Enhancing Loop-Mediated Isothermal Amplification through Nonpowered Nanoelectric Preconcentration.

The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by ∼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses.

Results of Eustachian tube balloon dilation measured using the nine-step test.

Suggested several decades ago, the nine-step test is an intuitive test of Eustachian tube function. However, studies employing the nine-step test to assess the results of Eustachian tube balloon dilation (EBD) are limited. We aimed to objectively evaluate the efficacy of EBD in opening failure patients with decreased maximal peak pressure difference (MPD) using the nine-step test. Patients who had MPD values ≤ 13 daPa in the nine-step test were enrolled. The patients were categorized into two groups according to treatment decisions after discussion with a clinician: an EBD group (N = 26) and a medication group (N = 30). One month after treatment, the seven-item Eustachian Tube Dysfunction Questionnaire (ETDQ7) and the nine-step test were administered to all participants and subgroups of symptomatic participants (ETDQ7 > 15). MPD improved (increased) in both the EBD group and the medication group. ETDQ7 values improved (decreased) in the EBD group, but not in the medication group. In subgroup analysis, MPD and ETDQ7 values improved only in the symptomatic EBD group. According to the nine-step test, EBD can normalize 53.8% of decreased MPD. Posttreatment MPD and ETDQ7 scores were significantly better in the EBD group than in the medication group. However, EBD in patients with abnormal nine-step test results seemed less efficacious when the treatment results of the medication group were considered.

Clinical Factors Associated With Prognosis of Tinnitus and Aural Fullness After Sudden Sensorineural Hearing Loss.

OBJECTIVES: We evaluated the long-term prognosis and risk factors associated with tinnitus and aural fullness, which occurred with sudden sensorineural hearing loss. STUDY DESIGN: Retrospective cross-sectional review. SETTING: Tertiary referral center. PATIENTS: Those who visited our clinic for sudden hearing loss from January 2016 to May 2020, diagnosed with sudden sensorineural hearing loss based on pure-tone audiometry, and underwent magnetic resonance imaging to rule out other cause of hearing loss. In total, 106 patients were enrolled in this study. INTERVENTION: All patients were treated with oral glucocorticoids. Salvage intratympanic dexamethasone injection therapy was performed for the patients whose hearing was not fully recovered. MAIN OUTCOME AND MEASURES: We scored the loudness of tinnitus and the intensity of aural fullness using the numerical rating scale. We used a mixed-effects model for repeatedly measured tinnitus and aural fullness scores. RESULTS: The time after the onset of sudden sensorineural hearing loss (SSNHL; ß = -0.07; 95% confidence interval, -0.09 to -0.05; p < 0.001) and hearing outcome after treatment (overall p = 0.003) were significant factors associated with the prognosis of tinnitus. Concerning aural fullness, the time after the onset of SSNHL was a significant prognosis factor ( ß = -0.08; 95% confidence interval, -0.09 to -0.06; p < 0.001), unlike hearing outcome (overall p = 0.261). Pretreatment pure-tone audiometry average threshold and mainly affected frequencies were not significant factors for tinnitus and aural fullness recovery, respectively. CONCLUSION: The persistence of tinnitus with SSNHL was significantly affected by hearing recovery after treatment, whereas aural fullness was not associated with hearing recovery. However, both symptoms were improved over time after SSNHL.

Origami paper-based sample preconcentration using sequentially driven ion concentration polarization.

Ion concentration polarization (ICP) is one of the preconcentration techniques which can acquire a high preconcentration factor. Still, the main hurdles of ICP are its instability and low efficiency under physiological conditions with high ionic strength and abundant biomolecules. Here, we suggested a sequentially driven ICP process, which enhanced the electrokinetic force required for preconcentration, enabling enrichment of highly ionic raw samples without increasing the electric field. We acquired a 13-fold preconcentration factor (PF) in human serum using a paper-based origami structure consisting of multiple layers for three-dimensional sequential ICP (3D seq-ICP). Moreover, we demonstrated a paper-based enzyme-linked immunosorbent assay (ELISA) by 3D seq-ICP using tau protein, showing a 6-fold increase in ELISA signals.

