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Background We aim to investigate the profile of acute antibody response in COVID-19 patients, and provide proposals for the usage of antibody test in clinical practice.Methods A multi-center cross-section study (285 patients) and a single-center follow-up study (63 patients) were performed to investigate the feature of acute antibody response to SARS-CoV-2. A cohort of 52 COVID-19 suspects and 64 close contacts were enrolled to evaluate the potentiality of the antibody test. ResultsThe positive rate for IgG reached 100% around 20 days after symptoms onset.The median day of seroconversion for both lgG and IgM was 13 days after symptoms onset. Seroconversion of IgM occurred at the same time, or earlier, or later than that of IgG. IgG levels in 100% patients (19/19) entered a platform within 6 days after seroconversion. The criteria of "IgG seroconversion" and "≥ 4-fold increase in the IgG titers in sequential samples" together diagnosed 82.9% (34/41) of the patients.Antibody test aided to confirm 4 patients with COVID-19 from 52 suspects who failed to be confirmed by RT-PCR and 7 patients from 148 close contacts with negative RT-PCR. ConclusionIgM and IgG should be detected simultaneously at the early phase of infection. The serological diagnosis criterion of seroconversion or the "≥ 4-fold increase in the IgG titer" is suitable for a majority of COVID-19 patients. Serologic test is helpful for the diagnosis of SARS-CoV-2 infection in suspects and close contacts.

Background: Marked progress is achieved in understanding the physiopathology of COVID-19 that caused global pandemics. However, CD4 + T cell population that is critical for antibody response in COVID-19 is poorly understood. Methods: In this study, we provided a comprehensive analysis of peripheral CD4 + T cells of 13 COVID-19 convalescent patients, as defined as confirmed free of SARS-CoV-2 for 2-4 weeks, using flow cytometry, magnetic chemiluminescence enzyme antibody immunoassay and correlated the data with clinical characteristics. Results: We observed that relative to healthy individuals, convalescent patients displayed an altered peripheral CD4 + T cell spectrum. Specifically, consistent with other viral infections, cTFH1 cell associated with SARS-CoV-2 targeting antibodies, which was found to skew with disease severity as more severe individuals showed higher frequency of TEM and TFH-EM cells but a lower frequency of TCM, TFH-CM and TNaive cells, relative to mild and moderate patients. Interestingly, higher frequency of cTFH-EM cells correlated with lower number of recorded admission blood oxygen level in convalescent patients. These observations might constitute residual effects by which COVID-19 can impact the homeostasis of CD4 + T cells in the long-term and explain the highest ratio of classswitched virus-specific antibody producing individuals found in our severe COVID-19 cohort. Conclusion: Together, our study demonstrated close connection between CD4 + T cells and antibody production in COVID-19 convalescents.

SummaryThe green-fluorescent protein (GFP) from Aequorea victoria has been shown to be a convenient and flexible reporter molecule within a variety of eukaryotic systems, including higher plants. It is particularly suited for applications in vivo, since the mechanism of fluorophore formation involves an intramolecular autoxidation and does not require exogenous co-factors. Unlike standard histochemical procedures of fixation and staining required for analysis of the cellular or tissue-specific expression of other popular reporter molecules, such as the I]-glucuronidase (GUS) marker, analysis of GFP can be done in living cells with no specific pretreatments. This implies that GFP might also be particularly suited for studies of intracellular protein targeting. In this paper, the use of GUS is compared with that of GFP for the analysis of nuclear targeting in tobacco. A novel oligopeptide motif from a tobacco protein is described which confers nuclear localization of GUS. The use of this oligopeptide and two from potyviral proteins to target GFP to the nucleus is examined. An essential modification of GFP is described, which specifically increases its molecular weight to eliminate its passive penetration into the nucleus. Three examples of the targeting of these enlarged GFP molecules to the nucleus are illustrated. GFP, in combination with confocal microscopy, offers significant advantages over traditional methods of studying nuclear targeting.

