A Pan-Dengue Virus Reverse Transcription-Insulated Isothermal PCR Assay Intended for Point-of-Need Diagnosis of Dengue Virus Infection by Use of the POCKIT Nucleic Acid Analyzer

Dengue virus (DENV) infection is considered a major public health problem in developing tropical countries where the virus is endemic and continues to cause major disease outbreaks every year. Here, we describe the development of a novel, inexpensive, and user-friendly diagnostic assay based on a reverse transcription-insulated isothermal PCR (RT-iiPCR) method for the detection of all four serotypes of DENV in clinical samples. The diagnostic performance of the newly established pan-DENV RT-iiPCR assay targeting a conserved 3′ untranslated region of the viral genome was evaluated. The limit of detection with a 95% confidence was estimated to be 10 copies of in vitro-transcribed (IVT) RNA. Sensitivity analysis using RNA prepared from 10-fold serial dilutions of tissue culture fluid containing DENVs suggested that the RT-iiPCR assay was comparable to the multiplex real-time quantitative RT-PCR (qRT-PCR) assay for DENV-1, -3, and -4 detection but 10-fold less sensitive for DENV-2 detection. Subsequently, plasma collected from patients suspected of dengue virus infection (n = 220) and individuals not suspected of dengue virus infection (n = 45) were tested by the RT-iiPCR and compared to original test results using a DENV NS1 antigen rapid test and the qRT-PCR. The diagnostic agreement of the pan-DENV RT-iiPCR, NS1 antigen rapid test, and qRT-PCR tests was 93.9%, 84.5%, and 97.4%, respectively, compared to the composite reference results. This new RT-iiPCR assay along with the portable POCKIT nucleic acid analyzer could provide a highly reliable, sensitive, and specific point-of-need diagnostic assay for the diagnosis of DENV in clinics and hospitals in developing countries

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Combining Weighted Contour Templates with HOGs for Human Detection Using Biased Boosting

This paper proposes a method to detect humans in the image that is an important issue for many applications, such as video surveillance in smart home and driving assistance systems. A kind of local feature called the histogram of oriented gradients (HOGs) has been widely used in describing the human appearance and its effectiveness has been proven in the literature. A learning framework called boosting is adopted to select a set of classifiers based on HOGs for human detection. However, in the case of a complex background or noise effect, the use of HOGs results in the problem of false detection. To alleviate this, the proposed method imposes a classifier based on weighted contour templates to the boosting framework. The way to combine the global contour templates with local HOGs is by adjusting the bias of a support vector machine (SVM) for the local classifier. The method proposed for feature combination is referred to as biased boosting. For covering the human appearance in various poses, an expectation maximization algorithm is used which is a kind of iterative algorithm is used to construct a set of representative weighted contour templates instead of manual annotation. The encoding of different weights to the contour points gives the templates more discriminative power in matching. The experiments provided exhibit the superiority of the proposed method in detection accuracy

Cloning of Zebrafish cDNA for 3 beta-hydroxysteroid dehydrogenase and P450scc

International audienceP450scc and 3 beta-HSD cDNA were isolated from a zebrafish lambda gt10 cDNA library using trout SCC and 3 beta-HSD cDNA as the probes. The zebrafish SCC cDNA encodes a protein of 509 amino acids, which shares a 78% similarities with the trout SCC and 58% with the human SCC. As for 3 beta-HSD, two forms of cDNA were isolated, termed HSD 5 and HSD 17, which may have resulted from alternative splicing. HSD 5 and HSD 17 encode proteins of 374 and 341 amino acids respectively. Both share 77% amino acid similarities with trout 3 beta-HSD and 53% similarities with the mouse 3 beta-HSD. Zebrafish has been increasingly used as a genetic model system to study organ development and to investigate human diseases. The cloning and the characterization of zebrafish P450scc and 3 beta-HSD should facilitate study of steroidogenesis and human disease associated with steroid imbalance

Modified Vaginal Mesh Procedure with DynaMesh^®-PR4 for the Treatment of Anterior/Apical Vaginal Prolapse

