Prevalence estimation of Tropheryma whipplei in duodenal biopsy tissues of Koreans.

Whipple's disease caused by Tropheryma whipplei is difficult to diagnose because of a broad spectrum of manifestations and non-specific clinical signs. In the current global era, the incidence of duodenal infection/inflammation caused by T. whipplei in Korea may has been underestimated. Here we estimated the prevalence of T. whipplei in duodenal biopsy tissues of Koreans using real-time PCRs (RT-PCRs). A total of 252 duodenal biopsy tissues were collected from Korean patients who underwent esophagogastroduodenoscopy and duodenal biopsy. DNA extracted from the duodenal biopsy tissues was analyzed using three RT-PCRs targeting T. whipplei-specific regions of the 16S-23S rRNA intergenic spacer, hsp65, and Dig15 in parallel. In the samples positive in RT-PCRs, direct sequencing was performed for each RT-PCR target. The prevalence of T. whipplei was estimated based on the RT-PCR and sequencing results. Among the analyzed samples, T. whipplei was not detected. The prevalence of T. whipplei in duodenal biopsy tissues of Koreans was estimated to be less than 0.4%. This is the first study to attempt to detect T. whipplei in duodenal biopsy tissues of Koreans and estimate its prevalence. Our findings infer that while T. whipplei carriers exist in Korea, the incidence of duodenal infection/inflammation caused by T. whipplei is extremely rare.

Bio-conjugated nanoarchitectonics with dual-labeled nanoparticles for a colorimetric and fluorescent dual-mode serological lateral flow immunoassay sensor in detection of SARS-CoV-2 in clinical samples.

Serological detection of antibodies for diagnosing infectious diseases has advantages in facile diagnostic procedures, thereby contributing to controlling the spread of the pathogen, such as in the recent SARS-CoV-2 pandemic. Lateral flow immunoassay (LFIA) is a representative serological antibody detection method suitable for on-site applications but suffers from low clinical accuracy. To achieve a simple and rapid serological screening as well as the sensitive quantification of antibodies against SARS-CoV-2, a colorimetric and fluorescent dual-mode serological LFIA sensor incorporating metal-enhanced fluorescence (MEF) was developed. For the strong fluorescence signal amplification, fluorophore Cy3 was immobilized onto gold nanoparticles (AuNPs) with size-controllable spacer polyethyleneglycol (PEG) to maintain an optimal distance to induce MEF. The sensor detects the target IgG with a concentration as low as 1 ng mL-1 within 8 minutes. The employment of the MEF into the dual-mode serological LFIA sensor shows a 1000-fold sensitivity improvement compared with that of colorimetric LFIAs. The proposed serological LFIA sensor was tested with 73 clinical samples, showing sensitivity, specificity, and accuracy of 95%, 100%, and 97%, respectively. In conclusion, the dual-mode serological LFIA has great potential for application in diagnosis and an epidemiological survey of vaccine efficacy and immunity status of individuals.

Establishing molecular pathology curriculum for pathology trainees and continued medical education: a collaborative work from the Molecular Pathology Study Group of the Korean Society of Pathologists.

BACKGROUND: The importance of molecular pathology tests has increased during the last decade, and there is a great need for efficient training of molecular pathology for pathology trainees and as continued medical education. METHODS: The Molecular Pathology Study Group of the Korean Society of Pathologists appointed a task force composed of experienced molecular pathologists to develop a refined educational curriculum of molecular pathology. A 3-day online educational session was held based on the newly established structure of learning objectives; the audience were asked to score their understanding of 22 selected learning objectives before and after the session to assess the effect of structured education. RESULTS: The structured objectives and goals of molecular pathology was established and posted as a web-based interface which can serve as a knowledge bank of molecular pathology. A total of 201 pathologists participated in the educational session. For all 22 learning objectives, the scores of self-reported understanding increased after educational session by 9.9 points on average (range, 6.6 to 17.0). The most effectively improved items were objectives from next-generation sequencing (NGS) section: 'NGS library preparation and quality control' (score increased from 51.8 to 68.8), 'NGS interpretation of variants and reference database' (score increased from 54.1 to 68.0), and 'whole genome, whole exome, and targeted gene sequencing' (score increased from 58.2 to 71.2). Qualitative responses regarding the adequacy of refined educational curriculum were collected, where favorable comments dominated. CONCLUSIONS: Approach toward the education of molecular pathology was refined, which would greatly benefit the future trainees.

