TiO2 is one of the most extensively researched topics in the field of resistance-switching random access memory (ReRAM). In this study, the subtle correlation between the formation and rupture of the Magnéli-based conducting filament (CF), which is the mechanism of non-polar thermochemical-reaction-based switching, and the electron trapping/detrapping at the defect centers, which is the mechanism of bipolar electronic switching, is examined in detail. The chemical interaction between the TiN top electrode and the TiO2 layer generates a stable and immobile electron trapping layer, which is called a "switching layer", whereas the thin region between the just-mentioned switching layer and the remaining Magnéli CF after the thermochemical reset comprises a non-switching layer. The seemingly very complicated switching behavior with respect to the bias polarity, compliance current, and detailed biasing sequence could be reasonably explained by the phenomenological model based on the combined motions of the CF, switching layer, and non-switching layer. Light-induced detrapping experiments further supplement the suggested switching model.

Dictyostelium discoideum Ax2 as an Assay System for Screening of Pharmacological Chaperones for Phenylketonuria Mutations.

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah(-)) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah- cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

Synthesis and anti-hepatitis C virus (HCV) activity of 3'-C-substituted-methyl pyrimidine and purine nucleosides.

On the basis of potent anti-hepatitis C virus (HCV) activity of 2'-C-hydroxymethyladenosine, 3'-C-substituted-methyl-ribofuranosyl pyrimidine and purine nucleosides were designed and synthesized from D-xylose. Among compounds tested, all adenine analogues, 4a, 4d, and 4g showed significant anti-HCV activity in a replicon-based cell assay irrespective of the substituent (Y=OH, N3, or F) at the 3'-C-substituted methyl position, among which 4g (Y=N3) was the most potent, but it is also cytotoxic. This study guarantees the 3'-C-substituted-methyl nucleoside serves as a new template for the development of new anti-HCV agents.

Synthesis and antitumor activity of 1-beta-4-selenoarabinofuranosyl cytosine (4'-seleno-ara-C).

4-Seleno analogue of 1-beta-arabinofuranosyl cytosine (ara-C) was synthesized via 4'-selenouridine as a key intermediate, which was easily prepared from D-ribose. The arabino configuration was achieved by chemoselective ring opening of the 2,2'-anhydro-4'-selenouridine. The synthesized 4'-seleno-ara-C showed potent antitumor activity (IC(50) = 1.5 microM) against stomach cancer cells (SNU638).

Synthesis of 3'-C-hydroxymethyl-substituted pyrimidine and purine nucleosides as potential anti-hepatitis C virus (HCV) agents.

On the basis of potent anti-hepatitis C virus (HCV) activity of 2'-C-hydroxymethyladenosine, 3'-C-hydroxymethyl-substituted pyrimidine and purine nucleosides as potential anti-HCV agents were designed and synthesized from D-xylose via stereoselective Grignard reaction and conversion of the vinyl into hydroxymethyl group as key steps.