Paper-Based Preconcentration and Isolation of Microvesicles and Exosomes.

Microvesicles and exosomes are small membranous vesicles released to the extracellular environment and circulated throughout the body. Because they contain various parental cell-derived biomolecules such as DNA, mRNA, miRNA, proteins, and lipids, their enrichment and isolation are critical steps for their exploitation as potential biomarkers for clinical applications. However, conventional isolation methods (e.g., ultracentrifugation) cause significant loss and damage to microvesicles and exosomes. These methods also require multiple repetitive steps  of ultracentrifugation, loading, and wasting of reagents. This article describes a detailed method to fabricate an origami-paper-based device (Exo-PAD) designed for the effective enrichment and isolation of microvesicles and exosomes in a simple manner. The unique design of the Exo-PAD, consisting of accordion-like multifolded layers with convergent sample areas, is integrated with the ion concentration polarization technique, thereby enabling fivefold enrichment of the microvesicles and exosomes on specific layers. In addition, the enriched microvesicles and exosomes are isolated by simply unfolding the Exo-PAD.

Multiple hypercoagulability disorders at presentation of non-small-cell lung cancer.

Hypercoagulability disorders are commonly encountered in clinical situations in patients with a variety of cancers. However, several hypercoagulability disorders presenting as first symptoms or signs in cancer patients have rarely been reported. We herein described a case of a woman with adenocarcinoma of the lung presenting with deep vein thrombosis, nonbacterial thrombotic endocarditis, recurrent cerebral embolic infarction, and heart failure.

Nanofluidic sustainable energy conversion using a 1D nanofluidic network.

We propose a 1-dimensional (1D) nanofluidic energy conversion device by implementing a surface-patterned Nafion membrane for the direct energy conversion of the pressure to electrical power. By implementing a -200-nm-thick nano-bridge with a 5-nm pore size between two microfluidic channels, we acquired an effective streaming potential of 307 mV and output power of 94 pW with 0.1 mM KCI under pressure difference of 45 MPa. The experimental results show both the effects of applied pressure differences and buffer concentrations on the effective streaming potential, and are consistent with the analytical prediction.

Erlotinib induced trichomegaly of the eyelashes.

Epithelial growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have been widely used for non-small-cell lung cancer patients. Its untoward cutaneous effects are largely well known and developed in many patients treated with EGFR TKIs. However trichomegaly of eyelash is rarely reported. Although trichomegaly is not a drug-limiting side effect, it could be troublesome of continuing the treatment because of cosmetic issue or eyeball irritation by long eyelashes. Therefore clinicians are needed to pay attention to this uncommon effect. We herein describe erlotinib induced trichomegaly of eyelashes in a woman with adenocarcinoma of the lung.

Gender differences in susceptibility to smoking among patients with lung cancer.

BACKGROUND/AIMS: To determine whether female smokers are more or less susceptible to the detrimental pulmonary-function effects of smoking when compared to male smokers among patients with lung cancer. METHODS: Pack-years and pulmonary function indices were compared between 1,594 men and women with lung cancer who were smokers or had a history of smoking. Differences in individual susceptibility to smoking were estimated using a susceptibility index formula. RESULTS: Of the patients, 959 (92.8%) men and 74 (7.2%) women were current smokers. Common histological types of lung cancer were squamous cell carcinoma, adenocarcinoma, and small cell carcinoma, among others. Women had a lower number of pack-years, forced expiratory volume in 1 second (FEV(1), liters), forced vital capacity (FVC, liters), and total lung capacity (TLC, liters) compared to those of men (25.0 ± 19.2 vs. 42.9 ± 21.7 for pack-years; 1.4 ± 0.5 vs. 2.0 ± 0.6 for FEV(1); 3.0 ± 0.7 vs. 2.0 ± 0.6 for FVC; 4.5 ± 0.8 vs. 5.7 ± 1.0 for TLC; all p < 0.001). The susceptibility index for women was significantly higher compared to that of men (1.1 ± 4.1 vs. 0.7 ± 1.1; p = 0.001). A significant inverse association was shown between the susceptibility index and TLC and FVC (r = -0.200 for TLC, -0.273 for FVC; all p < 0.001). CONCLUSIONS: The results suggest that the detrimental effects of smoking on pulmonary function are greater in women, as compared to those in men, among patients with lung cancer.