Enhanced biomass production through a repeated sequential auto-and heterotrophic culture mode in Chlorella protothecoides.

A repeated sequential auto-and heterotrophic (RSAH) culture mode was designed to enhancebiomass ofChlorella protothecoides. Based on the result that the photosynthesis system may receive damage if the light period is more than 16 h, autotrophy was applied in the 16 h of the light cycle and mixotrophy using acetic acid and glucose in the 8 h of dark cycle. In the dark cycle, an organic carbon source was added according to the Monod equation to maintain activation of the TCA cycle and organic carbon source-to-cell conversion. When acetic acid and glucose were used as organic carbon sources, this culture method was found to be 32.3% and 12.6% higher in biomass, 2.59 and 2.67 times higher in the organic carbon source-to-cell conversion factor, and 2.17 and 2.32 times higher in ATP/ADP ratio, respectively, compared to mixotrophy. Through this new culture method, economic feasibility and carbon reduction capabilities in large-scale cultures can be achieved.

A green decontamination technology through selective biomineralization of algicidal microorganisms for enhanced astaxanthin production from Haematococcus pluvialis at commercial scale.

Currently, there is a lack of an efficient, environmentally-benign and sustainable industrial decontamination strategy to steadily achieve improved astaxanthin production from Haematococcus pluvialis under large-scale outdoor conditions. Here, this study demonstrates for the first time that a CaCO3 biomineralization-based decontamination strategy (CBDS) is highly efficient in selectively eliminating algicidal microorganisms, such as bacteria and fungi, during large-scale H. pluvialis cultivation under autotrophic and mixotrophic conditions, thereby augmenting the astaxanthin productivity. Under outdoor AT and MT conditions, the average astaxanthin productivity of H. pluvialis using CBDS in a closed photobioreactor system was substantially increased by 14.85- (1.19 mg L-1 d-1) and 13.65-fold (2.43 mg L-1 d-1), respectively, compared to the contaminated H. pluvialis cultures. Given the exponentially increasing demand of astaxanthin, a natural anti-viral, anti-inflammatory, and antioxidant drug, CBDS will be a technology of interest in H. pluvialis-based commercial astaxanthin production which has been hindered by the serious biological contaminations.

Ileocolic Intussusception Accompanied with Inflamed Appendix: 2 Case Reports.

Intussusception and acute appendicitis are common emergency conditions in children. They should be promptly differentiated in pediatric patients presenting with suggestive symptoms. However, both diseases may occur simultaneously. Herein, we present two cases of intussusception of the appendix accompanied with appendicitis.

A case of mammary analogue secretory carcinoma of the parotid gland: A case report.

RATIONALE: Mammary analogue secretory carcinoma is a low grade salivary gland malignancy, first described by Skalova et al in 2010. The histological, immunohistochemical, and molecular characteristics of this tumor resemble those of secretory carcinoma of the breast. PATIENT CONCERNS: A 40-year-old male patient without any specific past history came with complaints of 4 cm-sized hard, fixed, right infra-auricular mass without tenderness. There was no enlarged or enhancing lymph node in both neck. DIAGNOSES: Fine needle aspiration biopsy was done for right parotid mass and pathologic report was "lymphocytes only" that suggested benign or low-grade malignant parotid tumor. INTERVENTIONS: Partial parotidectomy including mass excision was performed. Operative finding showed hard bluish mass located in deep lobe of right parotid gland. OUTCOMES: Final pathologic report revealed secretory carcinoma of parotid gland. Homogenous eosinophilic secretions were identified inside microcystic structure. The immunophenotype was positive for epithelial membrane antigen, vimentin, S-100 protein. After 15 months of the surgery, the patient showed negative evidence of disease state. LESSONS: We present here a case of mammary analogue secretory carcinoma of the parotid gland to help further characterize this rare tumor.

Repeated-batch production of omega-3 enriched biomass of Chlorella sorokiniana via calcium-induced homeoviscous adaptation.

This study aimed to improve valuable omega-3 fatty acids production in freshwater microalgae at normal temperature by inducing homeoviscous adaptation using CaCl2, which could have a role in decreasing the cellular membrane fluidity followed by increasing the rigidity of cell wall and membranes. At 10 mM CaCl2, simultaneous biomass and lipid production was obtained by Ca2+-based single strategy without considerable sacrifice of cellular logarithmic growth in Chlorella sorokiniana. The cells cultured at 10 mM CaCl2 (1-stage) showed relatively high levels of cellular membrane fluidity, caused by increased content in unsaturated fatty acids, compared to the conventional culture strategy (2-stage). Moreover, when this process was recycled by repeated-batch fermentation, the EPA productivity of 1-stage was 4.338 mg L-1 d-1, conspicuously increased by over 1300% compared to 2-stage. This strategy enhances the valuable omega-3 production, which can be commercially used for mass cultivation of omega-3-enriched biomass in the microalgae industry.