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Abstract. The eastern China seas are some of the largest marginal seas in the world, where high primary productivity and phytoplankton blooms are often observed. However, little is known about their systematic variation of phytoplankton blooms on large spatial and long temporal scales due to the difficulty of monitoring bloom events by field measurement. In this study, we investigated the seasonal and interannual variability and long-term changes in phytoplankton blooms in the eastern China seas using a 14 yr (1998–2011) time series of satellite ocean colour data. To ensure a proper satellite dataset to figure out the bloom events, we validated and corrected the satellite-derived chlorophyll concentration (chl a) using extensive in situ datasets from two large cruises. The correlation coefficients between the satellite retrieval data and the in situ chl a on the logarithmic scale were 0.85 and 0.72 for the SeaWiFS and Aqua/MODIS data, respectively. Although satellites generally overestimate the chl a, especially in highly turbid waters, both the in situ and satellite data show that the overestimation of satellite-derived chl a has an upper limit value (10 μg L−1), which can be used as a threshold for the identification of phytoplankton blooms to avoid the false blooms resulting from turbid waters. Taking 10 μg L−1 as the threshold, we present the spatial-temporal variability of phytoplankton blooms in the eastern China seas over the past 14 yr. Most blooms occur in the Changjiang Estuary and along the coasts of Zhejiang, with a maximal frequency of 20% (about 73 days per year). The coasts of the northern Yellow Sea and the Bohai Sea also have high-frequency blooms (up to 20%). The blooms show significant seasonal variation, with most occurring in spring (April–June) and summer (July–September). The study also revealed a doubling in bloom intensity in the Yellow Sea and Bohai Sea during the past 14 yr. The nutrient supply in the eastern China seas might be a major controlling factor in bloom variation. The time series in situ nutrient datasets show that both the nitrate and phosphate concentrations increased more than twofold between 1998 and 2005 in the Yellow Sea. This might be the reason for the doubling of the bloom intensity index in the Yellow Sea and Bohai Sea. In contrast, there has been no significant long-term increase or decrease in the Changjiang Estuary, which might be regulated by the Changjiang River discharge. These results offer a foundation for the study of the influence of phytoplankton blooms on the carbon flux estimation and biogeochemical processes in the eastern China seas.

Marine yeast strain 1, isolated from the surface of a marine alga, was found to secrete a large amount of inulinase into the medium. This marine yeast was identified as a strain of Pichia guilliermondii according to the results of routine yeast identification and molecular methods. The crude inulinase produced by this marine yeast worked optimally at pH 6.0 and 60 degrees C. The optimal medium for inulinase production was seawater containing 4.0% (w/v) inulin and 0.5% (w/v) yeast extract, while the optimal cultivation conditions for inulinase production were pH 8.0, 28 degrees C and 170 rpm. Under the optimal conditions, over 60 U ml(-1) of inulinase activity was produced within 48 h of fermentation in shake flasks. A large amount of monosaccharides and a trace amount of oligosaccharides were detected after the hydrolysis, indicating that the crude inulinase had a high exoinulinase activity.

Abstract. The East China Sea (ECS) and the South China Sea (SCS) are two major marginal seas of the North Pacific with distinct seasonal variations of primary productivity. Based upon field observations covering both the ECS and the northern SCS (NSCS) during December 2008-January 2009, we examined southward long-range transport of nutrients from the ECS to the northeastern SCS (NESCS) carried by the China Coastal Current (CCC) driven by the prevailing northeast monsoon in wintertime. These escaped nutrients from the ECS shelf, where primary production (PP) was limited in winter, might however refuel the PP on the NESCS shelf at lower latitude, where the water temperature remained favorable, but river-sourced nutrients were limited. By combining the field observation of nitrate+nitrite (NO 3 +NO 2 , DIN) with our best estimate of volume transport of the CCC, we derived a first-order estimate for DIN flux of 1430 ± 1024 mol s −1 . Under the assumption that DIN was the limiting nutrient, such southward DIN transport would have stimulated 8.84 ± 6.33 × 10 11 gC of new production (NP), accounting for 33-74 % of the NP or 14-22 % of PP in winter on the NESCS shelf shallower than 100 m.