Effect of BRCA1 haplotype on survival of non-small-cell lung cancer patients treated with platinum-based chemotherapy.

PURPOSE: To determine whether germ-line variations in BRCA1 affect outcome in non-small-cell lung cancer (NSCLC) patients treated with platinum combination chemotherapy. PATIENTS AND METHODS: We evaluated the associations of four tagging BRCA1 polymorphisms and their haplotypes with treatment outcome in 300 NSCLC patients at stages IIIA (16%), IIIB (31%), and IV (53%). RESULTS: The median age was 63 years (range, 28 to 89 years). Histologically, 139 (46.3%) of the patients had squamous cell carcinomas and 137 (45.7%) had adenocarcinomas. Patient median survival time (MST) was 13.0 months. We observed no significant association between any of the tagging polymorphisms [S1613G, IVS13-1893 (A>C), IVS12-1207 (C>T), and IVS12+112 (C>A)] and overall survival. Of the five haplotypes evaluated (AACC, AACA, GCTC, GATC, and AATC), the survival of patients with two copies of the AACC (wild-type) haplotype was significantly shorter than that of patients with zero to one copies (MST, 8.47 v 14.57 months; log-rank P = .0066), even after adjustment for body weight loss, performance status, stage, second-line treatment, and radiation therapy (hazard ratio = 2.097; 95% CI, 1.339 to 3.284). The survival of patients with squamous cell carcinoma and two copies was significantly shorter than that of other patients with squamous cell carcinoma (MST, 6.8 v 15.3 months; log-rank P = 3.6 x 10(-5)), whereas differences in survival between the two adenocarcinoma groups was not significant (log-rank P = .677). CONCLUSION: These findings suggest that the AACC haplotype of the BRCA1 gene is an important prognostic marker in NSCLC patients treated with platinum combination chemotherapy.

[The effect of smoking status upon occurrence of impaired fasting glucose or type 2 diabetes in Korean men].

OBJECTIVES: To investigate whether smoking and the smoking status are predictors of the incident impaired fasting glucose (IFG) or type 2 diabetes in Korean men. METHODS: A cohort of 1,717 Korean men without IFG or diabetes, who underwent annual periodic health examinations for 4 years (2002-2006), were retrospectively investigated. IFG and diabetes were defined as a serum fasting glucose concentration of 100-125 mg/dL and more than 126 mg/dL, respectively. Cox's proportional hazards model was used to evaluate the association between smoking and development of IFG or type 2 diabetes. RESULTS: A total of 558 cases (32.5%) of incident IFG and 50 cases (2.9%) of diabetes occurred. After controlling for the potential predictors of diabetes, the relative risk for IFG, compared with the never smokers, was 1.02 (95% CI=0.88 to 1.19) for the ever-smokers, 0.96 (95% CI=0.79-1.16) for those who smoked 1-9 cigarettes/d, 1.15 (95% CI=1.01 to 1.30) for those who smoked 10-19 cigarettes/d, and 1.31 (95% CI=1.10 to 1.57) for those who smoked 20 or more cigarettes/d (the P value for the current smokers was only p<0.002). The respective multivariate adjusted relative risks for type 2 diabetes, compared with the never-smokers, were 1.07 (95% CI=0.64 to 1.92), 1.47 (95% CI=0.71 to 3.04), 1.84 (95% CI=0.92-3.04), and 1.87 (95% CI=1.13-3.67), respectively (the P value for the current smokers was only p=0.004). CONCLUSIONS: The smoking status and the number of cigarettes smoked daily are associated with an increased risk for developing IFG or type 2 diabetes in Korean men.