Extrauterine Adenomyoma of the Liver Mimicking a Hepatic Adenoma: A Case Report.

Extrauterine adenomyoma of the liver is extremely rare. Only a few cases have been reported, and these reports have focused mainly on histopathology. Herein, we report the specific imaging findings of extrauterine adenomyoma of the liver in a 43-year-old woman with epigastric pain, which was initially diagnosed as a hepatic adenoma. CT and MRI revealed a solid and cystic mass with hemorrhagic foci and weak persistent enhancement, located in the subcapsular region of the right hepatic lobe.

Screening of oleaginous algal strains from Chlamydomonas reinhardtii mutant libraries via density gradient centrifugation.

Microalgae have a high potential to be utilized as feedstock for biofuels because they have high growth rates and do not compromise food production. Commercialized algae-based biofuel production relies on the development of strains with high lipid content. Based on the relatively low density of lipids compared to other cellular components, density gradient centrifugation was used to isolate high lipid content algal strains from Chlamydomonas reinhardtii mutant libraries. The correlation between cell density and lipid content was confirmed by analysis of Nile red fluorescence intensity, total lipids, and total fatty acid methyl ester content. A strain isolated by this screening method had 50% higher lipid content and 7% lower cell density than the parent wild-type strain. Consequently, we demonstrated that screening of algal strains with low cell density via continuous density gradient centrifugation allows simple, rapid, and inexpensive screening for high lipid content strains.

Split mixotrophy: A novel cultivation strategy to enhance the mixotrophic biomass and lipid yields of Chlorella protothecoides.

Mixotrophy (M) assumes sum of autotrophic (A) and heterotrophic (H) growths. In this study, a novel split-mixotrophic cultivation strategy (SMCS) developed as better mixotrophy via offering mutual-benefits through gas-exchange at both headspaces while splitting both trophic modes. To quantify synergistic-growth effects in combined-autotrophy and combined-heterotrophy (CA&CH) of SMCS, gross O2-evolution, DIC and DO concentrations were compared with A, H and M. Average 12-14% and 26-32% increase in DIC and DO concentrations were determined respectively in CA and CH than A, H and M. Biomass yield in CA + CH was increased approx.1.5-folds higher than yields of A + H and M regimes. These results show SMCS as better cultivation strategy than the M by increased biomass and lipid yields. Challenges associated with organic carbon can be solved by SMCS viz. chlorophyll loss, organic carbon uptake inhibition. SMCS could be a breakthrough to integrate bacterial process with algae for better bioprocess economy and energy recovery.

Enhanced biomass and lipid production of Neochloris oleoabundans under high light conditions by anisotropic nature of light-splitting CaCO3 crystal.

The aim of this work was to study the anisotropic effect of crystalline CaCO3 nanoparticles (CN)-driven multiple refraction/scattering from the CN-coated agglomerated cells on the rate of photosynthesis and the product yield under high light conditions in the freshwater microalgae Neochloris oleoabundans. The CN-coating via biomineralization significantly improved the biomass and lipid production of N. oleoabundans during second stage of autotrophic induction by sustaining relatively high rate of photosynthesis at high irradiance using the multiple-splitting effect of the anisotropic polymorphism. The CN were successfully produced, adsorbed and grown on the external cells under conditions of mild alkalinity (pH 7.5-8.0), mild CaCl2 concentration (0.05 M) and under nitrogen starvation with strong light (400 µE m-2 s-1). Consequently, lipid content and productivity of N. oleoabundans cells cultured with 0.05 M CaCl2 increased by 18.4% and 31.5%, respectively, compared to the cells cultured with 0.05 M CaCl2 and acetazolamide to inhibit calcification.

Effect of light conditions on mixotrophic cultivation of green microalgae.