(1) Background: Treating female pelvic organ prolapse (POP) is challenging. Surgical meshes have been used in transvaginal surgeries since the 1990s, but complications such as mesh exposure and infection have been reported. Polyvinylidene fluoride (PVDF) mesh, known for its stability and non-reactive properties, has shown promise in urogynecological surgeries. (2) Methods: A retrospective analysis was conducted on 27 patients who underwent a modified PVDF vaginal mesh repair procedure using DynaMesh®-PR4 and combined trans-obturator and sacrospinous fixation techniques. Additional surgeries were performed as needed. (3) Results: The mean operation time was 56.7 min, and the mean blood loss was 66.7 mL. The average hospitalization period was 4.2 days with Foley catheter removal after 2 days. Patients experienced lower pain scores from the day of the operation to the following day. Postoperative follow-up revealed that 85.2% of patients achieved anatomic success, with 14.8% experiencing recurrent stage II cystocele. No recurrence of apical prolapse was observed. Complications were rare, with one case (3.7%) of asymptomatic mesh protrusion. (4) Conclusions: The modified vaginal mesh procedure using DynaMesh®-PR4 showed favorable outcomes with a short operation time, low recurrence rate, rare complications, and improved functional outcomes. This surgical option could be considered for anterior and apical pelvic organ prolapse in women

Changes in Cystoscopic Findings after Intravesical Hyaluronic Acid Instillation Therapy in Patients with Interstitial Cystitis

(1) Background: Limited data showed changes in glomerulation in the bladder mucosa of patients with interstitial cystitis (IC) after intravesical hyaluronic acid (HA) bladder infusion. We aimed to investigate the above changes. (2) Methods: Medical records of IC patients were reviewed retrospectively, from January 2010 to October 2019. Patients who had received repeated cystoscopy after intravesical HA treatment were enrolled. The associations of multiple parameters, including the ages, symptoms, initial glomerulation stage, HA doses, and the interval period of repeated cystoscopy between the glomerulation change in the repeated cystoscopy were analyzed. (3) Results: Among the 35 patients, 9 cases (25.7%) showed better glomerulation grades in the repeated cystoscope (Group 1), 20 cases (57.1%) showed the same grades (Group 2), and 6 cases showed worse grades (Group 3). No difference was seen in the initial grades or treatment course among the three groups. The interval periods from the initial to the repeated cystoscopy of Group 1 were longer than Group 2 and Group 3 (p = 0.031). Group 3 presents an elder age trend than the other two groups. (4) Conclusion: Intravesical HA repaired bladder glomerulation in a small group of patients with IC. Prolonged treatment has potential benefits, while older age is possibly a negative factor. However, no strong correlation was found between the initial glomerulation grades or changes in glomerulation grades with clinical symptoms

Independent factors associate with hospital mortality in patients with acute exacerbation of chronic obstructive pulmonary disease requiring intensive care unit admission: Focusing on the eosinophil-to-neutrophil ratio.

BackgroundFactors associated with hospital mortality are unclear in patients with acute exacerbation of COPD (AECOPD) requiring intensive care unit (ICU) admission. We aimed to characterize these patients and identify factors associated with hospital mortality.Patients and methodsWe used a retrospective observational case-control design and recruited patients between January 2015 and March 2017. Of 146 patients enrolled, 24 (16.4%) died during their hospital stay, while 122 survived.ResultsMultivariate logistic regression analyses revealed factors associated with hospital mortality: age (adjusted odds ratio [AOR] 1.12, 95% CI: 1.03-1.23), C-reactive protein (CRP) level >7.5 mg/dL at the emergency room (AOR 4.52, 95% CI: 1.27-16.04), peak eosinophil-to-neutrophil ratio (ENR)×102 on days 8-14 of treatment (AOR 0.22, 95% CI: 0.08-0.63), and in-hospital complications (AOR 4.23, 95% CI: 1.12-15.98) (all P0.224 on days 8-14 and initial CRP 0.224 on days 8-14 and initial CRP >7.5 mg/dL; (G2) Peak ENR×102 7.5 mg/dL. For G2 and G3 patients, the AOR of mortality was significantly different from that of the reference group (G2: AOR 10.00, P = 0.020; G3: AOR 61.79, PConclusionOlder age, initial CRP >7.5 mg/dL, peak ENR on days 8-14, and in-hospital complications were associated with hospital mortality in patients with AECOPD requiring ICU admission. Patients with both biomarkers, initial CRP >7.5 mg/dL, and peak ENR×102 <0.224 on days 8-14 of treatment, had an increased risk of hospital mortality