Natural history and therapeutic implications of patients with catamenial hemoptysis.

Evidence as to whether clinician has to give specific treatment in all patients of catamenial hemoptysis is unclear. We considered that the current treatment such as long-term usage of hormonal agent or surgery might be excessive for the catamenial hemoptysis. Therefore, we developed prospective observation study with observation strategy and follow-up for the patients. In sequential four patients of catamenial hemoptysis between December 2000 and November 2001, physical examination, chest X-ray, pelvic ultrasonogram and chest CT scan were taken at both the diagnosis and last follow-up. All patients were only observed without specific treatment within the limit of the possibility and followed for average 58 months. Mean age of patients was 23.5years (range, 22-25years). All patients have a history of undertaking one or two dilatations and curettages before diagnosis. The chest CT scans of all patients presented with ground-glass opacities of peripheral location that were disappeared without any residual lesion at last follow-up. Hemoptysis of two patients was spontaneously disappeared after 6 months. In the other two patients, it was greatly lessened in amount and frequency, then clinically insignificant in one. It was disappeared after subsequent 2 months and then relapsed two times in the late of follow-up of another patient. This study suggests that observation only may be an alternative option in the treatment of catamenial hemoptysis.

Effect of ERCC1 polymorphisms and the modification by smoking on the survival of non-small cell lung cancer patients.

A few reports for ERCC1 SNPs were conducted in patients treated with cisplatin chemotherapy. The aim of this study is to determine whether the SNPs are a prognostic factor related to the treatment or not and if smoking level of the patients have any relationship to the SNPs' effect on the survival. Peripheral blood lymphocytes of 423 consecutive non-small cell lung cancer patients were examined: 245 of the patients received cisplatin combination chemotherapy and 178 received only conservative care. We examined ERCC1 SNPs (codon 118 C/T and 8092 C/A). Whereas ERCC1 118 SNP had no effect on the survival in patients' given no treatment, an effect of the SNP was observed in the treatment group, especially in stage III. When smoking was considered, the risk effect of the T allele was shown to be significantly associated with the group that had more than 50 pack yr (p=0.03). As for the ERCC1 8092 C/A, no significant effects were observed in the treated group and the non-treatment group. These findings may suggest that the ERCC1 118 SNP is an useful prognostic factor related to cisplatin therapy and the effect of this polymorphism appears to be modified by smoking.

A case of pulmonary cryptococcosis with non-small cell lung cancer in idiopathic CD4+ T-lymphocytopenia.

Cryptococcus neoformans commonly causes opportunistic infections in immunocompromised patients, especially in patients with AIDS. CD4+ T-lymphocytopenia in AIDS indicates an increased risk of opportunistic infection and a decline in immunological function. Idiopathic CD4 T-lymphocytopenia (ICL) is characterized by depletions in the CD4+ T-cell subsets, without evidence of HIV infection. Immunodeficiency can exist in the absence of laboratory evidence of HIV infection, and T-cell subsets should be evaluated in patients who present with unusual opportunistic infections. We report a case of pulmonary cryptococcosis and lung cancer in a patient with persistently low CD4+ cell counts, without evidence of HIV infection.

Acute respiratory distress syndrome induced by adenovirus in an otherwise healthy woman.

Acute respiratory distress syndrome (ARDS) caused by adenovirus is a rare event in healthy adults, especially in non- military settings. Although treatment with intravenous ribavirin has been reported, supportive care, including mechanical ventilation, is known to be the main stay in the treatment of ARDS caused by adenovirus, with high-dose steroid treatment having rarely been reported. We report our experience with a 41-year-old, otherwise healthy, woman with ARDS, treated with high-dose steroid and mechanical ventilatory support.