Current research aimed to increase mixotrophic biomass from various organic carbon sources by exploring best light conditions. Three substrates glucose, acetic acid and glycerol were studied for their effects on mixotrophic microalgae cultivation under four light conditions. Light irradiance exhibited variability in growth response and photosynthetic efficiency based on type of substrates used in mixotrophic growth. Each substrate showed variability in light requirements for their effective assimilations. From growth responses, glucose and acetic acid respectively exhibited heterotrophic and mixotrophic (better growth in light) natures. Continuous light-deficient condition was adequate for effective mixotrophic growth as well as energy saving for glucose. However, light-sufficient condition required for effective acetic acid supported mixotrophic growth. Mixotrophic benefits from glycerol and its uptake by Chlorella protothecoides was negligible in all light conditions. Investigation of heterotrophic biomass contribution by various substrates in overall mixotrophic yield, glucose offered maximum approx. 43% contribution.

High-efficiency cell disruption and astaxanthin recovery from Haematococcus pluvialis cyst cells using room-temperature imidazolium-based ionic liquid/water mixtures.

Energy-saving, high-efficiency cell disruption is a critical step for recovery of thermolabile antioxidant astaxanthin from Haematococcus pluvialis cyst cells of rigid cell-wall structure. In this study, as room-temperature green solvents, 10 types of 1-ethyl-3-methylimidazolium ([Emim])-based ionic liquids (ILs) were compared and evaluated for their abilities to disrupt H. pluvialis cyst cells for astaxanthin/lipid extraction. Among the 10 ILs tested, 3 [Emim]-based ILs with HSO4, CH3SO3, and (CF3SO2)2N anions were selected based on astaxanthin/lipid extraction performance and synthesis cost. When pretreated with IL/water mixtures, intact cyst cells were significantly torn, broken or shown to release cytoplasmic components, thereby facilitating subsequent separation of astaxanthin/lipid by hexane. However, excess IL pretreatments at high temperature/IL dosages and longer incubation times significantly deteriorated lipid and/or astaxanthin. Under optimized mild conditions (6.7% (v/v) IL in water solution, 30 °C, 60 min), almost complete astaxanthin recoveries (>99%) along with moderate lipid extractions (∼82%) could be obtained.

Performance and potential appraisal of various microalgae as direct combustion fuel.

Direct combustion of biomass is considered the most effective and simple means to contribute to CO2 reduction. In this context, the life-cycle potential of microalgal solid fuel, which has been overlooked so far, was comprehensively scrutinized ranging from cultivation to direct combustion. According to the quantitative data, using the raw fuel was confirmed to offer great benefits over the conventional lipid-targeted microalgal fuel systems through exploiting all of the biomass' energy potential, thereby being able to significantly increase the energy yield from biomass. The solid fuel is shown to exhibit diverse positive aspects owing to its remarkable calorific value, productivity and CO2 fixation ability. The combustion test reveals coal-microalgae co-combustion brings beneficial consequences on combustibility and environmental impacts with no notable thermal efficiency drop. This holistic appraisal shows microalgae patently possess high potential as a direct combustion fuel, even outperforming that of extensively used woody fuels.

Two-Dimensional Microfluidic System for the Simultaneous Quantitative Analysis of Phototactic/Chemotactic Responses of Microalgae.

Microalgae have been spotlighted as a renewable energy source to produce biofuels from CO2 by photosynthesis. However, their innate inefficiency of CO2 conversion using light energy has been a challenge to the commercialization of algae-based biofuel production. Photosynthetic organisms have evolved behavioral responses, including phototaxis and chemotaxis, to find optimal conditions for capturing light energy and inorganic carbon (Ci) sources for photosynthesis. In this context, investigation of phototaxis and chemotaxis to HCO3-, the predominant form of Ci in neutral aqueous solutions, is necessary to understand the physiological role of tactic responses in photosynthesis. In this study, a two-dimensional microfluidic system enabled efficient analysis of phototactic and chemotactic responses by investigation of cell distribution in the outlet chambers. From statistical analysis (skewness and kurtosis) of tactic responses of different algal strains to external stimuli, the preferred concentrations of HCO3- for Chlamydomonas reinhardtii CC125 (wild type), PTS42 (random insertional mutant of C. reinhardtii, high photosynthetic activity), and CC2702 ( cia5 mutant of C. reinhardtii, unable to acclimate to low CO2 concentration) were determined to be 27.22, 43.23, and 36.95 mM, respectively. From the analysis of tactic responses of wild type and 14 mutant strains, it was found that the photosystem II (PSII) operating efficiency and CO2 fixation rate were strongly correlated with the phototactic ( R2 = 0.931) and chemotactic response ( R2 = 0.857), respectively. Finally, this system can be applied to high-throughput screening strategies for the rapid isolation of high photosynthetically productive microalgal strains based on their tactic responses.

Rapid selection of astaxanthin-hyperproducing Haematococcus mutant via azide-based colorimetric assay combined with oil-based astaxanthin extraction.

The aim of this work was to develop a new approach for simple and high-throughput selection of astaxanthin-hyperproducing Haematococcus mutants through a sequential combination method of azide-based colorimetric assessment and oil-based astaxanthin quantification. Randomly mutagenized cells were spotted on solid culture medium containing 50 µM of sodium azide to accelerate the biosynthesis of astaxanthin. After 3 days, highly-induced mutants were preliminarily isolated by visual inspection and their astaxanthin accumulations were rapidly quantified by soybean oil-based extraction method. On the whole, the selected mutants showed reduced vegetative growth rates but eventually exhibited higher astaxanthin productions than the parental strain owing to their improved inductive growths. Among them, M13 showed 174.7 ±â€¯5.69 mg L-1 of the highest astaxanthin production, which is 1.59-times higher than that of wild-type. This wide-scope screening method expedites both upstream and downstream astaxanthin quantification, making it a useful tool for isolating microalgae with high astaxanthin production.

Improvement in modular scalability of polymeric thin-film photobioreactor for autotrophic culturing of Haematococcus pluvialis using industrial flue gas.

The aim of this study is investigate the effect of column diameter (D), height/diameter (H/D) ratio, and gas flow rate on microalgae cultivation, Haematococcus pluvialis. Bubble column reactors with various D and H/D ratio were tested to assess the hydrodynamic properties and biomass production performance. Then, H. pluvialis was cultured under outdoor autotrophic conditions using industrial flue gas. By optimizing the reactor module, reactor volume increased to 354% with minimized biomass loss. Compared to the control, developed module showed biomass and astaxanthin productivity of 0.052 versus 0.053 g/L/day, and 1.48 versus 1.47 mg/L/day, respectively. Consequently, biomass productivity was maintained with increased reactor scale, and the result is applicable to the scale up of overall microalgae cultivation process.

NanoString nCounter® Approach in Breast Cancer: A Comparative Analysis with Quantitative Real-Time Polymerase Chain Reaction, In Situ Hybridization, and Immunohistochemistry.

PURPOSE: Accurate testing for estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) is essential for breast cancer treatment. At present, immunohistochemistry (IHC)/florescence in situ hybridization (FISH) are widely accepted as the standard testing methods. To investigate the value of NanoString nCounter®, we performed its comparative analysis with IHC/FISH and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) for the assessment of ER, PR, and HER2. METHODS: Data on IHC/FISH results for ER, PR, and HER2 in 240 patients from a single tertiary hospital in Korea were collected and compared with NanoString nCounter® and qRT-PCR results at a single institution. RESULTS: Expression levels for each gene using NanoString nCounter® showed good correlation with the corresponding data for protein expression by IHC (p<0.001) and gene amplification status for HER2 (p<0.001). Comparisons between gene expression and IHC data showed good overall agreement with a high area under the curve (AUC) for ESR1/ER (AUC=0.939), PgR/PR (AUC=0.796), and HER2/HER2 (AUC=0.989) (p<0.001). CONCLUSION: The quantification of ER, PgR, and HER2 mRNA expression with NanoString nCounter® may be a viable alternative to conventional IHC/FISH methods.

Development of large-scale and economic pH control system for outdoor cultivation of microalgae Haematococcus pluvialis using industrial flue gas.

The aim of this study was to develop the economic and effective buffer system for microalgae mass cultivation using industrial flue gas. Due to the continuous flue gas supplement, culture media acidified, therefore cell growth inhibited. Although buffering agent was added, this result increase in cost for overall culture process. Therefore combined buffer system of bicarbonate and phosphate (BP) for large-scale use was investigated. The bicarbonate buffer system generated from CO2 dissolution, additionally phosphate buffer system improves the buffer performance under the continuous CO2 supplementation from flue gas. The microalgae Haematococcus pluvialis was cultivated under autotrophic outdoor conditions using these buffer solutions. As a result, the autotrophic BP buffer system enhanced the biomass and astaxanthin productivity of H. pluvialis to 105% and 103%, respectively. The results confirm that the BP buffer system reduces the cost of microalgal CO2 conversion process, particularly for the outdoor mass cultivation.